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Roberts, S. J. "Bacterial diseases of woody ornamental plants." Thesis, University of Leeds, 1985. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.375533.
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Akiew, E. B. "Potato diseases in South Australia : studies in leafroll, early blight and bacterial wilt /." Title page, contents and summary only, 1985. http://web4.library.adelaide.edu.au/theses/09PH/09pha315.pdf.
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Roberts, Daniel Paul. "Molecular mechanisms of pathogenesis incited by Erwinia carotovora subsp. carotovora." Diss., Virginia Polytechnic Institute and State University, 1985. http://hdl.handle.net/10919/71251.
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Erwinia carotovora subsp. Carotovora (Ecc) incites soft-rot on many plants. It is believed that soft-rot is due to the concerted activity of extracellular enzymes. Recombinant DNA techniques were used to study the molecular basis of pathogenesis incited by Ecc. Specifically, a clone library of Ecc strain EC14 DNA in plasmid pBR322 was constructed and transformed into Escherichia coli strain HB101. Some of the E. coli strains that contain these hybrid plasmids produce pectinases or cellulase(s). Plasmid pDR1 contains a 3.4 kilobase (kb) EC14 DNA fragment and mediates the production of endo-pectate lyases with isoelectric points (pI) of 9.5 and 7.5 in strain HB101. The pI 9.5 enzyme is believed to be the major extracellular pectolytic enzyme in soft-rot while the pI 7.5 enzyme has no documented counterpart in EC14. Subclone and transposon tn5 analyses of pDR1 indicate that 1.5 kb is necessary for the production of the pI 9.5 and pI 7.5 enzymes and that these enzymes are produced independently of other EC14 pectate lyase enzymes. Plasmid pDR30 contains a 2.1 kb EC14 DNA insert that mediates the production of an endo-polygalacturonase and an exo-pectate lyase in HB101. The exo-pectate lyase encoded by pDR30 produces an inducer of endo-pectate lyase synthesis as a reaction product. The endo-polygalacturonase encoded by pDR30 is thought to play a role in plant cell wall pectic polymer degradation. Restriction endonuclease and Southern hybrididizatian analyses indicate that the EC14 genes on plasmids pDR1 and pDR30 are not part of the same operon. Escherichia coli strain HB101 containing plasmid pDR1 or plasmid pDR30 is unable to macerate potato tuber slices. However, HB101 containing plasmids pDR1 and pDR30 can cause limited maceration of potato tuber slices. There appears to be a genetic interaction between plasmids pDR1 and pDR30 in maceration of potato tuber tissue. However, the EC14 gene(s) contained on plasmid pDR1 are transcribed independently of the EC14 genes contained on plasmid pDR30. It is possible that transcription of certain pectolytic enzymes independent of other pectolytic enzymes provides a flexible system for plant cell wall pectic polymer degradation.Ph. D.
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Proto, Mariarita <1990>. "Essential oils and hydrolates as potential tools for integrated prevention of bacterial diseases in plants." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2022. http://amsdottorato.unibo.it/10292/1/TESI%20PhD%20MRP.pdf.
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The present work aims to investigate the potential use of natural substances against bacterial plant pathogens. Microdilution tests were therefore carried out in vitro to identify the minimum inhibitory and bactericidal concentrations (MIC and MBC) of several EOs and Hys against selected bacterial pathogens. Commercial products based on a mixture of EOs were in addition assayed with macrodilution experiments against Erwinia amylovora (Ea-causal agent of fire blight). Subsequently, using selected EOs, Hys, and commercial products, ex vivo tests on disease incidence and Ea population dynamics were carried out; the latter experiment was followed by SEM observations. In addition, in vivo resistance induction test was carried out against bacterial leaf of tomato, caused by Xanthomonas vesicatoria (Xv). EOs and Hys showed high bactericidal activity in vitro (MBC <0.1 and <10% for the most active EOs and Hys: Origanum compactum and Thymus vulgaris EOs and Citrus aurantium var. amara Hy, respectively), but they were not effective ex vivo, while resulted very active when used in vivo as resistance inducers in the tomato-Xv pathosystem (relative protection >40%). Differently, commercial products resulted active in all tests, but not as resistance inducers against Xv. An open field trial with commercial products was carried out on strawberry plants naturally infected with Xanthomonas fragariae; the results showed discrete relative protection, concerning that provided by the conventional products based on copper; mostly, the disease severity reduction on those plants treated with EOs commercial products was significant when disease severity resulted high. The papers already published described in the present work investigate (1)the activity of Hys in comparison to EOs with respect to their active volatile content; (2) the potential use of EOs and Hys in cultural heritage; for the restoration of paintings; (3) the induction of resistance caused by plasma-activated water-based root treatments.
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DeYoung, Robyn Merrilee. "Gall formation by Erwinia species on Douglas-fir." Thesis, University of British Columbia, 1990. http://hdl.handle.net/2429/28980.
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Bacterial galls on Douglas-fir (Pseudotsuga menzeisii [Mirb.] Franco), collected from the southern tip of Vancouver Island, the Greater Vancouver area and the Hope region of British Columbia, were generally globose in shape with rough, irregular surfaces and measured between 0.5 and 2.0 cm in diameter. The galls were generally located on the tips of branches or twigs of 10- to 20-year old Douglas-fir trees.
The bacterial gall disease appeared to affect few Douglas-fir trees in the collection areas and bacterial galls were not found on any other coniferous species. Furthermore, there have been no reports of serious damage to natural forests in British Columbia due to bacterial gall disease. Young, greenhouse-grown Douglas-fir seedlings occasionally died if the tip of the main stem was artificially inoculated. Often new growing tips would be produced affecting the growth form of the seedlings.
Two types of gall-forming Erwinia spp. were isolated from Douglas-fir galls. Typical isolates, tentatively identified by fatty acid analysis as Erwinia salicis, produced galls which were rough and irregular in shape composed of multiple outgrowths marked by a single or cross-shaped fissure. The atypical isolate, tentatively identified by fatty acid analysis as Erwinia herbicola subsp. herbicola, produced galls which were smooth and generally round in shape with the surface cracking as the gall expanded.
Colonies of the typical isolates grown on casein-peptone-glucose media were characteristically round, slightly domed with
somewhat concentric ridging observed near the margins of the colonies. Three to 4 day old colonies of the atypical isolates grown on casein-peptone-glucose media were characteristically round and concave while older colonies produced an extracellular slime and were more irregular in shape. In Luria Broth, the typical isolates grew at temperatures of up to 32°C while the atypical isolate grew at temperatures of up 34°C. The typical isolate was resistant to a wider range of antibiotics than the atypical isolate.
Polyclonal antisera were produced against glutaraldehyde-fixed whole cells of both the typical T-2789 and atypical A-0181 gall-forming Erwinia isolates. The purified antisera were isolate specific as tested by immunodiffusion and an indirect ELISA against several different phytopathogenic bacteria including Pseudomonas syringae pv. syringae, Erwinia herbicola subsp. herbicola, Agrobacterium tumefaciens, Rhizobium leguminosarum and Erwinia carotovora subsp. carotovora.
Plasmid profiles of the typical Erwinia isolates contained one band while the atypical isolate characteristically contained 4 to 5 bands which appeared to be different forms of at least one plasmid. Restriction digests of the typical isolates suggested a size of approximately 50 kb while complex digestion profiles were obtained for the atypical isolates because of the difficulty in isolating individual plasmid types. From visual estimates against Hindlll-digested lambda DNA, a size of between 10 and 20 kb was suggested for the fastest moving plasmid band of the atypical isolate. No homology was observed between the different plasmid types characteristic of the two isolates. The
role of the plasmid DNA of the atypical isolate in pathogenesis was not determined because curing of the plasmid(s) was not successful using high temperature treatments plus chemical curing agents.
Heat treatment experiments, in which the pathogen was selectively killed at various times after inoculation, demonstrated that the bacteria are required to be present for gall induction and continued development of the gall for both of the gall-forming Erwinia isolate types.
Pathogenicity of the isolated bacteria was tested on 14 conifer species, other than Douglas-fir, including Abies, Chamaecyparis, Pinus and Thuja spp. The typical isolates were weakly pathogenic on Abies, Larix and Picea spp. The atypical isolate was weakly pathogenic on Abies, Chamaecyparis, Larix, Picea and Pinus spp. Due to the limited damage caused on the conifers tested and to their infrequent occurrence, these gall-forming pathogens do not appear to be of economic importance to the forestry industry.Land and Food Systems, Faculty of
Graduate
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Njom, Henry Akum. "Mechanism and synchronicity of wheat (Triticum aestivum) resistance to leaf rust (Puccinia triticina) and Russian wheat aphid (Duiraphis noxia) SA1." Thesis, University of Fort Hare, 2016. http://hdl.handle.net/10353/2700.
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Wheat (Triticum aestivum and T. Durum) is an extremely important agronomic crop produced worldwide. Wheat consumption has doubled in the last 30 years with approximately 600 million tons consumed per annum. According to the International Maize and Wheat Improvement Center, worldwide wheat demand will increase over 40 percent by 2020, while land as well as resources available for the production will decrease significantly if the current trend prevails. The wheat industry is challenged with abiotic and biotic stressors that lead to reduction in crop yields. Increase knowledge of wheat’s biochemical constitution and functional biology is of paramount importance to improve wheat so as to meet with this demand. Pesticides and fungicides are being used to control biotic stress imposed by insect pest and fungi pathogens but these chemicals pose a risk to the environment and human health. To this effect, there is re-evaluation of pesticides currently in use by the Environmental Protection Agency, via mandates of the 1996 Food Quality Protection Act and those with higher perceived risks are banned. Genetic resistance is now a more environmental friendly and effective method of controlling insect pest and rust diseases of wheat than the costly spraying with pesticides and fungicides. Although, resistant cultivars effectively prevent current prevailing pathotypes of leaf rust and biotypes of Russian wheat aphid from attacking wheat, new pathotypes and biotypes of the pathogen/pest may develop and infect resistant cultivars. Therefore, breeders are continually searching for new sources of resistance. Proteomic approaches can be utilised to ascertain target enzymes and proteins from resistant lines that could be utilised to augment the natural tolerance of agronomically favourable varieties of wheat. With this ultimate goal in mind, the aim of this study was to elucidate the mechanism and synchronicity of wheat resistance to leaf rust (Puccinia triticina) and Russian wheat aphid (Duiraphis noxia) SA1. To determine the resistance mechanism of the wheat cultivars to leaf rust infection and Russian wheat aphid infestation, a proteomics approach using two-dimensional gel electrophoresis was used in order to determine the effect of RWA SA1 on the wheat cultivars proteome. Differentially expressed proteins that were up or down regulated (appearing or disappearing) were identified using PDQuestTM Basic 2-DE Gel analysis software. Proteins bands of interest were in-gel trypsin digested as per the protocol described in Schevchenko et al. (2007) and analysed using a Dionex Ultimate 3000 RSLC system coupled to an AB Sciex 6600 TripleTOF mass spectrometer. Protein pilot v5 using Paragon search engine (AB Sciex) was used for comparison of the obtained MS/MS spectra with a custom database containing sequences of Puccinia triticina (Uniprot Swissprot), Triticum aestivum (Uniprot TrEMBL) and Russian wheat aphid (Uniprot TrEMBL) as well as a list of sequences from common contaminating proteins. Proteins with a threshold of ≥99.9 percent confidence were reported. A total of 72 proteins were putatively identified from the 37 protein spots excised originating from either leaf rust or Russian wheat aphid experiments. Sixty-three of these proteins were associated with wheat response to stress imposed by RWA SA1 feeding while 39 were associated with infection by Puccinia triticina. Several enzymes involved in the Calvin cycle, electron transport and ATP synthesis were observed to be differentially regulated suggesting greater metabolic requirements in the wheat plants following aphid infestation and leaf rust infection. Proteins directly associated with photosynthesis were also differentially regulated following RWA SA1 infestation and P.
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Mustafa, Bakray Nur Aqilah Binti. "Structure of root associated and soil fungal and bacterial communities in Southeast Asia tropical forest." Thesis, University of Aberdeen, 2018. http://digitool.abdn.ac.uk:80/webclient/DeliveryManager?pid=239153.
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The tropical rainforest has interested ecologists for hundreds of years because of its vast species diversity. The distribution and establishment of trees is related to soil properties and rootassociated microorganisms. The coexistence of hyper-diverse plant communities in tropical rainforests has resulted in associations being formed with belowground communities, mycorrhizas (particularly arbuscular mycorrhizal (AM), ectomycorrhizas (ECM)) and root associated bacterial communities. The rapid deforestation in Southeast Asia is causing the loss of the dominant and important tree species belonging to the family Dipterocarpaceae. It is important to understand whether different host species in the same environment maintain mycorrhizal and bacterial diversity, especially mycorrhizas with a restricted host range. In this study, I examine the ecology of mycorrhizas and bacteria associated with Dipterocarpaceae and also the plant community as a whole. The aim of this project is to understand the effect of host properties (e.g. species, size), soil factors (e.g. nutrient concentrations) and spatial factors on mycorrhizal fungi and bacterial diversity and community structure. The research took place in two Centre for Tropical Forest Science (CTFS) plots in Malaysia: Pasoh Forest Reserve (in Negeri Sembilan) and Danum Valley Conservation Area (in Sabah). Molecular protocols and a modern technique, Next Generation Sequencing (NGS), were adopted to quantify mycorrhizal and bacterial loads in tropical plants. ITS1 and ITS2 regions were used for ECM, 18S rRNA were used for AM, and 16S rRNA were used for bacteria. Mycorrhizas and bacteria present in the roots of Dipterocarpaceae from 60 individual plants belonged to 25 species within 6 genera were traced and sampled in 2015. To my knowledge, this study is the first attempt to study root-associated bacteria across multiple species within a single family, Dipterocarpaceae. Dipterocarpaceae's species was found to significantly influence root bacteria. Analyses showed that mycorrhizal communities are similar on the host, unlike the null model. Dipterocarpaceae was previously believed to solely host ECM, but this study disproves this. This study shows that Dipterocarpaceae can have dual colonization, as it iv can also associate with AM fungi. One soil core of 10 cm × 10 cm × 7.5 cm were collect randomly in three subplot and further divided at 2.5 cm each slice into 75 individual 'microcubes' of 2 cm × 2 cm × 2.5 cm depths enumerates a total of 192 fine root samples. Multivariate analysis revealed that AM fungi tend to associate with non-dipterocarp (as well as unidentified families) while ECM fungi tend to associate with dipterocarps. Data was also collected on host attributes, plant size, and root density. Dipterocarpaceae size does not influence the distribution of mycorrhizal or bacterial communities. The root density reduces as depth increases. Therefore, root density does have a significant influence on mycorrhizal community structure. The diversity of ECM and AM fungal communities within cubes decreased significantly with depth (p < 0.001), whereas the mycorrhizal communities did not change across horizontal distances within cubes. To investigate whether there is a relationship between belowground communities and soil properties, soil macro and micro nutrients were examined and a multivariate analysis was performed. The results showed that communities of belowground (mycorrhizal and bacterial) species correlate with soil parameters. Spatial scale also had an effect on community assembly, independent of environmental variation. These results demonstrate that mycorrhizal fungal communities can vary substantially over very fine spatial scales, and that the distribution of roots from different species do not reflect their proximity aboveground. This study clearly demonstrates the widespread presence of mycorrhizal fungi and root associated bacteria in tropical rainforest plants.
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Agustí, Alcals Lourdes. "Characterization and efficacy of bacterial strains for biological control of soil-borne diseases caused by Phytophthora cactorum and Meloidogyne javanica on Rosaceous plants." Doctoral thesis, Universitat de Girona, 2008. http://hdl.handle.net/10803/7930.
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S'avaluaren 58 soques de Pseudomonas fluorescens i Pantoea agglomerans per la seva eficàcia en el biocontrol de la malaltia causada per l'oomicet Phytophthora cactorum en maduixera i pel nematode formador de gal·les Meloidogyne javanica en el portaempelt GF-677.Es desenvolupà un mètode ex vivo d'inoculació de fulla amb l'objectiu de seleccionar soques bacterianes com a agents de control biològic de P. cactorum en maduixera. Tres soques de P. fluorescens es seleccionaren com a soques eficaces en el biocontrol del patogen en fulles i en la reducció de la malaltia en plantes de maduixera. La combinació de soques semblà millorar la consistència del biocontrol en comparació amb les soques aplicades individualment.
Tres soques de P. fluorescens es seleccionaren per la seva eficàcia en la reducció de la infecció de M. javanica en portaempelts GF-677. La combinació d'aquestes soques no incrementà l'eficàcia del biocontrol, però semblà reduir la seva variabilitat.
58 Pseudomonas fluorescens and Pantoea agglomerans strains were evaluated for their biocontrol efficacy against the oomycete Phytophthora cactorum in strawberry and the root-knot nematode Meloidogyne javanica in GF-677 rootstocks.
An ex vivo detached leaf inoculation method was developed to select bacterial strains as biological control agents of P. cactorum in strawberry. Three P. fluorescens strains were selected as effective in biocontrol of the pathogens on leaves and in disease reduction in strawberry plants. Combination of strains improved biocontrol consistency compared to strains applied individually.
Three P. fluorescens strains were selected for their efficacy in M. javanica infection reduction in GF-677 rootstocks. Combination of these strains did not increase biocontrol efficacy, but reduced its variability.
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Gumede, Halalisani. "The development of a putative microbial product for use in crop production." Thesis, Rhodes University, 2008. http://eprints.ru.ac.za/1352/.
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Pauw, Elizabeth Lita. "Evaluating the predictive value of a database of antimicrobial activities of leaf extracts of 537 southern African tree species against six important bacterial and fungal pathogens." Thesis, University of Pretoria, 2014. http://hdl.handle.net/2263/40266.
