Species Identification of Gram Wilt Complex through ITS Region by PCR-RFLP Analysis
Muhammad Jawad1*, Shahid Riaz Malik1, Rana Muhammad Atif2, Haris Ahmed2 and Muhammad Shahzad Afzal3
ABSTRACT
Chickpea is the most important pulse legume that serve as dietary protein source for poor farmers in the developing countries. Yield and production severely challenged by biotic stresses. Wilt complex is the major biotic factor that contribute significantly in yield loss. Wilt complex is caused by various pathogens, including diverse type of Fusarium species. Molecular approach is a useful technique for the identification of wilt complex pathogens. The study conducted to identify the polygenetic association of the pathogens of chickpea wilt complex. Universal Rice Primers based diversity analysis indicates high polymorphism within the collected wilt isolates. Primers ITS 1 and ITS 4 generated internal transcribed spacer fragments that were restricted four restriction enzymes (HhaeI, MboI, BsuRI, HinfI). PCR ITS-RFLP banding pattern identified two Fusarium species i.e. Fusarium oxysporum and Fusarium solani. In conclusion, PCR-ITS-RFLP proved to be an efficient and easy technique for the identification of wilt complex pathogens.
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