Academic literature on the topic 'Hygromycin A'

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Journal articles on the topic "Hygromycin A":

1

McCallum, B. D., C. C. Bernier, and L. Lamari. "Generation and utilization of chemical-resistant mutants in Pyrenophora tritici-repentis, the causal agent of tan spot of wheat." Canadian Journal of Botany 72, no. 1 (January 1, 1994): 100–105. http://dx.doi.org/10.1139/b94-014.

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Tan spot, caused by the fungal pathogen Pyrenophora tritici-repentis, is a major leaf spot disease of wheat worldwide. To facilitate genetic analysis of this homothallic fungus, mutants resistant to the fungicide iprodione or hygromycin B were created through ultraviolet light mutagenesis and used in sexual crosses. Conidia from two isolates of P. tritici-repentis, sensitive to both chemicals (iprodione-S hygromycin-S), were exposed to ultraviolet light to obtain four mutants resistant to iprodione but sensitive to hygromycin B (iprodione-R hygromycin-S) and three mutants resistant to hygromycin B but sensitive to iprodione (iprodione-S hygromycin-R). The mutants were paired in all combinations, and the markers allowed crossed progeny to be distinguished from selfed progeny. Crossed ascospore progeny from pairings between iprodione-R hygromycin-S isolates and iprodione-S hygromycin-R isolates and between iprodione-R hygromycin-R isolates and iprodione-S hygromycin-S segregated 1:1 for resistance–sensitivity to both iprodione and hygromycin B. These results indicate that one locus controls iprodione resistance and a second independent locus controls hygromycin B resistance. This study should facilitate further genetic research on the tan spot fungus by providing a simple marker system. Key words: genetics, inheritance, Drechslera, yellow spot, leaf spot.
2

Sultana, Shahanaz, Chai Ling Ho, Parameswari Namasivayam, and Suhaimi Napis. "Genotypic differences in response to Hygromycin effect on untransformed calli death and rice germination." Bangladesh Rice Journal 18, no. 1-2 (April 17, 2015): 38–43. http://dx.doi.org/10.3329/brj.v18i1-2.23001.

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Hygromycin is an efficient selective agent in transformation studies of wide ranges of crop. In this study, different concentrations of hygromycin were used to observe the effect on untransformed calli death, percent germination and seedling growth of three rice varieties (Oryza sativa L.) viz BRRI dhan29, MR219 and Taipei309. Hygromycin killed the untransformed calli and inhibited the germination of tested varieties in a concentration dependent manner. Among the tested varieties, the lowest and the highest calli death was observed in MR219 and Taipei309 respectively in all the concentrations of hygromycin. Whereas, the highest and the lowest percent germination were observed in MR219 and Taipei309 respectively. The minimal inhibitory concentration (MIC) for selection of calli were calculated as 42, 40 and 47 mg/L hygromycin for BRRI dhan29, MR219 and Taipei309 respectively. During germination, 35, 62 and 32 mg/L hygromycin were suitable for the selection of BRRI dhan29, MR219 and Taipei 309 respectively. Shoot and root growth reduction after germination was increased with the increased concentration of hygromycin. Besides, root growth was more sensitive to hygromycin than the shoot. These results suggest that hygromycin increases calli death, decreases percent germination, and shoot and root growth in all varieties with the increasing rate of hygromycin. But these characteristics vary with different degrees in different genotypes as well as different stages.Bangladesh Rice j. 2014, 18(1&2): 38-43
3

Pfister, P., M. Risch, D. E. Brodersen, and E. C. Böttger. "Role of 16S rRNA Helix 44 in Ribosomal Resistance to Hygromycin B." Antimicrobial Agents and Chemotherapy 47, no. 5 (May 2003): 1496–502. http://dx.doi.org/10.1128/aac.47.5.1496-1502.2003.

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ABSTRACT Hygromycin B is an aminoglycoside antibiotic active against prokaryotic and eukaryotic ribosomes. Ribosomal alterations in bacteria conferring resistance to hygromycin B have not been described, prompting us to use a single rRNA allelic derivative of the gram-positive bacterium Mycobacterium smegmatis for investigation of the molecular mechanisms involved in ribosomal resistance to hygromycin B in eubacteria. Resistance mutations were found to localize exclusively in 16S rRNA. The mutations observed, i.e., 16S rRNA U1406C, C1496U, and U1498C (E. coli numbering), are in close proximity to the hygromycin B binding site located in conserved helix 44 of 16S rRNA. The 16S rRNA positions involved in hygromycin B resistance are highly conserved in all three domains of life, explaining the lack of specificity and general toxicity of hygromycin B.
4

Mercuriani, Ixora Sartika, Aziz Purwantoro, Sukarti Moeljopawiro, Seonghoe Jang, and Endang Semiarti. "Selection of Phalaenopsis amabilis L. Blume Orchid Resistance to Hygromycin." Indonesian Journal of Biotechnology 17, no. 2 (November 9, 2015): 107. http://dx.doi.org/10.22146/ijbiotech.16000.

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Examination of Phalaenopsis amabilis orchid resistance to hygromycin antibiotic is an important step to doprior to Agrobacterium-mediated genetic transformation in this orchids using Hygromycin phosphotransferase(HPT) gene as a selection marker in the T-DNA that harboring a desired gene to be transfered. We exposedthe plant on hygromycin containing medium. The experiment was conducted using 6 weeks old P. amabilisprotocorms. These protocorms were subcultured onto NP medium supplemented with various concentrationof Hygromycin (0, 5, 10, 20, 1nd 40 mg/l). The number of survival protocorms were examined every week for4 weeks after subcultured (WAS). The resistancy of hygromycin was calculated as ratio of death protocormsper total protocorms). The result showed that 10 mg/l hygromycin with 1 weeks of application caused deathclose to LD 50. This data indicate that P. amabilis resistance to hygromycin treatment on the appropriateconcentration 10 mg/l, and this concentration can be used for other purposes in orchid system.
5

Hussain, Iqra. "INTRODUCTION OF RICE CHITINASE GENE IN POTATO BY AGROBACTERIUM-MEDIATED TRANSFORMATION." Pakistan Journal of Agricultural Sciences 56, no. 01 (January 1, 2019): 7–13. http://dx.doi.org/10.21162/pakjas/19.8154.

