Characterization of RAPD Markers,and the RFLP Marker Linked to Powdery Mildew Resistance Gene Derived from Different Accessions of H.villosa

The analysis was carried out on performance of the resistance gene from Haynaldia villosa accession of the former Soviet Union to different isolates of Bluemerie graminis. Polymorphisms were revealed between 6D/6V substitution line Pm930640 and its pedigree parents using five RAPD markers of OPAN031700, OPAI01700, OPAL03750, OPAD07480 and OPAG15580 screened out from 120 random 10-mers primers. Three RAPD markers of OPAN03, OPAI01 and OPAL03 were linked with the resistance gene by analysis of F2 population of Chancellor×Pm930640. Analysis of 29 wheat lines including part of lines conferring the known genes from Pm1 to Pm20 respectively, lines conferring resistance gene from two H. villosa accessions and the related wheat parents, were analyzed and the results showed that these markers not only linked to the gene resistant to powdery mildew from H. villosa, but also detected different genetic backgrounds. OPAL03750 can be used as the marker to distinguish the different resistant lines from two H. villosa accessions because it was only observed in the materials from H. villosa of the former Soviet Union. RFLP analysis also showed the polymorphisms between two H. villosa accessions and their derived resistant lines.

簇毛麦抗白粉病基因的RAPD及RFLP标记

分析了来自前苏联簇毛麦及其抗病衍生系的抗白粉病基因对不同生理小种的抗性反应。用 120 个随机引物对 6D/6V 代换系 Pm930640 进行 RAPD 分析,检测到 5 个引物 OPAN03、OPAI01、OPAL03、OPAD07 和 OPAG15,分别在大约 1 700、700、750、480 和 580 bp 处有区别于小麦亲本的多态性条带。对 Chancellor×Pm930640 F2群体进行 OPAN03、OPAI01 和 OPAL03 等 3 个 RAPD 标记与抗白粉病基因的连锁分析,表明这些标记同簇毛麦的抗白粉病基因是连锁的。对大部分分别含有 Pm1-Pm20 的已知抗病基因、含有簇毛麦抗病基因及其相关亲本的 29 个小麦品系进行RAPD标记分析。结果表明,这些标记不仅可以鉴定簇毛麦的抗病基因,而且可以判断其遗传背景。OPAL03750仅出现在含有前苏联簇毛麦 6VS 染色体的抗病材料中,可作为区别于 Pm21 的分子标记。RFLP 标记的结果也表明两个不同簇毛麦的 6VS 染色体有明显的多态性。

小麦-簇毛麦染色体代换系、易位系V染色体RAPD标记筛选

利用105条S系列和100条Operon随机引物,对小麦-簇毛麦代换系(6V／6A)、两个易位系(6VS／6AL,6VS／6DL)及亲本簇毛麦(VV)、硬粒小麦(AABB)、栽培小麦(AABBDD)的多态性进行了筛选分析。80.95％的S系列引物扩增出了结果,且条带较清晰;在100条OP系列引物中均扩增出结果,条带清晰可见。从205个随机引物中发现,只有1个引物OPW03在含有簇毛麦V染色体的4个材料中均扩增出1条约570 bp的谱带,而在栽培小麦和硬粒小麦中没有发现。因此,可以推测这个分子标记(OPW03-570)是位于簇毛麦V染色体短臂上的。

簇毛麦中谷胱甘肽硫转移酶基因的克隆与表达分析

利用抑制性消减杂交(SSH)方法克隆差异表达片段EST639,从簇毛麦cDNA文库中筛选到一个类谷胱甘肽硫转移酶基因(GST)的全长cDNA,命名为HvGSTF.该序列长1 232 bp,具完整的开放阅读框(ORF),编码一条含229个氨基酸残基的多肽,推导其分子量为25.6 kDa,等电点为4.89.该多肽的氨基酸序列与小麦、水稻、大麦、玉米、拟南芥的类GST分别具有83%,63%,61%,52%和47%的相似性.Northern杂交表明,HvGSTF在抗、感病簇毛麦叶片中都为组成型高表达,但在受白粉菌侵染的抗、感病簇毛麦中的表达模式不同,却与活性氧的积累模式存在着一致性,推测HvGSTF在簇毛麦细胞免受活性氧毒害方面起着重要作用.