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Infectious diseases are the world’s leading cause of premature deaths in humans and animals. The resistance to antibiotics and the emergence of new infectious diseases has increased the need for additional effective antimicrobial products. Despite numerous publications investigating antimicrobial activity of plant extracts it appears that no effective single product antimicrobial has yet been developed from plants. In many cases, however crude plant extracts have excellent activity and may provide useful products.
Plants are frequently selected based on traditional use. Traditional healers usually use aqueous extracts of plants which in our experience generally have very low activities and it may be one of the reasons why no new products were developed from plants. Another approach to select plants for research is to use the taxonomic approach based on the premises that: (1) there is a correlation between active chemical compounds and antimicrobial activity; and (2) species in a family or order may have similar activities if the chemical precursors are inherited from a common ancestor. Future screening programmes could then concentrate on close relatives of species within these promising families and orders.
The main aim of this study was to randomly screen leaf extracts of several hundred southern African tree species against important microbial pathogens to determine which taxa have the highest activity and may yield useful products to treat infections in human and animal health markets. A wide selection of plant species improved the possibility of finding promising extracts and has the advantage that active compounds may be discovered from plants that are not used traditionally. To ensure sustainable use only leaves of trees were examined. A spin off of this study would also indicate the susceptibility of different organisms, correlate the antimicrobial activities of the different organisms and determine what minimum inhibitory concentration (MIC) represents a good activity based on investigating many extracts against many microbes.
The antimicrobial activity was determined by using a sensitive serial dilution microplate method. Acetone extracts were tested against two Gram-positive bacteria, two Gram-negative bacteria and two fungi, i.e. Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Candida albicans and Cryptococcus neoformans.
Small and mostly insignificant differences were found between the susceptibility of the microbial pathogens to the extracts. E. faecalis was the most sensitive bacterium and C. neoformans the most sensitive fungal organism. The strongest correlations in activities among the pathogens were between C. albicans and C. neoformans, and among the pathogen classes between Gram-positive and Gram-negative bacteria. The tree extracts analysed in the present study had a wide range of activities against the different pathogens. Twenty six per cent of the extracts inhibited the pathogens at MIC levels of 0.16 mg/ml. This clearly shows that 0.16 mg/ml is not low enough to discriminate between promising species. Some of the extracts inhibited the growth of more than one pathogen while other extracts had selective activities and could be the most promising to follow up.
The study identified families and orders with either statistically significantly higher or lower antimicrobial activities. Among the large families, Combretaceae and Fabaceae had high mean activities against all test pathogens. The families Anacardiaceae and Moraceae had high activities against both Gram-positive and Gram-negative bacteria whereas the families Proteaceae and Meliaceae had higher antifungal activities. Among the large orders, Fabales had relatively high activities against all the pathogen classes. Considering that plants in related taxa often contain similar compounds and therefore similar activities, future studies could analyse more representative species in the promising taxa.
Many tree species, genera, families and orders, including well-known and lesser known medicinal taxa in southern Africa, were identified with promising activities. To evaluate the potential use of these results, additional cytotoxicity, phytochemical and pharmacological studies should be carried out. The study, although still exploratory, underlined the potential of southern African tree extracts as sources of antimicrobial products. Application of these results within the Phytomedicine Programme has led to patents and products that were as good as commercial products in animal and field trials. We hope that our results will provided a starting point for discovering new products with useful activities.Thesis (PhD)--University of Pretoria, 2014.
gm2014
Paraclinical Sciences
Unrestricted
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Badela, Andiswa Unathi. "Prevalence and pathogenicity of vibrios in treated final effluents of selected wastewater treatment plants in the Amathole District Municipality of Eastern Cape Province of South Africa." Thesis, University of Fort Hare, 2014. http://hdl.handle.net/10353/d1019774.
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Waterborne diarrhoeal infections continue to be a major health setback in developing countries, especially in rural areas which lack adequate supply of portable water and sanitation facilities. Globally, waterborne diarrhoeal infections occur with an estimated mortality rate of 10–25 million deaths per year, 95% of which are children under the age of 5 years. The Vibrio species is one of the major groups of enteric pathogens that are responsible for diarrhoeal infections. Many strains of these bacterial species continue to cause epidemics of diarrhoea throughout the world. In this study, the prevalence of Vibrio pathogens in wastewater final effluents was assessed. Wastewater final effluent and discharge point samples were collected monthly between September 2012 and August 2013. All samples were collected aseptically using sterile 1 L Nalgene bottles containing 0.5 ml of sterile sodium thiosulphate solution and transported on ice to the laboratory for analyses within 6 h of collection. The membrane filtration method was used for enumeration of presumptive Vibrio densities on thiosulfate citrate bile salt (TCBS) agar plates. Polymerase chain reaction (PCR) was then used to confirm the identities of the presumptive Vibrio species using the species-specific primers. The confirmed isolates were further subjected to molecular characterization to confirm their respective pathotypes. Presumptive Vibrio densities varied from 0 to 2.11 × 102 cfu/100 ml. Out of 300 confirmed Vibrio isolates; 13.3% (40/300) were Vibrio fluvialis, 22% (66/300) were confirmed to be Vibrio parahaemolyticus, and 24.7% (74/300) proved to be Vibrio vulnificus, and 40% (120/300) were other Vibrio species which were not assessed for in this study. The strains of Vibrio fluvialis were found to exhibit 100% resistance to Polymixin and Tetracycline. However, Gentamicin was active against all the three Vibrio species selected for the purpose of this research. The recovery of Vibrio species in the discharged effluents throughout the sampling period even in adequately disinfected effluents is not acceptable considering the fact that Vibrio is a pathogenic bacterium. The findings of this study underline the need for constant monitoring of the microbiological qualities of discharged effluents and might also be suggestive for a review of the disinfection methods used at the treatment works.
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Buhariwalla, Hutokshi Keki. "Bacterial L-form associations with plants." Thesis, University of Aberdeen, 1993. http://digitool.abdn.ac.uk/R?func=search-advanced-go&find_code1=WSN&request1=AAIU542133.
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L-form bacteria can be induced in vitro by the treatment of cell walled bacteria with compounds which suppress cell wall synthesis. It has previously been demonstrated that a variety of bacterial L-forms could invade plant-cells and establish a novel symbiotic association within the plant-host, with no apparent host limitations. Such associations were mainly recognised by direct microscopy and immunological detection techniques. The main aim of this project was to develop and optimise a reporter gene system for the detection of genetically modified Bacillus subtilis and Pseudomonas syringae pv. phaseolicola L-form associations in planta . Associations were attempted between B.subtilis L-forms chromosomally marked with -glucuronidase ( gus ) genes and Chinese cabbage. PCR detection indicated the lack of stable plant-L-form associations. Plasmid vectors encoding the genes for luciferase ( lux ) from Vibrio fischeri were introduced into the French bean pathogen Pseudomonas syringae pv. phaseolicola and L-forms were then induced. Symbiotic associations were established between these lux -modified L-forms and Chinese cabbage. Associations were detected at 7 days after germination by PCR amplification of a lux -gene fragment, using 100 ng of total plant DNA as a template. In addition the distribution of lux -modified P.syringae L-forms into various tissues of the plant was monitored using PCR amplification of the lux -gene and Staphylococcus agglutination, these techniques provided information on the presence of DNA and protein-antigens respectively. In contrast, bioluminescence, determined by luminometry, X-ray film imaging and charge-coupled device enhanced microscopy, permitted the detection of intact, active and viable populations of P.syringae L-forms. The bioluminescence based detection assays were non destructive and rapid (5 min) providing direct visualisation of microbial activity in planta . The data suggests that the distribution of lux -modified L-forms is non-uniform, predominantly in the roots and primary leaves of 14 day old Chinese cabbage.
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Seti, Nozuko Zukiswa. "Prevalence of pathogenic Escherichia coli strains in the final effluents of four wastewater treatment plants in the Eastern Cape Province of South Africa." Thesis, University of Fort Hare, 2014. http://hdl.handle.net/10353/d1019808.
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Water is an essential need that stimulates health and well being. Increase in population size and urbanization negatively affect water resources due to high demands of effluent outputs. Wastewater is an important reservoir for Escherichia coli and can present significant acute toxicity if released into receiving water sources without being adequately treated. E. coli is used as indicator organism for the detection of faecal contamination. These strains have been considered to be one of the primary causes of diarrhoeal infections worldwide. The present study was conducted between September 2012 and June 2013 to assess the prevalence of pathogenic E. coli strains in the final effluents of four wastewater treatment plants in Chris Hani and Buffalo City Municipalities in the Eastern Cape Province of South Africa. Standard membrane filtration technique was used for bacteriological analysis and molecular based technique was used for identification of E. coli pathotypes. The results were recorded in colony forming units/100 ml. Faecal coliforms ranged between 0-9.6×10³ CFU/100 ml for the wwtp-Q and E. coli densities ranged between 0-8.4×10³ CFU/100ml. Faecal coliforms ranged between 4×10²-9.7×10³ CFU/100 ml for wwtp-M and E. coli densities ranged between 1.2×10¹-8.4×10³ CFU/100 ml. The wwtp-E showed to have bacterial counts of faecal coliforms ranging between 4.0×10³-8.2×10³ CFU/100 ml and E. coli densities ranging between 3.5×10¹-7.1×10³ CFU/100 ml. The WWTP-K in this study was only assessed for the presence of E. coli. Faecal coliforms were assessed by the other members of the group. This plant showed to have E. coli densities ranging between 0-7.5×10²CFU/100 ml. A total of 200 presumptive E. coli isolates were subjected to screening by conventional PCR in which (29%) of the wwtp-M isolates were positively identified as E. coli, (16%) of the wwtp-K, (22%) of the wwtp-Q and (34%) of the wwtp-E isolates were positively confirmed as E. coli. A total of 100 randomly selected E. coli isolates were characterised into different pathotypes. (16%) of positive isolates were detected as EPEC and 11% were detected as UPEC strains. There was no detection for the ETEC strains. Antibiotic susceptibility patterns of E. coli strains showed high levels of resistance to Penicillin G, Erythromycin, Tetracycline and Sulfamethoxazole. High levels of Susceptibility were observed in antibiotics such as Chloramphenicol, Amoxicillin and Tetracycline. The results of this study reveal that the plants were above the recommended Standard limit of zero CFU/100 ml for effluents meant to be discharge into receiving water sources. This study reveals inadequacy of the plants studied to produce effluents of acceptable quality.
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Roselló, Prados Gemma. "Characterization and improvement of plant-associated Lactobacillus plantarum. Novel biocontrol agent for fire blight disease." Doctoral thesis, Universitat de Girona, 2016. http://hdl.handle.net/10803/403404.
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Fire blight is a disease caused by Erwinia amylovora. For the limited effectiveness of copper products and the new regulations for the sustainable use of pesticides, biological control is considered a good alternative for the control of fire blight. Therefore, this thesis focuses on the research of lactic acid bacteria strains as biocontrol agents of fire blight. Three out of one hundred strains, which were identified as Lactobacillus plantarum TC54, TC92 and PM411, were selected by their ability to inhibit E. amylovora infections in different performed assays. A multivariate statistical analysis, combining different phenotypic and genotypic properties, determined that strains TC92 and PM411 were good candidates as biocontrol agents. Finally, the combination of lactic acid and strains TC92 and PM411 was used to increase the biocontrol efficacy and reliability in the biocontrol of fire blightEl foc bacterià és una malaltia causada pel bacteri Erwinia amylovora. L’eficàcia limitada dels productes cúprics i les noves normatives d’ús sostenible dels productes fitosanitaris, fan que el control biològic es consideri una bona alternativa en el control d’aquesta malaltia. Aquesta tesi es centra en la recerca de soques de bacteris de l’àcid làctic com a agents de biocontrol del foc bacterià. Tenint en compte la seva eficàcia en el control de les infeccions causades per E. amylovora en diferents assajos realitzats es van seleccionar 3 soques de 100 inicials, identificades com a Lactobacillus plantarum TC54, TC92 i PM411. Un anàlisi estadístic multivariant, combinant diferents propietats genotípiques i fenotípiques, va confirmar que les soques TC92 i PM411 eren bones candidates a agents de biocontrol. Finalment, per incrementar la seva eficàcia i fiabilitat en el control del foc bacterià, es va utilitzar la combinació d’àcid làctic i les dues soques seleccionades
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Daranas, Boadella Núria. "Biological control of quarantine bacterial plant diseases with Lactobacillus plantarum strains. Improvement of fitness and monitoring." Doctoral thesis, Universitat de Girona, 2018. http://hdl.handle.net/10803/666181.
Full textAbstract:
The fruit production is threatened by several bacterial diseases, such as fire blight of apple and pear, bacterial canker of kiwifruit, bacterial spot of stone fruits, and angular leaf spot of strawberry. The conventional pesticides that are available for the control of these diseases are mainly copper compounds and they have a limited efficacy and negative impact on environment. Therefore, there is a need to develop alternative and sustainable management tools. This Ph.D. Thesis contributes to the development of a novel microbial biopesticide based on lactic acid bacteria. Two Lactobacillus plantarum strains were selected due to their broad-spectrum activity. In order to improve the epiphytic survival of both strains in plants and to get more consistency in their biocontrol efficacy, a physiological adaptive strategy was defined to increase the water-stress tolerance. Also, a monitoring method was developed to evaluate the population dynamics of a L. plantarum strain.La producció de fruita està afectada per diferents malalties bacterianes de quarantena com el foc bacterià de les pomeres i pereres, el xancre bacterià del kiwi, la taca bacteriana dels fruiters de pinyol i la taca angular de les fulles de maduixera. Els plaguicides disponibles pel seu control són principalment compostos cúprics els quals tenen una eficàcia limitada i un impacte negatiu en el medi ambient. Existeix la necessitat de desenvolupar eines de control alternatives i més sostenibles. Aquesta tesi contribueix en el desenvolupament d’un bioplaguicida microbià basat en bacteris de l’àcid làctic. Es van seleccionar dues soques de Lactobacillus plantarum amb activitat d’ampli espectre i es va definir una estratègia fisiològica d’adaptació per incrementar la tolerància a l’estrès per manca d’aigua i així millorar la supervivència epifítica a la planta. També es va desenvolupar un mètode de monitoratge per avaluar les dinàmiques poblacionals d’una soca de L. plantarum.
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Rastall, Robert A. "The cell surface biochemistry of Erwinia amylovora." Thesis, University of Greenwich, 1989. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.258366.
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Rose, Janna L. "Ant-diarrheal Plants of Central Anatolia: Do They Inhibit Diarrhea-causing Bacteria?" FIU Digital Commons, 2011. http://digitalcommons.fiu.edu/etd/430.
Full textAbstract:
Infectious diarrhea results in 2 to 5 million deaths worldwide per year, and treatments that are safe, effective, and readily available are under investigation. The field of medicinal ethnobotany focuses on plants that are used by different cultural groups for treating various diseases and evaluates these plants for efficacy and cytotoxicity. In the present study, ethnobotanical research was conducted with Central Anatolian villagers in Turkey. Folk concepts and etiologies surrounding diarrhea were analyzed, as were salient plant-based remedies for diarrhea. Reviewing the literature, 91 plant species were described as anti-diarrheal in all of Turkey. In Central Anatolia, villagers described 35 species. For continued research via bactericidal and bacteriostatic bioassays, 15 plants were selected. Methanolic and aqueous extracts of medicinally used plant parts were evaluated for inhibitory properties against 10 diarrhea-causing bacteria in the first bioassay, and later 21 bacteria in a second assay utilizing spectrophotometry. The cytotoxic properties were also evaluated in an Alamar Blue Assay using HepG-2, PC-3, and SkMEL-5 human cell lines. While several extracts showed bactericidal and bacteriostatic properties, the methanolic extract of R. canina galls inhibited the most bacteria at the lowest concentrations. They were not cytotoxic. Thus, R. canina methanolic gall extracts were selected for bio-assay guided fractionation. Antibacterial activity was maintained in the third fraction which was composed of almost pure ellagic acid. The bioassay was repeated with standard ellagic acid, and the polyphenol retained potency in inhibiting multiple bacterial strains. Several other extracts showed promise for safe, effective anti-bacterial remedies for diarrhea.
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Baró, Sabé Aina. "Experimental basis for the control of plant diseases caused by Xylella fastidiosa using synthetic antimicrobial peptides." Doctoral thesis, Universitat de Girona, 2021. http://hdl.handle.net/10803/673523.
Full textAbstract:
Xylella fastidiosa is one of the most damaging phytopathogenic bacteria in the world, causing a variety of diseases with a major economic impact on agriculture and the environment. Until now, no therapeutic solution has been found for its control in affected plants and trees. Antimicrobial peptides (AMPs) represent a promising alternative to traditional compounds due to their activity against a wide range of plant pathogens, low toxicity, mode of action, and availability to create sustainable pesticides as well as disease-resistant crops. Thus, the aim of the present Ph.D. thesis was focused on the study of AMPs as a sustainable alternative for the control of diseases caused by X. fastidiosa. In addition to identifying highly bactericidal AMPs, highlighting important parameters of their activity, the aggressiveness of Spanish isolates of X. fastidiosa was studied in order to design efficient treatment guidelines for its controlXylella fastidiosa és un dels bacteris fitopatògens que provoca més danys a nivell mundial, causant una varietat de malalties amb un gran impacte econòmic en l’agricultura i l’entorn. Fins al moment, no s’ha trobat cap solució terapèutica pel seu control en plantes i arbres afectats. Els pèptids antimicrobians (PAMs) representen una alternativa prometedora als compostos tradicionals degut a la seva activitat contra una àmplia gamma de patògens vegetals, baixa toxicitat, mode d’acció, i disponibilitat per crear pesticides sostenibles i cultius resistents a les malalties. Així doncs, l'objectiu de la present tesi doctoral es va centrar en l'estudi dels PAMs com a alternativa sostenible pel control de les malalties causades per X. fastidiosa. A més d'identificar PAMs altament bactericides, posant de manifest paràmetres importants de la seva activitat, es va estudiar l’agressivitat d’aïllats espanyols de X. fastidiosa per poder dissenyar pautes de tractament eficients pel seu control
Programa de Doctorat en Tecnologia
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Duval, Alice Maria Quezado. "Diversidade de Xanthomonas spp. associadas à mancha-bacteriana em tomateiro para processamento industrial no Brasil." Universidade de São Paulo, 2003. http://www.teses.usp.br/teses/disponiveis/11/11135/tde-07042003-150839/.