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Potato is an important food crop of the world. Different viral, bacterial and fungal pathogens cause heavy economic losses of this crop every year. Potato has complex genetic makeup due to which induction of disease resistance through conventional breeding is difficult. Genetic manipulation through different transformation techniques is more precise and successful tool. In the present study different factors were investigated, which have an influence on potato transformation. The optimal dose of cefotaxime was found 500 mg/l which did not affected the growth of the potato tissues. The explants treated with Agrobacterium in the presence of acetosyringone resulted in higher frequency of transformation as compared to the explant without it. Two minutes time for co infection was found appropriate for optimum transformation efficiency. The two days cocultivation period along with 7 days preselection was found suitable for potato transformation. The putative tranformants regenerated on MS medium supplemented with 20 mg/L hygromycine and 500 mg/L cefotaxime, from nodal explants while the non-transformed tissue turned brown and gradually died after two or three sub culturing on the selection media containing selective antibiotic hygromycin. The shoots obtained on selection media shifted on root induction media supplemented with similar concentrations of hygromycin and cefotaxime, resulted complete plantlet formation after 10 days. It was observed that all the hygromycin positive plants also exhibited positive bands of desired size of 823 bp for chitinase gene, suggesting cotransformation of both genes in transformed plants
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Dai, Qun, Zhihuan Sun, and Guido Schnabel. "Development of Spontaneous Hygromycin B Resistance in Monilinia fructicola and Its Impact on Growth Rate, Morphology, Susceptibility to Demethylation Inhibitor Fungicides, and Sporulation." Phytopathology® 93, no. 11 (November 2003): 1354–59. http://dx.doi.org/10.1094/phyto.2003.93.11.1354.

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Agrobacterium tumefaciens-mediated transformation with plasmids carrying the hygromycin B resistance gene hph frequently is being used for inserting genes into fungal spores and mycelial cells and for conducting insertional mutagenesis to identify genes connected to a particular phenotype. In this article, we report that stable hygromycin B resistance can develop spontaneously in germinating conidia from Monilinia fructicola and that the mutants exhibit altered phenotypes. One spontaneously developing hygromycin B-resistant colony developed per 2.5 × 105 germinating conidia. Mutants grew significantly slower on potato dextrose agar, were 2.4- to 3.1-fold more sensitive to demethylation inhibitor fungicides, lacked melanization, and did not produce spores. The mode of action of hygromycin B resistance in the mutants seemed to be different from the hph transgene-mediated hygromycin B resistance based on different phenotypic characters. The ability of M. fructicola and possibly other fungi to spontaneously develop hygromycin B resistance associated with an altered phenotype may interfere with the selection of true transformants if hygromycin B is used as selective agent. This is particularly confounding if the hph gene is used as selectable marker in insertional mutagenesis experiments conducted for the identification of genes involved in melanization, sporulation, or fungicide resistance.
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Ko, Moon Kyung, Hyunchul Soh, Kyung-Moon Kim, Young Soon Kim, and Kyunghoan Im. "Stable Production of Transgenic Pepper Plants Mediated by Agrobacterium tumefaciens." HortScience 42, no. 6 (October 2007): 1425–30. http://dx.doi.org/10.21273/hortsci.42.6.1425.

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The aim of this study was to establish a stable transformation method for hot pepper using the hygromycin phosphotransferase (hpt)/hygromycin selection strategy. Explants from aseptic pepper seedlings were inoculated with Agrobacterium tumefaciens carrying pCAMBIA1301. A number of calli were developed on the medium containing hygromycin to discriminate the induction of “false-positive buds,” and then shoots were successfully regenerated from the hygromycin-resistant calli. Southern and Northern hybridization analysis indicated that the hpt gene was integrated and expressed in the transgenic pepper plants (T0) and transmitted to the progeny (T1) without genetic modification. Most T1 progenies derived from self-pollination revealed a 3:1 segregation ratio for hygromycin resistance, indicating that one copy of the T-DNA was integrated into the respective transgenic lines. Both uidA and hpt genes were stably expressed in the T1 generation and coinherited in the progenies. Finally, homozygous progenies were identified in the T1 generation of the transgenic peppers, and the homozygous state was maintained in all progenies tested (T2). The results show the reliability and stability of the hpt/hygromycin selection protocol for pepper transformation.
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McGaha, Susan M., and W. Scott Champney. "Hygromycin B Inhibition of Protein Synthesis and Ribosome Biogenesis in Escherichia coli." Antimicrobial Agents and Chemotherapy 51, no. 2 (October 16, 2006): 591–96. http://dx.doi.org/10.1128/aac.01116-06.

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ABSTRACT The aminoglycoside antibiotic hygromycin B was examined in Escherichia coli cells for inhibitory effects on translation and ribosomal-subunit formation. Pulse-chase labeling experiments were performed, which verified lower rates of ribosomal-subunit synthesis in drug-treated cells. Hygromycin B exhibited a concentration-dependent inhibitory effect on viable-cell numbers, growth rate, protein synthesis, and 30S and 50S subunit formation. Unlike other aminoglycosides, hygromycin B was a more effective inhibitor of translation than of ribosomal-subunit formation in E. coli. Examination of total RNA from treated cells showed an increase in RNA corresponding to a precursor to the 16S rRNA, while mature 16S rRNA decreased. Northern hybridization to rRNA in cells treated with hygromycin B showed that RNase II- and RNase III-deficient strains of E. coli accumulated 16S rRNA fragments upon treatment with the drug. The results indicate that hygromycin B targets protein synthesis and 30S ribosomal-subunit assembly.
9

Liu, Hang, Zhongyi Zhang, Sijie Liang, Li Guo, Shiyang Sun, Kehou Pan, and Guanpin Yang. "Transcriptome Responses of Hygromycin B Resistance Gene-Transformed, Hygromycin B-Adaptive and Wild Nannochloropsis oceanica Strains to Hygromycin B." Journal of Ocean University of China 19, no. 2 (January 24, 2020): 453–58. http://dx.doi.org/10.1007/s11802-020-4252-4.

