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Identification And Differentiation Of Verticillium Species With PCR Markers And Sequencing Of Its Region
Poster Title: Identification And Differentiation Of Verticillium Species With PCR Markers And Sequencing Of Its Region
Submitted on 28 Jan 2015
Author(s): Taja Jeseničnik, Nataša Štajner, Jernej Jakše, Sebastijan Radišek and Branka Javornik
This poster was presented at PAGXXIII
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Poster Information
Abstract: We screened our Verticillium spp. collection with the new taxonomic PCR-markers using simplex and multiplex PCR assays. In addition, we also evaluated an alternative ITS-sequence based approach for the identification purposes.
The new PCR-marker analysis enabled us to confirm the identity of 88 isolates from various geographic locations in Europe, North America and Japan and from various host plants, including hop, potato, tomato, cotton, olive and alfalfa. According to the new taxonomic classification, 41 isolates were identified as V. nonalfalfae, 28 as V. dahliae, 6 as V. albo-atrum, 5 as V. alfalfa, 3 as V. isaacii, 2 as V. tricorpus, 1 as V. longisporum lineage A1/D1 and 1 as V. nubilum. Identification and differentiation of V. longisporum linegaes was performed by multiplex PCR assay.
We were also able to determine all isolates by means of sequence ITS analysis, reflecting two main groups of isolates, Flavexudans and Flavnonexudans. Moreover, based on ITS sequencing analysis, we were able to obtain additional information on the subgroups within the species V. albo-atrum and V. dahliae that were not revealed by PCR-marker analysis.
Summary: The genus Verticillium is a group of ascomycete fungi, including plant-pathogenic species capable of affecting the vasculature of many agricultural crops, and therefore causes major economic losses worldwide. In 2011, a new taxonomic classification of the genus was proposed, which is now referred to as Verticillium sensu stricto, comprising ten species: V. dahliae V. albo-atru, V. alfalfae, V. longisporum, V. nonalfalfae, V. tricorpus, V. zaregamsianum, V. nubilum, V. isaacii and V. klebahnii. <References: -Fradin E. F., Thomma B. P. H. J. 2006. Physiology and molecular aspects of Verticillium wilt diseases caused by V. dahliae and V. albo-atrum. Molecular Plant Pathology, 7, 2: 71–86
-Isaac I (1953) A further comparative study of pathogenic isolates of Verticillium: V.
-Inderbitzin P., Bostock R. M., Davis R. M., Usami T., Platt H. W., Subbarao K. V. 2011. Phylogenetics and taxonomy of the fungal vascular wilt pathogen Verticillium, with the descriptions of five new species. PLOS ONE, 6, 12: e28341, doi:10.1371/journal.pone.0028341: 21 str.
-Inderbiztin P., Davis R. M., Bostock R. M., Subbarao K. V. 2013. Identification and differentiation of Verticillum species and V. longisporum lineages by simplex and multiplex PCR sssay. PLOS ONE, 8, 6: e65990, doi:10.1371/journal.pone.0065990: 12 str.
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