yilmaz, gulenay tc gida, tarim ve hayvancilik bakanligi - zirai karantina mudurlugu, izmir, Turkiye , tosun, necip ege universitesi - ziraat fakultesi - bitki koruma bolumu, Bornova, Turkiye

The Studies on the Presence of Plasmopara halstedii (Farl.) Berl. De Toni) in Sunflower (Helianthus annuus L.) Seeds with Molecular Methods

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The fact that rapid diagnosing of especially seed-borne pathogens and their control is the first step to obtain healthy crop. The aims of the study were to optimize the analyzing methods in detecting presence of Plasmopara halstedii (Farl.) Berl. De Toni requiring analysis at the time of importation in the sunflower seed using current molecular methods that were much more advantageous than that of classical methods. Calculations related to classical PCR (agarose gel electrophoresis) were optimized by referring to Appendix 2 of the EPPO (European and Mediterranean Plant Protection Organisation) Standard Protocols while those related to real-time PCR (the hydrolyze probes) were optimized by referring to Appendix 3 of the EPPO Standard Protocols. Moreover, SyberGreen Mastermix Kit protocol was used for optimization for the study of real-time PCR (SyberGreen) that is not present in the EPPO Standard Protocols. Thus, it was provided that all three molecular protocols were confirmed for better applicable and practical ways for the quarantine laboratories in our country. Furthermore, the first molecular study for direct detection of the agent from sunflower seed through this optimization was conducted in quarantine laboratories of Turkey.

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Plasmopara halstedii , sunflower (Helianthus annuus) , PCR , hydrolysis probes , optimization

The Journal Of Ege University Faculty Of Agriculture

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