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Abdulmunim ‎, Zahraa, Rabah N. Jabba, and Abdulwahid B. Al-Shaibani. "Studying the Optimum Conditions of ‎Hygromycin B Production and Detect their ‎Toxicity." JOURNAL OF UNIVERSITY OF BABYLON for Pure and Applied Sciences 26, no. 2 (December 26, 2017): 119–30. http://dx.doi.org/10.29196/jub.v26i2.480.

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Hygromycin B was extracted with ethyl acetate, which separates organic phase from aqueous phase in the broth culture filtrate, only the aqueous phase showed significant antimicrobial activity by using agar well diffusion technique. At a concentration of 25mg/ml (as crude extract), this phase excreted its activity against the test microorganisms which include; one G(+) bacteria (Staphylococcus aureus), five G(–) bacteria (Pseudomonas aeruginosa , Proteus mirabilis, Escherichia coli , Klebsiella pneumoniae, Salmonella typhi) and one yeast (Saccharomyces cerevisiae). After detecting the aminoglycoside hygromycin B by the Thin Layer Chromatography (TLC) method to ensure presence of the antibiotic, same flow rate (Rf) value (0.357) as that of the standard hygromycin B was obtained. Results of the optimization conditions showed that the highest antimicrobial activity of hygromycin B was obtained at a medium pH of 8 and incubation temperature of 35°C for 10 days. When the toxicity of hygromycin B crude extract under such conditions was examined on mice liver, a mild effects were appeared

Dissertations / Theses on the topic "Hygromycin A":

1

Hill, David G. "A stereoselective approach towards hygromycin A." Thesis, Heriot-Watt University, 1994. http://hdl.handle.net/10399/1354.

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Kaminishi, Tatsuya, Andreas Schedlbauer, Attilio Fabbretti, Letizia Brandi, Lizarralde Borja Ochoa, Cheng-Guang He, Pohl Milon, Sean R. Connell, Claudio O. Gualerzi, and Paola Fucini. "Crystallographic characterization of the ribosomal binding site and molecular mechanism of action of Hygromycin A." Oxford University Press, 2015. http://hdl.handle.net/10757/608247.

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Hygromycin A (HygA) binds to the large ribosomal subunit and inhibits its peptidyl transferase (PT) activity. The presented structural and biochemical data indicate that HygA does not interfere with the initial binding of aminoacyl-tRNA to the A site, but prevents its subsequent adjustment such that it fails to act as a substrate in the PT reaction. Structurally we demonstrate that HygA binds within the peptidyl transferase center (PTC) and induces a unique conformation. Specifically in its ribosomal binding site HygA would overlap and clash with aminoacyl-A76 ribose moiety and, therefore, its primary mode of action involves sterically restricting access of the incoming aminoacyl-tRNA to the PTC.
Bizkaia:Talent and the European Union's Seventh Framework Program (Marie Curie Actions; COFUND; to S.C., A.S., T.K.); Marie Curie Actions Career Integration Grant (PCIG14-GA-2013-632072 to P.F.); Ministerio de Economía Y Competitividad (CTQ2014-55907-R to P.F., S.C.); FIRB Futuro in Ricerca from the Italian Ministero dell'Istruzione, dell'Universitá e della Ricerca (RBFR130VS5_001 to A.F.); Peruvian Programa Nacional de Innovación para la Competitividad y Productividad (382-PNICP-PIBA-2014 (to P.M. and A.F.)). Funding for open access charge: Institutional funding.
Revisión por pares
3

McGaha, Susan Mabe. "Antibiotics that Inhibit 30S or 50S Ribosomal Subunit Formation: Hygromycin B, Quinupristin-Dalfopristin and XRP 2868." Digital Commons @ East Tennessee State University, 2007. https://dc.etsu.edu/etd/2055.

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Several antibiotics that prevent translation by binding to ribosomal subunits have been shown to also inhibit ribosomal subunit assembly (Champney and Tober 2003). The aminoglycoside hygromycin B was examined in Escherichia coli cells for inhibitory effects on translation and ribosomal subunit assembly. The streptogramin antibiotics quinupristin-dalfopristin and XRP 2868 (NXL 103) were examined for similar effects on these 2 cellular functions in antibiotic-resistant strains of Haemophilus influenzae, Staphylococcus aureus, and Streptococcus pneumoniae. Pulse chase experiments were performed which verified slower rates of ribosomal subunit formation in drug treated cells. Hygromycin B exhibited a concentration dependent inhibitory effect on viable cell number, growth rate, protein synthesis and 30S and 50S subunit formation. 16S rRNA specific probes hybridized to rRNA fragments in cells treated with hygromycin B. RNase II and RNase III deficient strains of E. coli exhibited the most accumulation of 16S rRNA fragments upon treatment with hygromycin B. Examination of total RNA from treated cells showed an increase in RNA corresponding to precursor to the 16S rRNA while 16S rRNA decreased. There was also an increase in small fragment RNA. Hygromycin B was a more effective inhibitor of translation than ribosomal subunit formation in E. coli. Two streptogramin antibiotics were compared for inhibitory effects in antibiotic-resistant Haemophilus influenzae, Staphylococcus aureus, and Streptococcus pneumoniae. IC50 values for XRP 2868 were several fold lower than those of quinupristin-dalfopristin for inhibition of cell viability, protein synthesis, and ribosomal subunit formation. Both antibiotics revealed a concentration dependent inhibitory effect on cellular functions including 50S ribosomal subunit formation in the three organisms examined. XRP 2868 inhibited both 50S ribosomal subunit assembly and translation. XRP 2868 was effective against MRSA and was a better inhibitor in each of the antibiotic resistant strains examined compared with quinupristin-dalfopristin.
4

Cordesman, Alexander. "MECHANICAL ABRASION AND ELECTROPORATION IN THE TRANSFORMATION OF INTACT PLEUROTUS OSTREATUS HYPHÆ and NUTRIENT-DEPENDANT RESISTANCE TO HYGROMYCIN B." VCU Scholars Compass, 2008. http://scholarscompass.vcu.edu/etd/173.

