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Journal articles on the topic "ITS-RFLP"
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Lanoot, Benjamin, Marc Vancanneyt, Bart Hoste, Katrien Vandemeulebroecke, Margo C. Cnockaert, Peter Dawyndt, Zhiheng Liu, Ying Huang, and Jean Swings. "Grouping of streptomycetes using 16S-ITS RFLP fingerprinting." Research in Microbiology 156, no. 5-6 (June 2005): 755–62. http://dx.doi.org/10.1016/j.resmic.2005.01.017.
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KOFFI, YAO FULGENCE, CAMELIA DIGUTA, MIREILLE ALLOUE-BORAUD, LOUIS BAN KOFFI, MARCELLIN DJE, EVELINA GHERGHINA, and FLORENTINA MATEI. "PCR-ITS-RFLP identification of pineapple spoilage fungi." Romanian Biotechnological Letters 24, no. 3 (June 20, 2019): 418–24. http://dx.doi.org/10.25083/rbl/24.3/418.424.
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Midgley, David J., Susan M. Chambers, and John W. G. Cairney. "Spatial distribution of fungal endophyte genotypes in a Woollsia pungens (Ericaceae) root system." Australian Journal of Botany 50, no. 5 (2002): 559. http://dx.doi.org/10.1071/bt02020.
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Fungal endophytes were isolated from hair roots of Woollsia pungens Cav. (Muell.) and mapped according to the root portions from which they were isolated. A total of 119 isolates was obtained and restriction fragment length polymorphism (RFLP) analysis of the internal transcribed spacer (ITS) region indicated that the isolate assemblage comprised five RFLP types. ITS sequence comparison revealed that RFLP Types I and II had 99.6–99.8% sequence identity with known ericoid mycorrhizal endophytes from Australian epacrids. The remaining three RFLP types were most similar to non-mycorrhizal ascomycetes. Eighty-five per cent of isolates obtained were of RFLP Type I and these were widespread within the root system. Inter-simple sequence repeat PCR suggested that 94% of RFLP Type I isolates were of a single genotype that was widely distributed within the root system, with the remaining five isolates each representing a different genotype. Apparent spatial dominance of the root system by a single fungal genotype may indicate limited functional diversity in the mycorrhizal endophyte assemblage.
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Duttweiler, K. B., G. Y. Sun, J. C. Batzer, T. C. Harrington, and M. L. Gleason. "An RFLP-Based Technique for Identifying Fungi in the Sooty Blotch and Flyspeck Complex on Apple." Plant Disease 92, no. 5 (May 2008): 794–99. http://dx.doi.org/10.1094/pdis-92-5-0794.
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A restriction fragment length polymorphism (RFLP)-based technique was developed to identify members of the sooty blotch and flyspeck (SBFS) disease complex on apple because these fungi are difficult to identify using agar-plate isolation and morphological description. The method includes polymerase chain reaction (PCR) amplification of the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) using a fungal-specific forward primer (ITS1-F) and an SBFS-specific reverse primer (Myc1-R), followed by digestion of the PCR product by the HaeIII restriction enzyme. When applied to previously identified isolates of 24 SBFS-causing species in nine genera, the PCR-RFLP assay produced 14 unique banding patterns. Different genera never shared the same RFLP pattern. To evaluate performance in vivo, the technique was applied to DNA extracted directly from SBFS colonies on apple fruit from three Iowa orchards. The primers amplified the rDNA of only SBFS fungi, with the exception of a Cladosporium sp.; however, its RFLP banding pattern was distinct from those of SBFS fungi. The majority (60%) of SBFS colonies in the in vivo trial were identified to genus by RFLP analysis. The PCR-RFLP assay greatly streamlined the identification process by minimizing the need for culturing, indicating its value as a tool for field studies of the SBFS complex.
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Hazlianda, Cut, Kamaliah Muis, and Isma Lubis. "A Comparative Study of Polymerase Chain Reaction-Restriction Fragment Length Polymorphism and Fungal Culture for the Evaluation of Fungal Species in Patients with Tinea Cruris." Open Access Macedonian Journal of Medical Sciences 5, no. 7 (November 21, 2017): 844–47. http://dx.doi.org/10.3889/oamjms.2017.197.
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BACKGROUND: Tinea cruris is the second most common dermatophytosis in the world and the most common in Indonesia. The conventional laboratory tests for dermatophyte infection are slow and less specific. Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) is a PCR method with the addition of enzyme after amplification, therefore enabling for more specific results.AIM: This study aimed to find whether the PCR-RFLP test could yield the same fungal species result as a fungal culture.METHODS: The specimens were skin scrapings from thirty-one patients suspected tinea cruris. The tools and materials that were used were Sabaroud’s dextrose agar media, primer ITS 1 and ITS 4 and MvaI.RESULTS: The equation percentage of the test result species between PCR-RFLP and fungal culture was 50% of 12 subjects whose the test results were both positive from the fungal culture and PCR-RFLP. The percentage of the test result with fungal culture the fungal species were found, but in the PCR-RFLP test which the fungal species was not found, the percentage was 50% of 12 subjects which the test results were both positive as fungi from the culture and PCR-RFLP test.CONCLUSIONS: The species from PCR-RFLP examination was the same with the fungal culture.
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Burgermeister, Wolfgang, Helen Braasch, Kai Metge, Jianfeng Gu, Thomas Schröder, and Elvira Woldt. "ITS-RFLP analysis, an efficient tool for differentiation of Bursaphelenchus species." Nematology 11, no. 5 (2009): 649–68. http://dx.doi.org/10.1163/156854108/399182.
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Abstract Restriction analysis of amplified ribosomal ITS sequences has provided species-specific fragment patterns for nematodes of several genera, including Bursaphelenchus. We used restriction enzymes RsaI, HaeIII, MspI, HinfI and AluI to produce ITS-RFLP reference profiles of 44 Bursaphelenchus species, including two intraspecific types in each of B. mucronatus and B. leoni. In addition, reference profiles of Aphelenchoides stammeri and Ruehmaphelenchus asiaticus were produced. Reference profiles of six species are shown here for the first time. Identical ITS-RFLP patterns were usually obtained from different isolates and from individual specimens of the same species. However, in the case of B. 'corneolus', B. lini, B. singaporensis and B. sexdentati, additional bands in the patterns of certain isolates or individual nematodes were observed which may be explained by ITS sequence microheterogeneity, i.e., the presence of ITS sequence variants within the number of rDNA tandem repeats. Since these 'extra' bands appeared only with one out of the five restriction enzymes employed, they did not seriously impair identification of species based on the overall reference patterns. ITS-RFLP analysis has proved valuable for differentiation of the pathogenic pine wood nematode, B. xylophilus, from related species. In many recent descriptions of new Bursaphelenchus species, ITS-RFLP profiles have been used as additional species identification criteria. Comparison of profiles from isolates of many different origins has provided new information on intraspecific types or genetically distinct provenances of several Bursaphelenchus species.
