IDENTIFICATION AND DIFERENTIATION OF Verticillium SPECIES WITH PCR MARKERS AND SEQUENCING OF ITS REGION
Poster Jan 28, 2015
Taja Jesenicnik, Nataša Štajner, Jernej Jakše, Sebastijan Radišek and Branka Javornik
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.
Despite the developments in conventional PCR, the complexity of multiplex Real Time PCR is still limited due to the lack of sufficient detection channels. To achieve high-end multiplexing capacity on standard Real Time PCR machines, Anapa Biotech has developed the MeltPlex® technology (see box on right).READ MORE