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ABSTRACT 1: Abrasive treatment and subsequent electroporation of the basidiomycete Pleurotus ostreatus in a hypertonic buffer was investigated as a potential method of transforming of intact hyphæ. Mycelia, which are not capable of being transformed via electroporation due to interference by the cell wall, were abraded in an attempt to mechanically degrade the cell wall prior to electroporation. An electroporation field strength of 12,500 V/cm for 500 μs to 1.25 ms was found to be optimal based upon mortality effects. A 32 μm carborundum abrasive was initially evaluated but was quickly found to be inappropriately large so a corundum abrasive with an average size of 300 nm was focused on. Vortexing as well as low and medium power ultrasonic agitation with the corundum abrasive were investigated for their potential to cause mechanical degradation of the cell walls. Vortexing and low power sonication were found to be ineffective at causing adequate degradation while medium power sonication was found to be both ineffective and super lethal. While the possibility of mechanical abrasion facilitating transformation via electroporation remains, it is unlikely that conventional methods of agitation will be effective. ABSTRACT 2: The dose response of the basidiomycetes Pleurotus ostreatus, Agrocybe aegerita and Cordyceps millitaris to the antibiotic hygromycin B was tested on two common and one in-house solid growth media. The three species were grown on Potato dextrose agar, malt extract agar and an agar containing maltose, yeast-extract, peptone and glucose with concentrations of hygromycin B from zero to 100 μg/ml for 11 to 14 days. Micrometer measurements were used to determine the growth rate of each species on each media. Significant differences in hygromycin B tolerance for each species between the three media types were evident (p-value < .0001 by ANOVA for all). Neither the media type nor the growth rate on hygromycin B free plates were useful predictors of effective hygromycin B doses so optimization should be performed on every strain and media type used for selection.
5

Chowdhury, Shukti Rani. "Establishment of regeneration and transient transformation systems for Australian native resurrection plant Tripogon loliiformis." Thesis, Queensland University of Technology, 2019. https://eprints.qut.edu.au/132682/1/Shukti%20Rani_Chowdhury_Thesis.pdf.

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This thesis contributed to unlock the regeneration and transient transformation systems for Tripogon loliiformis, an Australian native extremophile. This resurrection grass has suitable structural, physiological and molecular mechanisms to survive in extreme desiccation. The regeneration and transient transformation systems will assist in establishing a stable transformation system for Tripogon which will enable more understanding of its unique genes for coping abiotic stresses. This may contribute to the bioengineering of these genes to important cereal crops like rice, wheat, maize and sorghum for the improvement of abiotic stress tolerance.
6

Locken, Kristopher Michael. "Super hypersensitivity to hygromycin B (S-HHY) gene functions converge at the trans-golgi and late endosome interface have a role in tor1p location to the vacuole." Thesis, California State University, Long Beach, 2014. http://pqdtopen.proquest.com/#viewpdf?dispub=1527392.

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The vacuole in Saccharomyces cerevisiae serves as a model for the mammalian lysosome. In a genome wide screen for mutants with severe growth hypersensitivity to hygromycin B, our lab identified 14 HHY genes. Each of the hhy mutants is defective in vacuolar trafficking and/or function and also sensitive to rapamycin and caffeine, suggesting a compromised target of rapamycin (TOR) kinase pathway. My research divides the hhy mutants into two groups based on quantitative growth analyses in the presence of hygromycin B. (1) a super affected group (s-hhy's) and (2) a dose-dependent group (d-hhy's). The s-HHY genes include CHC1, DRS2, SAC1, VPS1, VPS34, VPS45, VPS52, and VPS54. Evaluation of the known functions of s-HHY gene products reveals vesicular trafficking function at the trans-Golgi and late endosome interface to be a common factor. In yeast, the TORC1 complex localizes to the vacuole. Because a compromised TORC1 complex signaling is suggested in hhy mutant strains due to their caffeine or rapamycin sensitivity, I hypothesized that compromised TORC1 signaling in HHY's may be due to defects in the vacuolar localization of Tor1 kinase. To assess Tor1 kinase localization, we utilized a strain expressing endogenously tagged Tor1-GFP and assayed localization to the vacuolar membrane in each of the s-hhy mutants using confocal microscopy. In wild-type cells, Tor1-GFP co-localizes with the vacuolar membrane marker FM4-64 while s-hhy deletion strains fail to localize Tor1-GFP to the vacuolar membrane when treated with hygromycin B. Our results implicate that Tor1p is transported to the vacuole membrane via the late endosome (CPY pathway) and not the ALP (Vam3) pathway. Additionally, s-hhy mutants are unable to recover growth after a 4-hour treatment with hygromycin B, similar to EG0 mutants, which fail to exit from G0 after treatment with the TORC1 inhibiting drug rapamycin. Based on our data, we propose a model in which the s-HHY gene functions in vesicular trafficking at the trans-Golgi/late endosome interface are involved in recruitment and subsequent transport of Tor1p to the vacuolar membrane, and that interface is hypersensitive to hygromycin B. We also propose that Tor1 kinase localization at the vacuole is essential for its cell cycle regulatory function.

7

Dere, Madhavi Suresh. "Genetic Transformation of Switchgrass (Panicum Virgatum L.) with Endoglucanase Gene and Characterization of Plants with Endoglucanase Transgene." Thesis, Virginia Tech, 2012. http://hdl.handle.net/10919/76816.