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Wang, Qin, and Liang-Dong Guo. "Ectomycorrhizal community composition of Pinus tabulaeformis assessed by ITS-RFLP and ITS sequences." Botany 88, no. 6 (June 2010): 590–95. http://dx.doi.org/10.1139/b10-023.
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Ectomycorrhizal (ECM) fungal composition was examined in a Pinus tabulaeformis Carr. forest. A total of 28 root samples of P. tabulaeformis were collected in June and September. Thirty-five ECM morphotypes were identified according to ECM morphological characters, and 26 ECM fungi were identified based on the analyses of ITS-RFLP and ITS sequences. Tomentella , Sebacina , and Tuber were common genera, and Atheliaceae sp., Lactarius deliciosus , Tomentella ferruginea , and Tomentella sp. 3 were dominant species. Of these ECM fungi, 13 were found in June, 19 in September, and 6 during both sampling times. Atheliaceae sp. and T. ferruginea were the dominant fungi both in June and September. Lactarius deliciosus was dominant in June, but rare in September. Tomentella sp. 3 was dominant in September but rare in June.
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Wurzburger, Nina, Martin I. Bidartondo, and Caroline S. Bledsoe. "Characterization of Pinus ectomycorrhizas from mixed conifer and pygmy forests using morphotyping and molecular methods." Canadian Journal of Botany 79, no. 10 (October 1, 2001): 1211–16. http://dx.doi.org/10.1139/b01-079.
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We used morphotyping and molecular methods to characterize ectomycorrhizas of bishop pine (Pinus muricata D. Don) and Bolander pine (Pinus contorta ssp. bolanderi (Parl.) Critchf.) from mixed conifer and hydric pygmy forests on the northern California coast. Sixteen ectomycorrhizal morphotypes were described, producing 15 internal transcribed spacer restriction fragment length polymorphism (ITS-RFLP) types, and 12 were identified via ITS sequencing. From a given site, all root tips of a specific morphotype produced identical ITS-RFLP patterns. However, sometimes two morphotypes produced the same ITS-RFLP type, and sometimes samples of the same morphotype from two different sites produced two different ITS-RFLP types. These results indicate that surveys of ectomycorrhizal fungi based on morphology alone are not sufficient, and that grouping morphotypes prior to molecular analysis can expedite the process. Ectomycorrhizas from mixed conifer included Russuloid sp., Tomentella sublilacina (Ellis & Holw.) Wakef., Tuber sp., and two Thelephoroid species. Ectomycorrhizas from hydric pygmy included two Dermocybe spp., a Cortinarius sp., two Thelephoroid spp., and Suillus tomentosus (Kauffman) Singer. Both plant communities contained Cenococcum geophilum Fr.:Fr. The hydric pygmy sites were more similar to each other than to the mixed conifer site (Jaccard similarity). The presence of ectomycorrhizal taxa in one plant community type may reflect biotic (host specificity) or abiotic (soil fertility or hydrology) adaptation.Key words: ectomycorrhiza, bishop pine, Pinus muricata, Bolander pine, Pinus contorta ssp. bolanderi, morphotyping, ITS-RFLP.
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Diao, Ying, Xian-Ming Lin, Chao-Lin Liao, Chun-Zi Tang, Zhong-Jian Chen, and Zhong-Li Hu. "Authentication ofPanax ginsengfrom its Adulterants by PCR-RFLP and ARMS." Planta Medica 75, no. 05 (February 2, 2009): 557–60. http://dx.doi.org/10.1055/s-0029-1185321.
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Viaud, Muriel, Aymeric Pasquier, and Yves Brygoo. "Diversity of soil fungi studied by PCR-RFLP of ITS." Mycological Research 104, no. 9 (September 2000): 1027–32. http://dx.doi.org/10.1017/s0953756200002835.
Full textDissertations / Theses on the topic "ITS-RFLP"
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Anderson, Ian C., of Western Sydney Nepean University, of Science Engineering and Technology Faculty, and School of Science. "Inter- and intraspecific variation in Pisolithus from central and eastern mainland Australia." THESIS_FST_SS_Anderson_I.xml, 2000. http://handle.uws.edu.au:8081/1959.7/237.
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Pisolithus is an important ectomycorrhizal genus world-wide, however to date we remain largely ignorant of the genetic and functional variation that exists within isolates of this genus. Fifty-three isolates of Pisolithus were obtained from various locations in central and eastern Australia and genetic variation within the isolates was assessed using ITS-RFLP and ITS sequencing analyses. RFLP analysis initially grouped the isolates into eight RFLP types. Neighbour-joining analysis of ITS sequences with Pisolithus ITS sequences available in databases clustered the majority of isolates into four groups within two major clades, each comprising isolates of similar basidiospre characteristics. Most Australian isolates correspond with recent provisional descriptions of P. albus or P. marmoratus. One isolate (LJ30) had low sequence identity (61.6-78.0%) to the other isolates and probably represents a separate undescribed Australian species. Significant intraspecific variation was observed in ITS-RFLP profiles for the putative P. albus isolates, suggesting that the sole use of RFLP analysis in diversity assessment may over-estimate Pisolithus species richness. Investigations were also initiated to identify if a relationship exists between genetic and physiological diversity in Australian Pisolithus. It is, however, clear that extensive physiological variation exists in Australian Pisolithus isolates. The size and distribution of genets of Australian Pisolithus species I and II ( putative P. albus and P. marmoratus) was also assessed using microsatellite-primed PCR to gain a better understanding of the likely distribution of underground mycelial networks and possible reproduction strategies in native soils. The data demonstrate that both species have the ability to be long-lived and extend for significant distances in native soils in undisturbed conditions. The field site for Pisolithus species I, however, also contained of a large number of small individuals suggesting that this species may employ a life-history strategy combining r-, C and S characteristics depending on local soil conditionsDoctor of Philosophy (PhD)
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Diguta, Filofteia Camelia. "Ecologie des moisissures présentes sur les baies de raisin." Phd thesis, Université de Bourgogne, 2010. http://tel.archives-ouvertes.fr/tel-00597399.
