發(fā)布時(shí)間：2018-09-10 08:16

【摘要】：Fusarium graminearum sensu stricto和F.asiaticum是中國主要的禾谷鐮刀菌復(fù)合種(FGSC)成員,為了提高這2個(gè)種的鑒定效率,選取48個(gè)引起小麥赤霉病和玉米莖腐病的FGSC菌株,比較了系統(tǒng)發(fā)育譜系分析、PCR-RFLP(聚合酶鏈?zhǔn)椒磻?yīng)-限制性片段長度多態(tài)性)和多重PCR 3種分子鑒定方法。系統(tǒng)發(fā)育譜系分析首先擴(kuò)增菌株的EF-1α和Tri101兩個(gè)基因的部分序列,測序之后利用MEGA 6.06軟件構(gòu)建系統(tǒng)發(fā)育樹,結(jié)果表明,有11個(gè)菌株與F.asiaticum聚在一起,37個(gè)菌株與F.graminearum sensu stricto聚為一枝,此法繁瑣、復(fù)雜、成本高,但鑒定結(jié)果準(zhǔn)確。PCR-RFLP基于組蛋白H3基因特殊的酶切位點(diǎn),設(shè)計(jì)引物H3d StyⅠ/H3R1,通過PCR和酶切反應(yīng)進(jìn)行鑒定,電泳檢測發(fā)現(xiàn),有11個(gè)菌株為F.asiaticum,37個(gè)菌株為F.graminearum sensu stricto,該法工作量大且對操作過程要求高。多重PCR基于CYP51A基因設(shè)計(jì)特異引物Fa F/FaR和Fg F/FgR進(jìn)行鑒定,電泳檢測結(jié)果表明,有11個(gè)菌株為F.asiaticum,37個(gè)菌株為F.graminearum sensu stricto,此法操作簡單、效率高。以上3種分子鑒定方法結(jié)果一致,但多重PCR方法具有快速、準(zhǔn)確、高效的特點(diǎn)。
[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 偽 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，

本文編號(hào)：2233881