簇毛麦6VS上4个新分子标记的鉴定及与抗白粉病基因Pm21的连锁分析

Identification and Analysis of Four Novel Molecular Markers Linked to Powdery Mildew Resistance Gene Pm21 in 6VS Chromosome Short Arm of Haynaldia villosa

Pm21具有强抗白粉病特性,位于簇毛麦6V染色体短臂(6VS)上。染色体分析和白粉病抗性检测表明,Pm21在受体小麦内是稳定遗传的。筛选与Pm21相连锁分子标记,在小麦抗白粉病辅助选择育种上有重要意义。利用RAPD和AFLP分子标记方法,对簇毛麦6V染色体和含有6V的小麦-簇毛麦代换系、6VS的易位系6AL/6VS、6DL/6VS进行分子标记筛选,以分析位于6VS上的分子标记及其与Pm21的关系。RAPD检测表明,OPK08910特异片段存在于含簇毛麦6V染色体的代换系(6A/6V)、易位系(6AL/6VS,6DL/6VS)和簇毛麦中。AFLP检测显示,PstⅠ+AGG/MseⅠ+CAC、PstⅠ+ACC/MseⅠ+CCT和PstⅠ+AAG/MseⅠ+CGC共3对引物分别可以在6A/6V、6AL/6VS、6DL/6VS和VV中扩增出264bp、218bp和232bp特异带,并与抗白粉病基因Pm21共分离,因此,上述来源于6VS上的4个新的分子标记,可作为源自簇毛麦Pm21基因的选择标记,用于小麦抗病育种。

The Pm21 gene, originating from Haynaldia villosa （6AL/6VS）, is one of the effective powdery mildew resistance genes. It was observed from chromosome analysis and resistant testing to powdery mildew that Pm21 on 6VS could be stably inherited when transferred to wheat. It is valuable for identification of the molecular markers linked to wheat powdery mildew resistance gene Pm21 in marker-assisted selection in wheat breeding practice. In this study, wheat-Haynaldia chromosome substitution hne （6A/6V）, translocation hnes （6AL/6VS, 6DL/6VS） were used to identify molecular markers located in chromosome short ann 6VS by using RAPD, AFLP techniques, and to investigate the relationship between the molecular markers and Pm21. RAPD test indicated that a Haynaldia villosa chromosome 6VS-specific pelymorphic DNA segment OPK08910 was amplified in 6A/6V substitution hne, 6AL/6VS and 6DL/6VS translocation lines of wheat-H. villosa. AFLP analysis showed three H. villosa chromosome 6VS-specific polymorphic DNA sequences （254 bp, 218 bp, and 232 bp） could be amplified by three AFLP primer pairs, PstⅠ ＋ AGG / Mse Ⅰ ＋ CAC, Pst Ⅰ ＋ ACC / Mse Ⅰ ＋ CCT and Pst Ⅰ ＋ AAG / Mse Ⅰ ＋ CGC in substitution line （6A/6V）, translocation hnes （6AL/6VS and 6DL/6VS） and H. villosa （VV）. These AFLP markers of Pst Ⅰ ＋ AGG / Mse Ⅰ ＋ CAC-264, Pst Ⅰ ＋ ACC / Mse Ⅰ ＋ CCT-218 and Pst Ⅰ ＋ AAG / Mse Ⅰ ＋ CGC-232 which derived from 6VS, were found to be cesegregated with powdery mildew resistance gene Pm21. These molecular markers can be used as novel DNA markers for selecting powdery mildew resistance gene Pm21 in wheat breeding program.