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La microflore des raisins est importante d'un point de vue technologique car elle conditionne en partie la qualité du vin. Or, la diversité des flores fongiques présentes sur baies de raisin ainsi que leur potentiel de contamination du produit final ne sont pas encore pleinement connus. Dans ce cadre, la caractérisation des flores fongiques cultivables présentes sur baies de raisin a été réalisée par PCR ITS-RFLP. 41 espèces de moisissures différentes sur les 43 étudiées appartenant à 11 genres différents ont été caractérisées de façon fiable. Seules les espèces Penicillium thomii et Penicillium glabrum ont présenté le même profil. Ainsi 96.3% des souches étudiées ont été caractérisées avec au maximum 4 enzymes de restriction et 41.5% des souches ont pu l'être avec seulement 2 enzymes de restriction. Ces résultats ont permis d'enrichir les bases de données, moyennement pourvues en séquences ITS caractéristiques de genres ou d'espèces de moisissures présentes sur baies de raisin. De plus, une étude exhaustive des moisissures présentes sur baies de raisin en Bourgogne a permis, par PCR ITS-RFLP, d'identifier 199 souches au niveau de l'espèce et ce quelque soit le genre. Penicillium spinulosum est l'espèce majoritaire isolée pour le millésime 2008 en Bourgogne. Parallèlement, la quantification de Botrytis cinerea, choisi comme micro-organisme modèle, a été réalisée par qPCR. La technique qPCR décrite dans ce travail présente (i) une bonne sensibilté avec une limite de détection de 6.4 pg d'ADN correspondant à 540 spores, (ii) l'originalité de travailler en échantillons naturellement contaminés et la fiabilité d'utiliser un standard interne. L'évaluation de l'efficacité de différentes stratégies de traitements anti-Botrytis a confirmé l'importance de la prophylaxie (effeuillage) dans la lutte contre Botrytis cinerea.
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Keriuscia, Gokul Jarishma. "Eukaryotic diversity of miers valley hypoliths." Thesis, University of the Western Cape, 2012. http://hdl.handle.net/11394/4031.
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>Magister Scientiae - MScThe extreme conditions of Antarctic desert soils render this environment selective towards a diverse range of psychrotrophic microbial communities. Cracks and fissures in translucent quartz rocks permit an adequate amount of penetrating light, sufficient water and nutrients to support cryptic microbial development. Hypolithons colonizing the ventral surface of these quartz rocks have been classified into three types: cyanobacterial dominated (Type I),moss dominated (Type II) and lichenized (Type III) communities. Eukaryotic microbial communities were reported to represent only a minor fraction of Antarctic communities. In this study, culture independent techniques (DGGE, T-RFLP and clone library construction) were employed to determine the profile of the dominant eukaryotes, fungi and microalgae present in the three different hypolithic communities. The 18S rRNA gene (Euk for eukaryotes), internal transcribed spacer (ITS for fungi) and microalgal specific regions of the 18S rRNA gene, were the phylogenetic markers targeted for PCR amplification from hypolith metagenomic DNA. Results suggest that the three hypolith types are characterized by different eukaryotic, fungal and microalgal communities, as implied by nMDS analysis of the DGGE and T-RFLP profiles. Sequence analysis indicates close affiliation to members of Amoebozoa, Alveolata, Rhizaria (general eukaryote), Ascomycota (fungal) and Streptophyta (microalgal). Many of these clones may represent novel species. This study demonstrates that Dry Valley hypolithons harbour higher eukaryote diversity than previously recognised.Each hypolithon is colonized by specialized microbial communities with possible keystone species. The ecological role of the detected microorganisms in the hypolith environment is also theorized, and a trophic hierarchy postulated.
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Diguta, Camelia Filofteia. "Ecologie des moisissures présentes sur les baies de raisin." Thesis, Dijon, 2010. http://www.theses.fr/2010DIJOS045/document.
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La microflore des raisins est importante d’un point de vue technologique car elle conditionne en partie la qualité du vin. Or, la diversité des flores fongiques présentes sur baies de raisin ainsi que leur potentiel de contamination du produit final ne sont pas encore pleinement connus. Dans ce cadre, la caractérisation des flores fongiques cultivables présentes sur baies de raisin a été réalisée par PCR ITS-RFLP. 41 espèces de moisissures différentes sur les 43 étudiées appartenant à 11 genres différents ont été caractérisées de façon fiable. Seules les espèces Penicillium thomii et Penicillium glabrum ont présenté le même profil. Ainsi 96.3% des souches étudiées ont été caractérisées avec au maximum 4 enzymes de restriction et 41.5% des souches ont pu l’être avec seulement 2 enzymes de restriction. Ces résultats ont permis d’enrichir les bases de données, moyennement pourvues en séquences ITS caractéristiques de genres ou d’espèces de moisissures présentes sur baies de raisin. De plus, une étude exhaustive des moisissures présentes sur baies de raisin en Bourgogne a permis, par PCR ITS-RFLP, d’identifier 199 souches au niveau de l’espèce et ce quelque soit le genre. Penicillium spinulosum est l’espèce majoritaire isolée pour le millésime 2008 en Bourgogne. Parallèlement, la quantification de Botrytis cinerea, choisi comme micro-organisme modèle, a été réalisée par qPCR. La technique qPCR décrite dans ce travail présente (i) une bonne sensibilté avec une limite de détection de 6.4 pg d’ADN correspondant à 540 spores, (ii) l’originalité de travailler en échantillons naturellement contaminés et la fiabilité d’utiliser un standard interne. L’évaluation de l’efficacité de différentes stratégies de traitements anti-Botrytis a confirmé l’importance de la prophylaxie (effeuillage) dans la lutte contre Botrytis cinereaMicrobial population of grapes is important from a technological point of view because it determines the quality of wine. But few studies have focused on fungal populations of grapes. A better knowledge of the fungal diversity on grapes, particularly as concerns species responsible for wine defects, may help efforts to control their development.We report the development of a PCR ITS-RFLP method as a fast and easy technique for identifying species of fungal genera present on grapes. By this methode, 41 different fungal species among 43 studied species belonging to 11 genera were characterized at the species level. Only P. thomii remained indistinguishable from P. glabrum. Using this PCR-ITS-RFLP, 96.3% strains tested could be differentiated to the species level with only four enzymes and 41.5% only with two enzymes. Moreover this work has contributed to the enriching of the database of fungal ITS sequences.Thus 199 isolated strain were on grapes in Burgundy vineyard were chacacterized at species level indepdantly of the genus by this method. P. spinolusum was the most frequently isolated species of Penicillium in Burgundy for 2008 vintage. Paralelly, the quantification of Botrytis cinerea, used as model, was developped by qPCR. The assay contained an internal amplification control to compensate for variations in DNA extraction and the various compounds from grapes, had high efficiency and the limit of detection was estimated to be 6.3 pg DNA (corresponding to 540 spores). This method was applied to assess the effects of various treatment strategies against Botrytis in the vineyard and demonstrates the importance of the prophylactic method
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Saraiva, Greice Kelle Viegas. "Diversidade genética de leveduras isoladas indústria de leite da Zona da Mata Mineira por RAPD e PCR-RFLP da região ITS do rDNA." Universidade Federal de Viçosa, 2002. http://www.locus.ufv.br/handle/123456789/10643.
