【作者】 王黎明；

【导师】 王洪刚；

【作者基本信息】 山东农业大学 ， 作物遗传育种， 2006， 博士

【摘要】 中间偃麦草(Thinopyrum intermedium,E1E1E2E2StSt,2n=6x=42)是小麦的三级基因源,含有丰富的优良基因,在小麦的遗传改良中具有重要的利用价值。本研究利用普通小麦烟农15与中间偃麦草杂交获得F1,以烟农15作轮回亲本,与杂种F1回交2次,再经自交3代,从BC2F4选出山农0095,在对山农0095主要农艺性状鉴定的基础上,综合采用细胞遗传学、A-PAGE、SDS-PAGE、RAPD、SSR、GISH等方法对其进行了鉴定;对山农0095中外源染色体同源群归属、传递规律及其对杂交后代的影响进行了分析;运用CellCut?激光显微切割系统对山农0095与普通小麦杂种F1花粉母细胞单价体、山农0095根尖染色体进行切割的条件和方法进行了初步探索;对从烟农15与中间偃麦草杂交后代中选出的部分种质系进行了初步鉴定。获得以下主要研究结果:1.对山农0095进行主要农艺性状鉴定结果表明,与亲本烟农15相比,山农0095茎杆粗壮,繁茂性好,穗子变长、穗粒数增多,推测中间偃麦草中含有控制产量相关性状的主效基因。细胞遗传学鉴定结果表明,山农0095根尖细胞染色体数目2n=42,PMC MⅠ染色体构型为2n=21Ⅱ,与普通小麦的杂种F1 PMC MⅠ染色体构型为2n=20Ⅱ+2Ⅰ;以中间偃麦草基因组DNA为探针,中国春基因组DNA为封阻对山农0095的根尖染色体GISH,结果显示,山农0095中含有2条中间偃麦草染色体。这表明,山农0095是一个小麦-中间偃麦草二体异代换系。2.对山农0095及其亲本的种子醇溶蛋白进行A-PAGE分析结果表明,山农0095与烟农15的电泳带型并不完全一致,在β区含有来自于中间偃麦草特异谱带,进而说明山农0095含有中间偃麦草的遗传物质;HMW-GS鉴定结果表明,山农0095可能含有一个双亲所没有的新的谷蛋白亚基。RAPD、SSR分子标记鉴定结果表明,供试的180个RAPD引物中筛选出2个特异引物(S186、S32),在山农0095及其亲本间能揭示出多态性差异, S186900、S32400可以作为山农0095中特异RAPD标记;525对SSR引物中筛选出1对引物BARC159能在山农0095及其双亲之间扩增多态性差异,BARC159240可以作为山农0095中中间偃麦草的SSR标记。3.运用假鹅冠草(StSt,2n=2x=14)、二倍体长穗偃麦草(EE,2n=2x=14)基因组DNA为探针对山农0095根尖进行GISH结果表明,山农0095可能含有2条中间偃麦草的E基因组染色体;利用定位于不同小麦染色体上的SSR引物对一套中国春的缺体-四体系、山农0095及其双亲进行PCR扩增,结果表明,山农0095缺少3D染色体,进而说明小更多还原

【Abstract】 Thinopyrum intermedium (E1E1E2E2StSt,2n=6x=42) is the third gene pool for wheat, conferring many valuable genes for wheat, and has been widely used in wheat genetic improvement. A novel wheat germplasm, Shannong 0095, was developed from BC2F4 , the self-crossing progenies of common wheat Yannong15 backcrossed two times with hybrid F1 between Yannong 15 and Thinopyrum intermedium, and was characterized by using of cytological genetics, A-PAGE, SDS-PAGE, RAPD, SSR, and GISH before its major agronomic traits were identified. Homoeologous group of alien chromosome in Shanong 0095 was identified, its genetic laws in different wheat background were observed, and its genetic effects on different yield compmonent traits of hybrids F1were statistically analyzed. The two univalents of F1 between Shannong 0095 and common wheat at PMC MI and chromosomes in root cell of Shannong 0095 were micro-dissected by using of CellCut? laser micro-dissection machine. Several new germplasms derived from the progenies of common wheat Yannong15 and Thinopyrum intermedium were identified. The main results were as follows:1. Major agronomic traits of Shanong 0095 were identified. The results showed, comparison with Yannong 15, that Shannong 0095 has stronger stem, longer spike, much more spikelets, more luxuriant than that of Yannong15, which indicated that major genes controlling yield characteristics relative, derived from Thinopyrum intermedium, has been transferred into wheat background.Cytogenetics results showed that the chromosome number of Shannong 0095 in root tip cell was 2n=42 and its chromosome configuration at PMC MI was 2n=21Ⅱ. The mean chromosome configuration of hybrids F1 of the crossing between Shannong 0095 and Yannong 15 at PMC MI was 2n=20Ⅱ+2Ⅰ. The result of GISH with the probe of Thinopyrum intermedium genomic DNA and the blocker of Chinese Spring (C.S) genomic DNA, further conformed that Shannong 0095 contained 2 chromosomes of Thinopyrum intermedium. The results proved that Shannong 0095 was a wheat- Thinopyrum更多还原