scholarly journals Characterization of the Colletotrichum Species Causing Anthracnose in Andean Blackberry in Colombia

Plant Disease ◽  
2014 ◽  
Vol 98 (11) ◽  
pp. 1503-1513 ◽  
Author(s):  
Lucía Afanador-Kafuri ◽  
Alonso González ◽  
Lederson Gañán ◽  
Juan Fernando Mejía ◽  
Nadya Cardona ◽  
...  
Keyword(s):  
Colletotrichum Gloeosporioides ◽  
Morphological Characteristics ◽  
High Sensitivity ◽  
Chain Reaction ◽  
Internal Transcribed Spacer Region ◽  
Colletotrichum Spp ◽  
Polymerase Chain ◽  
Polymerase Chain Reaction Primers ◽  
Multilocus Phylogeny

Since 1992, anthracnose of Andean blackberry (Rubus glaucus) has generated losses as high as 40% for farmers in Colombia. In this study, our goal was to characterize 240 Colletotrichum isolates from Andean blackberry in eight areas of Colombia. These isolates were evaluated according to morphological characteristics, sensitivity to benomyl, pathogenicity, and genetic variability. Identification of the genus Colletotrichum was achieved by using species complex-specific polymerase chain reaction primers. A multilocus phylogeny approach was used to identify isolates to the species level with sequences from the ribosomal internal transcribed spacer region and partial sequences of the actin, β-tubulin 2, calmodulin, chitin synthase 1, glutamine synthetase, and glyceraldehyde-3-phosphate dehydrogenase genes. Most of the isolates were identified as Colletotrichum gloeosporioides sensu lato, were associated with the Castilla ecotype, showed high sensitivity to benomyl, and were highly aggressive. Isolates identified as C. acutatum sensu lato were found mainly on the Thornless ecotype, were highly resistant to benomyl, and showed intermediate aggressiveness. Only three isolates were identified as C. boninense sensu lato. The species identified included C. fructicola, C. kahawae subsp. ciggaro, C. godetiae, C. karstii, C. brassicicola, and undetermined Colletotrichum spp. This study is the first report of these species associated with anthracnose in Andean blackberry.

scholarly journals Etiology and Population Genetics of Colletotrichum spp. Causing Crown and Fruit Rot of Strawberry

2002 ◽  
Vol 92 (11) ◽  
pp. 1245-1252 ◽  
Author(s):  
A. R. Ureña-Padilla ◽  
S. J. MacKenzie ◽  
B. W. Bowen ◽  
D. E. Legard
Keyword(s):  
Random Amplified Polymorphic Dna ◽  
Crown Rot ◽  
Fruit Rot ◽  
Chain Reaction ◽  
Internal Transcribed Spacer Region ◽  
Low Diversity ◽  
Diseased Fruit ◽  
Colletotrichum Spp ◽  
Polymerase Chain ◽  
Polymerase Chain Reaction Primers

Isolates of Colletotrichum spp. from diseased strawberry fruit and crowns were evaluated to determine their genetic diversity and the etiology of the diseases. Isolates were identified to species using polymerase chain reaction primers for a ribosomal internal transcribed spacer region and their pathogenicity was evaluated in bioassays. Isolates were scored for variation at 40 putative genetic loci with random amplified polymorphic DNA and microsatellite markers. Only C. acutatum was recovered from diseased fruit. Nearly all isolates from crowns were C. gloeosporioides. In crown bioassays, only isolates of C. gloeosporioides from strawberry caused collapse and death of plants. A dendrogram generated from the genetic analysis identified several primary lineages. One lineage included isolates of C. acutatum from fruit and was characterized by low diversity. Another lineage included isolates of C. gloeosporioides from crowns and was highly polymorphic. The isolates from strawberry formed distinctive clusters separate from citrus isolates. Evaluation of linkage disequilibrium among polymorphic loci in isolates of C. gloeosporioides from crowns revealed a low level of disequilibrium as would be expected in sexually recombining populations. These results suggest that epidemics of crown rot are caused by Glomerella cingulata (anamorph C. gloeosporioides) and that epidemics of fruit rot are caused by C. acutatum.

scholarly journals Identification and Characterization of Benomyl-Resistant and -Sensitive Populations of Colletotrichum gloeosporioides from Statice (Limonium spp.)

2006 ◽  
Vol 96 (5) ◽  
pp. 542-548 ◽  
Author(s):  
Marcel Maymon ◽  
Aida Zveibil ◽  
Shimon Pivonia ◽  
Dror Minz ◽  
Stanley Freeman
Keyword(s):  
Colletotrichum Gloeosporioides ◽  
Sequence Analyses ◽  
Specific Primers ◽  
Tubulin Genes ◽  
Colletotrichum Spp ◽  
Polymerase Chain ◽  
Species Specific ◽  
Identification And Characterization ◽  
Propagation Material

Sixty-four isolates of Colletotrichum gloeosporioides were isolated from infected Limonium spp. cultivated in 12 different locations in Israel. All isolates were identified as belonging to the C. gloeosporioides complex by species-specific primers. Of these isolates, 46 were resistant to benomyl at 10 μg/ml and 18 were sensitive to this concentration of fungicide. Based on arbitrarily primed polymerase chain reaction of all isolates and internal transcribed spacer-1 sequence analyses of 12 selected isolates, the benomyl-resistant and -sensitive populations belong to two distinct genotypes. Sequence analyses of the β-tubulin genes, TUB1 and TUB2, of five sensitive and five resistant representative isolates of C. gloeosporioides from Limonium spp. revealed that the benomyl-resistant isolates had an alanine substitute instead of a glutamic acid at position 198 in TUB2. All data suggest that the resistant and sensitive genotypes are two independent and separate populations. Because all Limonium plant propagation material is imported from various geographic regions worldwide, and benomyl is not applied to this crop or for the control of Colletotrichum spp. in Israel, it is presumed that plants are bearing quiescent infections from the points of origin prior to arrival.