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
A diversidade genética de vinte e sete isolados de leveduras coletadas em laticínios, utilizando como referências dos gêneros Kluyveromyces e Debaryomyces, foi averiguada por meio de RAPD (Randomly Amplified Polymorphic DNA). A amplificação resultou em um total de oitenta e oito fragmentos polimórfico de DNA, utilizando treze oligonucleotídeos decâmeros aleatórios. As distâncias genéticas variaram de 6,5 a 71%, gerando na análise gráfica, cinco grupos geneticamente divergentes. Nas avaliações da região ITS do rDNA foi observado um polimorfismo de tamanho que variou de 380 a 710 pb. A análise de agrupamento utilizando valores da distância genética resultou na formação de oito grupos, sugerindo a existência de pelo menos oito espécies de leveduras. Na análise por PCR-RFLP da região ITS do rDNA, os produtos das amplificações foram hidrolisados com diferentes endonucleases de restrição evidenciando o padrão polimórfico, e os valores das distâncias genéticas foram utilizados para o agrupamento, resultando na formação de quinze grupos. Os agrupamentos obtidos com os marcadores moleculares possibilitaram a diferenciação genética dos isolados. Os resultados sugerem que quatro dos vinte e sete isolados pertencem à espécie Kluyveromyces lactis.
The genetic diversity of twenty-seven yeasts collected at dairies, was evaluated by RAPD using Kluyveromyces and Debaryomyces reference genera. Amplification using thirteen random oligonucleotides resulted in eighty-eight DNA polymorphic fragments. The Genetic distances varied from 6,5 to 71%, and generated a Dendrogram with five different genetic groups. The amplification of the ITS 18SrDNA region from the yeast resulted in DNA fragments with length polymorphism (380 and 710 bp). The clustering analysis using the genetic distance value produced eight groups, reflecting at least eight yeasts species. The amplification products of the rDNA ITS region using PCR-RFLP analyses were cleaved with different restriction endonucleases showing polymorphic patterns. The values of the genetic distances were used for the clustering resulted in fifteen groups. The clusters obtained using the molecular markers showed genetic difference between you isolates. The results suggest that four of the twenty-seven isolates can be identified as the yeast Kluyveromyces lactis.
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Guérin, Alexis. "Les Lactaires à lait rouge : mycorhization controlée des pins et caractérisation moléculaire : Application à l'étude de la compétence écologique et de la compétitivité d'isolats de lactarius deliciosus." Montpellier, ENSA, 1998. http://www.theses.fr/1998ENSA0004.
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Anderson, Ian C. "Inter- and intraspecific variation in Pisolithus from central and eastern mainland Australia." Thesis, View thesis, 2000. http://hdl.handle.net/1959.7/uws:237.
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Pisolithus is an important ectomycorrhizal genus world-wide, however to date we remain largely ignorant of the genetic and functional variation that exists within isolates of this genus. Fifty-three isolates of Pisolithus were obtained from various locations in central and eastern Australia and genetic variation within the isolates was assessed using ITS-RFLP and ITS sequencing analyses. RFLP analysis initially grouped the isolates into eight RFLP types. Neighbour-joining analysis of ITS sequences with Pisolithus ITS sequences available in databases clustered the majority of isolates into four groups within two major clades, each comprising isolates of similar basidiospre characteristics. Most Australian isolates correspond with recent provisional descriptions of P. albus or P. marmoratus. One isolate (LJ30) had low sequence identity (61.6-78.0%) to the other isolates and probably represents a separate undescribed Australian species. Significant intraspecific variation was observed in ITS-RFLP profiles for the putative P. albus isolates, suggesting that the sole use of RFLP analysis in diversity assessment may over-estimate Pisolithus species richness. Investigations were also initiated to identify if a relationship exists between genetic and physiological diversity in Australian Pisolithus. It is, however, clear that extensive physiological variation exists in Australian Pisolithus isolates. The size and distribution of genets of Australian Pisolithus species I and II ( putative P. albus and P. marmoratus) was also assessed using microsatellite-primed PCR to gain a better understanding of the likely distribution of underground mycelial networks and possible reproduction strategies in native soils. The data demonstrate that both species have the ability to be long-lived and extend for significant distances in native soils in undisturbed conditions. The field site for Pisolithus species I, however, also contained of a large number of small individuals suggesting that this species may employ a life-history strategy combining r-, C and S characteristics depending on local soil conditions
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Marques, Cálita Pollyanna. "Padronização da análise dos produtos da PCR/RFLP que amplifica os genes Ribossomal Internal transcribed spacer (ITS) e Glucose-6- phosphate dehydrogenase (G6PD) para identificação de Leishmania spp. em gel de poliacrilamida." Universidade Federal de Goiás, 2017. http://repositorio.bc.ufg.br/tede/handle/tede/7789.
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Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG
American Cutaneous Leishmaniasis (LTA) is caused by protozoa of the genus Leishmania and it attacks the skin and/or mucous membranes. It is an endemic zoonosis in Brazil being notified in all the Regions. There is a relation between the species of parasite and the clinical manifestations, contributing to the diagnostic complexity, which up to the present, did not exist species-specific diagnosis. In the present work, the proposal was to standardize the analysis of the PCR/RFLP products to the ITS and G6PD genes in polyacrylamide gel. In order to standardize PCR/RFLP analysis on the ITS and G6PD genes, DNA from the reference Leishmania spp. Sample was used and 5%, 6%, 7% 8% and 10% polyacrylamide were tested. The protocol design considered the size of the amplicons generated by the PCR / RFLP, correlating the concentration of the polyacrylamide. To standardize the electrophoresis time the migration of the dyes used in the DNA sample buffer was monitored. To validate the technique, we selected 521 samples from Tocantins patients fixed on slides and 43 samples of parasite isolates stored in Leishbank. The samples from the patients from Tocantins were analyzed by PCR/RFLP to the kDNA and were selected to those positive for the validation. The concentration of the matrix for analysis of the PCR and RFLP products to the ITS gene was standardized in 6% and 8%, respectively, and distinguished the species L. (L.) amazonensis, but the species L. (V.) braziliensis, L. (V.) guyanensis and L. (V.) lainsoni were not distinguished among themselves, occurring the same between L. (V.) naiffi and L. (V.) shawi. The concentration of the matrix for analysis of the PCR products to the G6PD gene was standardized in 6% and distinguished L. (V.) braziliensis from the others. The validation of the polyacrylamide gel analysis of the PCR products to the ITS target from samples of patients fixed in slides was not performed, since of the 256 samples analyzed only 4 amplified. The validation of the analysis of the products of the PCR to the G6PD target in polyacrylamide gel from 48 samples fixed in slides characterized 81% as L. (V.) braziliensis and, characterized 63% of the isolates of parasites as L. (V.) braziliensis. The PCR/RFLP analyzes performed to discriminate Leishmania spp. are complementary and thus contribute to the species-specific diagnosis of LTA.