Full textAbstract:
A mancha-bacteriana, causada por bactérias do gênero Xanthomonas, é uma das doenças mais importantes da cultura do tomate (Lycopersicon esculentum Mill.) para processamento industrial no Brasil. A etiologia dessa doença é complexa e a ocorrência das espécies envolvidas nas epidemias em lavouras da cultura no país é notavelmente pouco conhecida. Os objetivos desta tese foram identificar as espécies/grupos genéticos e as raças presentes em campos comerciais nas macro-regioes produtoras do Brasil-Central e Nordeste, determinar a sensibilidade in vitro de isolados ao cobre e aos antibióticos estreptomicina e oxitetraciclina. Inicialmente, 447 isolados foram analisados quanto aos perfis de eletroforese de campo pulsado (PFGE) e quanto às atividades amidolítica e pectolítica. Após identificação dos baplótipos PFGE, grupos de isolados foram caracterizados através de testes de patogenicidade em tomateiro e pimentão, presença das proteínas a ou b, utilização de fontes de carbono, sensibilidade in vitro ao cobre, estreptomicina e oxitetraciclina, reação provocada em hospedeiras diferenciais de tomateiro e Capsicum e presença dos genes de avirulência avrRxv e avrXv3. As análises da diversidade foram efetuadas a partir de índices de diversidade de Nei e pela análise de variância molecular (AMOVA). As três espécies de Xanthomonas relatadas na literatura como associadas à mancha-bacteriana em tomateiro, foram detectadas nos campos comerciais amestrados. Porém, no Brasil-Central as epidemias foram causadas principalmente por 'X. gardneri', enquanto que no Nordeste, exclusivamente por X. axonopodis pv. vesicatoria. Este é o primeiro relato de epidemias causadas por 'X gardneri' no mundo. A diversidade baplotípica foi baixa dentro de populações. Não obstante, menor diversidade intra-em variedades híbridas importadas do que PA nacionais. Da mesma forma, populações do Brasil-Central foram menos variáveis do que as do Nordeste. Esses fatos se explicam pela predominância em variedades PA nacionais e macro-região Nordeste de isolados pertencentes ao grupo PFGE 1, que apresentou-se mais pofimórfico, em oposição aos grupos 3 e 4, menos variáveis que predominaram em variedades híbridas e no Brasil-Central. A maior proporção da variação total por sua vez, foi atribuída ao componente inter-populacional, tanto para macro-regiões como para tipos de variedade. No entanto, considerável variação genética foi atribuída ao efeito da macro-geografia (30,6%) ou tipo de variedade (24,4%). Em relação às raças, relatou-se a ocorrência no país pela primeira vez das raças T3 e TIP8, no Nordeste e T2P7 e T2P8, no Brasil-Central. Esse fato indica que as resistências derivadas de 'NIL 216', L. pennelli e 'Hawaii 7998' são promissoras para o Nordeste. Quanto à sensibilidade aos agentes de controle químico, não foram encontrados isolados resistentes à oxitetraciclina (25 mg/ml). No entanto, foram encontrados isolados resistentes ao cobre (50 ml/ml) e à estreptomicina (25 mg/ml) em alta freqüência principalmente entre isolados de 'X. gardneri', o que explica a baixa eficiência do controle químico muitas vezes observada no campo. Este trabalho representa o primeiro relato sobre a composição populacional em larga escala de Xanthomonas spp. associadas à mancha-bacteriana em tomateiro rasteiro no país.Bacterial spot caused by bacteria of the genus Xanthomonas is one of the most important diseases of tomatoes for processe (Lycopersicon esculentum Mill.) in Brazil. The etiology of this disease is complex and there is notable lack of knowledge about the species involved in epidemics on this crop in Brazil. The objectives of this thesis were to identify the species/genetic groups, as well as races present in commercial fields in in Central and Northeastern Brazil, to determine the in vitro sensitivity of strains to copper and to streptomycin and oxytetracyclin. Initially, 447 strains were characterized according to their pulsed-field electrophoresis haplotypes and also for their amilolytic and pectolytic activities. After these analyses, selected strains were then tested for their pathogenicity on tomato and pepper, for the presence of a or b proteíns, utilization of carbon sources, in vitro sensitivity to copper, streptomycin and oxitetracyclin, reactions on differential tomato and pepper hosts and presence of the avirulence genes avrRxv and avrXv3. Diversity analysis of bacterial populations were done with Nei's diversity index and analysis of molecular variance (AMOVA). The three Xanthomonas species reported in the literature as associated with tomato bacterial spot, were detected in the sampled fields. However, the populations from Central Brazil were mainly composed by 'X. gardneri', whereas in the Northeast, X axonopodis pv. vesicatoria was the only species found. The haplotipic diversity was considerably low within populations. However, intra-populational diversity was lower among itnported hybrids than among national OP varieties. Similarly, populations from Central-Brazil were less variable than the ones from Northeast. These facts are explained by the predominance of strains from the more variable PFGE group 1 in OP varieties and in the Northeast region as opposed to the less variable groups 3 and 4 which predominated in hybrids and in Central Brazil. The greatest proportion of genetic in both analyses. However, considerable genetic variation was conferred by macrogeography (30.6%) or type of variety (24.4%) effects. Inoculation of selected strains on differential cultivars of pepper and tomato revealed, for the first time in Brazil, the occurrence of races T3 and TIP7, in the Northeast and T2P7 and T2P8, in Central Brazil. This fact indicates the promissing use of derived resistances from 'NIL 216', L. pennelli and 'Hawaii 7998' for the Northeast. No strains were resistant to oxitetracyclin (25 mg/ml) but a high frequency of strains resistant both to copper (50 ml/ml) and streptomycin (25 mg/ml) was found within 'X. gardneri'. This fact may explain the low efficiency of chemical control sometimes observed in the field. This work represents the first report about a large scale populational composition Xanthomonas spp. associated with processing tomatoes bacterial spot in Brazil.
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Bredenkamp, Jane. "Characterization of tolerance to bacterial wilt in the model plant Arabidopsis." Diss., University of Pretoria, 2014. http://hdl.handle.net/2263/40348.
Full textAbstract:
Ralstonia solanacearum, the causal agent of bacterial wilt disease, has been found to affect
numerous economically important plants. Understanding the molecular basis of resistance,
tolerance and susceptibility of plants to pathogens such as R. solanacearum is a major goal
of molecular plant pathologists. Prior to this study it was thought that Arabidopsis accession
Kil-0 shows gene-for-gene “resistance” to an African Eucalyptus isolate of R. solanacearum,
BCCF402. However, a subsequent preliminary study indicated that Kil-0 may exhibit
“tolerance” which is defined as the plant’s ability to support high pathogen numbers without
displaying disease symptoms or a reduction in host fitness. The aim of this study was to
determine if Kil-0 was tolerant to R. solanacearum BCCF402. The bacterial load of
R. solanacearum was quantified in accessions Kil-0 and Be-0 using dilution plating and
quantitative PCR methods. The cytC gene region was used to quantify R. solanacearum in
Arabidopsis plants and the amount of bacterial DNA was normalized to “alien” DNA that was
spiked into each sample. High bacterial concentrations of BCCF402 were found in Kil-0 but
plants exhibited no wilting symptoms. Additionally, Kil-0 plants inoculated with BCCF402
showed no significant reduction in fitness compared to control Kil-0 plants. In contrast, high
bacterial numbers and severe disease symptoms were observed in the susceptible Be-0
plants, whereas Nd1 plants contained a low number of bacteria and no disease symptoms
indicative of a resistance response. These results illustrated that Kil-0 is tolerant to
R. solanacearum isolate BCCF402. A tool for the visualization of R. solanacearum in
Arabidopsis plants was designed. R. solanacearum isolate BCCF402 was tagged with two
mCherry-containing plasmids under the constitutive expression of the tac promoter. The
expression levels of mCherry were suitable for successful visualization in planta. BCCF402
cells transformed with the mCherry-containing plasmids were not affected in terms of
virulence or disease progression compared to wildtype BCCF402 cells. A plasmid loss of
30-35% was observed in mCherry-tagged BCCF402 cells at later stages of Arabidopsis
infection. mCherry-tagged BCCF402 was successfully visualized in Kil-0 leaves at early
infection stages.Dissertation (MSc)--University of Pretoria, 2014.
gm2014
Plant Science
unrestricted
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Sanders, Gina Mercia. "Detection of Xanthomonas Campestris PV. magniferaeindicae in mango plants." Diss., University of Pretoria, 1993. http://hdl.handle.net/2263/39792.
Full textAbstract:
The yearly losses incurred by bacterial blackspot disease are high. Often trees are
asymptomatic, with the pathogen either in the resident phase or latent stage of infection.
Detection of the pathogen in these asymptomatic trees is one of the most important
means of controlling the disease. Isolates which consistently differed in virulence were
isolated from symptomatic mango plants. These isolates could be categorised into four
groups based upon differences in virulence. Monoclonal antibodies (mAbs) were
successfully raised using separate and pooled isolates for immunisation. MAbsraised were
of the lgG class and reacted with a proteinaceous epitope. These monoclonal antibodies
could distinguish between different virulence groups of Xanthomonas campestris pv.
mangiferaeindicae by means of Western Blot analysis. These antibodies were used along
with a selective medium, BVGA for detection of epiphytic populations as well as latent
infections in mango. An enrichment step prior to the enzyme- linked immunosorbent assay
(ELISA) is important, since bacterial counts on trees with latent infections are too low to
result in a positive signal. These techniques in combination are thus useful for detection
and monitoring of the pathogen, which may play an important role in controlling the
spread of the disease.Dissertation (MSc Agric)--University of Pretoria, 1993.
gm2014
Microbiology and Plant Pathology
Unrestricted
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González, Hernández Ana Isabel. "New approaches to control Pseudomonas syringae disease in tomato plants." Doctoral thesis, Universitat Jaume I, 2019. http://hdl.handle.net/10803/666683.
Full textAbstract:
Las plagas y enfermedades son factores a tener en cuenta ya que producen elevadas pérdidas en la producción de los cultivos. Actualmente, la búsqueda de compuestos no perjudiciales para el medioambiente y que sean capaces de inducir los mecanismos de defensa en las plantas frente a estreses es un tema prioritario. Por ello, el objetivo de esta tesis ha sido estudiar el mecanismo de inducción de defensa de los compuestos heptagluconato de cobre, 1-metiltriptófano (1-MT) y amonio, en el patosistema tomate-P. syringae. De forma general, el heptagluconato de cobre induce compuestos fenólicos, el 1-MT induce el cierre estomático y afecta al contenido del apoplasto y a la motilidad de la bacteria y, el amonio produce una reprogramación del metabolismo primario. Además, se ha estudiado como se modifica la arquitectura radicular bajo distintas fuentes de N y C, observando que la adición de nitrato o compuestos carbonados al tratamiento con amonio alivia la toxicidad causada por éste.Pests and diseases are important factors to take into account in order to avoid losses in crop production. Nowadays, the search of new enviro-friendly compounds to induce defence mechanisms in plants to fight against plant diseases is a priority. Thus, the goal of this thesis has been to study the action mode of the enviro-friendly inducers of resistance copper-heptagluconate, 1-methyltryptophan (1-MT) and ammonium in tomato-P. syringae pathosystem. In general terms, Cu-heptagluconate induces phenolic compounds, 1-MT induces stomatal closure and affects apoplastic content and bacterial motility, and ammonium nutrition produces a primary metabolism reprogramming. Moreover, the study of changes at root level under nitrate and ammonium sources has been carried out, showing that the addition of nitrate or C compounds to ammonium treatment alleviates its mild toxicity.
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Gasparoto, Maria Cândida de Godoy. "Epidemiologia comparativa entre \'Candidatus Liberibacter asiaticus\' e \'Candidatus Liberibacter americanus\' em ambiente controlado." Universidade de São Paulo, 2010. http://www.teses.usp.br/teses/disponiveis/11/11135/tde-20102010-164555/.
Full textAbstract:
Huanglongbing é a mais destrutiva doença da citricultura mundial. Seu recente relato no Brasil, em 2004, e a detecção de duas espécies bacterianas associadas à doença -Candidatus Liberibacter asiaticus e Candidatus Liberibacter americanus resultaram em poucos estudos do comportamento do Huanglongbing nas condições dos pomares brasileiros. Os objetivos deste trabalho foram (i) comparar a eficiência de infecção por Candidatus Liberibacter asiaticus e Candidatus Liberibacter americanus em plantas cítricas inoculadas por meio de enxertia com as duas espécies bacterianas, separadamente, e mantidas sob diferentes temperaturas, e (ii) comparar o progresso temporal e espacial do Huanglongbing causado por ambas as espécies, a partir da infecção natural por Diaphorina citri, tendo plantas de citros ou Murraya paniculata (murta) como fonte primária de inóculo. Para alcançar o primeiro objetivo, dois experimentos foram conduzidos em câmaras de crescimento com temperatura e fotofase controladas. A incidência da doença foi determinada por Real-Time PCR de amostras foliares. As temperaturas mais amenas foram favoráveis à infecção por Candidatus Liberibacter americanus, enquanto que altas temperaturas não afetaram a infecção de Candidatus Liberibacter asiaticus. A eficiência de infecção para esta última espécie foi favorecida nas temperaturas de 17/22°C e 22/27°C (noturna/diurna, respectivamente). Para cumprir com o segundo objetivo, um experimento foi conduzido com diferentes fontes de inóculo, que corresponderam aos seguintes tratamentos: (i) plantas de laranjeira doce da variedade Natal infectadas por Candidatus Liberibacter asiaticus ou Candidatus Liberibacter americanus, (ii) unicamente plantas de Murraya paniculata infectadas por Candidatus Liberibacter americanus e (iii) unicamente plantas de laranjeira doce da variedade Natal infectadas por Candidatus Liberibacter americanus. Cada tratamento foi aplicado em um compartimento isolado de uma casade- vegetação. O primeiro tratamento foi repetido. Em todos os compartimentos, 156 a 158 plantas de laranjeira doce da variedade Valência foram dispostas ao redor das plantas fonte de inóculo. Psilídeos livres das bactérias foram periodicamente confinados em plantas fonte de inóculo e, posteriormente foram liberados na casa-de-vegetação para livre movimentação e multiplicação. Quando as duas espécies bacterianas estavam presentes nas diferentes plantas cítricas utilizadas como fonte de inóculo, houve maior detecção de Candidatus Liberibacter asiaticus e maior eficiência de aquisição desta espécie bacteriana por psilídeos. Quando somente Candidatus Liberibacter americanus estava presente nas plantas fonte de inóculo, a eficiência de transmissão foi maior quando Murraya paniculata foi utilizada como fonte de inóculo, mostrando que Diaphorina citri pode adquirir naturalmente Candidatus Liberibacter 12 americanus de Murraya paniculata e transmiti-la para plantas cítricas. A eficiência de aquisição da espécie Candidatus Liberibacter americanus também foi maior tendo Murraya paniculata como planta fonte de inóculo e não citros.Huanglongbing is the most destructive disease of citrus worldwide. There are few studies on the behavior of Huanglongbing in the Brazilian orchards conditions due to the recent report in Brazil, in 2004. Two bacterial species were associated with the disease in Brazil: Candidatus Liberibacter asiaticus and Candidatus Liberibacter americanus. The aims of this work were (i) to compare the efficiency of infection by Candidatus Liberibacter asiaticus and Candidatus Liberibacter americanus in citrus graft-inoculated plants with both bacterial species, separately, and kept under different temperatures, and (ii) to compare the spatio-temporal progress of Huanglongbing caused by both species, naturally transmitted by Diaphorina citri, having infected citrus plants or Murraya paniculata (jasmine orange) as primary inoculum sources. To achieve the first objective, two experiments were carried out in plant growth chambers with controlled temperature and light. Disease incidence in the inoculated plants was determined by Real-Time PCR of the leaf samples. The milder temperatures were favorable to Candidatus Liberibacter americanus infection, while the higher temperatures did not affect the infection by Candidatus Liberibacter asiaticus. The infection by this latter species was favored by temperatures of 17/22ºC and 22/27°C (night/day, respectively). To achieve the second objective, an experiment was carried out with different sources of inoculum, corresponding to the following treatments: (i) Natal sweet orange plants infected by Candidatus Liberibacter asiaticus or Candidatus Liberibacter americanus, (ii) only Murraya paniculata plants infected by Candidatus Liberibacter americanus and (iii) only Natal sweet orange plants infected by Candidatus Liberibacter americanus. Each treatment was applied in an isolated compartment of a screenhouse. The first treatment was repeated. In all of the compartments, from 156 to 158 Valencia sweet orange plants were located around the source of inoculum plants. Psyllids free of the bacteria were periodically confined in source of inoculum plants and, after this period, they were set free, and allowed to the free movement and multiplication inside the screenhouse. When both bacterial species were present in the different citrus plants as source of inoculum, there was higher detection of Candidatus Liberibacter asiaticus than Candidatus Liberibacter americanus and higher acquisition efficiency of the former bacterial species than the later by psyllids. When only Candidatus Liberibacter americanus was present in the source of inoculum plants, the higher transmission efficiency was reached when Murraya paniculata served as source of inoculum, showing that Diaphorina citri can naturally acquire Candidatus Liberibacter americanus from Murraya paniculata and transmit it to citrus plants. The acquisition efficiency of the 14 Candidatus Liberibacter americanus species was higher with Murraya paniculata as source of inoculum than with citrus plants was the source of inoculum.