Genetic variability and molecular identification of Brazilian Biomphalaria species (Mollusca: Planorbidae)

Parasitology ◽  
2001 ◽  
Vol 123 (7) ◽  
pp. 197-209 ◽  
Author(s):  
O. S. CARVALHO ◽  
R. L. CALDEIRA ◽  
A. J. G. SIMPSON ◽  
T. H. D. A. VIDIGAL
Keyword(s):  
Polymerase Chain Reaction ◽  
Morphological Characteristics ◽  
Neotropical Region ◽  
Snail Host ◽  
Molecular Techniques ◽  
Its2 Region ◽  
Intermediate Hosts ◽  
Chain Reaction ◽  
Internal Transcribed Spacer Region ◽  
Polymerase Chain

Freshwater snails belonging to the genus Biomphalaria are intermediate hosts of the trematode Schistosoma mansoni in the Neotropical region and Africa. In Brazil, one subspecies and ten species of Biomphalaria have been identified: B. glabrata, B. tenagophila, B. straminea, B. occidentalis, B. peregrina, B. kuhniana, B. schrammi, B. amazonica, B. oligoza, B. intermedia and B.t. guaibensis. However, only the first three species are found naturally infected with S. mansoni. The classical identification of these planorbids is based on comparison of morphological characteristics of the shell and male and female reproductive organs, which is greatly complicated by the extensive intra-specific variation. Several molecular techniques have been used in studies on the identification, genetic structure as well as phylogenetic relationships between these groups of organisms. Using the randomly amplified polymorphic DNAs (RAPD) analysis we demonstrated that B. glabrata exhibits a remarkable degree of intra-specific polymorphism. Thus, the genetics of the snail host may be more important to the epidemiology of schistosomiasis than those of the parasite itself. Using the simple sequence repeat anchored polymerase chain reaction (SSR-PCR) in intra-populational and intra-specific studies we have demonstrated that snails belonging to the B. straminea complex (B. straminea, B. kuhniana and B. intermedia) clearly presented higher heterogeneity. Using the low stringency polymerase chain reaction (LS-PCR) technique we were able to separate B. glabrata from B. tenagophila and B. tenagophila from B. occidentalis. To separate all Brazilian Biomphalaria species we used the restriction fragment length polymorphism (PCR-RFLP) of the internal transcribed spacer region (ITS) of the DNA gene. The method also proved to be efficient for the specific identification of DNA extracted from snail eggs. Recently we have sequenced the ITS2 region for phylogenetic studies of all Biomphalaria snails from Brazil.

scholarly journals Molecular Characterization of Isolates of Fusarium spp. Associated With Wilt in Capsicum spp.

2019 ◽  
Vol 11 (6) ◽  
pp. 519
Author(s):  
Isabela V. dos Anjos ◽  
Suelene S. de Melo ◽  
Thiago A. S. Gilio ◽  
Jessé P. Kreitlow ◽  
Sandra M. A. da S. Neves ◽  
...  
Keyword(s):  
Fusarium Species ◽  
Morphological Characteristics ◽  
Fusarium Spp ◽  
Chain Reaction ◽  
Tissue Samples ◽  
Mato Grosso ◽  
Capsicum Spp ◽  
Identification Of Species ◽  
Polymerase Chain

Fusarium is a diverse and heterogeneous fungi genus. Its wide genetic variability and similarity in morphological characteristics hinder the identification of species of this genus. Identifying Fusarium species is difficult due to the genus. Several molecular methods have been useful for differentiating these species, and the amplification of internal transcribed spacer (ITS) regions of the fungus ribosomal DNA has been successfully used, since ITS are preserved regions of the DNA that assists in distinguishing species. The objective of this work was to collect and characterize isolates of Fusarium spp. associated with wilt symptoms in Capsicum spp. in the biomes of the state of Mato Grosso, Brazil. Were collected tissue samples of plants with wilt symptoms. The DNAs of the Fusarium spp. found were extracted, and subjected to polymerase chain reaction, using the primers ITS1 (5’TCCGTAGGTGAACCTGCGG-3’) and ITS4 (5’-TCCTCCGCTTATTGATATGC-3’). Subsequently, the sequencing was performed. The resulting sequences were, five Fusarium species were found F. solani, F. oxysporum, F. equiseti, F. incarnatum, F. chlamydosporum, predominating F. solani and F. equiseti.

scholarly journals Characterization of Xanthomonas axonopodis pv. phaseoli isolates

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
Keyword(s):  
Xanthomonas Campestris ◽  
Random Amplified Polymorphic Dna ◽  
Hypersensitive Reaction ◽  
Chain Reaction ◽  
Xanthomonas Axonopodis ◽  
Paulo State ◽  
Total Dna ◽  
Polymerase Chain ◽  
Bacterial Plant Pathogen

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.

Characterization of Giardia duodenalis by polymerase-chain-reaction fingerprinting

1998 ◽  
Vol 84 (9) ◽  
pp. 707-714 ◽  
Author(s):  
Wieger L. Homan ◽  
Margriet Gilsing ◽  
Hafida Bentala ◽  
Louis Limper ◽  
Frans van Knapen
Keyword(s):  
Polymerase Chain Reaction ◽  
Giardia Duodenalis ◽  
Chain Reaction ◽  
Polymerase Chain
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