Molecular identification for some Pythium aphanidermatum isolates from different fields of An-Najaf Al-Ashraf governorate

Abstract

The aim of the study included the isolation and identification of Pythium species depend on phenotypic biotechnique on culture media and test it's pathogenicity and identification isolates of P. aphanidermatum by molecular test and genotype by using PCR technique from soils of some locations of Najaf province. Five fungal genera were isolated and identified from: Baher Al-Najaf, Al-Hasaweya, Al-Barakia, Al-Haiderya, Khan Al-Rube', Al-Mushkhab, Al-Abbassia and extension farm in Kufa/Al-Najaf for spring 2013. The results of the polymerase chain reaction (PCR) by using a pair primers specialist for P.aphanidermatum and analyze the results of immigration electric using agarose concentration of 1.5% the appearance of bands of isolates A, B, C, D, E, F, G, H, I, J, K, L which ensure the identification of P.aphanidermatum . It turns out the result of the migration to the electric gel Agarose that all primers have interacted with isolates of P. aphanidermatum and succeeded each primers to identify all isolates where appeared polymorphic band and 100% with all of the isolates. Random Amplified Polymorphic DNA (RAPD) adopted to identify the fingerprint DNA of twelve isolates of P. aphanidermatum having been exposing DNA to four primers (P1, P2, P3 and P4) was able to produce a number of bands reached to 131 where recorded 18, 32, 40 and 41 bands in P1, P2, P3 and P4 primers respectively.