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小麦6VS·6DL易位系Pm97033和6VS·6AL易位系92R137中的6VS染色体臂来自不同的簇毛麦种质,均表现良好的白粉病抗性,本研究利用分子标记对这2个易位系所包含抗病基因的异同进行了鉴定。利用与Pm21抗白粉病相关的丝氨酸/苏氨酸蛋白激酶Stpk-V基因(GenBank登录号为HQ864471.1)的基因组和cDNA序列为基础,在包含至少1个内含子的2个编码区设计引物,从Pm97033中扩增获得特异的多态性片段。为进一步提高特异性和扩增的稳定性,对特异扩增片段测序并重新设计引物,扩增筛选获得2个引物对,其中PK-F1/PK-R可专一扩增6VS·6DL易位系Pm97033及其抗病亲本,而PK-F2/PK-R可同时特异扩增2个不同来源的簇毛麦6VS染色体,但二者间的特异片段具有多态性。利用这2对引物,对系谱中包含6V(6D)和6VS·6AL、抗白粉病的小麦品系CB037进行检测,发现仅出现与6VS·6AL易位系相同的簇毛麦扩增片段,不存在簇毛麦No.1026(Pm97033的6VS供体)的扩增片段。基因组原位杂交结果表明,CB037仅含1对小麦-簇毛麦的易位染色体,用已报道的分子标记检测证明易位涉及的小麦染色体为6A,与本研究开发的分子标记检测结果相吻合,表明CB037携带的白粉病抗性基因来自6VS·6AL易位系92R137,其白粉病抗性可能与Pm97033具有不同的遗传基础。
The 6VS chromosome arms of wheat 6VS · 6DL translocation lines Pm97033 and 6VS · 6AL translocation lines 92R137 were all from different germplasms of Hamster, which showed good resistance to powdery mildew. In this study, The similarities and differences between the genes involved in the resistance were identified. Based on the genomic and cDNA sequences of the serine / threonine protein kinase Stpk-V gene (GenBank accession number HQ864471.1) related to Pm21-resistant powdery mildew, two coding regions comprising at least one intron were designed Primer, amplified from Pm97033 specific polymorphic fragments. In order to further improve the specificity and stability of amplification, specific primers were designed and redesigned. Two primer pairs were obtained by amplification and screening. Among them, PK-F1 / PK-R could specifically amplify the 6VS · 6DL translocation Pm97033 and its parental parents, while PK-F2 / PK-R could specifically amplify 6VS chromosomes from two different origins, but the specific fragments between them were polymorphic. The two pairs of primers were used to detect CB037, a 6V (6D) and 6VS · 6AL resistant powdery mildew resistant wheat line in the pedigree and found that only the same amplified fragment of H. rhamnoides was present in the 6VS · 6AL translocation line Amplified fragments of Rumel # 1026 (6VS donor for Pm97033). Genomic in situ hybridization results showed that CB037 contained only one pair of translocation chromosomes of wheat-cluster-wool, and the molecular marker proved that the translocation involved in chromosome 6 A was consistent with the results of molecular markers developed in this study , Indicating that the powdery mildew resistant gene carried by CB037 is derived from the 6VS · 6AL translocation line 92R137 and the powdery mildew resistance may have a different genetic basis with that of Pm97033.