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As a warm season grass native to the North American continent, switchgrass is considered as one of the most promising biofuel crops in the USA. It is a C4 plant that makes it energy efficient. Switchgrass has a deep root system that allows it to grow on marginal land with low water and nutrient input. Switchgrass has been used as a forage crop and its use for biofuel will not affect food security. Biofuels are more environment-friendly than fossil fuels as they do not produce net greenhouse gases. However, the problem of high cost of production per unit for biofuel has to be overcome if we want to replace fossil fuels with biofuels. One of the major factors related to the high cost of biofuel are the expensive cellulase enzymes used in the pretreatment of feedstock. Endoglucanase is the key enzyme used for breaking down cellulose before fermentation. Currently, endoglucanase is produced from engineered E. coli or yeast strains, which is still expensive for enzyme production and purification of industrial scales. Expression of endoglucanase in plants has been previously reported. However, there are no reports of transgenic switchgrass producing cellulase enzyme. In this study, the catalytic domain of beta-endoglucanase gene was codon-optimized and synthesized based on the cDNA cloned from Hypocrea jecorina. Rice RuBisCO small subunit targeting signal peptide was fused to the N-terminus of the beta-endoglucanase gene, which was expected to target the fusion protein to chloroplast. This subcellular compartment targeting could minimize negative effects on cell function and plant development. The endoglucanase gene was cloned with maize ubiquitin promoter in a modified binary vector pCambia 1305-2 and transformed into switchgrass genotype HR8 by using Agrobacterium tumefaciens. In this study, I generated five independent transgenic switchgrass lines and they were confirmed by growing on the selection agent hygromycin, GUS assay, PCR amplification, southern blotting hybridization, for the presence of hygromycin and endoglucanase genes. However, based on RT-PCR analysis, only two transgenic lines were confirmed to produce mRNAs of the endoglucanase gene. These two transgenic lines were further characterized for their agronomic traits and the chlorophyll contents. Our results suggested that expression of endoglucanase in switchgrass could reduce chlorophyll content and affect plant development. Nevertheless, in this study, we demonstrated that a fungal endoglucanase gene could be expressed in switchgrass transgenic plants, though the gene expression level and the subcellular localization need to be carefully regulated in order to minimize the toxic effect of endoglucanase on plant cells.
Master of Science
8

Ruggieri, Francesca. "Putting nature back into drug discovery : selection, design and synthesis of bioinspired chemical libraries for the discovery of new antibacterials." Electronic Thesis or Diss., Université de Lille (2022-....), 2024. http://www.theses.fr/2024ULILS013.

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Les produits naturels (PNs) ont perdu en popularité depuis l'introduction des petites molécules synthétiques il y a plusieurs années. De nombreuses raisons expliquent ce choix, telles que les difficultés d'accès et d'approvisionnement, la complexité de la chimie des PNs et l'avènement de la chimie combinatoire. Cependant, les PNs offrent de nombreuses propriétés intéressantes par rapport aux molécules synthétiques conventionnelles, ce qui leur confère à la fois des avantages et des inconvénients dans le contexte de la recherche de principes actifs. En général, les PNs se caractérisent par un plus grand nombre de carbones sp3 et de centres stéréogènes, une grande diversité de squelettes et une grande complexité structurelle. La moitié des médicaments approuvés par la FDA depuis 1994 étant des PNs ou des dérivés hémisynthétiques, et étant donnée la récente stagnation de la recherche et du développement de nouveaux médicaments, il devient de plus en plus évident que les produits naturels doivent être utilisés dans le processus de découverte de médicaments en tant que source d'inspiration.Par conséquent, de nombreuses stratégies émergent aujourd'hui pour la construction de chimiothèques inspirées par la nature, selon les stratégies "top-down" et "bottom-up". Dans les approches "bottom-up", la complexité est créée à partir de réactifs simples. En revanche, les approches "top-down" consistent à apporter des modifications structurelles à un produit naturel déjà complexe.Le travail présenté décrit deux approches différentes pour enrichir la chimiothèque de notre unité de recherche avec des composés dérivés de PNs. Une stratégie hémisynthétique "top-down" a été planifiée pour obtenir des dérivés de la lactucine et de la 11β,13-dihydrolactucine, deux lactones sesquiterpéniques extraites des racines de chicorée. 36 dérivés esters ont été synthétisés en trois étapes (synthèse classique), ainsi que deux bibliothèques de dérivés aminés (en utilisant la synthèse parallèle). Tous les composés ont ensuite été testés contre Mycobacterium tuberculosis et certaines molécules ont montré des activités prometteuses (CMI < 1,2 µM).D'autre part, une stratégie "bottom-up" a permis la synthèse de deux analogues de l'antibiotique naturel hygromycine A. Pour cette approche nous sommes partis de réactifs simples disponibles dans le commerce et avons appliqué une stratégie de déaromatisation dans le processus de synthèse.L'ensemble de ces travaux nous a permis d'explorer un espace chimique plus large, d'accroître la diversité structurelle de notre chimiothèque et de découvrir de nouveaux "hits". Nous pourrons également ainsi identifier de nouvelles cibles antibactériennes
Natural products (NPs) have declined in popularity since the introduction of synthetic small molecules several years ago. Many are the reasons behind this choice, such as difficulties in access and supply, complexities of NP chemistry and the advent of combinatorial chemistry. However, NPs offer many interesting properties compared to conventional synthetic molecules, which confer both advantages and challenges for the drug discovery process. Usually, NPs are characterized by a higher number of sp3 carbons and stereogenic centres, large scaffold diversity and structural complexity. With half of the drugs approved by the FDA since 1994 being NPs or hemisynthetic derivatives and the recent stagnation in new drug research and development, it is becoming more and more evident that NPs should be reintroduced in the drug discovery process as a source of inspiration.Therefore, many strategies are now emerging for the construction of nature-inspired chemical libraries, such as “top-down” and “bottom-up” strategies. In “bottom-up” approaches, complexity is created starting from simple building blocks. On the other hand, “top-down” approaches are assumed to make structural modifications to an already complex NP.Our presented work describes two different approaches to enrich the chemical library of our research unit with NP-derived compounds. A “top-down” semisynthetic strategy was planned to obtain derivatives of lactucin and 11β,13-dihydrolactucin, two sesquiterpene lactones extracted from chicory roots. Thirty-six ester derivatives were synthesized in three steps (classical synthesis), together with two amine derivative libraries (using parallel synthesis). All the compounds were then tested against Mycobacterium tuberculosis and some promising hits were found (MICGFP < 1.2 μM). On the other hand, a “bottom-up” strategy allowed the synthesis of two analogues of the known natural antibiotic hygromycin A. This approach started from simple commercially available building blocks and employed a dearomatization strategy in the synthetic process.Together, we explored a broader chemical space, increased the structural diversity of our chemical library and discovered new potential antibacterial hits. Moreover, this work paves the way for the discovery of new antibacterial targets
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Oliveira, Jonas Ivan Nobre. "Modelagem molecular na caracteriza??o eletr?nica de oligopept?deos e na descri??o qu?ntica da intera??o f?rmaco-receptor." Universidade Federal do Rio Grande do Norte, 2012. http://repositorio.ufrn.br:8080/jspui/handle/123456789/13075.