A Leishmaniose Tegumentar Americana (LTA) é causada por protozoários do gênero Leishmania e acomete pele e/ou mucosas. É uma zoonose endêmica no Brasil sendo notificada em todas as Regiões. Há uma relação entre a espécie de parasito e as manifestações clínicas, contribuindo à complexidade diagnóstica, que até o presente, não existi o diagnóstico espécie-específico. No presente trabalho, a proposta foi padronizar a análise dos produtos da PCR/RFLP aos genes ITS e G6PD em gel de poliacrilamida. Para padronizar as análises dos produtos da PCR/RFLP aos genes ITS e G6PD, foram utilizadas DNA da amostra de Leishmania spp., de referência, sendo testada a poliacrilamida na concentração de 5%, 6%, 7% 8% e 10%. O delineamento do protocolo considerou o tamanho dos amplicons gerados pela PCR/RFLP, correlacionando-o a concentração da poliacrilamida. Para padronizar o tempo da eletroforese foi monitorada a migração dos corantes usados no tampão da amostra de DNA. Para validar a técnica, foram selecionadas 521 amostras de pacientes tocantinenses fixadas em lâminas e 43 amostras de isolados de parasitos armazenados no Leishbank. As amostras dos pacientes tocantinenses foram analisadas pela PCR/RFLP ao kDNA e foram selecionadas àquelas positivas para a validação. A concentração da matriz para analise dos produtos da PCR e RFLP ao gene ITS ficou padronizada respectivamente em 6% e 8% e distinguiu a espécie L.(L.) amazonensis, porém as espécies L. (V.) braziliensis, L. (V.) guyanensis e L. (V.) lainsoni não foram distinguidas entre si, ocorrendo o mesmo entre L. (V.) naiffi e L. (V.) shawi. A concentração da matriz para analise dos produtos da PCR ao gene G6PD ficou padronizada em 6% e distinguiu L. (V.) braziliensis das demais. A validação da análise em gel de poliacrilamida dos produtos da PCR ao alvo ITS a partir de amostras de pacientes fixadas em lâminas não ser realizada, pois das 256 amostras analisadas somente 4 amplificaram. A validação da análise dos produtos da PCR ao alvo G6PD em gel de poliacrilamida a partir de 48 amostras fixadas em lâminas caracterizou 81% como L. (V.) braziliensis e, caracterizou 63% dos isolados de parasitos como L. (V.) braziliensis. As análises pela PCR/RFLP realizadas para discriminar Leishmania spp. se complementam e, assim, contribuem para o diagnóstico espécie-específico da LTA.
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Yamoah, Emmanuel. "A model system using insects to vector Fusarium tumidum for biological control of gorse (Ulex europaeus)." Phd thesis, Lincoln University. Bio-Protection and Ecology Division, 2007. http://theses.lincoln.ac.nz/public/adt-NZLIU20080131.114607/.
Full textAbstract:
The overall objective of this study was to test the hypothesis that insects can vector F.
tumidum conidia to infect gorse plants with the aim of developing an alternative approach to mycoherbicide delivery to control weeds. Four potential insect species (Apion ulicis, Cydia ulicetana, Epiphyas postvittana and Sericothrips staphylinus) were assessed for their ability to vector F. tumidum conidia. To achieve this, the external microflora (bacteria and fungi) and the size and location of fungal spores on the cuticle of these insect species were determined. In addition, the ability of the insects to pick up and deposit F. tumidum conidia on agar was studied. Based on the results from these
experiments, E. postvittana was selected for more detailed experiments to determine transmission of F. tumidum to infect potted gorse plants. The factors promoting pathogenicity of F. tumidum against gorse and the pathogen loading required to infect and kill the weed were also determined.
The external microflora of the four insect species were recovered by washing and plating
techniques and identified by morphology and polymerase chain reaction restriction
fragment length polymorphism (PCR-RFLP) and sequencing of internally transcribed
spacer (ITS) and 16S rDNA. A culture-independent technique (direct PCR) was also used
to assess fungal diversity by direct amplification of ITS sequences from the washings of the insects. All insect species carried Alternaria, Cladosporium, Nectria, Penicillium,
Phoma, Pseudozyma spp. and entomopathogens. Ninety four per cent of the 178 cloned
amplicons had ITS sequences similarity to Nectria mauritiicola. E. postvittana carried the
largest fungal spores (mean surface area of 125.9 ìm2) and the most fungal CFU/insect.
About 70% of the fungi isolated from the insects were also present on the host plant
(gorse) and the understorey grass. The mean size of fungal spores recovered from the
insect species correlated strongly with their body length (R² = 85%). Methylobacterium
aquaticum and Pseudomonas lutea were common on all four insect species.
Pseudomonas fluorescens was the most abundant bacterial species.
In the pathogenicity trials, the effectiveness of F. tumidum in reducing root and shoot
biomass of 16 and 8 wk old gorse plants was significantly increased with wounding of the
plants. Older plants (32 wk old) which were wounded and inoculated were significantly
shorter, more infected and developed more tip dieback (80%) than plants which were not
wounded (32%). This indicates that damage caused by phytophagous insect species
present on gorse through feeding and oviposition may enhance infection by F. tumidum.
Wounding may release nutrients (e.g. Mg and Zn) essential for conidia germination and
germ tube elongation and also provide easier access for germ tube penetration. Conidial
germination and germ tube length were increased by 50 and 877%, respectively when
incubated in 0.2% of gorse extract solution for 24 h compared with incubation in water.
Inoculum suspensions amended with 0.2% of gorse extract caused more infection and
significantly reduced biomass production of 24 wk old gorse plants than suspensions
without gorse extract. A minimum number of about 900 viable conidia/infection site of F.
tumidum were required to infect gorse leaves. However, incorporation of amendments
(which can injure the leaf cuticle) or provision of nutrients (i.e. gorse extract or glucose)
in the formulation might decrease the number of conidia required for lesion formation.
Scanning electron micrographs showed that germ tube penetration of gorse tissue was
limited to open stomata which partly explain the large number of conidia required for
infection. The flowers and leaves were more susceptible to F. tumidum infection than the
spines, stems and pods. An experiment to determine the number of infection sites
required to cause plant mortality showed that the entire plant needs to be inoculated in
order for the pathogen to kill 10 wk old plants as F. tumidum is a non systemic pathogen.
The number of infection sites correlated strongly with disease severity (R² = 99.3%). At
least 50% of the plant was required to be inoculated to cause a significant reduction in
shoot dry weight. F. tumidum, applied as soil inoculant using inoculated wheat grains in three separate experiments, significantly suppressed gorse seedling emergence and biomass production.
