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.
When there is a need to quickly analyze samples using a number of different PCR assays, it is likely that optimal conditions for each assay will not be the same. First, different assays often will require different annealing temperatures for their primers. In addition, amplicons may be designed to be of different lengths and therefore require varying durations of the extension step.READ MORE