Quantitative measurment of Verticillium dahliae in roots of four olive cultivars using Real-time PCR

Khanmohammadi, Sahar; Jafary, Hossein; Salari, Mohammad; Javadi, Mehri

doi:10.22055/ppr.2015.11229

Quantitative measurment of Verticillium dahliae in roots of four olive cultivars using Real-time PCR

Document Type : Research paper-Persian

Authors

¹ Former M.Sc.student, Department of plant pathology, Faculty of Agriculture, university of zabol, Iran.

² Scientific board member of agricultural and Natural Resources Research Center of Zanjan, Iran

³ Associate Proferror of Plant protection, Faculty of Agriculture, university of zabol, Iran.

⁴ Department of Agriculture Biotecnology, Academic centerfor Education, Culture and research, Zanjan, Iran

10.22055/ppr.2015.11229

Abstract

Verticilium wilt of olive, caused by Verticillium dahliae, is a devastating disease of olive which can reduce the yield and may kill eventually the infected olive trees by destructing of their vascular system. In this research, some wood sampels of olive trees in Tarom showing wilting symptoms were collected. The causal fungus was isolated on Czapek medium. Total DNA of V.dahliae isolates was extracted using CTAB method with minor modifications. For evaluation of reaction of olive cultivars to the fungus, 5-months old roots saplings of four olive cultivars including: Zard, Arbequin, Oblonga and Koroneiky were inoculated with suspension of 107 spores/ml of a D isolate using root deep inoculation. Total DNA of inoculated plants were isolated immediately after inoculation with the fungus and then 2,7,15,27,41,56,77 days after inoculation. The real-time PCR with the SYBR Green, was applied on the total DNA extracted from infected olives at different stages of infection. Results of quantification of V.dahliae genomic DNA in total genomic DNA isolated from infected olives sampled at different times showed that the amounts of DNA of pathogen in the roots of Zard and Arbequin cultivars increased faster than in Oblonga and Koroneiky. The fungus had more restrictions with colonizing of roots of Koroneiky, rather than other cultivars. In the root system of "Zard" cultivar however, fungus colonized the roots faster than other cultivars. The results of this research also showed that Real time PCR can be used as an efficient way of screening olive genotypes for resistance to Verticillium wilt.

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