Fusarium graminearum sensu stricto和F.asiaticum3种分子鉴定方法的比较
发布时间：2018-09-10 08:16
【摘要】：Fusarium graminearum sensu stricto和F.asiaticum是中国主要的禾谷镰刀菌复合种(FGSC)成员,为了提高这2个种的鉴定效率,选取48个引起小麦赤霉病和玉米茎腐病的FGSC菌株,比较了系统发育谱系分析、PCR-RFLP(聚合酶链式反应-限制性片段长度多态性)和多重PCR 3种分子鉴定方法。系统发育谱系分析首先扩增菌株的EF-1α和Tri101两个基因的部分序列,测序之后利用MEGA 6.06软件构建系统发育树,结果表明,有11个菌株与F.asiaticum聚在一起,37个菌株与F.graminearum sensu stricto聚为一枝,此法繁琐、复杂、成本高,但鉴定结果准确。PCR-RFLP基于组蛋白H3基因特殊的酶切位点,设计引物H3d StyⅠ/H3R1,通过PCR和酶切反应进行鉴定,电泳检测发现,有11个菌株为F.asiaticum,37个菌株为F.graminearum sensu stricto,该法工作量大且对操作过程要求高。多重PCR基于CYP51A基因设计特异引物Fa F/FaR和Fg F/FgR进行鉴定,电泳检测结果表明,有11个菌株为F.asiaticum,37个菌株为F.graminearum sensu stricto,此法操作简单、效率高。以上3种分子鉴定方法结果一致,但多重PCR方法具有快速、准确、高效的特点。
[Abstract]:Fusarium graminearum sensu stricto and F.asiaticum are the main (FGSC) members of Fusarium graminearum complex species in China. In order to improve the identification efficiency of the two species, 48 FGSC strains were selected to cause wheat scab and maize stem rot. Three molecular identification methods, PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) and multiplex PCR, were compared. Phylogenetic analysis first amplified the partial sequences of EF-1 伪 and Tri101 genes of the strain. After sequencing, the phylogenetic tree was constructed by MEGA 6.06 software. The results showed that 11 strains were clustered with F.asiaticum and 37 strains were clustered with F.graminearum sensu stricto. The method was complicated, complicated and costly, but the results were accurate. PCR-RFLP was based on the specific restriction site of histone H3 gene, designed primers H3D Sty 鈪，
本文编号：2233881
[Abstract]:Fusarium graminearum sensu stricto and F.asiaticum are the main (FGSC) members of Fusarium graminearum complex species in China. In order to improve the identification efficiency of the two species, 48 FGSC strains were selected to cause wheat scab and maize stem rot. Three molecular identification methods, PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) and multiplex PCR, were compared. Phylogenetic analysis first amplified the partial sequences of EF-1 伪 and Tri101 genes of the strain. After sequencing, the phylogenetic tree was constructed by MEGA 6.06 software. The results showed that 11 strains were clustered with F.asiaticum and 37 strains were clustered with F.graminearum sensu stricto. The method was complicated, complicated and costly, but the results were accurate. PCR-RFLP was based on the specific restriction site of histone H3 gene, designed primers H3D Sty 鈪，
本文编号：2233881
本文链接：https://www.wllwen.com/kejilunwen/nykj/2233881.html