In experiments to determine the loading capacity of the insect species, E. postvittana, the
largest insect species studied, carried significantly more (68) and deposited significantly more (29) F. tumidum conidia than the other species. Each E. postvittana, loaded with 5,000 conidia of F. tumidum, transmitted approximately 310 conidia onto gorse plants but
this did not cause any infection or affect plant growth as determined by shoot fresh
weight and shoot height. E. postvittana on its own did not cause any significant damage
to gorse and did not enhance F. tumidum infection. It also failed to spread the pathogen
from infected plants to the healthy ones. There was no evidence of synergism between the
two agents and damage caused by the combination of both E. postvittana and F. tumidum
was equivalent to that caused by F. tumidum alone.
This study has shown that E. postvittana has the greatest capacity to vector F. tumidum
since it naturally carried the largest and the most fungal spores (429 CFU/insect).
Moreover, it naturally carried Fusarium spp. such as F. lateritium, F. tricinctum and
Gibberella pulicaris (anamorph Fusarium sambucinum) and was capable of carrying and
depositing most F. tumidum conidia on agar. Coupled with the availability of pheromone
for attracting the male insects, E. postvittana may be a suitable insect vector for
delivering F. tumidum conidia on gorse using this novel biocontrol strategy. Although it
is a polyphagous insect, and may visit non-target plants, F. tumidum is a very specific
pathogen of gorse, broom and a few closely related plant species. Hence, using this insect
species to vector F. tumidum in a biological control programme, should not pose a
significant threat to plants of economic importance. However, successful control of gorse
using this "lure-load-infect" concept would depend, to a large extent on the virulence of
the pathogen as insects, due to the large size of F. tumidum macroconidia, can carry only
a small number of it.
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Kolenda, Magdalena. "The polymorphism in GDF8 gene and its association with meat performance in the chosen sheep breeds." Rozprawa doktorska, Uniwersytet Technologiczno-Przyrodniczy w Bydgoszczy, 2016. http://dlibra.utp.edu.pl/Content/937.
Full textBooks on the topic "ITS-RFLP"
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Hering, Olaf. Charakterisierung und Differenzierung bei Fusarium Link mittels RAPD und ITS-RFLP. Berlin: Parey, 1997.
Find full textBook chapters on the topic "ITS-RFLP"
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Rousseaux, Sandrine, and Michèle Guilloux-Bénatier. "PCR ITS-RFLP for Penicillium Species and Other Genera." In Methods in Molecular Biology, 321–33. New York, NY: Springer New York, 2016. http://dx.doi.org/10.1007/978-1-4939-6707-0_21.
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Croft, J. H., V. Bhattacherjee, and K. E. Chapman. "RFLP Analysis of Nuclear and Mitochondrial DNA and its Use in Aspergillus Systematics." In Modern Concepts in Penicillium and Aspergillus Classification, 309–20. Boston, MA: Springer US, 1990. http://dx.doi.org/10.1007/978-1-4899-3579-3_28.
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Maccaferri, Marco, Martina Bruschi, and Roberto Tuberosa. "Sequence-Based Marker Assisted Selection in Wheat." In Wheat Improvement, 513–38. Cham: Springer International Publishing, 2022. http://dx.doi.org/10.1007/978-3-030-90673-3_28.
Full textAbstract:
AbstractWheat improvement has traditionally been conducted by relying on artificial crossing of suitable parental lines followed by selection of the best genetic combinations. At the same time wheat genetic resources have been characterized and exploited with the aim of continuously improving target traits. Over this solid framework, innovations from emerging research disciplines have been progressively added over time: cytogenetics, quantitative genetics, chromosome engineering, mutagenesis, molecular biology and, most recently, comparative, structural, and functional genomics with all the related -omics platforms. Nowadays, the integration of these disciplines coupled with their spectacular technical advances made possible by the sequencing of the entire wheat genome, has ushered us in a new breeding paradigm on how to best leverage the functional variability of genetic stocks and germplasm collections. Molecular techniques first impacted wheat genetics and breeding in the 1980s with the development of restriction fragment length polymorphism (RFLP)-based approaches. Since then, steady progress in sequence-based, marker-assisted selection now allows for an unprecedently accurate ‘breeding by design’ of wheat, progressing further up to the pangenome-based level. This chapter provides an overview of the technologies of the ‘circular genomics era’ which allow breeders to better characterize and more effectively leverage the huge and largely untapped natural variability present in the Triticeae gene pool, particularly at the tetraploid level, and its closest diploid and polyploid ancestors and relatives.
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Krause, D., R. Szibor, W. Kuchheuser, and R. Brückner. "The Practical Significance of Human Genetic RFLP-Systems in Paternity Testing." In DNA — Technology and Its Forensic Application, 153–56. Berlin, Heidelberg: Springer Berlin Heidelberg, 1991. http://dx.doi.org/10.1007/978-3-642-76632-9_20.
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Scheithauer, R., and H. J. Weisser. "DNA Extraction and RFLP Analysis of Bloodstains on a Variety of Textiles — Investigation of Various Extraction Procedures." In DNA — Technology and Its Forensic Application, 181–86. Berlin, Heidelberg: Springer Berlin Heidelberg, 1991. http://dx.doi.org/10.1007/978-3-642-76632-9_26.
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"Appendix IV. Protocols For Identification Of Cyst Nematode Species With PCR-ITS-RFLP Using AB28 And TW81 Primers." In Systematics of Cyst Nematodes (Nematoda: Heteroderinae), Part A, 299–301. BRILL, 2010. http://dx.doi.org/10.1163/ej.9789004162259.i-352.72.
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Sana, Sadia, Naheed Akhter, Fozia Amjum, Samreen Gul Khan, and Muhammad Akram. "Genetic Diversity in Almond (Prunus dulcis)." In Prunus - Recent Advances [Working Title]. IntechOpen, 2021. http://dx.doi.org/10.5772/intechopen.99249.
Full textAbstract:
Almond (Prunus dulcis), a stone fruit belonging to a family Rosaceae (rose) is broadly cultivated for ornament and fruit. Within this genus, the almond is very much associated with the peach, and these two fruits share the same subgenus the Amygdalus. About 430 species are spread all through the northern temperate regions of the world. The Mediterranean climate region of the Middle East like Turkey and Pakistan eastward to Syria is native to the almond and its related species. Almond is one of the ancient fruit trees known to the Asian as well as European regions with the most primitive proof of cultivation dating about 2000 B.C. Prunus dulcis (Almond) is a nutrient-loaded nut crop. Almond possesses a great genetic diversity due to the genetically controlled self-incompatibility system which can be estimated by a morphological characteristic including molecular markers and isoenzymes with a wide range of marker techniques. Simple sequence repeats (SSR) involving RFLP or SNP are the most commonly used molecular techniques among the DNA-based molecular symbols. Particular agronomic characters, e.g. kernel bitterness or self-compatibility can also be traced by these molecular markers. The direct association between the level of diversity and the basis of the germplasm cannot be understood by the studies of genetic diversity. Genetic diversity cannot be seriously lost by self-compatibility in almonds. The breeding, conservation, and cultivation of wild-growing almonds may similarly advantageous after the genetic diversity research studies (especially those applying molecular markers).