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Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior
In this dissertation, the theoretical principles governing the molecular modeling were applied for electronic characterization of oligopeptide α3 and its variants (5Q, 7Q)-α3, as well as in the quantum description of the interaction of the aminoglycoside hygromycin B and the 30S subunit of bacterial ribosome. In the first study, the linear and neutral dipeptides which make up the mentioned oligopeptides were modeled and then optimized for a structure of lower potential energy and appropriate dihedral angles. In this case, three subsequent geometric optimization processes, based on classical Newtonian theory, the semi-empirical and density functional theory (DFT), explore the energy landscape of each dipeptide during the search of ideal minimum energy structures. Finally, great conformers were described about its electrostatic potential, ionization energy (amino acids), and frontier molecular orbitals and hopping term. From the hopping terms described in this study, it was possible in subsequent studies to characterize the charge transport propertie of these peptides models. It envisioned a new biosensor technology capable of diagnosing amyloid diseases, related to an accumulation of misshapen proteins, based on the conductivity displayed by proteins of the patient. In a second step of this dissertation, a study carried out by quantum molecular modeling of the interaction energy of an antibiotic ribosomal aminoglicos?dico on your receiver. It is known that the hygromycin B (hygB) is an aminoglycoside antibiotic that affects ribosomal translocation by direct interaction with the small subunit of the bacterial ribosome (30S), specifically with nucleotides in helix 44 of the 16S ribosomal RNA (16S rRNA). Due to strong electrostatic character of this connection, it was proposed an energetic investigation of the binding mechanism of this complex using different values of dielectric constants (ε = 0, 4, 10, 20 and 40), which have been widely used to study the electrostatic properties of biomolecules. For this, increasing radii centered on the hygB centroid were measured from the 30S-hygB crystal structure (1HNZ.pdb), and only the individual interaction energy of each enclosed nucleotide was determined for quantum calculations using molecular fractionation with conjugate caps (MFCC) strategy. It was noticed that the dielectric constants underestimated the energies of individual interactions, allowing the convergence state is achieved quickly. But only for ε = 40, the total binding energy of drug-receptor interaction is stabilized at r = 18A, which provided an appropriate binding pocket because it encompassed the main residues that interact more strongly with the hygB - C1403, C1404, G1405, A1493, G1494, U1495, U1498 and C1496. Thus, the dielectric constant ≈ 40 is ideal for the treatment of systems with many electrical charges. By comparing the individual binding energies of 16S rRNA nucleotides with the experimental tests that determine the minimum inhibitory concentration (MIC) of hygB, it is believed that those residues with high binding values generated bacterial resistance to the drug when mutated. With the same reasoning, since those with low interaction energy do not influence effectively the affinity of the hygB in its binding site, there is no loss of effectiveness if they were replaced.
Nessa disserta??o, os princ?pios te?ricos que regem a Modelagem Molecular foram aplicados na caracteriza??o eletr?nica do oligopept?deo α3 e seus variantes (5Q,7Q)-α3, como tamb?m na descri??o qu?ntica da intera??o do aminoglicos?deo higromicina B e a subunidade 30S do ribossomo bacteriano. No primeiro estudo, os dipept?deos lineares e neutros constituintes das biomol?culas mencionados foram modelados e posteriormente otimizados at? uma estrutura de menor energia potencial e ?ngulos diedros adequados. No caso, tr?s processos de otimiza??o geom?trica, baseados subsequentemente na teoria cl?ssica newtoniana, na semi-emp?rica e na teoria do funcional da densidade (DFT), varreram a paisagem de energia de cada dipept?deos na busca de uma estrutura de energia m?nima ideal. Por fim, os conf?rmeros ?timos foram descritos quanto ao potencial eletrost?tico, energia de ioniza??o (amino?cidos), orbitais de fronteira HOMO/HOMO-1 e termo de hopping. A partir dos termos de hopping descritos nesse trabalho, foi poss?vel, em estudos subsequentes, caracterizar as propriedades de transporte de cargas destes modelos pept?dicos. Vislumbra-se uma nova tecnologia de biosensores capaz de diagnosticar doen?as amiloides, relacionadas ao ac?mulo de pept?deos disformes, a partir do perfil de condutividade el?trica apresentado pelas prote?nas do paciente. Em um segundo momento dessa disserta??o, realiza-se um estudo qu?ntico por modelagem molecular da energia de intera??o de um antibi?tico aminoglicos?dico em seu receptor riboss?mico. Sabe-se que a higromicina B (higB) ? um antibi?tico aminoglicos?deo que afeta a transloca??o ribossomal pela intera??o direta com a subunidade menor do ribossomo bacteriano (30S), especificamente com nucleot?deos da h?lice 44 do RNA riboss?mico 16S (rRNA 16S). Devido ao forte car?ter eletrost?tico desta conex?o, foi proposta a investiga??o energ?tica do mecanismo de liga??o da higB no 30S usando diferentes valores de constantes diel?tricas (ε=0, 4, 10, 20 e 40), as quais s?o amplamente utilizadas no estudo das propriedades eletrost?ticas de biomol?culas. Para isso, foram medidos raios crescentes centralizados no centr?ide da higB tendo por base a estrutura cristalina higB-30S (1HNZ.pdb), e apenas a energia de intera??o individual de cada nucleot?deo englobado foi calculada quanticamente utilizando a estrat?gia de fracionamento molecular com capuzes conjugados (MFCC). Percebeu-se que as constantes diel?tricas subestimam as energias de intera??o individuais, permitindo que o estado de converg?ncia energ?tica seja alcan?ado rapidamente. Por?m apenas para ε=40, a energia de intera??o total droga-receptor se estabilizou em r=18?, o que se constituiu como um adequado s?tio de liga??o, pois englobou os res?duos do 16S que interagem mais fortemente com a higB - C1403, C1404, G1405, A1493, G1494, U1495, C1496 e U1498. Assim, a constante diel?trica ≈40 ? ideal para o tratamento de sistemas com muitas cargas. Confrontando as energias de liga??o individuais dos nucleot?deos 16SrRNA com ensaios experimentais para determina??o da concentra??o inibit?ria m?nima (MIC) da higB, acredita-se que esses res?duos com elevados valores de intera??o gerariam resist?ncia bacteriana ? droga quando mutados. Com o mesmo racioc?nio, visto que aqueles com baixa energia n?o influenciariam de forma eficaz a afinidade da higB em seu s?tio de liga??o, n?o ocorreria perda de efic?cia caso fossem substitu?dos.
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Chen, Jong-Shenq, and 陳忠勝. "Transformation system of Trichoderma harzianum with the hygromycin B resistance gene, hph." Thesis, 1996. http://ndltd.ncl.edu.tw/handle/49887372835077302152.

