向日葵白锈病菌巢式PCR检测方法的研究

Study on the Nest-PCR to Detect the Albugo tragopogi (Persoon) Schruter var Helianthi Novotlenova

【目的】建立向日葵白锈病菌(Albugo tragopogi (Persoon) Schruter var helianthi Novotlenova)的巢式PCR检测方法。【方法】用通用引物NL1/NL4对采自伊犁的向日葵白锈病菌进行PCR扩增、克隆、酶切和测序,设计该病菌的特异性引物,用巢式PCR法检测供试样品。【结果】序列测定和比对结果表明,该病菌的核酸序列与GeneBank公布的向日葵白锈病菌核酸序列同源率为97%。针对该段基因设计向日葵白锈病菌的特异性引物ATHP3/ATHP4,用巢式PCR法对供试样品进行扩增,扩增片段为370 bp。【结论】成功建立了向日葵白锈病菌巢式PCR检测方法。

[ Objective] Establish the nest - PCR method to detect the Albugo tragopogi （Persoon） Schruter var helianthi Novotlenova. [Method]The primer NL1/NL4 was used to detect the pathogen which was collected from sunflower fields in Yili, Xinjiang Uyghur Autonomous Region by PCR amplification and cloning, enzyme-cutting and sequence analysis. And it designed specificity primer for the pathogen, and detected the samples by nest - PCR. [ Result] The sequence analysis and eontrastive result show that the pathogen＇ s nocleotide sequence was similar to the Albugo tragopogi （Persoon） Schruter var helianthi Novotlenova. The homology of the pathogen reaches 97% compared with the Albugo tragopogi （Persoon） Schruter in GeneBank. The specific primer ATHP3/ ATHP4 was designed and nest - PCR was used to detect the samples. The amplified product was 370 bp. [ Conclusion]The nest- PCR was established successfully.