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Maher, Mary. "Effects of Plant Growth-Promoting Bacteria and Fungi on Strawberry Plant Health, Fruit Yield, and Disease Susceptibility." DigitalCommons@CalPoly, 2021. https://digitalcommons.calpoly.edu/theses/2335.
Full textAbstract:
Studies on plant growth-promoting rhizobacteria (PGPR) and fungi (PGPF) as biostimulants have shown significant positive effects on plant health, fruit yield, or pest management. However, very few published studies to date have been specific regarding their effects on strawberries (Fragaria × ananassa), particularly on soilborne disease prevalence in organically grown strawberries. Empirical data on the results of using these products in commercial growing applications under various conditions would be highly valuable, especially for organic growers who have limited synthetic chemical pesticides, herbicides and fertilizers registered for use. The objective of this study is to evaluate the efficacy of biostimulant supplementation on strawberries for improving fruit yield, fruit quality, and plant health in both high-tunnel, open-sided ‘hoophouse’ and field conditions.
This study consisted of two research projects. The first project investigated the effects of commercially available PGPR-based biostimulant products on strawberry plant health. The three products contained differing proprietary combinations of PGPR, primarily from the Bacillus and Lactobacillus genera. Plants were grown in two different soil types: sandy and clay, in order to investigate the effects of biostimulant supplementation in different soil conditions. In fall of 2018, 160 ‘Monterey’ strawberry plants were grown in an outdoor hoophouse in 3-gallon pots. Plants were either treated monthly with a single bacterial biostimulant product (EM-1, Accomplish LM, or Armory), or left untreated as a control. Plants were grouped into 20 blocks, each block comprised of 8 plants (each of the four treatments replicated in both soil types). Fruit yield (g), fruit sugar content (Brix), and leaf SPAD absorbance levels were measured weekly from January 27 to June 26, 2019. The treatments tested had no significant effects on fruit yield, leaf SPAD absorbance or Brix; soil type, however, did significantly impact fruit yield, with higher yields in sandy soil.
The second project was a field trial beginning in spring of 2020, in collaboration with Rutiz Farms in Arroyo Grande, CA, involving a total of 480 ‘Chandler’ strawberry plants. The farm is organically managed and has a history of soilborne diseases, including Verticillium dahliae. These plants were either treated monthly with one of three microbial biostimulant products: a product containing a proprietary strain of Trichoderma harzianum biocontrol fungus (TrichoSym), and two of the same PGPR-based products used the previous year (Accomplish LM and Armory); or left untreated as a control. The experiment was laid out in a randomized complete block design with four blocks, with each block consisting of 4 plots for each of the 4 treatments; each plot contained 30 plants. Fruit yield (g) per plot was measured weekly throughout the 2020 growing season and phenotypic disease incidence was measured biweekly. Soil samples were taken at three different points throughout the season, cultured on selective media, and analyzed to obtain estimates of V. dahliae colony-forming units (CFU) per gram soil. The treatments tested had no significant effect on fruit yield, phenotypic disease incidence, or V. dahliae CFU/g soil. The results are inconclusive as to whether this lack of effect is due to viability of the products themselves, ineffective application techniques resulting in lack of rhizosphere colonization, or some combination of these. Further research is needed to determine whether or not supplementation with microbial biostimulants can produce reliable, beneficial results in strawberries.
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Coaker, Gitta Laurel. "Genetic and biochemical characterization of resistance to bacterial canker of tomato caused by Clavibacter michiganensis subsp. michiganensis." Connect to this title online, 2004. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=osu1069188955.
Full textAbstract:
Thesis (Ph. D.)--Ohio State University, 2003.Title from first page of PDF file. Document formatted into pages; contains xvii, 163 p.; also includes graphics (some col.). Includes bibliographical references (p. 148-163).
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Peng, Zhao. "Contribution of TAL effectors in Xanthomonas to diseases of rice and wheat." Diss., Kansas State University, 2015. http://hdl.handle.net/2097/20443.
Full textAbstract:
Doctor of PhilosophyPlant Pathology
Frank F. White
Rice and wheat are two major crops that suffer losses from the diseases of bacterial blight and bacterial leaf streak, which are caused by Xanthomonas oryzae pv. oryzae (Xoo) and Xanthomonas translucens pv. undulosa (Xtu), respectively. Transcriptional-Activator Like (TAL) effectors, a special family of type III effector proteins from Xanthomonas, have been demonstrated as critical virulence factors that act by inducing corresponding susceptibility (S) genes in several disease complexes of plants. In this study, I analyzed the contributions of TAL effectors from Xoo and Xtu to virulence and in modulating host gene expression to enhance susceptibility. Specifically, the TalC effector from the African Xoo strain AXO1947 was identified as a critical virulence factor, which functions by promoting expression of the gene OsSWEET14 in rice. TalC is interchangeable with other major TAL effectors from Asian strains of Xoo on the basis of functional complementation. The TAL effector PthXo2 from the Asian Xoo strain JXO1 is a major virulence factor and contains 21.5 repeats in the central repetitive region that targets OsSWEET13 in indica rice varieties but not in japonica rice varieties. A one repeat deletion in the PthXo2 effector enabled effector specificity to switch from indica rice to japonica rice. TAL effector genes from a genomic analysis of the Xtu strain XT4699 and related strains were characterized with regards to their involvement in virulence and the modulation of host gene expression in the Chinese Spring wheat cultivar. The identification of TAL effectors with virulence contributions and their target S genes is important for understanding the virulence mechanisms of Xanthomonas bacteria and promises to provide new strategies for disease control.
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Silva, Ricardo Ferrari. "Indução de resistência em plantas de berinjela e tomate por Lentinula edodes e Agaricus blazei contra bactérias causadoras de murcha (Ralstonia solanacearum) e cancro (Clavibacter michiganensis subsp. michiganensis)." Universidade de São Paulo, 2007. http://www.teses.usp.br/teses/disponiveis/11/11135/tde-05062007-123711/.
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Devido ao aumento da preocupação com o impacto dos agrotóxicos no meio ambiente e na saúde humana, busca-se uma agricultura sustentável. É no âmbito dessa questão que a resistência induzida torna-se uma ferramenta fundamental no manejo integrado de doenças e indispensável para uma nova agricultura, mais racional e sustentável. Dentre os diversos agentes bióticos e abióticos, utilizados em trabalhos de indução de resistência de plantas a patógenos, os cogumelos Lentinula edodes e Agaricus blazei vem sendo pesquisados. Desse modo, este trabalho teve como objetivos avaliar o efeito de diferentes isolados de L. edodes e A. blazei e do acibenzolar-S-metil (aSm) in vitro contra as bactérias e o controle de doenças de importância econômica para as culturas do tomate e da berinjela, em casa-de-vegetação. Depois de obtida a proteção, estudar os possíveis mecanismos bioquímicos ativados nas plantas através do uso dos extratos dos cogumelos e buscar a purificação parcial destes extratos, a fim de identificar o(s) princípio(s) ativo(s). No patossistema berinjela/Ralstonia, os extratos aquosos dos cogumelos não exerceram nenhum efeito direto sobre o patógeno, sendo que os isolados Abl-11 e Abl-28 de de A. blazei reduziram significativamente a ocorrência de folhas murchas das plantas em casa-devegetação, em relação aos demais tratamentos. Ocorreu um aumento na atividade da peroxidase, fenilalanina amônia-liase e polifenoloxidase nas folhas tratadas. O preciptado 60-80% obtido pela precipitação com sulfato de amônia e a fração 4 da cromatografia de troca aniônica (CTA) de Abl-28 reduziram a ocorrência de folhas murchas, sendo que a separação eletroforética revelou a presença de uma banda no gel com aproximadamente 29 kDa nesta fração. Em tomate, os extratos aquosos dos isolados dos cogumelos e o acibenzolar-S-metil não exerceram nenhum efeito inibitório in vitro no crescimento de Ralstonia solanacearum e Clavibacter michiganensis subsp. michiganensis. Porém, os isolados Abl-26 de A. blazei, Le-96/17 de L. edodes e aSm foram os que conferiram maior proteção das plantas de tomates contra os patógenos, diminuindo a ocorrência de folhas murchas, proporcionando um aumento na atividade da peroxidase no patossistema tomate/Ralstonia e um aumento na atividade de peroxidase, quitinase, fenilalanina amônia-liase e polifenoloxidase no patossistema tomate/Clavibacter. O preciptado 40-80% de Le-96/17 foi submetido à CTA, obtendo-se seis frações protéicas. As frações 3 e 4, junto com aSm e o extrato aquoso de Le-96/17 reduziram a ocorrência de folhas murchas. A separação eletroforética destas frações da CTA, do preciptado 40-80% e do extrato aquoso de Le-96/17 revelaram a presença de mais de uma banda no gel na fração 3 e 4 da CTA, no preciptado 40-80% e no extrato aquoso bruto de Le-96/17. Com base nos resultados, os cogumelos A. blazei e L. edodes apresentam compostos que induziram resistência em plantas berinjela e tomate, podendo auxiliar no controle de doenças.Because the increase of the impact of chemical products in the environment and in human health, a search by sustainable agriculture is needed. It is in the scope of this problem that the induced resistance becomes a tool in the integrated management of pests and diseases and indispensable for a new agriculture, more rational and sustainable. Among the biotic and abiotic agents used to induce resistance, the mushrooms Lentinula edodes and Agaricus blazei have being studied. Thus, the objectives of the present work were evaluate the effects of different isolates of L. edodes and A. blazei and of the acibenzolar-S-methyl (aSm) on in vitro bacterial growth and the control of the diseases in tomato and eggplant under greenhouse conditions. The studies also tried to elucidate the mode of action of the extracts from the fruiting bodies and partially purify them. In eggplant plants, the aqueos extracts from the different mushroom isolates did not have any direct effect on the pathogen. The isolates Abl-11 and Abl-28 of A. blazei reduced the wilt in eggplant leaves, under greenhouse conditions, and increased peroxidase, phenylalanine ammonia-lyase and polyphenoloxidase activities in treated leaves. The fraction of aqueous extract of A. blazei (Abl-28) obtained with ammonium sulfhate and fraction 4 from anion exchange chromatography reduced bacterial wilt and a protein fraction exhibiting molecular mass around 29 kDa was obtained. In tomato plants, the aqueos extracts from the different mushrooms and the acibenzolar-S-methyl did not inhibit in vitro growth of Ralstonia solanacearum and Clavibacter michiganensis subsp. michiganensis. However, the isolates Abl-26 of A. blazei, Le-96/17 of L. edodes and aSm protected tomato plants against the bacterial pathogens, reducing the wilt and causing an increase in peroxidase activity in the tomato/Ralstonia interaction and an increase in peroxidase, chitinase, phenylalanine ammonialyase and polyphenoloxidase activities in the tomato/Clavibacter interaction. The ammonium sulphate fraction of Le-96/17 was submitted to anion exchange chromatography, and the proteins from fractions 3 and 4, aSm and the aqueous extract of Le-96/17 reduced the occurrence of wilt in the leaves. A protein fraction exhibiting proteins with molecular mass around 29, 37 and 45 kDA was obtained in fractions 3 and 4. Thus, the results showed that the mushrooms A. blazei and L. edodes edodes have substances that induce resistance in eggplant and tomato plants.
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Ma, Xing. "Characterization and Management of Bacterial Leaf Spot of Processing Tomato in Ohio." The Ohio State University, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=osu1440386548.
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Rezende, de Castro Moretti Fernanda. "Identification of candidate resistance metabolites to Leifsonia xyli subsp. xyli in sugarcane through metabolomic profiling." The Ohio State University, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=osu1512478087354334.
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Chidburee, Siripun. "Biological control of soil-borne disease in soybean by denitrifying antagonistic bacteria : the possible role of reduced nitrogen compounds for control of plant pathogens." Thesis, University of Aberdeen, 1998. http://digitool.abdn.ac.uk/R?func=search-advanced-go&find_code1=WSN&request1=AAIU602299.
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A number of experiments were carried out to study the potential of denitrifying bacteria and reduced nitrogen compounds for control of soil-borne damping-off pathogens. Measurement of the rhizosphere pH of growing soybean roots was carried out in soil adjusted to different pH states and packed into sheet microcosms. The results showed that the rhizosphere pH of soybean was lower than the bulk soil. Nitrate reductase activity and nitrite production was then characterised for the rhizosphere of intact 14 day-old soybean roots that were incubated in nitrate substrates adjusted to different pH values under water-logged conditions. The results showed that the rate and the quantity of nitrite production increased with increasing nitrate concentration and pH in the solution. A growth room experiment was carried out to determine root colonization by denitrifying bacteria in relation to disease caused by soil-borne pathogens, which are favoured by high soil moisture (approximately -5 KPa) and low oxygen levels. Nitrite producing bacteria were isolated from soybean roots grown in Grampian (Insch) soils which had not been cropped with soybean and Thai (Phitsanulok) soils which previously had been cropped with soybean. In the first pot experiment, the nitrite producing bacteria were isolated from different root sections of 12 and 19 day-old soybean plants after 8 weeks of continuous cropping of soils with soybean (a new crop was planted every week), and using different isolation media in order to determine the genus/species composition of the denitrifying bacteria on the rhizoplane. The results showed that continuous cropping of Thai soil and Insch soil with soybean increased pre-emergence damping-off disease and decreased fresh weight yields in seedlings that did emerge. ANOVA showed significant differences between root sections for most bacterial groups monitored {Bacillus spp., Pseudomonas and Enterobacteriaceae), with regression analysis generally showing densities increasing with root age or toward the shoot base. All nitrite producing bacterial isolates were screened for antifungal activity against Macrophomina phaseolina on agar plates and between 10 and 25% of nitrite producing bacteria were found to show in vitro antagonism. In a second pot experiment, the nitrite producing bacteria were isolated from root tissue below the crown (5 cm in length) every 2 weeks of continuous cropping of soils with soybean (a new crop was planted every 2 weeks). Plate-counting was carried out to determine the population of nitrite producing bacteria while a liquid culture MPN method was used for determination of NO, N2O and N2 producing bacteria. Linear regression analysis of the incidence of pre-emergence damping-off and soybean yields in seedling that did emerge showed a highly significant negative correlation between these parameters for both soils. ANOVA showed that there was a significant difference between soil type, with the Thai soil showing higher population densities of antagonistic bacteria on soybean roots. All nitrite producing bacterial isolates were screened for antifungal activity, but the plant pathogenic fungus, Pythium ultimum, was used in this experiment. The results showed that between 10 and 40% of nitrite producing bacteria showed in vitro antagonism. However, regression analysis showed that there was no significant increase or decrease in the nitrite producing antagonistic bacterial population with continuous soybean cropping. All 900 isolates of nitrite producing bacteria isolated from the soybean rhizoplane were screened for antagonistic activity towards Pythium ultimum based on a pot trial assay in the greenhouse. As expected, very low numbers of nitrite producing bacteria showed activity against P. ultimum and only one isolate gave a significant reduction in disease incidence in pot trials. The interactive effects of nitrite producing antagonist and an Arbuscular Mycorrhizal fungus (Glomus mosseae) and Bradryrhizobium japonicum, on control of the fungal pathogens, P. ultimum or M. phaseolina were investigated in the greenhouse. The results showed that improved plant growth was obtained with certain combined inocula involving nitrite producing bacterial antagonists, Glomus mosseae and Bradryrhizobium japonicum.
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Frare, Vanessa Cristina. "Tratamento de sementes de melão (Cucumis melo L.) para o controle de Acidovorax avenae subsp. citrulli." Universidade de São Paulo, 2010. http://www.teses.usp.br/teses/disponiveis/11/11135/tde-25052010-092252/.
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O melão é uma fruta de grande importância para o comércio de exportação do Brasil e, embora restrito a um pequeno número de estados produtores, o cultivo dessa fruta ampliou-se de forma significativa nos últimos anos. Um dos maiores problemas para essa cultura é a presença de patógenos, como a bactéria Acidovorax avenae subsp. citrulli (Aac), responsável por perdas de até 100%. O uso de sementes garantidamente sadias é a principal e mais efetiva medida de controle preconizada para essa bacteriose. Este trabalho teve como objetivo selecionar, in vitro e posteriormente in vivo, produtos capazes de erradicar a bactéria Aac de sementes de melão. Sessenta produtos fitossanitários comerciais foram selecionados para a avaliação do controle in vitro de quatro isolados, sendo nove bactericidas e 51 fungicidas, correspondendo a 6 e 45 ingredientes ativos distintos, respectivamente. Além desses produtos, avaliou-se também o efeito do óleo de melaleuca e do ácido peroxiacético sobre o crescimento dos isolados. Nos ensaios in vivo, sementes inoculadas e naturalmente infectadas foram tratadas, de maneira convencional e a vácuo, com os produtos e doses selecionados no ensaio in vitro. Todas as plântulas foram avaliadas quanto à presença de sintomas característicos de mancha-aquosa aos 21 dias após a emergência. Os antibióticos casugamicina (100 e 200 ug/L), oxitetraciclina (10, 100 e 200 ug/L), oxitetraciclina+sulfato de cobre (10, 100 e 200 ug/L), os fungicidas captana, carboxina+tiram, cloreto de benzalcônio, mancozebe+oxicloreto de cobre, metiram, metiram+piraclostrobina e tebuconazol, nas doses de 100 e 200 ug/L e o ácido peroxiacético a partir da dose de 300 ug/L apresentaram resultados satisfatórios na avaliação in vitro. No ensaio in vivo nenhum tratamento foi eficiente na erradicação da bactéria das sementes.Melon is a fruit of great importance for the export trade of Brazil. Although cultivated in a small number of states, the cropping of this fruit has expanded significantly in recent years. Pathogens represent the most limiting factors, among which stands out the bacterium Acidovorax avenae subsp. citrulli (Aac), responsible for up to 100% losses. The use of healthy seed is the main and the most effective control measure of this bacterium. This study aimed to select first in vitro and subsequently in vivo, erradicant chemicals of Aac in melon seeds. Sixty commercial products were were tested in vitro in the control of four isolates, being nine bactericides and 51 fungicides, corresponding to six and 45 distinct active ingredients, respectively. Besides these products the effects of oil of the melaleuca tea tree and peroxyacetic acid were also evaluated. In the in vivo tests, inoculated and naturally infected seeds were treated by conventional manner and under vacuum, with the products and doses selected in the in vitro test. All seedlings were evaluated for the presence of characteristic bacterial fruit blotch symptoms at 21 days after emergence. The antibiotics kasugamycin (100 and 200 ug/L), oxytetracycline (10, 100 and 200 ug/L), oxytetracycline+copper sulphate (10, 100 and 200 ug/L), the fungicides captan, carboxin+thiram, benzalkonium chloride, mancozeb+copper oxychloride, metiran, metiran+pyraclostrobin and tebuconazole at doses of 100 and 200 ug/L and peroxyacetic acid at the dose of 300 ug/L showed satisfactory results in the in vitro control of the bacterium. However, no chemical efficiently eradicated the bacterium from the seeds in the \"in vivo\" tests.
