Genetic diversity among the different isolates of Nucleopolyhedrovirus in Mazandaran and Golestan provinces using RAPD molecular marker
Subject Areas : Plant PestsMarzieh Shazdehahmadi 1 , Hoda Assemi 2 , Zein-Al-Abedin Shahadatimoghadam 3 , Seyed Afshin Sajjadi 4
1 - Researchers, Tirtash Tobacco Research & Education Center, Behshahr, Mazandaran، Iran
2 - Researchers of Tirtash Tobacco Research and Education Center, Behshahr, Iran
3 - Researchers of Tirtash Tobacco Research and Education Center, Behshahr, Iran.
4 - Researchers of Tirtash Tobacco Research and Education Center, Behshahr, Iran
Keywords: genetic diversity, RAPD marker, NPV, Tobacco bud worm,
Abstract :
Tobacco budworm (Helicoverpa armigera) is one of the key pests of tobacco farms. Nucleopolyhedrovirus is one of the most important biological control agents of budworm and a number of other species. In this study the genetic diversity of different isolates of Nucleopolyhedrovirus belonging to 16 natural populations in Mazandaran and Golestan provinces, were investigated. The tobacco budworm samples were collected from tobacco fields and transferred to biotechnology Laboratory of Tirtash Research and Education Center. Purification of NPV virus and DNA extraction was done using CTAB method. Among ten pair of primers which we used for amplification of NPV, two pair of RAPD primers that showed the highest Polymorphism were selected. Sixteen DNA samples isolated from different geographical regions of Golestan and Mazandaran provinces were checked with these selected RAPD primers and they created countable bands. Totally, 87 Countable bands were generated following the PCR products. Among them,75 bands(86%) were polymorphic. Data analysis was done using the PyElph Software. Population grouping using UPGMA method based on genetic distance of 20, revealed five major clusters. Using the underlying RAPD marker, the isolates of Nucleopolyhedrovirus were in a one gene cluster, showed the greatest genetic affinity.
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