小麦叶锈菌特异分子标记建立

Development of specific molecular markers in Puccinia triticina

利用21对EST-SSR引物、30对SSR引物及40对小麦叶锈菌EST引物,以小麦条锈菌为对照,对小麦叶锈菌基因组DNA进行PCR扩增筛选。三类引物在小麦叶锈菌和小麦条锈菌基因组中分别扩增出55、77、53个稳定条带,其中多态性条带分别为25、36、12个,平均多态性百分率分别为45.45%,46.75%,22.64%。每对引物等位基因数为1～9,3种引物平均等位基因数分别为2.6、2.6、1.3。2对小麦叶锈菌中单一等位基因的EST引物EST-6和EST-23可在小麦叶锈菌中分别扩增出625和384bp的特异片段。进一步用其他6种锈菌和小麦散黑穗病菌对这2个特异引物进行检测,均未扩增出其特异性片段,表明引物EST-6和EST-23在小麦叶锈菌中具有特异性。

PCR was conducted to screen the genomic DNA of Pucciniatriticinausing 21 pairs of EST-SSR primers,30 pairs of SSR primers and 40 pairs of EST primers,with P.striiformis as a control.55,77 and 53stable bands were amplified by three types of primers respectively in both P.triticinaand P.striiformis,which contained 25,36 and 12polymorphic bands with an average percentage of polymorphism as 45.45%,46.75%and 22.64%,respectively.The allele numbers of each primer were 1to 9,and the average number of the alleles was 2.6,2.6and 1.3of the three types of primers.Two specific bands with the length of 620 bp and 384 bp were amplified in P.triticina with the primer EST-6and EST-23 respectively.Additionally,the two specific primers were used to amplify other six kinds of rust fungi and Ustilago tritici.The specific bands of the two primers were only detected in P.triticina which indicated the specificity of the two primers,EST-6and EST-23 in P.triticina.