scholarly journals Variation in Genotype and Aggressiveness of Ralstonia solanacearum Race 1 Isolated from Tomato in Taiwan

1999 ◽  
Vol 89 (4) ◽  
pp. 320-327 ◽  
Author(s):  
Thierry X. Jaunet ◽  
Jaw-Fen Wang

A population of Ralstonia solanacearum race 1 from tomato (Lycopersicon esculentum) was analyzed for genetic polymorphism and aggressiveness on tomato. The 46 strains were collected from main tomato-growing areas in Taiwan. Genetic analysis was achieved by two polymerase chain reaction (PCR)-based methods: REP-, ERIC-, and BOX-PCR (collectively as rep-PCR) and random amplified polymorphic DNA (RAPD) techniques. RAPD (with three 10-mers) and rep-PCR revealed 35 and 30 haplotypes, respectively, that were grouped in 14 clusters and 3 clusters, respectively. Distribution of strains into genetic clusters did not appear related to biovar or geographic origin in considering RAPD, rep-PCR, or composite data. Although strains were more dissimilar based on RAPD data than on rep-PCR data, the two techniques gave complementary results for strain clustering. A set of 40 strains representing the main haplotypes was inoculated on six tomato cultivars differing in their bacterial wilt resistance. Six groups differing in general level of aggressiveness and cultivar specificity were detected. Although populations were highly diverse in both genotype and aggressiveness, no association was found between the two characteristics. Although the sample sizes in this study were not adequate to draw definite conclusions about population structure, these results will be valuable for future population genetic studies on R. solanacearum.

1970 ◽  
Vol 35 (2) ◽  
pp. 313-322 ◽  
Author(s):  
M Maniruzzaman ◽  
ME Haque ◽  
MM Haque ◽  
MA Sayem ◽  
M Al-Amin

A polymerase chain reaction (PCR) based approach, namely random amplified polymorphic DNA (RAPD) analysis was applied to l0 varieties of onion (Allium cepa) in order to assess the degree of polymorphism within the genes and to investigate if this approach was suitable for genetic studies of onion. For this study, ten cultivars of onion were evaluated for variability using a set of 15 random l0-mer primers. The polymorphisms in PCR amplification products were subjected to the unweighed pair group method for arithmetic averages (UPGMA) and plotted in a phenogram. The dendogram constructed from the similarity data showed that all the cultivars analyzed were related. Among them, 12 of the primers revealed scorable (168 bands) polymorphisms between cultivars of A. cepa and the rest did not show polymorphism in their genetic level. In this study, it was found that Bermis and India-2 were more dissimilar and on the other hand, Faridpuri and Bhati were the most similar in their genetic level. Keywords: RAPD; onion; genetic diversity; polymorphism. DOI: 10.3329/bjar.v35i2.5894Bangladesh J. Agril. Res. 35(2) : 313-322, June 2010


2001 ◽  
Vol 91 (4) ◽  
pp. 399-407 ◽  
Author(s):  
Mitsuo Horita ◽  
Kenicki Tsuchiya

The genetic diversity of 74 Japanese strains of Ralstonia solanacearum was assessed by pathogenicity tests and the repetitive sequencebased polymerase chain reaction (rep-PCR) fingerprint method. Based on their genomic fingerprints, biovar N2 strains were divided into two distinct groups, one consisting of potato isolates belonging to race 3, and the other consisting of tomato, eggplant, pepper, and tobacco isolates belonging to race 1. Biovar 3 strains had low average similarity and were divided into five groups that differed in original host or pathogenicity. Biovar 4 strains consisted of only one group at the 80% similarity level. Comparative analysis of the rep-PCR fingerprints of 78 strains, including six biovars from Japan and various countries, revealed two main clusters. Cluster 1 comprised all biovar 3, 4, and 5 strains, biovar 1 strains from Reunion, and some biovar N2 strains from Japan. Cluster 2 included most of the biovar 1, 2, and N2 strains. The fingerprints showed low average similarity with biovar N2 strains from Japan and Brazil.


