【作者】 张祥林； 乾义柯； 马春春； 王翀；

【Author】 ZHANG Xiang-lin~1,QIAN Yi-ke~(1,2),MA Chun-chun~2,WANG Chong~1 (1.Xinjiang Entry and Exit Inspection and Quarantine Bureau,Urumqi 830063,China;2.College of Agronomy,Xinjiang Agricultural University,Urumqi 830052,China)

【机构】 新疆出入境检验检疫局； 新疆农业大学农学院；

【摘要】 [目的]对多种植原体16SrDNA基因序列进行限制性片段长度多态性(RFLP)分析比较,以期区分葡萄金黄化植原体不同亚种,从而为葡萄金黄化植原体的鉴定提供新依据。[方法]用植原体通用引物R16mF2/R16mR1扩增7种不同植原体16SrDNA基因序列,得到约1.5kb的DNA片段,将此片段克隆到PGM-T载体,并通过酶切鉴定,对重组阳性克隆进行核酸序列测定及同源性分析,利用限制性内切酶XmnⅠ、XspⅠ、TaqⅠ和RsaⅠ对目的片段进行酶切电泳分析。[结果]所扩增两个不同亚种的葡萄金黄化植原体D型和C型序列同源性最高,达99.72%,但利用限制性内切酶可以将上述两个亚种从植原体榆树黄化组中区分。[结论]利用限制性内切酶分析通用引物R16mF2/R16mR1所扩增的基因片段,可以将葡萄金黄化植原体区分。更多还原

【Abstract】 [Objective]The phytoplasma 16SrDNA gene sequences were compared through RFLP analysis,based on the similarity of RFLP patterns,Flavescence doree would be differentiated from other strains,provide a new thereunder for detecting Flavescence doree.[Method]A 1.5kb DNA fragment was amplified by PCR with primer R16mF₂/R16mR₁ from 16SrDNA of 7 different phytoplasmas.This amplified fragment was ligated into PGM-T vector and the clones were verified by restriction enzyme digestion,sequenced and homology analysis.The amplified fragments were analyzed with XmnⅠ、XspⅠ、TaqⅠor RsaⅠand separated by electrophoresis.[Result]The result showed that the Flavescence doree-D and Flavescence doree-C strains had the highest similarity（99.72%）,but this tow strains could be differentiated from Elm Yellows group by restriction enzyme digestion.[Conclusion]Phytoplasma Flavescence doree could be differentiated through RFLP analysis of 16SrDNA amplified with primer R16mF₂/R16mR₁.更多还原