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Dorighello, Dalton Vinicio 1985. "Versatilidade de Bacillus spp. no controle biológico de doenças de plantas e na promoção de crescimento de soja /." Botucatu, 2017. http://hdl.handle.net/11449/151203.
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Orientador: Wagner BettiolBanca: Antonio Carlos Maringoni
Banca: Carlos Gilberto Raetano
Banca: Regina Maria Villas Boas de Campos Leite
Banca: Marcelo Augusto Boechat Morandi
Resumo: O mercado de produtos de controle biológico, principalmente os que possuem isolados de Bacillus spp. em sua formulação, tem aumentado significativamente, devido às exigências de diminuição do uso de agrotóxicos e maior sustentabilidade do segmento agrícola. Assim, o presente trabalho teve por objetivos: 1- Avaliar o potencial de bactérias do gênero Bacillus no controle de diferentes doenças de plantas; 2- Selecionar isolados de Bacillus spp. promotores de crescimento da soja e antagonistas a Sclerotinia sclerotiorium; 3- Avaliar o potencial de um produto comercial à base de B. subtilis QST 713 e outro à base de óleo de café torrado para compor um programa de manejo da ferrugem-asiática da soja (Phakopsora pachyrhizi) em aplicação sequencial ou alternada com um fungicida. Inicialmente foi avaliado o efeito de B. subtilis, B. firmus e B. amyloliquefaciens no controle dos patógenos Erysiphe diffusa, Podosphaera xanthii, Sclerotinia sclerotiorum, P. pachyrhizi, Fusarium solani, Fusarium oxysporum f. sp. phaseoli, Colletotrichum lindemuthianum e Corynespora cassiicola em condições controladas. Neste estudo, nenhum dos isolados controlou Erysiphe difusa e Podosphaera xanthii. Em folhas destacadas, todos os isolados reduziram o diâmetro da lesão de S. sclerotiorum e a severidade de P. pachyrhizi, sendo mais efetivos com aplicação preventiva. B. subtilis, B. firmus e B. amyloliquefaciens também inibiram o desenvolvimento de F. oxysporum f. sp. phaseoli, C. cassiicola e C. lindemuthia... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: The market of products for biological control, especially formulations using Bacillus spp., has increased significantly, due to the requirements for the reduction of the use of agrochemicals in crop protection and sustainability of the agricultural sector. The objectives of this thesis were: 1- To evaluate the potential of Bacillus in plant disease control; 2- To select isolates of Bacillus spp. for growth promotion of soybean and antagonists to Sclerotinia sclerotiorium; 3- To evaluate the potential of a commercial product based on B. subtilis QST 713 and another product based on roasted coffee oil in order to compose a management program for asian soybean rust (Phakopsora pachyrhizi) in sequential or alternating application with a fungicide. Initially, the effect of B. subtilis, B. firmus and B. amyloliquefaciens on the control of pathogens Erysiphe diffusa, Podosphaera xanthii, Sclerotinia sclerotiorum, P. pachyrhizi, Fusarium solani, Fusarium oxysporum f. sp. phaseoli, Colletotrichum lindemuthianum and Corynespora cassiicola was evaluated under controlled conditions. In this study, Bacillus isolates did not controlled Erysiphe diffusa and Podosphaera xanthii. All isolates were effective in reducing the diameter of the S. sclerotiorum lesion and the severity of P. pachyrhizi, being more effective by preventive application in detached leaf. B. subtilis, B. firmus and B. amyloliquefaciens inhibited the mycelial growth of F. oxysporum f. sp. phaseoli, C. cassiicola and C. lindemuthianum ...
Doutor
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Traore, Sy M. "Characterization of Effector Genes in Acidovorax citrulli the Causing Agent of Bacteria Fruit Blotch Disease of Cucurbits." Diss., Virginia Tech, 2014. http://hdl.handle.net/10919/64519.
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Bacterial fruit blotch (BFB) of cucurbits is caused by Acidovorax citrulli, a Gram-negative seedborne bacterium that can cause up to 100% fruit yield losses in the field. Currently, BFB is a major problem for the cucurbits industry worldwide. Thus far, attempts to identify resistance in cucurbit germplasm for controlling BFB have been unsuccessful. Despite the importance of the disease, little is known about the molecular mechanisms of A. citrulli pathogenicity, due to a lack of molecular tools for studying the A. citrulli/cucurbit interaction. The genomic sequence of A. citrulli strain AAC00-1 has been determined, and the components of type III secretion system have been identified. The goal of this research was to develop molecular tools for studying the BFB disease. Nineteen putative type III effector genes were cloned from two representative A. citrulli strains (AAC00-1 and M6). The distribution of 19 type III effectors among A. citrulli strains, collected worldwide, was studied. A novel Gateway-compatible binary vector was developed for transient expression of A. citrulli type III effectors genes in planta. A set of modified vectors for marker-exchange mutagenesis in A. citrulli were constructed. The model plant species Nicotiana benthamiana was found to be susceptible to A. citrulli, while Nicotiana tabacum was resistance to A. citrulli, so therefore could carry nonhost resistance genes. Two T3S effectors, Aave1548 and Aave2166, triggered water soaking-like cell death in N. benthamiana, but HR-like cell death in N. tabacum. Bacterial mutagenesis and in planta disease assay confirmed that both Aave1548 and Aave2166 have significant virulence contributions to A. citrulli in N. benthamiana plant and melon seeds. Aave2166 encodes a putative acetyltransferase that belongs to the YopJ super family, which is conserved in both animal and plant pathogenic bacteria. Wild type but not the putative catalytic mutant (C232A) of Aave2166 can trigger cell death phenotype in N. benthamiana and N. tabacum. N. benthamiana yeast two-hybrid cDNA library screening using Aave2166 identified six N. benthamiana proteins/peptides which specifically interacted with Aave2166. Further characterization of these Aave2166 interactors may allow us to understand the virulence mechanism provided by Aave2166. The identification of nonhost resistance genes that can recognize Aave2166 and other type III effectors may help to develop novel strategies to control BFB disease of cucurbit.Ph. D.
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Bressan, Dayanne Fabrício. "Uso dos eliciadores acibenzolar-s-metílico e harpina na indução de resistência em plantas de feijoeiro comum ao crestamento bacteriano." Universidade Tecnológica Federal do Paraná, 2011. http://repositorio.utfpr.edu.br/jspui/handle/1/232.
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CAPESO crestamento bacteriano comum, causado por Xanthomonas axonopodis pv. phaseoli, é uma das principais doenças da cultura do feijoeiro. Em busca por alternativas no controle de doenças em plantas, o uso de eliciadores/indutores de resistência tem sido explorado. Desta forma, o objetivo deste trabalho foi verificar a eficiência dos eliciadores harpina e acibenzolar-S-metílico na indução de mecanismos de defesa em plantas de feijoeiro (Phaseolus vulgaris L.) ao crestamento bacteriano comum, causado por X. axonopodis pv. phaseoli, avaliandose possíveis alterações no processo de trocas gasosas e ativação das principais enzimas envolvidas na defesa vegetal - peroxidase, superóxido dismutase, além de proteínas e fenóis. Foram utilizadas sementes dos cultivares IAPAR 31 e IPR Colibri. Depois de emergidas as primeiras folhas trifoliadas na fase vegetativa (V3) as plantas, foram tratadas com os eliciadores acibenzolar-S-metílico, harpina e água destilada (controle). Sete dias após a primeira aplicação dos eliciadores, foi inoculado o patógeno nas folhas do feijoeiro. A avaliação da severidade foi realizada a cada 10 dias sendo a primeira avaliação realizada 15 dias após a inoculação do patógeno. As leituras de fotossíntese foram realizadas 40 dias após a inoculação quando os sintomas e danos da doença demonstravam-se estabelecidos. Para a realização das análises bioquímicas, amostras de tecidos foliares de cada tratamento foram coletadas 24 horas antes, 24, 48 e 72 horas após a aplicação dos eliciadores. Também, foram determinados o rendimento e seus componentes em ambas as cultivares.A aplicação de ASM e harpina proporcionou redução na severidade da doença do crestamento bacteriano comum nas plantas de feijoeiro da cultivar IPR-Colibri e a mesma tendência foi observada para a cultivar IAPAR 31 tratada com ASM. As plantas da cultivar IAPAR 31 tratadas com o eliciador ASM, apresentaram maiores taxas de assimilação de CO2 quando comparadas ao controle e a mesma tendência foi observada para a cultivar IPR-Colibri. A aplicação do ASM promoveu o aumento da atividade da enzima antioxidativa peroxidase e a indução da síntese de compostos fenólicos e especialmente na cultivar suscetível IPRColibri. Para o rendimento de grãos e seus componentes, não foram observadas diferenças significativas entre o controle e os eliciadores ASM e harpina. Os eliciadores ASM e harpina induziram os mecanismos de resistência em feijoeiro contra X. axonopodis pv. phaseoli, indicando a potencialidade do uso dos eliciadores na cultura do feijoeiro.
The common bacterial blight, caused by Xanthomonas axonopodis pv. phaseoli is one of the major diseases of the bean crops. In search for alternatives to control the plant diseases, the use of the elicitors / inductors of resistance has been explored. Thus, the objective of this work was to verify the effectiveness of the elicitors harpina and acibenzolar-S-methyl in the induction of the defense mechanisms in bean plants (Phaseolus vulgaris L.) against common bacterial blight caused by X. axonopodis pv. phaseoli, evaluating the possible changes in the process of gas exchange and the activation of the key enzymes involved in the plant defense - peroxidase, superoxide dismutase, besides proteins and phenols. It was used seeds from IAPAR 31 and IPR Colibri cultivars. After emerged the first trifoliate leaves in the vegetative stage (V3) the plants were treated with the elicitors Acibenzolar-S-methyl and harpina, and distilled water (control). Seven days after the first application of the elicitors, the pathogen was inoculated on the leaves of bean plants. The severity assessment was performed every 10 days and the first evaluation 15 days after the pathogen inoculation. The measurement photosynthesis were performed 40 days after the inoculation when the symptoms and the damages of the disease showed themselves established. To carry out the biochemical analysis, samples of leaf tissue from each treatment were collected 24 hours before, 24, 48 and 72 hours after the application of the elicitors. Also, it was determined the yield and its components in both cultivars. The application of ASM and harpina provided the reduction in the severity of the common bacterial blight disease of the bean plants of IPR-Colibri cultivars and the same trend was observed for the IAPAR 31 cultivars treated with the elicitor ASM. Plants by IAPAR 31 treated with the elicitor ASM showed higher rates of assimilation of CO2 when compared to the control and the same trend was observed for cultivar IPR-Colibri. The application of ASM promoted the increase of the antioxidative enzyme peroxidase activity, and the induction of the phenolic compounds synthesis and, especially in the susceptible cultivar IPR-Colibri. For grain yield and its components, significant differences were not observed between the control and the elicitors ASM and harpina. The elicitors ASM and harpina induced the resistance mechanisms in bean plant against X. axonopodis pv. phaseoli, indicating the potentiality of the elicitor utilization in bean crops.
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Rankin, Lynda. "Evaluation of native rhizosphere bacteria for use as biological control agents against Pythium aphanidermatum root rot of European greenhouse cucumbers." Thesis, McGill University, 1992. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=56660.
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Thirty-two isolates of rhizosphere bacteria, selected for their ability to inhibit zoospore germination and/or mycelial growth of Pythium aphanidermatum (Pa) in vitro, were evaluated in a test tube bioassay using cucumber c.v. 'Straight 8'. These isolates were identified as Psedudomonas corrugata (Pc13 or 35) and P. fluorescens (Pf15, 16 or 27). All but one of the five isolates effectively colonised the roots of cucumber plants in short term studies. Isolates 15 and 35 were found to maintain high population densities throughout the time period. Pa-inoculated plants treated with the Pc13 or Pf15 produced fruit yields equal to 92 and 74% respectively of the control (no Pa, no bacteria). Pa-inoculated plants without bacteria yielded only 46% of the control. In the fall crop, Pa-inoculated plants treated with Pc13 or Pf15 yielded 52 and 47% of the control compared to Pa-only treatment, which yielded 12.5% of the control. In both crops, treatment with any of the bacterial isolates resulted in significantly reduced cull rates compared to the Pa-only treatment.
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Matta, Frederico de Pina. "Mapeamento de QRL para Xanthomonas axonopodis pv. passiflorae em maracujá-amarelo (Passiflora edulis Sims f. flavicarpa Deg.)." Universidade de São Paulo, 2005. http://www.teses.usp.br/teses/disponiveis/11/11137/tde-19052005-154945/.
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Dando continuidade aos trabalhos realizados no Departamento de Genética da ESALQ/USP, referentes aos estudos de mapeamento de genes de resistência à bacteriose em maracujá-amarelo, foram realizados dois novos experimentos de inoculação envolvendo uma população obtida a partir do cruzamento entre IAPAR-06 e IAPAR- 123, ambos acessos pertencentes ao Instituto Agronômico do Paraná (IAPAR). Essa população F1, composta de 160 indivíduos, foi utilizada para a construção dos mapas de ligação com base em marcadores AFLP, utilizando tanto marcas com segregação 1:1 quanto marcas bi-parentais 3:1, as quais serviram de ponte para a integração dos mapas construídos para cada genitor. Para a avaliação fenotípica, 104 indivíduos, além dos genitores, foram inoculados por dois isolados de Xanthomonas axonopodis pv. passiflorae, em dois experimentos distintos. Foi constatado que a resistência à bacteriose é poligênica e há, pelo menos, três locos quantitativos (QRL) envolvidos. Testes quanto à metodologia de mensuração das lesões foliares também foram realizados. Foi verificado que, independente da metodologia de avaliação fenotípica, os resultados foram equivalentes quanto ao mapeamento de QRL. De acordo com as diferentes datas de avaliação e/ou idades das folhas, QRL com diferentes efeitos foram detectados ou, até mesmo, QRL foram alocados em diferentes grupos de ligação. Os dados oriundos do presente estudo corroboram resultados obtidos em trabalho anterior envolvendo a mesma população de mapeamento. A utilização de um segundo isolado bacteriano possibilitou a detecção de um novo QRL. As limitações do uso de marcadores dominantes para fins de mapeamento de locos quantitativos são discutidas.Continuing the studies carried out in the Genetics Department at ESALQ/USP concerning the mapping of resistance genes for the bacterial disease in yellow passion fruit, two new assays were conducted involving a population derived from a cross between IAPAR-06 and IAPAR-123, both accessions belonging to Instituto Agronômico do Paraná (IAPAR). This F1 population, composed of 160 individuals, was used for constructing genetic maps based on AFLP markers, using both markers with segregation 1:1 and with biparental 3:1, which act as a bridge to integrate the maps constructed for each of the parents. For phenotypic evaluation, 104 individuals plus the parents were inoculated with two isolates of Xanthomonas axonopodis pv. passiflorae in two different assays. It was seen that the resistance to bacterial disease is polygenic, and there are at least three quantitative loci (QRL) involved. Tests regarding methods for measuring leaf lesions were also carried out. It was seen that, irrespective of the methodology used for phenotypic evaluation, the results were identical for QRL mapping. Different evaluation times and/or leaf ages resulted in detection of QRL with different effects, and even QRL allocated on different linkage groups. Data derived from the present study corroborate results obtained previously involving the same mapping population. The use of a second bacterial isolate did allow a new QRL to be detected. The limitations of using of dominant markers for mapping quantitative loci are discussed.
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Mano, Emy Tiyo. "Identificação de genes de Burkholderia sp. associados ao controle biológico de Pectobacterium carotovora." Universidade de São Paulo, 2011. http://www.teses.usp.br/teses/disponiveis/87/87131/tde-23032011-143651/.
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A bacteria Pectobacterium carotovora causa danos a diferentes hospedeiros por meio da produção de enzimas pectinolíticas que degradam o pectato de cálcio da lamela media próximo a parede celular, causando extravasamento do conteúdo celular, sintomas da podridão mole. Bactérias do gênero Burkholderia tem se mostrado capazes em controlar a podridão mole em orquídeas, no entanto, os aspectos moleculares envolvidos neste controle ainda não foram estudados. Neste trabalho, foram avaliados 602 transformantes quanto a sua habilidade em inibir os sintomas da podridão mole, onde foram observados 16 mutantes defectivos no controle da doença. Destes, foram identificados sete diferentes genes inativados pelo transposon Tn5, e estes genes podem estar envolvidos em processos de síntese de aleloquímicos, competição por nutrientes, adaptação a condições ambientais, e na interação com o hospedeiro e/ou entre microrganismos. No entanto, o envolvimento destes genes na perda da capacidade em controlar a podridão mole deve ser melhor estudado.The bacterium Pectobacterium carotovora cause damage to different hosts and by production of pectic enzymes that degrade calcium pectate of the middle lamella near of the cell wall, causing overflow of cell content and consequently the soft rot. Burkholderia genus has proven able to control the soft rot in Orchids, however, the molecular aspects involved in the control have not been studied. In this work, 602 transformants were characterized for their ability to inhibit soft rot caused by P. carotovora. We identified 16 mutants showing shifts in inhibition pattern or lost of the ablitity to inhibit soft rot symptoms. Among these mutants, we identified 7 genes related to disease inhibition,and this genes may be involved in process of allelochemicals synthesis, competition for nutrients, adapting to environmental conditions, and interaction between the host and microorganisms. However, the involvement of these genes in loss of ability to control the soft rot disease is being further studied in details.
