【作者】 王宏英；

【导师】 沈银柱； 黄占景；

【作者基本信息】 河北师范大学 ， 细胞生物学， 2002， 硕士

【摘要】 我国的盐渍土壤约有5亿亩，其中盐渍耕地1亿多亩，还有3亿多亩盐渍荒地有待开垦利用。土壤盐渍化是影响农业生产和生态环境的重要问题，因此耐盐植物的培育、耐盐机理的研究及耐盐基因的定位分离受到了人们越来越多的关注。本实验以“一粒传”后代中耐盐性不同的RH8706-49（一级耐盐）、H8706-34（四级耐盐）以及两者杂交得到的F₁、F₂群体为研究材料。采用SSR技术结合BSA方法对其中的耐盐相关基因进行定位。共筛选了246对引物，其中只有10对在两个材料间有多态性，多态性指数为4.07％，这说明我们用来杂交的两个材料间的遗传背景非常相似，利用多对引物进行筛选得到的差异具有较高的可靠性。这10对引物中只有WMS299在耐盐池和敏感池中存在差异。将WMS299在F₂群体的80个单株中进行扩增，发现绝人部分耐盐株与耐盐亲本的带型一致，而大部分敏感单株与敏感亲本的带型一致，表明WMS299与耐盐突变体中的突变位点存在连锁关系，并将该位点定位于3B染色体的长臂上。利用Jionmap1.4软件进行作图分析，WMS299与该位点间的遗传距离为14.003cM。为本材料中可能的耐盐相关基因的进一步分离打下了基础。 本实验室用RH8706-49与另一个不耐盐的材料进行正反交，发现F₁各生育期耐盐指数均高于不耐盐亲本，以RH8706-49为母本的组合F₁耐盐指数高于反交组合，这一现象说明耐盐性遗传可能在一定程度上与细胞质基因有关。本实验第二部分，以RH8706-49、H8706-34及它们的母本濮农3665为材料，利用RAPD方法研究它们的线粒体DNA间的差异。随机选取了53个随机引物，其中有两个引物在RH8706-49中有特异扩增产物，将特异条带回收、克隆、测序分析。并对其中的一个设计特异引物，进行特异扩增。更多还原

【Abstract】 Salinization of soil is a severe problem that has negative effects on farm production and ecological environment. Much attention is paid to raising salt-tolerant plants, studying salt-tolerance mechanism, locating and isolating salt-tolerance genes. In this experiment, F2 population was used as the population of mapping genes. The F2 population was derived from the hybrids of wheat salt-tolerant mutant RH8706-49 and salt-sensitive mutant H8706-34 (both of them are derived from a single seed). The relative salt-tolerance gene of wheat salt-tolerant mutant was mapped by using microsatellite marker and BSA. Among the 246 pairs of microsatellite primers, there were 10 pairs had polymorphism between RH8706-49 and H8706-34 (the polymorphic index is 4.07%). When further verification was taken in the salt-tolerant pool and the salt-sensitive pool, only the PCR products of WMS299 showed difference in the two pools. After WMS299 was amplified in the F2 population, most of the salt-tolerant plants had the same band as that of RH 8706-49, and most of the salt-sensitive plants had the same band as that of H8706-34. This suggested that there be a linkage between microsatellite marker WMS299 and the mutant site which had been located on the 3BL. By mapping the gene with Joinmap 1.4 (software), the genetic distance between the site and WMS299 is 14.003cM. This result had established the fundament for isolating salt-tolerance genes from the experiment materials.In our lab, RH8706-49 and a salt-sensitive material had been chose as parent plants to be hybridized and anti-hybridized. The F1 of the two hybridized combinations had higher salt-tolerant index than the salt-sensitive parent’s. FI of the combination that took RH8706-49 as female parent had higher salt-tolerant index than that of FI of the anti-hybridized combination. This indicated that the heredity of plants’ salt-tolerance might be relevant to cytoplasmic genes. In the second part of the experiment, RH8706-49^ H8706-34 and their female parent Punong3665 were used as experimental materials, and RAPD was used to find the differences among their mtDNA. Out of the 53 random primers, there were 2 primers whose products had specific bands in RH8706-49. The specific bands of RH8706-49 were collected cloned and sequenced.更多还原