摘要
从马铃薯块茎上分离纯化得到晚疫病菌、银腐病菌、癌肿病菌、红腐病菌、黑点病菌和镰刀菌等并提取它们的全基因组DNA,用引物08-3和08-4对这些真菌的DNA进行PCR扩增,以检验该引物扩增晚疫病菌DNA的特异性。取马铃薯种薯的芽眼、芽及其他不同部位并用NaOH法提取晚疫病菌DNA,再用引物08-3和08-4进行特异性扩增以检验种薯是否带晚疫病菌。结果表明:(1)引物08-3和08-4有很强的特异性,只能扩增出大小为257bp的马铃薯晚疫病菌DNA片段;(2)用该引物能扩增出种薯芽眼组织中大小为257bp的晚疫病菌DNA,说明通过PCR特异扩增,能直接检测出马铃薯种薯中潜伏的晚疫病菌。(3)本次检测的本地感病品种米拉有10%的种薯带晚疫病菌。该研究为种薯传播晚疫病菌的分子检测提供了依据和方法。
The fungi Phytophthora infestans,Helminthosporium solani,Synchtrium endobioticum,Phytophthora erythroseptica,Colletotrichum atramenttamiu and Fusarium were isolated from potato tubers,and incubated in different medium.The genomic DNA of these fungi were extracted and used for PCR amplified by primer 08-3&08-4 to test its specifity of amplifying the DNA of Phytophthora infestans.The sprout eye site,sprout and the other part tissues of potato tubers were taken and the genomic DNA of Phytophthora infestans in these potato tissues were extracted by method of NaOH.The genomic DNA of Phytophthora infestans extracted from the potato tissue were PCR amplied with primer 08-3&08-4.The results show that the primer 08-3&08-4 have definite specifity of amplifying the DNA of Phytophthora infestans,and only amplified 257 bp DNA fragment of P.infestans.The primer 08-3&08-4 can amplify 257 bp DNA fragment of P.infestans extracted from those tissues of sprout eye site of potato tubers,suggesting that the PCR amplification with primer 08-3&08-4 can be used for detection of Phytophthora infestans in potato tuber.In this research,10% of potato tubers of the cultivar Mira were infected by Phytophthora infestans according to PCR amplification with primer 08-3&08-4.
出处
《中国马铃薯》
2004年第2期73-77,共5页
Chinese Potato Journal
基金
云南省自然科学基金(97CO23M)
马铃薯晚疫病综合防治技术研究与示范(2002NG9)。