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Takahashi, Elizabete Keiko. "Transferência do gene atacina A para plantas de maracujá amarelo (Passiflora edulis Sims f. flavicarpa Deg.) por biobalística." Universidade de São Paulo, 2002. http://www.teses.usp.br/teses/disponiveis/11/11137/tde-03122002-083637/.
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O Brasil é o principal produtor de maracujá amarelo. Entretanto, a produtividade é baixa, cerca de 1 0.000 t por hectare. A produção de frutos varia com o cultivar, condições climáticas, manejo e outros fatores, principalmente doenças causadas por bactérias e vírus. Metodologias de transformação genética são alternativas modernas para obter plantas resistentes. A proteína derivada de inseto, atacina A atua como bactericida, e tem sido utilizada para conferir resistência a espécies vegetais. Os objetivos do presente estudo foram (i) obter a regeneração de brotos in vitro, (ii) testar a eficiência de agentes seletivos durante o processo organogênico, (iii) construir o cassete contendo o gene atacína A, e (iv) determinar as condições físicas e biológicas para a transformação genética de plantas de maracujá amarelo utilizando o método de biobaiística. Em relação a estudos in vitro, três recipientes de cultura foram avaliados como também diferentes concentrações de benzylaminopurina (BA) e água de coco que foram adicionadas ao meio basal. Phytagei e agar também foram testados como agentes solidificantes. As culturas foram avaliadas quanto à resposta morfogênica dos discos foliares. O gene atacina A foi sequenciado e clonado para receber o promotor CAMV 35S com um enhancer duplicado e o terminador 35S. Este vetor foi denominado pFFatacina. O cassete de expressão foi cionado nos vetores pcambia 1300 e pcambia 2300 que contêm os genes higromicina (hpt) e canamicina (nptll), respectivamente. Discos foliares, assim como segmentos entrenodais e hipocotiledonares que induzem calos, foram usados nos experimentos de biobaiística. A expressão do gene uida foi avaliada para testar os parâmetros de bombardeamento, pressão de gás Hélio (psi) e a distância da tela de retenção até o tecido alvo (cm). A resposta organogênica dos discos foliares não diferiu quando placas de petrí, tubos ou frascos foram usados, embora os tubos (2,4 x 8,5 cm, 30 mi) mostraram uma resposta ligeiramente melhor. O meio MS (Murashige & Skoog, Physiologia Plantarum, 15, 1962) solidificado com agar (0,6%) e suplementado com 0,5 mg/L BA e 5% de água de coco (w/v) provou ser eficiente na indução de organgênese. Brotos foram obtidos após 30 dias. Segmentos entrenodais e hipocotiledonares produziram 300 estruturas semelhantes a gemas por explante. Microscopia de varredura e análises histológicas demonstraram ser estruturas foliares, as quais evoluíram em brotos após 50 dias em Y2 MS. Higromicina a 5 mg/L provou ser um agente seletivo apropdado, inibindo organogênese em 60% dos explantes. Canamicina a 50 mg/L foi também efetiva. Calos morfogênicos de até 10 dias e discos foliares de 3 dias de cultivo mostraram elevados níveis de expressão transiente sob 80016,5 ou 100019,5 (psi de gás Héliolcm de distância de võo dos microprojéteis). Foram realizados experimentos de co-transformação com pB[426 (8,7 kb) que contém o gene npdi e pFFatacina (5,25 kb), como também utilizando-se um único vetor (pcatacina 1300), Freqüência de transformação estável de 0,85% foi obtida. A integração do transgene foi confirmada por PCR para o gene atacina A. Este é o primeiro trabalho que relata a transferência de um gene de interesse para plantas de maracujá amarelo por biobalística.Brazil is the leading producer of the yellow passion fruit. However, the productivity is fairly low, about 10,000 t per hectare. Fruit yields vary with cultivars, climatic conditions, management and other factors, namely bacterial and virus díseases. Genetic transformation methodologies are modern alternatives to obtain plant resistance. The insectderived protein, attacin A acts as bacterícide, and it has been used to confer resistance to plant species. The objectives of the present study were (i) to obtain in vitro shoot regeneration, (ii) to test the efficiency of certain selective agents during the organogenesis process, (iií) to construct the cassette containing the attacin A gene, and (iv) to determine the physical and biological conditions for genetic transformation of passion fruit plants by using the biolistic approach. Regarding the ín vítro studies, three culture recipients were evaluated as well as different concentrations of benzylaminopurine (BA) and coconut water that suppiemented the basal medium. Phytagei and agar were aiso tested as solidifying agents. Cultures were evaluated with respect to leaf dises morphogenic responses. The attacín A gene was sequenced, and cioned to receive the CAMV 35S promoter with a duplicated enhancer sequence, and the 35S terminator. This vector was denoted pFFatacina. The cassette was cioned in pcambia 1300 and pcambia 2300 vectors that contain the hygromícin (hpt) and kanamycin (nptil) genes, respectively. Leaf discs, as well as internodal segments and hypocotyl-derived sections chosen to índuce calii, were used in the biolistic experiments. The uida gene expression was evaluated for testing bombardment parameters, namely the helium pressure (psi) and the distance from the stopping screen to the target tissue (cm). The organogenic response of the leaf discs did not differ when petri dishes, tubes or culture vesseis were used although the tubes (2,4 x 8,5 cm, 30 ml capacity) showed to be slightly better. The agar (0.6%)-solidifíed MS (Murashige & Skoog, Physíología Plantarum, 15, 1962) medium suppiemented with 0.5 mg/L BA and 5% (wlv) coconut water proved to be efficient to induce organogenesis. Shoots were obtained after 30 days. Internada[ and hypocotyl-derived segments produced 300 bud-like structures per explant. Scanning microscopy and histologícal anaiyses provided evidences that they were leaf structures, which last 50 days in 1/2 MS to evolve into shoots. Hygromicin at 5 mg/L proved to be proper as selective agent, inhibiting organogenesis in 60% of the explants. Kanamycin at 50 mg/L was also effective. Morphogenic calli up to 10 days old and 3 d-leaf discs showed high levels of transient uida gene expression under 80016.5 or 1000/9.5 helium pressureldistance from the stopping screen to the target tissue. Cotransformation experiments with pBI426 (8.7 kb) that contain the nptil gene and pFFatacina (5.25 kb) were carried on as well singre vector transformation tríals (pcatacina 1300). Stable transformation frequency of 0.85% was obtained. Transgene integration was confirmei by PCR for the attacin A gene. This is the first report on agronomicaily useful-gene transfer to yeilow passion fruit plants by biolistics.
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Gonçalves, Fabrício Packer. "Progresso poliético e quantificação de danos da Clorose Variegada dos Citros em laranjeiras \'Natal\' submetidas a déficits hídricos." Universidade de São Paulo, 2010. http://www.teses.usp.br/teses/disponiveis/11/11135/tde-19042010-151206/.
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A Clorose Variegada dos Citros (CVC), causada pela bactéria Xylella fastidiosa, é atualmente a doença de maior incidência no Estado de São Paulo. A incidência e a severidade da doença são maiores nas regiões Norte e Noroeste do Estado, onde o clima é predominantemente quente e com déficit hídrico, principalmente no inverno. Embora não haja nada comprovado, alguns estudos indicam que o déficit hídrico aumenta a severidade dos sintomas nas plantas afetadas pela CVC. Além disso, até hoje não foi estabelecida uma função que relacione doença e dano. Dessa forma, o estudo presente objetivou avaliar o efeito de diferentes níveis de irrigação no progresso poliético da CVC, avaliar métodos de avaliação da doença e determinar uma função de dano para a CVC. Para isso, um experimento foi instalado no município de Bebedouro, região Norte do Estado de São Paulo sob o delineamento blocos ao acaso no esquema fatorial 3 x 2, com os seguintes tratamentos: sem irrigação, irrigado com 50% e 100% da evapotranspiração da cultura (ETc), combinados com inoculação natural e artificial de X. fastidiosa. Cada parcela foi constituída de 6 plantas úteis, havendo 4 repetições de cada tratamento, totalizando 144 plantas úteis. As variáveis quantificadas em cada árvore do experimento foram: (a) produção, considerando frutos assintomáticos e sintomáticos, no período de 2006 a 2008 e produção total referente às safras 2001 a 2005; (b) intensidade da doença, através da incidência de ramos com sintomas da CVC e utilizando uma escala descritiva; (c) vigor da planta, estimado pela medida da altura da planta, diâmetro da copa, diâmetro do tronco, volume de copa e do índice de área foliar (IAF). O modelo monomolecular foi utilizado para descrever o progresso poliético da incidência da CVC, no período de 2006 a 2008, para todos os tratamentos. A partir desse modelo foi possível observar que plantas sem irrigação apresentam maior incidência de ramos com sintomas que plantas irrigadas a 100% da ETc, principalmente quando inoculadas de forma artificial. Esse resultado ficou ainda mais evidenciado quando se avalia o número de frutos com sintomas (ø < 50mm) da CVC por tratamento. Também foi possível observar que os sintomas nas plantas são distribuídos de forma irregular, apresentando um gradiente decrescente significativo da parte superior para inferior. As variáveis relacionadas ao vigor da planta, na sua maioria, não apresentaram relação alguma com a produção em todos os tratamentos. De modo geral, as plantas do tratamento irrigado a 100% da ETc combinado com inoculação natural apresentaram maior produção total, considerando as safras de 2001 a 2008. A relação entre CVC e dano pode ser descrita pela equação (y=114,067*exp(-0,017555)*x) onde y é a produção da planta em Kg e x a incidência de ramos com CVC em porcentagem. A escala descritiva não permitiu avaliação precisa e apresentou baixa correlação entre os avaliadores, o contrário foi observado realizando a avaliação da CVC através da incidência de ramos com sintomas.The Citrus Variegated Chlorosis (CVC) caused by the bacteria Xylella fastidiosa is nowadays the disease with the most incidence in the state of São Paulo. The incidence and severity of the disease are greater in the Northern and Northwestern regions of the state where the climate is predominately warm and presents a water deficits especially in the winter. Although there is no definite proof, some studies indicate that the water deficits increases the severity of the symptoms in plants affected by the CVC. Besides, up to the present there has not been a function which relates the damage to an intensity of the disease. In that manner, the present study has been conducted in order to evaluate the effect of different irrigation levels on the polyetic progress of the CVC as well as to study methods of assessment of the disease and determine the relationship between disease intensity and yield. In order to carry out this study, an experiment was conducted in the city of Bebedouro north of São Paulo state. The experimental design was in randomized blocks in a 3 x 2 factorial scheme with the following treatments: no irrigation, irrigated with 50% and 100% of the evapotranspiration of the crop (ETc), combined with natural and artificial inoculation of X. fastidiosa. Each group was made of 6 useful plants, with 4 repetitions for each treatment, totalizing 144 useful plants. The variables quantified in each tree of the experiment were: (a) (a) yield, considering asymptomatic and symptomatic fruit in the period 2006 to 2008 and total yield on the crops from 2001 to 2005; (b) intensity of the disease through incidence of branches with symptoms of CVC and using a descriptive scale; (c) plant vigor, estimated through measure of height, crown diameter, trunk diameter, canopy volume and of the leaf area index (LAI). The monomolecular model was used to describe the polyetic progress of CVC incidence between 2006 and 2008 for all treatments. From this model it was possible to observe that plants without irrigation present greater incidence of branches with symptoms than the ones irrigated at 100% of ETc, especially when inoculated in an artificial manner. This result was even more evident when the number of fruit with symptoms is evaluated (ø < 50mm) of CVC per treatment. It was also possible to observe that the symptoms in the plants are scattered in an irregular way, presenting a significant decreasing rate from the upper to the lower part. Most of the variables related to the plant vigor did not present any relation to the production in all treatments. In general, the plants from the irrigated treatment at 100%of ETc combined with natural inoculation presented more total yield considering the harvests from 2001 to 2008. The relationship between CVC and damage can be described by the equation (y=114,067*exp(-0,017555)*x) where y is the plant yield in Kg and x is the incidence in percentage of branches with CVC. The descriptive scale did not allow accurate assessment and showed a low correlation between the raters, the opposite was observed performing the assessment of CVC through the incidence of branches with symptoms.
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Felipe, Rafaella Teles Arantes. "Avaliação da resistência à Candidatus Liberibacter asiaticus em laranja doce expressando o gene attA ou hrpN." Universidade de São Paulo, 2012. http://www.teses.usp.br/teses/disponiveis/11/11144/tde-23032012-093114/.
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Huanglongbing (HLB), considerada uma das mais graves doenças dos citros, está associada a Candidatus Liberibacter spp., bactérias endógenas restritas ao floema e de difícil cultivo em meio de cultura. Diferentemente de outras doenças que afetam plantas cítricas, ainda não foram encontradas dentro do gênero Citrus espécies resistentes ao HLB. Genes de interesse agronômico têm sido empregados na transformação genética de citros visando a resistência a doenças. Dentre estes, destacam-se os que conferem resistência a bactérias incluindo attA, que codificam os peptídeos antibacterianos atacinas, e hrpN, que ativam a produção de proteínas harpinas, relacionadas ao sistema de defesa. O objetivo deste trabalho foi avaliar a resistência de plantas de laranja doce contendo o gene da atacina A (attA) ou o gene da harpina (hrpN) à Candidatus Liberibacter asiaticus (CLas) utilizando duas formas de inoculação: borbulhões infectados e o inseto vetor da bactéria, Diaphorina citri. Para as plantas contendo o gene hrpN, apenas o segundo método foi utilizado. Os principais sintomas do HLB foram observados aos quatro e oito meses após a inoculação por borbulhões. A infecção das plantas foi confirmada com a detecção de CLas por PCR (quatro e oito meses) e Rt-qPCR (oito meses). Em plantas inoculadas por D. citri, os sintomas foram observados oito e doze meses após a inoculação, assim como a detecção da bactéria por PCR. Após 15, 17 e 18 meses, foi realizada uma nova avaliação por Rt-PCR a partir de spots (imprints em membrana) de folhas. Rt-PCR foi empregado também em spots dos psilídeos utilizados na inoculação. Não foi possível avaliar a resistência ao HLB em plantas contendo o gene attA ou hrpN a partir da inoculação por D. citri. Os resultados de detecção da bactéria nas plantas e nos psilídeos utilizados para inoculação indicam que, possivelmente, não ocorreu a inoculação devido ao baixo percentual de psilídeos que continham CLas utilizados. Dentre as plantas transgênicas contendo o gene attA inoculadas por borbulhões infectivos, oito eventos (cinco de laranja Pera, dois de laranja Hamlin e um de laranja Valência) apresentaram menores títulos bacterianos e algumas também demonstraram redução dos sintomas do HLB quando comparadas com plantas não transgênicas, oito meses após a inoculação, indicando uma possível ação do peptídeo atacina A contra o agente causal do HLB.Haunglongbing (HLB), considered one of the most serious diseases of citrus, is associated to Candidatus Liberibacter spp., endogenous and phloem-inhabiting bacteria not easily grown in culture medium No species within the genus Citrus is known to resist this bacterial infection. The use of genes of agronomic interest for genetic transformation aiming disease resistance in citrus has been reported. Among these genes, attA that codes for the antibacterial peptides attacin, and hrpN, that codes for proteins harpin that activate the plant defense system may have potential in searching for HLB resistance. The objective of this study was to evaluate the resistance of sweet orange containing attA or hrpN to Candidatus Liberibacter asiaticus (CLas) inoculated through infected budstick grafting or the insect vector, Diaphorina citri. For the plants containing hrpN, only the second method was used. The most obvious HLB symptoms were observed four and eight months after inoculation by infected budstick when CLas also was detected by PCR (four months) and RT-qPCR (eight months). For those inoculated with D. citri, symptoms were observed and bacteria detected eight and twelve months after inoculation. Fifteen, 17 and 18 months after inoculation, a new attempt was made for CLas detection, now through Rt-PCR from leaf and psyllids imprinting spots on membrane. It was not possible to evaluate the HLB resistance in plants containing attA or hrpN gene from D. citri inoculation. The results of CLas detection in plants and psyllids indicate that possibly there was no inoculation due the low rate of psyllids contained CLas used. Among the plants containing attA, five, two and one event of, respectively, Pera, Hamlin, and Valencia sweet orange had lower bacterial titers than those non transgenic plants and some also showed milder HLB symptoms, eight months after inoculation, suggesting a possible effect of attacin A against the causal agent of HLB.
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Rodrigues, Italo Sarto Carvalho 1983. "Análise da diversidade bacteriana associada ao biofilme dentário por polimorfismo de comprimento de fragmentos terminais de restrição (T-RFLP)." [s.n.], 2011. http://repositorio.unicamp.br/jspui/handle/REPOSIP/290618.
