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Abstract Detail



Mycological Section

Barasubiye, Tharcisse [1], Mack, Jonathan [1], Seifert, Keith A. [2], Patrick, Susan [3], Sivagnanam, Kumaran [3], Gräfenhan, Tom [3].

Canadian taxa of the Alternaria species complex identified by multilocus DNA sequence analysis.

Species of the genus Alternaria cause a wide range of economically important diseases on a variety of crops and are difficult to identify based on morphology. We report a DNA method based on multilocus (Adenosine triphosphatase (ATPase), Chitinase, Translation elongation factor 1-alpha (EF1-alpha), Endo-polygalacturonase (Endo-PG), Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and RNA polymerase II (RPB2) sequence analysis to characterize and identify Alternaria species collected in Canada from different crops. Genomic DNA was extracted from a total of 160 cultures of Alternaria species using the Qiagen DNeasy Plant Mini kit. We successfully amplified amplicons of uniform sizes from genomic DNA of all the Alternaria isolates using primers targeting the six selected genes. No amplification was obtained from genomic DNA of large-spored species with primers designed to amplify a portion of Endo-PG gene. With a primer pair designed to amplify EF1-alpha, we consistently obtained an amplicon (330 bp) that discriminates the group with small conidia from the cultures of Alternaria producing large conidia. Using a multilocus DNA sequencing approach, we demonstrated the potential of Endo-PG gene to discriminate Alternaria gaisen, a quarantine species in the Alternaria alternata species complex. Phylogenetic analysis based on RPB2 sequences, revealed 8 lineages within a clade harboring Alternaria species producing small conidia. These results could have practical application in reliable detection and identification of Alternaria species in agricultural important crops.


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1 - Agriculture and Agri-Food Canada, 960 Carling Avenue, Ottawa, ON, K1A0C6, Canada
2 - Eastern Cereal and Oilseed Research Centre, Agriculture & Agri-Food Canada, 960 Carling Ave., Ottawa, ON, K1A0C6, Canada
3 - Canadian Grain Commission, 303 Main Street, Winnipeg, MB, R3C3G8, Canada

Keywords:
Alternaria
Sequencing
identification
DNA diagnostic.

Presentation Type: Oral Paper:Papers for Sections
Session: 64
Location: Salon 1/The Shaw Conference Centre
Date: Wednesday, July 29th, 2015
Time: 11:45 AM
Number: 64007
Abstract ID:1068
Candidate for Awards:None


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