腐烂茎线虫种内群体特异性检测的研究.pdfVIP

浙江大学学报( ) 33( 5) : 490~ 496, 2007 Journal of Zhejiang University ( Agric Life Sci) : 2007) 刘 斌, 梅圆圆, 郑经武 ( , 310029) : 对腐烂茎线虫(D ity lenchus d estructor Thorne) 7 个不同地理群体 rDNA 中的IT S 区进行PCR RFLP 和序列测定, 发 种内群体间无论在序列长度还是酶切图谱均存在 一定的变异. 根据其序列特 征设计出种特异性引物, 对所供试的腐烂茎线虫群体及属间不同种群体样品的rDNA IT S 区相应区段 扩增以验证其特异性, 同时对分别由1~ 5 条腐烂茎线虫成虫和幼虫提取的DNA 进行特异性扩增以验 证其灵敏度. 结果表明: 所有供试的腐烂茎线虫群体均可以特异性扩增一个346 bp 的片段, 而所有供 试的属外种均没有扩增片段; 由1~ 5 条腐烂茎线虫成虫和 4~ 5 条幼虫进行的特异性扩增也获得稳定 的目的片段. 显示该特异性引物具有较高的稳定性特异性和灵敏度, 能快速准确地检测出腐烂茎 线虫. : 腐烂茎线虫; rDNA IT S 区; 特异性检测; 灵敏性 : Q78; S435313 : A LIU Bin, MEI Yu nyu n, ZH ENG Jingw u ( Institute of Biotechnology , Zhej iang University , H angz hou 31002 , China) Speciesspecific detection of interpopulations of Dity lenchus destructor . Journ l of Zheji ng University ( Agric Life Sci) , 2007, 33( 5) : 490496 Abstract: PCRRFLP of rDNA IT S regions of seven geogr phic popul tions of D. destructor were m de, nd the v ri tion in the sequence length nd digest ion p tterns mong intr species popul tions w s detected. According to the sequence ch r cter of the IT S region of rDNA of D. destructor, p ir of specific primers w ere designed, nd effectiveness w s proved by specif ic mplifying the corresponding IT S region of ll the D. d estructor popul tions. T o test the sensitivit y of the primers, one to five dult nd juvenile specimens of D. destructor were chosen for specific mplific tion. All t he D. destructor popul tions y ielded specific fr gment wit h the length of 346 bp, nd the intergenus species h d no specific mplific tion. T he t rget fr gment w s lso mplified w ith 1 to 5 dult specimens nd 4 to 5 l rv specime