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Orientadores: Daniel Saito, José Francisco HoflingDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba
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Resumo: As comunidades bacterianas presentes na cavidade bucal desempenham papel importante no equilíbrio saúde/doença em seres humanos. Nesse sentido, o estudo da composição de microambientes orais pode contribuir para uma melhor precisão no diagnóstico de doenças infecciosas e, consequentemente, para o desenvolvimento de abordagens terapêuticas mais eficazes. A periodontite é uma doença dos tecidos de suporte do dente, caracterizada por resposta imunológica exacerbada do hospedeiro, frente à presença bacteriana no biofilme dentário. A grande diversidade bacteriana observada na cavidade bucal, aliada aos variados quadros clínicos da periodontite, ressaltam a necessidade de investigações mais aprofundadas sobre a composição bacteriana periodontopatogênica. O objetivo deste trabalho é a caracterização da comunidade bacteriana associada ao biofilme periodontopatogênico, pelo uso da técnica de Polimorfismos de Comprimento de Fragmentos Terminais de Restrição (T-RFLP). Amostras de biofilme dentário supragengival e subgengival foram coletadas de 11 pacientes portadores de periodontite. O DNA amostral foi extraído e submetido à Reação em Cadeia da Polimerase (PCR) direcionada ao gene ribossomal 16S, utilizando-se iniciador senso marcado com molécula repórter fluorescente. Os produtos da PCR foram digeridos pelas endonucleases tetraméricas HhaI, MspI e RsaI e os fragmentos terminais de restrição (T-RFs) resultantes foram analisados em um sequenciador automatizado de DNA, gerando diferentes perfis de fragmentos para cada amostra. Ao todo, 19 T-RFs distintos foram detectados com a enzima RsaI (média = 6,4), 61 com MspI (média = 19,3) e 63 com HhaI (média = 16,3). Uma grande variabilidade nos perfis de restrição foi observada, sendo que 51% a 63% dos T-RFs foram detectados em menos de quatro amostras. A predição taxonômica de T-RFs in silico demonstrou a presença de gêneros bacterianos reconhecidamente periodontais, incluindo Actinomyces, Eubacterium, Fusobacterium, Haemophilus, Porphyromonas, Prevotella e Propionibacter. Embora alguns gêneros tenham sido encontrados em todas as amostras avaliadas, as análises de clusterização e estatística multivariada não demonstraram agrupamentos de perfis T-RFLP conforme paciente ou sítio amostral. Os resultados do estudo permitem concluir que a comunidade bacteriana do biofilme periodontopatogênico é bastante variável entre indivíduos, embora possua alguns gêneros predominantes
Abstract: The bacterial communities present in the oral cavity play an important role in maintaining healh/disease equilibrium. In this sense, studying the microbial compostition of the oral environment may contribute to attain a better precision in diagnosis of infectious diseases and, also, to develop efficient therapeutic approaches. Periodontitis is a disease that affects the supporting tissues of the tooth, characterized by a heightened immunologic response to bacteria present in dental biofilm. The broad microbial diversity present in the oral cavity, along with the various clinical features of periodontitis, highlight the necessity of further investigations on the composition of periodontopathic biofilm. The objective of this study is the characterization of the bacterial communities associated with periodontopathic biofilm, by use of the Terminal Restriction Fragment Length Polymorphism (T-RFLP) techique. Supra and subgingival biofilm samples were collected from 11 periodontitis subjects. Total DNA was extracted from the samples and submitted to the Polymerase Chain Reaction (PCR) targeting the 16S rRNA gene, with a fluorescently labeled forward primer. PCR products were digested with the tetrameric endonucleases HhaI, MspI and RsaI, and the resulting terminal restriction fragments (T-RFs) were analyzed in an automated DNA sequencer. In all, 67 T-RFs (mean = 17.3) were detected with RsaI endonuclease, 61 T-RFs with MspI (mean = 19.3) and 19 T-RFs (mean = 6.4) with HhaI. A great variability in the restriction patterns was observed, since 51% to 63% of T-RFs were detected in less than 4 samples, and two T-RFs were exclusively found in supragingival biofilm (p = 0.018). In silico taxonomical prediction of T-RFs demonstrated the presence of well-known periodontal species belonging to the Acinomyces, Eubacterium, Fusobacterium, Haemophilus, Porphyromonas, Prevotella and Propionibacter genera. Although some species were found in all samples, clustering and multivariate statistical analysis did not reveal evident groupings of T-RFLP profiles according to patient or sampling site. The results of this study indicate that the microbiota of periodontopathic biofilm is highly variable among subjects, albeit a core microbial community may be observed
Mestrado
Microbiologia e Imunologia
Mestre em Biologia Buco-Dental
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Vrisman, Claudio M. "Real-time imaging and characterization of colonization of cucurbit hosts by Erwinia tracheiphila, the impact of intra-specific competition, and the discovery and characterization of novel approaches to manage bacterial wilt of cucurbits." The Ohio State University, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=osu1542889416739096.
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Aquino, Ana Rafaela Luz de. "Detec??o de bact?rias periodontopatog?nicas cultiv?veis e n?o cultiv?veis em placas ateromatosas." Universidade Federal do Rio Grande do Norte, 2009. http://repositorio.ufrn.br:8080/jspui/handle/123456789/17045.
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Periodontal infections consist of a group of inflammatory conditions caused by microorganisms that colonize the tooth surface through the formation of dental biofilm. Chronic infections such as periodontitis have been associated to the development and progression of atherosclerosis. AIM: Detect cultivatable and non-cultivatable periodontopathogenic bacteria in atheromatous plaques; search for factors associated to the presence of these bacteria in the atheromatous plaques and characterize the presence of cultivatable and non-cultivatable bacteria in these plaques. METHODOLOGY: A cross-sectional study was performed with a sample of 30 patients diagnosed with atherosclerosis in the carotid, coronary or femoral arteries and surgically treated with angioplasty and stent implant, bypass or endarterectomy. The plaques were collected during surgery and analyzed using the PCR molecular technique for the presence of the DNA of the cultivatable bacteria Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Treponema denticola and of the non-cultivatable Synergistes phylotypes. The patients were examined in the infirmary, after the surgery, where they also responded to a questionnaire aimed at determining factors associated to the presence of periodontopathogenic bacteria in the atheromatous plaques. RESULTS: All patients with tooth (66,7%) possessed disease periodontal, being 95% severe and 65% widespread. No periodontopathogenic bacteria were found in the atheromatous plaques. However, four samples (13.3%) were positive for the presence of bacteria. Of these, three participants were dentate, being two carriers of widespread severe chronic periodontite and one of located severe chronic periodontitis. None of them told the accomplishment of procedures associated to possible bacteremia episodes, as treatment endodontic, extraction the last six months or some procedure surgical dental. CONCLUSION: The periodontopathogenic bacteria studied were not found in the atheromatous plaques, making it impossible to establish the prevalence of these pathogens or the factors associated to their presence in plaques, the detection of positive samples for bacteria suggests that other periodontal and non-periodontal pathogens be studied in an attempt at discovering the association or not between periodontal disease and/or others infections and atherosclerosis, from the presence of these bacteria in atheromas
As infec??es periodontais compreendem um grupo de condi??es inflamat?rias, causadas por microrganismos que colonizam a superf?cie dent?ria, atrav?s da forma??o do biofilme dent?rio. Atualmente, infec??es cr?nicas como as periodontais t?m sido associadas ao desenvolvimento e progress?o da aterosclerose. OBJETIVOS: Detectar bact?rias periodontopatog?nicas cultiv?veis e n?o cultiv?veis em placas ateromatosas; buscar fatores associados ? presen?a destas bact?rias nas placas ateromatosas e caracterizar a presen?a de bact?rias cultiv?veis e n?o cultiv?veis em placas ateromatosas. METODOLOGIA: Foi realizado um estudo seccional, utilizando uma amostra de 30 pacientes que apresentaram o diagn?stico de aterosclerose nas art?rias car?tidas, coronarianas ou femorais, e foram tratados cirurgicamente atrav?s da realiza??o dos procedimentos de angioplastia com implante de stent, bypasse ou endartarectomia. As placas foram coletadas durante as cirurgias realizadas e analisadas atrav?s da t?cnica molecular de PCR para a presen?a de DNA das bact?rias periodontais cultiv?veis Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis e Treponema denticola e o filotipo n?o-cultiv?vel Synergistes species. Os pacientes foram examinados no leito da enfermaria, ap?s as cirurgias, onde tamb?m responderam a um question?rio cujo objetivo foi buscar fatores associados ? presen?a de bact?rias periodontopatog?nicas nas placas ateromatosas. RESULTADOS: Todos os pacientes dentados (66,7%) possu?am doen?a periodontal, sendo 95% severa e 65% generalizada.Nenhuma bact?ria periodontopatog?nica pesquisada foi encontrada nas placas ateromatosas. No entanto, quatro amostras (13,3%) foram positivas para a presen?a de bact?rias. Destas, tr?s participantes eram dentados, sendo dois portadores de periodontite cr?nica severa generalizada e um de periodontite cr?nica severa localizada. Nenhum deles relatou a realiza??o de procedimentos associados a poss?veis epis?dios de bacteremia, como tratamento endod?ntico, exodontia nos ?ltimos seis meses ou algum procedimento cir?rgico odontol?gico. CONCLUS?O: As bact?rias periodontopatog?nicas estudadas n?o foram encontradas nas placas ateromatosas, n?o pudendo ser estabelecida a preval?ncia destes pat?genos e nem os fatores associados ? presen?a deles nas placas, mas, a detec??o das amostras positivas para bact?rias abre caminhos para que outros pat?genos periodontais e n?o periodontais sejam estudados na tentativa de elucidar de maneira definitiva a associa??o ou n?o entre a doen?a periodontal e/ou outras infec??es e a aterosclerose, atrav?s da presen?a destas bact?rias nos ateromas
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Dore, Dalin Shelley. "Grapevine rhizosphere bacteria : influence of diversity and function on two root diseases : a thesis submitted in fulfilment of the requirements for the degree of Master of Science at Lincoln University /." Diss., Lincoln University, 2009. http://hdl.handle.net/10182/1305.
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The overall goal of this research was to determine what, if any, role grapevine rhizosphere bacteria play in the differing susceptibilities of New Zealand grown rootstocks to Cylindrocarpon black foot disease. The size and diversity of bacterial populations associated with the rhizospheres of grapevine rootstocks: 101-14, 5C, Schwarzmann and Riparia Gloire were evaluated. Dilution plating showed that total bacterial (P=0.012, P=0.005 for NA and KB, respectively) and fluorescent Pseudomonad (P=0.035) rhizosphere counts differed between rhizosphere and bulk soils but did not correlate with the differing susceptibilities of the rootstock varieties to black foot. No varietal differences were found for spore forming bacteria (P=0.201). SSCP banding patterns showed that species diversity was similar for most rootstocks, but that there were some differences in the composition of bacterial populations, probably attributable to vigour. Some functional characteristics of the bacteria isolated from the rhizospheres of the most and least susceptible rootstock varieties were assessed to investigate their potential to suppress the pathogen. In dual culture, bacteria from Riparia Gloire, 101-14 and the control soil all had little ability to antagonise Cylindrocarpon destructans. However, they differed in their degrees of activity for glucanase (P=0.000), protease (P=0.001) and siderophores (P=0.000). In all tests, bacterial isolates from the rhizosphere of 101-14 had the largest number of active isolates (P≤0.002); however, those from Riparia Gloire had the greatest degree of positive responses for the glucanase and siderophore assays. Bacterial isolates from the control soil produced few glucanases and no siderophores, but had the highest degree of protease activity. Bands excised and sequenced from SSCP gels frequently matched to other ‘uncultured bacteria’ in GenBank, as well as to other bacterial phyla, classes and genera commonly isolated from soil and sediment samples. These included members of the Firmicutes, Proteobacteria (α, δ, γ), Verrucomicrobia, Acidobacteria and Chromatiales. The pathogenicity of C. destructans and Fusarium oxysporum was investigated by inoculating soil containing wounded ungrafted rootstocks of 101-14, 5C, Schwarzmann and Riparia Gloire. Results indicated that F. oxysporum might be a more aggressive pathogen than C. destructans. Inoculation with F. oxysporum or C. destructans increased disease severity, P=0.018 and P=0.056, respectively at 0 cm. Rootstock variety influenced disease severity caused by C. destructans (P<0.001) and F. oxysporum (P=0.090), with rootstocks 101-14 and 5C being most susceptible to C. destructans, and Riparia Gloire and Schwarzmann most susceptible to F. oxysporum. There was also an indication that inoculation with one pathogen increased plant susceptibility to the other, with increased F. oxysporum infection in the C. destructans inoculated treatments of Riparia Gloire and Schwarzmann (P<0.05). The effect of carbohydrate stress (leaf trimming) and inoculation on C. destructans disease severity, incidence, and rootstock rhizosphere bacterial populations was evaluated by inoculating the soil containing one year old plants of Sauvignon Blanc scion wood grafted to rootstocks 101-14 and Schwarzmann. Disease severity and incidence was similar for both Schwarzmann (8.4% and 29.3%, respectively) and 101-14 (14.9% and 31.0%, respectively). When data for the moderate and no stress treatments were combined, because their effects were similar, the disease severity was significantly higher for the highly stressed plants(P=0.043). Stress did not influence disease incidence (P=0.551). Infection occurred in the non-inoculated plants, but disease severity was higher in the plants inoculated with C. destructans than those that were not. Root dry weight of highly stressed plants was lower than in both the moderately stressed (P=0.000) and unstressed plants (P=0.003). An interaction between inoculation and stress (P=0.031) showed that inoculated and highly stressed plants had the lowest root dry weight but there was no effect of rootstocks (P=0.062). There was no significant effect of carbohydrate stress (P=0.259) or inoculation (P=0.885) on shoot dry weight. SSCP banding patterns showed that bacterial diversity was generally similar between rootstocks, but stress and inoculation altered rhizosphere bacterial communities. This study has demonstrated that functionality of grapevine rhizosphere bacteria do differ between grapevine rootstock varieties that have different susceptibilities to black foot disease, but that this role needs to be further investigated if more accurate and practically relevant conclusions are to be drawn.
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Ros, Patricia Benites. "Avaliação da resistência de variedades de cana-de-açúcar ao raquitismo-da-soqueira com base na taxa de colonização dos colmos por Leifsonia xyli subsp. xyli." Universidade de São Paulo, 2004. http://www.teses.usp.br/teses/disponiveis/11/11144/tde-14122004-111712/.
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O método sorológico \"tissue blot enzyme immunoassay\" foi utilizado para determinar a taxa de colonização dos vasos do xilema por Leifsonia xyli subsp. xyli (Lxx) em colmos de 28 variedades de cana-de-açúcar, incluindo a CB47-355, CB41-76 e CB49-260 como padrão resistente, intermediário e suscetível, respectivamente. Para cada variedade, 16 gemas foram tratadas termicamente a 50,5ºC por 2 horas e, em seguida, submetidas a banho fungicida a base de benomyl na dose de 6,5 g do princípio ativo por 10 litros de água. Oito gemas foram imersas por 10 minutos em caldo da CB49-260 sabidamente colonizada por Lxx diluído em água destilada na proporção 1:5 (\"INOCULADO\") e a outra metade em água (\"SADIO\"). Após a inoculação, foram plantados três vasos (repetições) para o tratamento, sendo as plantas mantidas em casa-de-vegetação até a colheita durante 7 meses, quando então 2 colmos de cada vaso foram amostrados para determinar a taxa de colonização dos vasos. A variedade CB47-355 não apresentou nenhum vaso colonizado, enquanto na CB41-76 e CB49-260 a taxa de colonização foi igual a 17% e 76%, sendo classificadas como resistente, intermediária e suscetível, respectivamente, concordando com literatura prévia. Das variedades em teste, 12 comportaram-se como intermediárias e 13 como suscetíveis. No primeiro caso, a %VC variou de 7 a 25 e, nas suscetíveis, de 25,7 a 76. Portanto, o aumento da taxa de colonização é praticamente linear da mais resistente para a mais suscetível, o que é característico de caráter quantitativo. Considerando que nenhuma variedade comercial mostrou-se resistente e que Lxx causa perdas significativas em todos os países produtores de cana-de-açúcar, sugere-se que os programas de melhoramento devem dirigir programas para a seleção de genitores e progênies para obter variedades resistentes. Enquanto isso não ocorre, atenção dever ser dada para o uso de mudas indexadas.The tissue blot immunoassay (TBIA) for detecting Leifsonia xyli subsp. xyli (Lxx) colonized vascular bundles (CVB) were used in determining the reaction of 28 brazilian sugarcane cultivars to ratoon stunting disease (RSD), including CB47-355, CB41-76 e CB49-260 as resistant, intermediate and susceptible standards. For each cultivar, 16 buds were hot water treated at 50,5 oC for 2 hours and protected against root rots with benomyl. Eight buds of each cultivar were than inoculated with infected sap of the CB49-260 diluted 1:5 in water, and the remaining 8 buds were inoculated with distilled water for planting the control plots. The reps of each cultivar were planted in plastic plots at green house. At harvesting, seven months after planting, two stalks were sampled from each plot. The rate of colonization of vascular bandles by Lxx in stalks of each cultivar was determined. Statistical analysis by the univariate clustering method (Scott-Knott) was reliable for grouping genotypes according to their levels of Lxx resistance by the percentage of CVB. The cultivar CB47-355 did not present any CVB, while CB41-76 and CB49-260 presented 17% and 76%, being classified as resistant, intermediate and susceptible, in agreement with previous literature. Among the varieties in test, 12 were intermediates and 13 susceptibles. The percent of CVB among the intermediate and susceptible cultivars varied from 7% to 25% and 25,7 to 76%, respectively. The knowledge of resistance of each cultivar is essential for advising the use of complementary phytossanitary measures to control the disease. Since all cultivars are intermediate or susceptible, the sugarcane breeding programs need specific works for selection and releasing of resistant cultivars. Before this, schemes of nurseries and the use clean seed cane are recommended.
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Tomaseto, Arthur Fernando. "Capacidade de dispersão de Diaphorina citri Kuwayama (Hemiptera: Psyllidae)." Universidade de São Paulo, 2013. http://www.teses.usp.br/teses/disponiveis/11/11146/tde-21022013-160927/.