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Budakva, Ye, K. Pochernyaev, S. Korinnyi, and M. Povod. "The use of mitochondrial genome polymorphism to establish pro-maternal breeds in the final hybrids of pigs." In Technological innovation engineering manufacturing agricultural complex and zoology (1st ed ), 34–41. Primedia eLaunch LLC, 2020. http://dx.doi.org/10.36074/ti:emacaz.ed-1.03.
Full textAbstract:
The purpose of this work was to determine the origin of pigs (Large White × Landrace) × Maxgro of the final hybrid using mitochondrial DNA markers. As a genetic material used bristles from the auricle of commercial pigs (large white × landrace) × Maxgro. DNA secretion was carried out according to the method of Korinnyi S.M., etc.2005, using Chelex ion exchange resin – 100. For the analysis of the mitochondrial genome, the method of polymorphism of the lengths of restriction fragments amplified in PCR was used (Polymerase chain reaction-restriction fragment of polymorphism – PCR-RFLP). The D-loop of the mitochondrial genome of a pig of 428 pairs is subject to analysis of nucleotide with Tasl recognition sites in positions 15558, 15580, 15616, 15714, 15758). Comparison of inherited on the maternal line restrict fragments in the breed allowed get reliable information about the origin of the experimentally studied sample of pigs of the final hybrid (n=15) from RPE "Globinsky Pig Farm", Globyno, town Poltava region, Ukraine allowed getting reliable information of their origin. Laboratory research was carried out based at the Institute of Pig Breeding and Agricultural Production NAAS in the Laboratory of Genetics. By analyzing nucleotide sequences, one monomorphic site was identified and experimentally investigated in position 15558 p.n. and three polymorphic sites in positions 15580, 15616, 15714, 15758 p.n. for endonuclease TasI (↓AATT). To systematize combinations of mitochondrial DNA restriction fragments, three breed-specific mitochondrial haplotypes were identified in an experimentally studied sample of hybrid pigs (n=15). With its use, mitochondrial haplotypes of pigs of the final hybrid were determined: 4 animals with haplotype C, 6 with haplotype N, and 5 with haplotype O. According to many authors pieces of research, these haplotypes characterize different breeds, namely C – Landrace, N – Large White (Asian type) and O – Landrace. The obtained data on the origin of animals of the final hybrid suggests that two-breed sows were the result of direct (Large White × Landrace) and reciprocal crossing (Landrace × Large White). Genetic examination in the establishment of maternal breeds of hybrid pigs with the help of markers of the mitochondrial genome and the search for a polymorphic area of the X and Y-chromosome to determine the ancestral line has become an urgent issue in continuing our research in modern pig breeding technologies of commercial lines. The work was done with the support of the National Academy of Agrarian Sciences of Ukraine 31.01.00.07. F. “Investigate the pleiotropic effect gens that the SNP use in marker-associated pig breeding”. DR № 0121U109838.
Conference papers on the topic "ITS-RFLP"
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Sharma, Vineeta, Pallavi Singhal, Anoop Kumar, V. G. Ramachandran, Shukla Das, and Mausumi Bharadwaj. "Association of TNF-α–rs 281865419 polymorphism with reproductive tract infections in Indian population." In 16th Annual International Conference RGCON. Thieme Medical and Scientific Publishers Private Ltd., 2016. http://dx.doi.org/10.1055/s-0039-1685357.
Full textAbstract:
Aim: To investigate the presence of reproductive tract infections (RTIs) in symptomatic and asymptomatic women in North India and association of SNPs in TNF? gene (rs-281865419 C/T) with susceptibility to these RTIs. Methods: We collected 100 symptomatic (cases) and 100 asymptomatic women (controls) samples and screened them for RTIs. Then genotyping of TNF-? gene was performed by PCR-RFLP. Results: Among cases the frequencies of RTIs infection is higher than control. The prevalence of HPV, C. trachomatis, T. vaginalis, Bacterial vaginosis and N. gonorrhoeae are 28% and 6%; 11%, 32% respectively while in controls it was 5%, 2%, 1% and 8% and 1%. In the present study we found that the frequency of wild homozygous genotype (TT) was lower in cases 30% (6/20) as compared to controls 60% (12/20). The frequency of the heterozygous polymorphic genotype (CT) was higher in cases 65% (65/100) as compared to controls 32% (32/100). It was interesting to note that the frequency of the polymorphic homozygous genotype (CC) was higher in cases 15% (15/100) than controls 2% (2/100). While the frequency of the carrier genotype (CT + TT) was found to be more in cases 70% (70/100) than in controls 40/100 (40%). This study shows that T allele may be risk factor for reproductive tract infections as its percentage is higher in cases as compare to normal controls. Conclusion: TNF-? rs-281865419 locus may serve as an important biomarker for RTIs predisposition in Indian population though larger sample size is needed to validate the findings.
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Sharma, Vineeta, Pallavi Singhal, Anoop Kumar, V. G. Ramachandran, Shukla Das, and Mausumi Bharadwaj. "Association of TNF-α rs-281865419 polymorphism with reproductive tract infections in Indian population." In 16th Annual International Conference RGCON. Thieme Medical and Scientific Publishers Private Ltd., 2016. http://dx.doi.org/10.1055/s-0039-1685270.
Full textAbstract:
Aim: To investigate the presence of reproductive tract infections (RTIs) in symptomatic and asymptomatic women in North India and association of SNPs in TNFα gene (rs-281865419 C/T) with susceptibility to these RTIs. Methods: We collected 100 symptomatic (cases) and 100 asymptomatic women (controls) samples and screened them for RTIs. Then genotyping of TNF-α gene was performed by PCR-RFLP. Results: Among cases the frequencies of RTIs infection is higher than control. The prevalence of HPV, C. trachomatis, T. vaginalis, Bacterial vaginosis and N. gonorrhoeae are 28% & 6%; 11%, 32% respectively while in controls it was 5%, 2%, 1% and 8% & 1%. In the present study we found that the frequency of wild homozygous genotype (TT) was lower in cases 30% (6/20) as compared to controls 60% (12/20). The frequency of the heterozygous polymorphic genotype (CT) was higher in cases 65% (65/100) as compared to controls 32% (32/100). It was interesting to note that the frequency of the polymorphic homozygous genotype (CC) was higher in cases 15% (15/100) than controls 2% (2/100). While the frequency of the carrier genotype (CT + TT) was found to be more in cases 70% (70/100) than in controls 40/100 (40%). This study shows that T allele may be risk factor for Reproductive tract infections as its percentage is higher in cases as compare to normal controls. Conclusion: TNF-? rs-281865419 locus may serve as an important biomarker for RTIs predisposition in Indian population though larger sample size is needed to validate the findings.