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Abstract:
碩士
國立中興大學
植物學系
84
Trichoderma harzianum was transformed with two different plasmids: pAN7-1 and pUCH1. Both plasmids carry hph gene ( encoding hygromycin B phosphotransferase) as the selectable marker. Transformation frequency were 124 and 77 transformants per ug DNA for pAN7-1 and pUCH1 respectively. In stability test, 30.3±3.7% conidia of pAN7-1 transformants was HygR and 34.8±3.4% for pUCH1 transformants. All transformants were stable after several subculture without any selective pressure. The stable transformants were analyzed by Southern hybridization, showed several copies of pAN7-1 were integrated into the chromosomes of stable transformants with non-homologous recombination, apparently at the same site in different transformants. There was no correlation between the growth and copy number in the pAN7-1 transformants.

Book chapters on the topic "Hygromycin A":

1

Palaniappan, Nadaraj, and Kevin A. Reynolds. "Hygromycin A Biosynthesis." In ACS Symposium Series, 16–32. Washington, DC: American Chemical Society, 2007. http://dx.doi.org/10.1021/bk-2007-0955.ch002.

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Schomburg, Dietmar, and Dörte Stephan. "Hygromycin-B kinase." In Enzyme Handbook, 81–84. Berlin, Heidelberg: Springer Berlin Heidelberg, 1997. http://dx.doi.org/10.1007/978-3-642-59025-2_14.

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Schomburg, Dietmar, and Ida Schomburg. "hygromycin B 4-O-kinase 2.7.1.163." In Class 2–3.2 Transferases, Hydrolases, 371–80. Berlin, Heidelberg: Springer Berlin Heidelberg, 2013. http://dx.doi.org/10.1007/978-3-642-36240-8_84.

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Hung, Chiung-Yu, Hua Zhang Wise, and Garry T. Cole. "Gene Disruption in Coccidioides Using Hygromycin or Phleomycin Resistance Markers." In Host-Fungus Interactions, 131–47. Totowa, NJ: Humana Press, 2012. http://dx.doi.org/10.1007/978-1-61779-539-8_9.

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Zhang, Haiyan, and Jinjin Li. "From Cytosol to the Apoplast: The Hygromycin Phosphotransferase (HYGR) Model in Arabidopsis." In Unconventional Protein Secretion, 81–90. New York, NY: Springer New York, 2016. http://dx.doi.org/10.1007/978-1-4939-3804-9_5.

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Gravelat, Fabrice N., David S. Askew, and Donald C. Sheppard. "Targeted Gene Deletion in Aspergillus fumigatus Using the Hygromycin-Resistance Split-Marker Approach." In Host-Fungus Interactions, 119–30. Totowa, NJ: Humana Press, 2012. http://dx.doi.org/10.1007/978-1-61779-539-8_8.

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van der Straten, A., H. Johansen, R. Sweet, and M. Rosenberg. "Efficient Expression of Foreign Genes in Cultured Drosophila Melanogaster Cells Using Hygromycin B Selection." In Invertebrate and Fish Tissue Culture, 131–34. Berlin, Heidelberg: Springer Berlin Heidelberg, 1988. http://dx.doi.org/10.1007/978-3-642-73626-1_32.

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Majumder, Shuvobrata, Karabi Datta, and Swapan Kumar Datta. "Agrobacterium tumefaciens-Mediated Transformation of Rice by Hygromycin Phosphotransferase (hptII) Gene Containing CRISPR/Cas9 Vector." In Methods in Molecular Biology, 69–79. New York, NY: Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1068-8_5.

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Niklitschek, Mauricio, Marcelo Baeza, María Fernández-Lobato, and Víctor Cifuentes. "Generation of Astaxanthin Mutants in Xanthophyllomyces dendrorhous Using a Double Recombination Method Based on Hygromycin Resistance." In Microbial Carotenoids From Fungi, 219–34. Totowa, NJ: Humana Press, 2012. http://dx.doi.org/10.1007/978-1-61779-918-1_15.

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Punt, Peter J., and Cees A. M. J. J. van den Hondel. "[39] Transformation of filamentous fungi based on hygromycin b and phleomycin resistance markers." In Methods in Enzymology, 447–57. Elsevier, 1992. http://dx.doi.org/10.1016/0076-6879(92)16041-h.

