不同簇毛麦6VS染色体臂的白粉病抗性特异功能标记的开发及应用  

Development and Application of Functional Markers Specific to Powdery Mildew Resistance on Chromosome Arm 6VS from Different Origins of Haynaldia villosa

作　　者：张云龙[1,2] 王美蛟[1] 张悦[1] 褚翠萍[2] 林志珊[1] 徐琼芳[1] 叶兴国[1] 陈孝[1] 张宪省[2] 

机构地区：[1]中国农业科学院作物科学研究所/农作物基因资源与基因改良国家重大科学工程/农业部麦类生物学与遗传育种重点实验室,北京100081 [2]山东农业大学/作物生物学国家重点实验室,山东泰安271018

出　　处：《作物学报》2012年第10期1827-1832,共6页Acta Agronomica Sinica

摘　　要：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。Lines Pm97033 and 92R137 with good resistance to powdery mildew are wheat–Haynaldia villosa translocation lines carrying 6VS from different H. villosa germplasms. In this study, we identified the similarity of powdery mildew resistance genes in both lines using molecular markers. Based on the sequence of Stpk-V gene （GenBank accession number HQ864471.1）, we designed three primer pairs in the coding regions containing at least one intron. Using one pair of primers, a polymorphic fragment of Pm97033 pattern was amplified, which was specific to the alien chromosome arm 6VS. According to the sequencing result of this specific fragment, two new primer pairs were designed for better stability. Primer pair A amplified a specific band specific to 6VS·6DL translocation line Pm97033 and its resistance donor H.v#2. Primer pair B amplified two polymorphic fragments corresponding to 6VS from different H. villosa donors. The effectiveness of primer pairs A and B was then verified in wheat line CB037 with strong powdery mildew resistance, which was developed using both 6V（6D） substitution and 6VS·6AL translocation lines. The banding pattern in CB037 was identical to that in 92R137; however, the amplification fragment of Pm97033 was not observed in CB037. The result of genomic in situ hybridization revealed only one pair of wheat–H. villosa translocation chromo-somes in CB037, and the translocation chromosome arm was identified as 6A by molecular markers NAU/xibao15F and NAU/xibao15R. The GISH analysis confirmed the result based on molecular markers designed in this study. Thus, we infer that the powdery mildew resistance in CB037 is derived from 92R137, and CB037 and Pm97033 probably have different genetic bases on resistance to powdery mildew.

关 键 词：6VS·6AL易位系 6VS·6DL易位系 簇毛麦 白粉病抗性 功能标记 

分 类 号：S512.1[农业科学—作物学]