Reports on the topic "ITS-RFLP"
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Dubcovsky, Jorge, Tzion Fahima, and Ann Blechl. Positional cloning of a gene responsible for high grain protein content in tetraploid wheat. United States Department of Agriculture, September 2003. http://dx.doi.org/10.32747/2003.7695875.bard.
Full textAbstract:
High Grain Protein Content (GPC) is a desirable trait in breadmaking and pasta wheat varieties because of its positive effects on quality and nutritional value. However, selection for GPC is limited by our poor understanding of the genes involved in the accumulation of protein in the grain. The long-term goal of this project is to provide a better understanding of the genes controlling GPC in wheat. The specific objectives of this project were: a) to develop a high-density genetic map of the GPC gene in tetraploid wheat, b) to construct a T. turgidum Bacterial Artificial Chromosome (BAC) library, c) to construct a physical map of the GPC gene and identify a candidate for the GPC gene. A gene with a large effect on GPC was detected in Triticum turgidum var. dicoccoides and was previously mapped in the short arm of chromosome 6B. To define better the position of the Gpc-B1 locus we developed homozygous recombinant lines with recombination events within the QTL region. Except for the 30-cM region of the QTL these RSLs were isogenic for the rest of the genome minimizing the genetic variability. To minimize the environmental variability the RSLs were characterized using 10 replications in field experiments organized in a Randomized Complete Block Design, which were repeated three times. Using this strategy, we were able to map this QTL as a single Mendelian locus (Gpc-B1) on a 2.6-cM region flanked by RFLP markers Xcdo365 and Xucw67. All three experiments showed that the lines carrying the DIC allele had an average absolute increase in GPC of 14 g/kg. Using the RFLP flanking markers, we established the microcolinearity between a 2.l-cM region including the Gpc-B1 gene in wheat chromosome 6BS and a 350-kb region on rice chromosome 2. Rice genes from this region were used to screen the Triticeae EST collection, and these ESTs were used to saturate the Gpc-B1 region with molecular markers. With these new markers we were able to map the Gpc-B1 locus within a 0.3-cM region flanked by PCR markers Xucw83 and Xucw71. These flanking markers defined a 36-kb colinear region with rice, including one gene that is a potential candidate for the Gpc-B1 gene. To develop a physical map of the Gpc-B1 region in wheat we first constructed a BAC library of tetraploid wheat, from RSL#65 including the high Gpc-B1 allele. We generated half- million clones with an average size of l3l-kb (5.1 X genome equivalents for each of the two genomes). This coverage provides a 99.4% probability of recovering any gene from durum wheat. We used the Gpc-BI flanking markers to screen this BAC library and then completed the physical map by chromosome walking. The physical map included two overlapping BACs covering a region of approximately 250-kb, including two flanking markers and the Gpc-B1 gene. Efforts are underway to sequence these two BACs to determine if additional wheat genes are present in this region. Weare also developing new RSLs to further dissect this region. We developed PCR markers for flanking loci Xucw79andXucw71 to facilitate the introgression of this gene in commercial varieties by marker assisted selection (httQ://maswheat.ucdavis.edu/ orotocols/HGPC/index.hlm). Using these markers we introgressed the Gpc-B1 gene in numerous pasta and common wheat breeding lines.
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Feldman, Moshe, Eitan Millet, Calvin O. Qualset, and Patrick E. McGuire. Mapping and Tagging by DNA Markers of Wild Emmer Alleles that Improve Quantitative Traits in Common Wheat. United States Department of Agriculture, February 2001. http://dx.doi.org/10.32747/2001.7573081.bard.
Full textAbstract:
The general goal was to identify, map, and tag, with DNA markers, segments of chromosomes of a wild species (wild emmer wheat, the progenitor of cultivated wheat) determining the number, chromosomal locations, interactions, and effects of genes that control quantitative traits when transferred to a cultivated plant (bread wheat). Slight modifications were introduced and not all objectives could be completed within the human and financial resources available, as noted with the specific objectives listed below: 1. To identify the genetic contribution of each of the available wild emmer chromosome-arm substitution lines (CASLs) in the bread wheat cultivar Bethlehem for quantitative traits, including grain yield and its components and grain protein concentration and yield, and the effect of major loci affecting the quality of end-use products. [The quality of end-use products was not analyzed.] 2. To determine the extent and nature of genetic interactions (epistatic effects) between and within homoeologous groups 1 and 7 for the chromosome arms carrying "wild" and "cultivated" alleles as expressed in grain and protein yields and other quantitative traits. [Two experiments were successful, grain protein concentration could not be measured; data are partially analyzed.] 3. To derive recombinant substitution lines (RSLs) for the chromosome arms of homoeologous groups 1 and 7 that were found previously to promote grain and protein yields of cultivated wheat. [The selection of groups 1 and 7 tons based on grain yield in pot experiments. After project began, it was decided also to derive RSLs for the available arms of homoeologous group 4 (4AS and 4BL), based on the apparent importance of chromosome group 4, based on early field trials of the CASLs.] 4. To characterize the RSLs for quantitative traits as in objective 1 and map and tag chromosome segments producing significant effects (quantitative trait loci, QTLs by RFLP markers. [Producing a large population of RSLs for each chromosome arm and mapping them proved more difficult than anticipated, low numbers of RSLs were obtained for two of the chromosome arms.] 5. To construct recombination genetic maps of chromosomes of homoeologous groups 1 and 7 and to compare them to existing maps of wheat and other cereals [Genetic maps are not complete for homoeologous groups 4 and 7.] The rationale for this project is that wild species have characteristics that would be valuable if transferred to a crop plant. We demonstrated the sequence of chromosome manipulations and genetic tests needed to confirm this potential value and enhance transfer. This research has shown that a wild tetraploid species harbors genetic variability for quantitative traits that is interactive and not simply additive when introduced into a common genetic background. Chromosomal segments from several chromosome arms improve yield and protein in wheat but their effect is presumably enhanced when combination of genes from several segments are integrated into a single genotype in order to achieve the benefits of genes from the wild species. The interaction between these genes and those in the recipient species must be accounted for. The results of this study provide a scientific basis for some of the disappointing results that have historically obtained when using wild species as donors for crop improvement and provide a strategy for further successes.