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Conference papers on the topic "Hygromycin A":

1

Taipova, R. M., and B. R. Kuluev. "Agrobacterium-mediated transformation of Amaranthus cruentus L. epicotyls by the ARGOS-LIKE transgene." In 2nd International Scientific Conference "Plants and Microbes: the Future of Biotechnology". PLAMIC2020 Organizing committee, 2020. http://dx.doi.org/10.28983/plamic2020.244.

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The present study describes the results of our research in Agrobacterium-mediated transformation of epicotyl segments of Amaranthus cruentus variety “Bagryanyi” by the ARGOS-LIKE transgene of Arabidopsis thaliana controlled by the 35S promoter. For shoot regeneration from epicotyl segments after Agrobacterium-mediated transformation, Murashige-Skoog (MS) medium containing 13 μM 6-benzylaminopurine and 1 μM α-naphthylacetic acid was used. For the selection of transgenic shoots, 10 mg/L of hygromycin B was added to the MS medium.
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Susiyanti, Winda Wati, Nurmayulis Nurmayulis, Sulastri Isminingsih, and Sjaifuddin Sjaifuddin. "The Influence of Magnetic Fields Supply and Hygromycin Concentration to the Optimization of Folate Gene Putative Transformant Selection in Rice (cv. Fatmawati)." In 2nd and 3rd International Conference on Food Security Innovation (ICFSI 2018-2019). Paris, France: Atlantis Press, 2021. http://dx.doi.org/10.2991/absr.k.210304.046.

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Reports on the topic "Hygromycin A":

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Bostock, Richard M., Dov Prusky, and Martin Dickman. Redox Climate in Quiescence and Pathogenicity of Postharvest Fungal Pathogens. United States Department of Agriculture, May 2003. http://dx.doi.org/10.32747/2003.7586466.bard.

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Monilinia fructicola causes brown rot blossom blight and fruit rot in stone fruits. Immature fruit are highly resistant to brown rot but can become infected. These infections typically remain superficial and quiescent until they become active upon maturation of the fruit. High levels of chlorogenic acid (CGA) and related compounds occur in the peel of immature fruit but these levels decline during ripening. CGA inhibits cutinase expression, a putative virulence factor, with little or no effect on spore germination or hyphal growth. To better understand the regulation of cutinase expression by fruit phenolics, we examined the effect of CGA, caffeic acid (CA) and related compounds on the redox potential of the growth medium and intracellular glutathione (GSH) levels. The presence of CA in the medium initially lowered the electrochemical redox potential of the medium, increased GSH levels and inhibited cutinase expression. Conidia germinated in the presence of CA, CGA, or GSH produced fewer appressoria and had elongated germ tubes compared to the controls. These results suggest that host redox compounds can regulate fungal infectivity. In order to genetically manipulate this fungus, a transformation system using Agrobacterium was developed. The binary transformation vector, pPTGFPH, was constructed from the plasmid pCT74, carrying green fluorescent protein (GFP) driven by the ToxA promoter of Pyrenophora tritici-repentis and hygromycin B phosphotransferase (hph) under control of the trpC promoter of from Aspergillus nidulans, and the binary vector pCB403.2, carrying neomycin phosphotransferase (nptII) between the T-DNA borders. Macroconidia of M. fructicola were coincubated with A. tumefaciens strain LBA 4404(pPTGFPH) on media containing acetosyringone for two days. Hygromycin- and G418-resistant M. fructicola transformants were selected while inhibiting A. tumefaciens with cefotaxime. Transformants expressing GFP fluoresced brightly, and were formed with high efficiency and frequency of T-DNA integration frequency. The use of these transformants for in situ studies on stone fruit tissues is discussed.
2

Prusky, Dov, Noel T. Keen, and Stanley Freeman. Elicitation of Preformed Antifungal Compounds by Non-Pathogenic Fungus Mutants and their Use for the Prevention of Postharvest Decay in Avocado Fruits. United States Department of Agriculture, January 1996. http://dx.doi.org/10.32747/1996.7570573.bard.

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C. gloeosporioides attacks unripe avocado fruits in the orchard. Germinated spores produce appressoria that germinate and breach the cuticle, but the resultant subcuticular hyphae become quiescent and do not develop further until fruit is harvested and ripens. Resistance of unripe avocado to attach by C. gloeosporioides is correlated with the presence of fungitoxic concentrations of the preformed antifungal compound, 1-acetoxy-2-hydroxy-4-oxoheneicosa-12, 15 diene in the pericarp of unripe fruits. The objective of this proposal was to study the signal transduction process by which elicitors induce resistance in avocado. It was found that abiotic elicitors, infection of avocado fruit with C. gloeosporioides or treatment of avocado cell suspension with cell-wall elicitor induced a significant production of reactive oxygen species (ROS). Ripe and unripe fruit tissue differ with regard to the ROS production. The unripe, resistant fruit are physiologically able to react and to produce high levels of ROS and increased activity of H+ATPase that can enhance the phenylpropanoid pathway ad regulate the levels of the antifungal compound-diene, inhibit fungal development, resulting in its quiescence. Interestingly, it was also found that growth regulators like cytokinin could do activation of the mechanism of resistance. Postharvest treatments of cytokinins strongly activated the phenylpropanoid pathway and induce resistance. We have developed non-pathogenic strains of C. gloeosporioides by Random Enzyme Mediated Integration and selected a hygromycin resistance, non-pathogenic strain Cg-142 out of 3500 transformants. This non-pathogenic isolate activates H+ATPase and induces resistance against Colletotrichum attack. As a basis for studying the importance of PL in pathogenicity, we have carried out heterologous expression of pel from C. gloeosporioides in the non-pathogenic C. magna and determine the significant increase in pathogenicity of the non-pathogenic strain. Based on these results we can state that pectate lyase is an important pathogenicity factor of C. gloeosporioides and found that fungal pathogenicity is affected not by pel but by PL secretion. Our results suggest that PH regulates the secretion of pectate lyase, and support its importance as a pathogenicity factor during the attack of avocado fruit by C. gloeosporioides . This implicates that if these findings are of universal importance in fungi, control of disease development could be done by regulation of secretion of pathogenicity factors.

To the bibliography