2005 ◽  
Vol 47 (5) ◽  
pp. 295-300 ◽  
Author(s):  
Jorge Fraga ◽  
Jinnay Rodriguez ◽  
Omar Fuentes ◽  
Aymé Fernandez-Calienes ◽  
Mayda Castex

Random amplified polymorphic DNA (RAPD) technique is a simple and reliable method to detect DNA polymorphism. Several factors can affect the amplification profiles, thereby causing false bands and non-reproducibility of assay. In this study, we analyzed the effect of changing the concentration of primer, magnesium chloride, template DNA and Taq DNA polymerase with the objective of determining their optimum concentration for the standardization of RAPD technique for genetic studies of Cuban Triatominae. Reproducible amplification patterns were obtained using 5 pmoL of primer, 2.5 mM of MgCl2, 25 ng of template DNA and 2 U of Taq DNA polymerase in 25 µL of the reaction. A panel of five random primers was used to evaluate the genetic variability of T. flavida. Three of these (OPA-1, OPA-2 and OPA-4) generated reproducible and distinguishable fingerprinting patterns of Triatominae. Numerical analysis of 52 RAPD amplified bands generated for all five primers was carried out with unweighted pair group method analysis (UPGMA). Jaccard's Similarity Coefficient data were used to construct a dendrogram. Two groups could be distinguished by RAPD data and these groups coincided with geographic origin, i.e. the populations captured in areas from east and west of Guanahacabibes, Pinar del Río. T. flavida present low interpopulation variability that could result in greater susceptibility to pesticides in control programs. The RAPD protocol and the selected primers are useful for molecular characterization of Cuban Triatominae.


HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 660b-660
Author(s):  
Robert L. Jarret

Patterns of diversity among thirty diploid clones of banana (Musa acuminata Colla.), collected in Papua New Guinea and the surrounding islands between 1987 and 1989, were examined genetically using the polymerase chain reaction (PCR) and random primers, to detect random amplified polymorphic DNA (RAPDs). PCR products were visualized on ethidium bromide stained agarose gels. Twenty of 60 random primers examined detected RAPDS in CTAB-extracted genomic DNA. Banding patterns ranged from very simple (1 or 2 bands/gel) to very complex (more than 20 bands/gel). All 30 Musa clones were distinguishable from each other based on their unique RAPD banding pattern. Principal component analysis (PCA) revealed several clusters of closely related clones within the materials examined. However, these clusterings were not correlated with either the geographic origin or the morphological characteristics of the clones. A role of the use of RAPDs in germplasm characterization is discussed.


Beverages ◽  
2021 ◽  
Vol 7 (2) ◽  
pp. 27
Author(s):  
Dimitrios Kontogiannatos ◽  
Vicky Troianou ◽  
Maria Dimopoulou ◽  
Polydefkis Hatzopoulos ◽  
Yorgos Kotseridis

Nemea and Mantinia are famous wine regions in Greece known for two indigenous grape varieties, Agiorgitiko and Moschofilero, which produce high quality PDO wines. In the present study, indigenous Saccharomyces cerevisiae yeast strains were isolated and identified from spontaneous alcoholic fermentation of Agiorgitiko and Moschofilero musts in order to evaluate their oenological potential. Random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) recovered the presence of five distinct profiles from a total of 430 yeast isolates. The five obtained strains were evaluated at microvinifications trials and tested for basic oenological and biochemical parameters including sulphur dioxide and ethanol tolerance as well as H2S production in sterile grape must. The selected autochthonous yeast strains named, Soi2 (Agiorgitiko wine) and L2M (Moschofilero wine), were evaluated also in industrial (4000L) fermentations to assess their sensorial and oenological characteristics. The volatile compounds of the produced wines were determined by GC-FID. Our results demonstrated the feasibility of using Soi2 and L2M strains in industrial fermentations for Agiorgitiko and Moschofilero grape musts, respectively.


Forests ◽  
2021 ◽  
Vol 12 (2) ◽  
pp. 222
Author(s):  
Bartosz Ulaszewski ◽  
Joanna Meger ◽  
Jaroslaw Burczyk

Next-generation sequencing of reduced representation genomic libraries (RRL) is capable of providing large numbers of genetic markers for population genetic studies at relatively low costs. However, one major concern of these types of markers is the precision of genotyping, which is related to the common problem of missing data, which appears to be particularly important in association and genomic selection studies. We evaluated three RRL approaches (GBS, RADseq, ddRAD) and different SNP identification methods (de novo or based on a reference genome) to find the best solutions for future population genomics studies in two economically and ecologically important broadleaved tree species, namely F. sylvatica and Q. robur. We found that the use of ddRAD method coupled with SNP calling based on reference genomes provided the largest numbers of markers (28 k and 36 k for beech and oak, respectively), given standard filtering criteria. Using technical replicates of samples, we demonstrated that more than 80% of SNP loci should be considered as reliable markers in GBS and ddRAD, but not in RADseq data. According to the reference genomes’ annotations, more than 30% of the identified ddRAD loci appeared to be related to genes. Our findings provide a solid support for using ddRAD-based SNPs for future population genomics studies in beech and oak.