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O Greening ou Huanglongbing (HLB) é a doença mais devastadora dos citros na atualidade. Levantamentos mostram que a enfermidade está presente em cerca de 60% dos talhões do parque citrícola paulista, demostrando sua grande capacidade de disseminação. O psilídeo-asiático-dos-citros, Diaphorina citri Kuwayama (Hemiptera: Psylidae) é um importante vetor dos agentes causais associados ao HLB. Sua capacidade de dispersão está intimamente relacionada ao progresso da doença no campo. Não se tem conhecimento dos principais fatores que possam interferir no deslocamento do inseto até plantas cítricas. Sendo assim, a presente pesquisa teve como objetivo investigar a influência de alguns fatores ambientais sobre a dispersão do psilídeo D. citri, discutindo-os em relação à epidemiologia do HLB dos citros. Inicialmente foram realizados testes em laboratório para adaptação da metodologia de marcação, liberação e recaptura usando pó fluorescente. Grupos de psilídeos adultos foram marcados e sua sobrevivência avaliada, verificando a influência de quatro cores distintas de pó fluorescente (amarelo, azul, rosa e verde). Posteriormente realizaram-se estudos de campo com quatro liberações de psilídeos, com marcadores distintos, em quatro áreas experimentais circulares e adjacentes, sendo formadas por círculos concêntricos localizados a 18, 24 e 30 m de cada centro de liberação. Os insetos foram monitorados visualmente e por armadilhas adesivas amarelas. Nas 1ª e 2ª liberações, foi verificado o efeito das coberturas vegetais milho (Zea mays L.) e capim-braquiária (Brachiaria decumbens Stapf.) na dispersão de D. citri. Já nas 3ª e 4ª liberações, foi estudado o efeito de brotações no deslocamento do inseto. A influência dos ventos foi avaliada em ambas as situações. A análise de sobrevivência indicou que não houve interferência das quatro cores de pó fluorescente em D. citri, sendo a marcação visível no corpo dos insetos durante os 48 dias de avaliação. De acordo com as análises fatoriais, não foi observada qualquer interferência da cobertura vegetal milho, medindo 2 m de altura, na dispersão do inseto. Na presença de brotações, adultos de D. citri dispersaram menos em busca de novos hospedeiros, acumulando-se nas primeiras linhas de plantas cítricas. Após 6 horas da liberação, a capacidade de dispersão máxima de D. citri foi três vezes maior na condição de ausência de brotações. A dispersão média e a área total abrangida variaram assimptoticamente com o tempo, ou seja, o inseto possui inicialmente grande velocidade de dispersão, porém esta tende a se estabilizar com o incremento do tempo. Esses resultados deixam claro que D. citri realiza movimentos dependendo das condições fenológicas de seus hospedeiros, ressaltando a grande importância de se intensificar métodos de controle regionais em épocas em que o fluxo vegetativo das plantas cítricas não é tão intenso.Greening or Huanglongbing (HLB) is the most devastating disease of citrus today. Surveys show that the disease is present in about 60% of the park stands citrus Sao Paulo, demonstrating its great ability to spread. The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psylidae) is an important vector of the causative agents associated with HLB. Its ability to spread is closely related to disease progression in the field. There is no knowledge of the main factors that can affect the movement of the insect to citrus plants. Thus, the present study aimed to investigate the influence of some environmental factors on the dispersal of psyllid D. citri, discussing them in relation to the epidemiology of HLB of citrus. Initially tests were performed in the laboratory to adapt the methodology of marking, release and recapture using fluorescent powder. Groups of adult psyllids were marked and their survival assessed by checking the influence of four different colors of fluorescent powder (yellow, blue, pink and green). Subsequent studies were conducted with four field releases psyllids, with distinct markers on four experimental areas adjacent circular and being formed by concentric circles located at 18, 24 and 30 m for each center release. The insects were monitored visually and by yellow sticky traps. In the 1st and 2nd releases, was to determine the effect of cover crops maize (Zea mays L.) and grass (Brachiaria decumbens Stapf.) in the dispersal of D. citri. In the 3rd and 4th releases, were studied the effect of shootings in insect movement. The influence of the wind has been evaluated in both situations. Survival analysis indicated that there was no interference of the four colors of fluorescent powder on D. citri, and marking was visible on the body of insects during the 48 days of evaluation. According to the factor analysis, no interference of vegetation corn (measuring 2 m in height) was observed in the dispersal of the insect. In the presence of shoots, adult D. citri less dispersed looking for new hosts and it accumulate in the first lines of citrus plants. After 6 hours of release, the D. citri maximum dispersal was three times higher in the condition of absence of shoots. The dispersal medium and the total area covered varied asymptotically with time, ie the insect has initially high speed dispersal, but this tends to stabilize with increasing time. These results clearly show that D. citri performs movements depending on the phenology of its hosts, emphasizing the importance of intensifying regional control methods at times when the flow of plant citrus is not intense.
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Balbinotte, Juliana. "Desenvolvimento de sistemas de aquisição de Bacillus thuringiensis por Diaphorina citri Kuwayama (Hemiptera:Psyllidae) para estudos de patogenicidade." Universidade de São Paulo, 2011. http://www.teses.usp.br/teses/disponiveis/11/11146/tde-22032012-111908/.
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Bacillus thuringiensis (Bt) é uma bactéria entomopatogênica utilizada como biopesticida contra insetos, principalmente Lepidoptera, Coleoptera e Diptera, e na produção de organismos geneticamente modificados. Com a descoberta da capacidade de Bt se movimentar sistemicamente em plantas, potencialmente atingindo insetos sugadores, surge uma nova possibilidade para o controle de Diaphorina citri Kuwayama, que transmite bactérias associadas ao Huanglongbing, uma séria doença da citricultura. O objetivo do trabalho foi desenvolver sistemas de aquisição de Bt por D. citri, in vitro e in planta, para estudos de patogenicidade. Uma estirpe de Bt transformada com o gene green fluorescent protein (Btk-gfp), cultivada em meio NYSM, foi usada como marcador de aquisição pelo inseto e movimentação nas plantas. Para o sistema de aquisição in vitro, selecionaram-se dietas com base na sobrevivência e atividade alimentar de D. citri. Btk-gfp foi adicionada à dieta selecionada, composta por uma solução de sacarose a 30% em água mineral com corantes alimentícios (verde 0,1% e amarelo 0,4%). A dieta foi acondicionada em um sachê formado por duas membranas de Parafilm®, sobre a parte inferior de uma placa de Petri de 40 mm de diâmetro (gaiola de alimentação). Dez insetos foram confinados em cada gaiola para períodos de acesso à aquisição (PAA) de até 48 h, estabelecendo-se 10 gaiolas por tratamento; como controle, utilizou-se a dieta sem Btk-gfp. Para testes de aquisição in planta, suspensões de Btk-gfp foram inoculadas em ramos novos cortados e no solo, próximo à haste de seedlings de laranja doce [Citrus sinensis (L.) Osbeck] e de murta [Murraya paniculata (L.) Jack], e em diferentes concentrações, avaliando-se a movimentação da bactéria após um período de 48 h. Posteriormente, adultos e ninfas de D. citri foram confinados nestas plantas para avaliar a aquisição de Btk-gfp, usando-se plantas inoculadas apenas com água como controle. Para os bioensaios de patogenicidade a D. citri, 21 estirpes de Bt foram testadas no sistema de aquisição em ramos cortados de murta e 5 estirpes em ramos cortados de laranja para ninfas de 3º. ínstar. In vitro, testaram-se 3 estirpes contra ninfas de 3º. ínstar e 9 contra adultos. Btgfp foi absorvido pelas raízes e ramos novos cortados de seedlings de laranja doce e de murta, e translocado até as folhas, mostrando movimentação sistêmica. Esta estirpe foi adquirida por adultos e ninfas de D. citri que se alimentaram nos ramos novos cortados, mantidos em suspensão bacteriana. O sistema de alimentação in vitro permitiu a aquisição de Bt-gfp (pellet ressuspendido na dieta de sacarose com corantes) por adultos de D. citri com apenas 12 h de PAA, mas 30 h é um período ótimo para exposição do inseto ao sistema. Nenhuma das nove estirpes testadas no sistema in vitro foi patogênica aos adultos de D. citri. Das 21 estirpes testadas contra ninfas de 3º. ínstar em ramos novos de murta, cinco causaram mortalidade de 24 a 45%, em 5 dias. Essas cinco estirpes também foram testadas contra ninfas dem ramos de citros cortados, causando mortalidade de 35 a 75% em 48 h.Bacillus thuringiensis (Bt) is an entomopathogenic bacterium widely used as a biopesticide against insect pests, mainly Coleoptera, Lepidoptera and Diptera, or for engineering genetically-modified plants. The recent finding that Bt is able to move systemically within plants, potentially targeting piercing-sucking insects, suggests that the bacterium may be effective for microbial control of Diaphorina citri Kuwayama, the vector of Huanglongbing-associated bacteria, a serious citrus disease. The goal of this research was to develop in vitro and in planta acquisition systems of Bt by D. citri for pathogenicity assays. A transformed Bt strain with the green fluorescent protein gene (Btk-gfp), grown in NYSM medium, was used as a marker to demonstrate bacterial acquisition by the insect and movement within the plants. Artificial diets were selected for the in vitro acquisition system based on insect survival and feeding activity. Btk-gfp was added to the selected diet, a 30% sucrose solution in mineral water with green (0.1%) and yellow (0.4%) food coloring. The diet was placed inside a Parafilm® membrane sachet, covering the opening of the lower half of a 40-mm diameter culture plate, and forming the feeding cage. Ten D. citri adults or nymphs were introduced in each cage and allowed acquisition access periods (AAP) of up to 48 h on the diet; as a control, a diet without Btk-gfp was used. For testing the in planta acquisition system, Btk-gfp suspensions were inoculated in young stem cuttings or in the soil, near the stem of Citrus sinensis (L.) Osbeck and Murraya paniculata (L.) Jack seedlings, as well as in different concentrations, and bacterial movement was assessed after 48 h; plants inoculated with water were used as controls. D. citri adults and nymphs were confined on inoculated plants to verify Btk-gfp acquisition. In pathogenicity assays, 21 Bt strains were tested against 3rd instars of D. citri using the acquisition system with stem cuttings of M. paniculata and 5 strains were tested using citrus stem cuttings. The in vitro acquisition system was used to test pathogenicity of 3 and 9 Bt strains against 3rd-instar nymphs and adults, respectively. Bt-gfp absorbed by roots or young stem cuttings of inoculated C. sinensis and M. paniculata was detected in the leaves, showing systemic movement. Bt-gfp was isolated from groups of nymphs and adults that were fed on inoculated stem cuttings or on artificial diets with bacterial suspension, showing that both in planta and in vitro acquisition take place. D. citri adults can acquire Bt-gfp within 12 h of AAP to the artificial diet with bacterium inoculum, but 30 h is the optimum AAP. None of the nine Bt strains assayed in vitro were pathogenic to D. citri adults. Of 21 strains tested for pathogenicity against nymphs using inoculated stem cuttings of M. paniculata, five caused mortality rates varying from 24 to 45% mortality within 5 days. These five Bt strains were also tested against D. citri nymphs using young stem cuttings of C. sinensis, causing mortality rates of 35 to 75% within 48 h.
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Goumas, Dimitrios. "Possibilites de detection d'erwinia chrysanthemi pv. Dianthicola (hellmers) dickey 1979-agent de la bacteriose du dahlia sp. Evaluation des methodes immunoenzymatiques pour le controle sanitaire du materiel de propagation." Paris 6, 1987. http://www.theses.fr/1987PA066405.
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La bacteriose a e. Chrysanthemi pv. Dianthicola (echr) facteur limitant de la production du dahlia est transmise par la multiplication vegetative. Afin de proposer une methode de diagnostic plus precise que la detection visuelle, les methodes immunoenzymatiques ont ete etudiees et adaptees pour la detection d'echr dans les tissus du dahlia. La methode das-elisa (double antibody sandwich) est evaluee par rapport aux methodes de diagnostic de reference (isolement et immunofluorescence). Son utilisation, pour l'analyse sanitaire du materiel de propagation vis-a-vis d'echr seul et associe eventuellemnt a la mosaique du dahlia (damy), est etudiee en vue d'une selection sanitaire. Les etudes effectuees pour optimiser les reactifs, pour determiner les parametres pouvant modifier la reaction antigene-anticorps
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Ben, Chobba Kadri Inès. "Élaboration et mise en oeuvre d'une approche de caractérisation systémique d'un agent étiologique émergent à fort impact économique et de moyens de lutte biologique : application à la maladie de la feuille cassante du palmier dattier (Phoenix dactylifera L.)." Thesis, Montpellier 2, 2013. http://www.theses.fr/2013MON20077.
Full textAbstract:
La Maladie de la Feuille Cassante du Palmier dattier (Phoenix dactylifera L.) constitue un cas d'émergence d'une maladie à fort impact économique causée par un agent étiologique inconnu. Notre stratégie a visé à élaborer une approche sans à priori de l'émergence pouvant être transposée à n'importe quelle situation de ce type. En nous appuyant sur des caractérisations successives des compartiments viraux, bactériens et fongiques de tissus sains et malades, nous avons cherché à mettre en évidence des différences de composition spécifiques et de distribution de ces flores sur 2 campagnes de prélèvements réalisées en 2010 et 2012. Alors que la microscopie électronique à transmission nous a permis de visualiser des structures d'origine virale probable au niveau des chloroplastes du parenchyme chlorophyllien, une étude moléculaire de séquençage de gènes ribosomaux nous a permis de corréler l'apparition de ces structures a de profondes modifications qualitative et quantitative de la microflore endophyte. Ainsi il nous est apparu que la symptomologie de la maladie était corrélée à une modification profonde de la distribution spécifique de la microflore endophyte, visible à la fois au niveau du compartiment fongique et bactérien, suggérant la complète disparition de la pression de sélection exercée par le palmier sain sur sa flore et mise en évidence dans les 2 cas, par un shift d'une répartition de type Poisson vers une répartition normale. Dans le compartiment fongique, une claire déplétion des Pleosporaceae, associées à la plante saine pouvait ainsi être liée à une apparition de nouvelles familles (Trichocomaceae et Mycosphaerellaceae). De même, parmi les bactéries, une disparition des Rhizobium et Ensifer sp associés au compartiment racinaire de la plante saine a ainsi pu être mise en évidence, ces espèces pouvant servir ultérieurement d'indicatrices de bonne santé des palmiers. Dans une deuxième partie de notre travail nous avons cherché à utiliser des éléments de la flore endophyte mais également de substances naturelles dans la lutte biologique contre d'autres pathogènes du palmier. Ainsi, un antagonisme a été mis en évidence entre une souche endophyte d'Arthrobacter agilis et un pathogène majeur, Fusarium oxysporum sp AlbedinisThe Brittle Leaf Disease of the Date Palm Tree (Phoenix dactylifera L.) constitutes a case study of an emerging disease of economic impact caused by a yet uncharacterized etiologic agent. Our strategy was to develop an approach that could be indistinctly transposed to any situation of this type. While basing our investigations on the successive characterization of the diversity of viral, bacterial and fungal endophytic compartments of healthy and diseased Palm trees, we aimed at enlightening differences in species composition but also distribution over two sampling campaigns performed in 2010 and 2012. While transmission electronic microscopy allowed us to visualize structures of probable viral origin affecting chloroplasts of the chlorophyllic cell layer, a molecular approach based on ribosomal gene sequencing allowed us to evidence correlations between the occurrence of such structures and deep modifications of the structure of the palm date tree associated endophytic flora suggesting a strong depletion of the ability of the palm to control its associated endophytes. This was evidenced in both fungal and bacterial compartments by a shift from a Poisson like diversity distribution towards a Gaussian distribution in the flora associated to MFC affected palms. In the fungal compartment, Pleosporaceae, that dominated in healthy palms were replaced by an opportunistic flora of Trichocomaceae and Mycosphaerellaceae. Among bacteria, the disappearance of Rhizobium and Ensifer species, typically associated to the root compartment of healthy palms was enlighten, suggesting that these species could indeed be used as biomarkers of healthy plant status. In a second part of this study, we investigated the potential use of cultivable palm endophytes, but also natural compounds for biocontrol applications. In particular, we evidence the antagonistic potential of Arthrobacter agilis, a palm endophyte, against a major palm date disease agent, Fusarium oxysporum sp. Albedinis
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Thomas, Matthew Robert. "Functional analysis of the ALS/FTD associated gene FUS using a novel in vitro genomic DNA expression system." Thesis, University of Oxford, 2013. http://ora.ox.ac.uk/objects/uuid:176cbd1b-623e-41cb-9237-cd6c82fc0ad4.
Full textAbstract:
Aggregations of fused in sarcoma (FUS), a multifunctional RNA processing protein, define a pathological subtype of both frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS), whilst mutations in the FUS gene are causative for ALS. To model the impact of FUS mutations, expression vectors containing the entire genomic sequence of FUS, up and downstream regions, and native promoter sequences have been generated. The constructs have been tagged with an mCherry fluorescent tag, and three separate pathological mutations (R244C, R521C, and P525L) have been separately inserted. Transgenic mice have been generated using the WT and P525L FUS vectors to provide a highly physiological model of FUS in disease. Within transfected HEK293 cells, insertion of the P525L and R521C FUS mutations leads to relocalisation of FUS from the nucleus to the cytoplasm. R521C and P525L mutant FUS incorporates into cytoplasmic aggregations of untranslated mRNA and RNA binding proteins known as stress granules. The strong relocalisation seen with P525L-FUS is associated with a gain of cytotoxicity. Reversal of this cytoplasmic relocalisation by demethylation of FUS rescues this cytotoxicity, suggesting a toxic gain of cytoplasmic function in the majority of FUS mutations. By contrast, insertion of the R244C mutation leads to neither relocalisation, stress granule association, nor cytotoxicity. Notably the R244C mutation, located away from the nuclear localization domain in which the majority of FUS mutations are found, leads to the presence of smaller FUS fragments in western blot analyses. These fragments appear not to be due to splicing defects in FUS but rather are due to post-translational modifications or aberrant protein cleavage. These data suggest an alternative pathway for FUS toxicity based upon a nuclear loss of function.