【作者】 李光蓉；

【导师】 杨足君；

【作者基本信息】 四川农业大学 ， 作物遗传与育种， 2005， 硕士

【摘要】 中间偃麦草（Thinopyrum intermedium）是偃麦草的一个异源六倍体种（2n=42,染色体组为E₁E₂St或JJ^sS）,对条、叶、秆三锈及白粉、黑穗等病害免疫,高抗黄矮病、根腐与叶枯性病等对小麦改良十分重要的性状,它与小麦杂交易成功,后代类型比较丰富,为创制各种多抗、优质小偃麦新种质和优异小麦品种奠定了基础。茸毛偃麦草（Thinopyrum intermedium ssp.trichophorum）是中间偃麦草的一个新亚种,我们用远缘杂交和染色体工程方法将茸毛偃麦草遗传物质转移到小麦,创制了一个部分双二倍体TE-3以及转移了偃麦草抗病性Y176株系材料。本文综合利用醇溶蛋白和谷蛋白、RAPD、SCAR（sequence characterized amplified regoin）、根尖染色体计数观察、C-分带和基因组原位杂交（GISH）分析,对这些小偃麦新种质进行了鉴定,结果如下: 1.醇溶蛋白和谷蛋白分析表明,TE-3和部分Y176株系导入了源自茸毛偃麦草特异蛋白带。该偃麦草染色体醇溶蛋白特征带的存在和发现,为特定的偃麦草染色体或染色体片段导入小麦后的鉴定提供了生化标记。 2.用80个RAPD引物（H、M、P和O组）进行PCR扩增,得到3个（H18,H20,M04）偃麦草特异引物。对其中一个引物OPMO4₁₄₃₀扩增的特异片段分析,初步表明该片段是偃麦St基因组区域的特异标记,将该特异片段分离回收、克隆、测序,其序列为1423bp（序列登录号:AY618664）,根据测序结果重新设计引物一对引物,成功地转化RAPD标记为SCAR标记,并利用SACR标记对我们选育的小偃麦新种质进行了鉴定。更多还原

【Abstract】 Thinopyrum intermedium （Host） B.D., a hexaploid species （2n=6X=42,E₁E₂St orJJJ^sJ^sSS）, is an impotant source for improving genetic variability of cultivated wheat. Desirable characteristics of Th. intermedium of potential valuable for wheat improvement include perennial habit, stress tolerance and disease resistance （rusts and viral diseases）. There was lack of studies on the pubescent subspecies, Thinopyrum intermedium ssp. trichophorum. To broaden the genetic diversity of wheat, new wheat Th. -Trichophorum germplasms including partial amphiploid TE-3 and a wheat - Th. trichophorum derivatives Y176 were produced by chromosomal manipulation. In the present study, Gliad in and glutenin electrophoresis, Random amplified polymorphic DNA （RAPD）, Sequence characterized amplified region （SCAR）, Chromosome C-banding and Genomoic in situ Hybridization （GISH） analysis were used to characterize the new dermplasm. and the main results are showed as following:1. Analyzed by the composition of gliadin and glutenin, Th. Trichophorum partial amphiploid TE-3, Y176 and Chinese Spring. It was indicatede that some characteristic gliadin bands from Th. Trichophorum were expressed in the TE-3 and some of Y176. The specific protein bands of showed the useful biochemical marker for tracing the incorporation of the alien chromatin.2. Three specific primers OPM04, OPH20 and OPH18 showing the Th. Trichophorum specific bands were selected from 80 random primers by RAPD analysis. One of the primer OPM04, can amplify a specific band from Th. Tichophorum, which was furtherly confirmed to amplify the St genome of Th. Trichophorum. This specific band of RAPD marker from OPM04 was isolated and cloned, and showed the size of 1430bp （Gene bank number: AY618664）. Based onthe sequence, one pair of primers PI and P2 was designed, synthesized and used to amplify the above germplasms, which resulted in the similar results of RAPD analysis. It is indicated the RAPD marker OPM04i423 was successfully transferred to SCAR marker. The present SCAR were consequently used to identify our v/heat-Thinopyrum derivatives.3. One of advanced line Y176-3 with high resistance to stripe rust was obtained to subsequently cytogical descriptoion based on the existence of the specific protein bands and SCAR marker. Cytogenetic analysis showed that the Y176-3 has the chromosome number of is 44 in moititic root-tip cell, including four satellite chromosomes. The observation of meiotic chromosome configuration of PMCs was 22 bivalents. Giemsa-C banding showed the two smaller chromosomes of Th. Trichophorum with tolemeric bands.4. In order to reveal the genomic distribution of the added chromosomes of Y176-3, the Genomic in situ hybridization （GISH） was carried out by using the St genome a probe, and the common wheat genomic DNA as a blocker. The results indicated that the introduced Th. Trichophorum chromosomes are belongs to St genomes. Terefore, it is Concluded that the target St chromosomes of Th. Trochophorum was responsible for the specific protein bands and the rust resistance.更多还原