2008 ◽  
Vol 34 (3) ◽  
pp. 228-231 ◽  
Author(s):  
Willian Mário de Carvalho Nunes ◽  
Maria Júlia Corazza ◽  
Silvana Aparecida Crestes Dias de Souza ◽  
Siu Mui Tsai ◽  
Eiko Eurya Kuramae

A simple, quick and easy protocol was standardized for extraction of total DNA of the bacteria Xanthomonas axonopodis pv. phaseoli. The DNA obtained by this method had high quality and the quantity was enough for the Random Amplified Polymorphic DNA (RAPD) reactions with random primers, and Polymerase Chain Reaction (PCR) with primers of the hypersensitivity and pathogenicity gene (hrp). The DNA obtained was free of contamination by proteins or carbohydrates. The ratio 260nm/380nm of the DNA extracted ranged from 1.7 to 1.8. The hrp gene cluster is required by bacterial plant pathogen to produce symptoms on susceptible hosts and hypersensitive reaction on resistant hosts. This gene has been found in different bacteria as well as in Xanthomonas campestris pv. vesicatoria (9). The primers RST21 and RST22 (9) were used to amplify the hrp gene of nine different isolates of Xanthomonas axonopodis pv. phaseoli from Botucatu, São Paulo State, Brazil, and one isolate, "Davis". PCR amplified products were obtained in all isolates pathogenic to beans.


Genome ◽  
2001 ◽  
Vol 44 (6) ◽  
pp. 995-999 ◽  
Author(s):  
H I Amadou ◽  
P J Bebeli ◽  
P J Kaltsikes

Random amplified polymorphic DNA (RAPD) markers were used to assess genetic diversity in Bambara groundnut (Vigna subterranea L.) germplasm using 25 African accessions from the collection in the International Institute for Tropical Agriculture, Ibadan, Nigeria. Fifty random decamer primers were screened to assess their ability to detect polymorphism in bambara; 17 of them were selected for this study. Considerable genetic diversity was found among the V. subterranea accessions studied. The relationships among the 25 accessions were studied by cluster analysis. The dendrograms showed two main groups of accessions mainly along the lines of their geographic origin. It is concluded that RAPD can be used for germplasm classification in bambara groundnut and hence for improving this crop.Key words: germplasm, PCR, RAPD, Vigna subterranea.


Plant Disease ◽  
2005 ◽  
Vol 89 (4) ◽  
pp. 380-384 ◽  
Author(s):  
Rick D. Peters ◽  
Rod J. Clark ◽  
Albert D. Coffin ◽  
Antony V. Sturz ◽  
David H. Lambert ◽  
...  

Pink rot of potato (Solanum tuberosum), caused by Phytophthora erythroseptica, is found wherever potatoes are grown, and in the last decade, it has reemerged as an economically important disease in Canada and the United States. A selection of isolates of P. erythroseptica from major potato-growing regions in North America, namely Prince Edward Island and New Brunswick, Canada, and Maine and Idaho, U.S.A., was assessed for genetic diversity with randomly chosen decanucleotide primers which were used to amplify regions of DNA to reveal polymorphisms among templates (random amplified polymorphic DNA [RAPD]). The isolates varied in their geographic origin as well as in their sensitivity to mefenoxam, as determined by an in vitro assay. In three separate RAPD screens (I, II, and III) with 23 isolates of P. erythroseptica chosen from a larger collection, 1,410, 369, and 316 robust, scorable bands were amplified, respectively. However, among the bands amplified in screens I, II, and III, only 3, 1, and 3 bands, respectively, were polymorphic. When three primers yielding polymorphisms were used to screen 106 isolates from Prince Edward Island and New Brunswick, or a representative collection of 32 isolates from Prince Edward Island, New Brunswick, Maine, and Idaho, no major variation was discovered. RAPD markers were not correlated with geographic origin or mefenoxam sensitivity of the isolates. From an evolutionary standpoint, the absence of genetic diversity among the isolates of P. erythroseptica we examined may be attributable to the relatively recent introduction of a small founding population of the pathogen in North America.


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