【作者】 曾丽萍；

【导师】 孟金凌；

【作者基本信息】 华中农业大学 ， 作物遗传育种， 2011， 硕士

【摘要】 本实验室以欧洲甘蓝型油菜冬性品种Tapidor和中国半冬性栽培种宁油7号为亲本,构建TN DH群体,包含202个株系；并以该群体为母本,Ningyou7为父本,经过4次回交构建了导入系群体,称之为T-BC4群体。本研究目的为构建甘蓝型油菜双向导入系,因此以Tapidor为父本,构建了世代平衡的回交导入系群体,称之为N-BC4群体。同时,对双导入系群体进行了初步评估,并利用N-BC4群体对A9硫甙性状相关QTL进行了分析和研究。主要结果如下：完成甘蓝型油菜双向导入系的构建。以Tapidor为父本,构建N-BC4导入系群体。经过回交和自交的方法,得到包含大于4000个单株的BC4F1和BC4F2群体,完成了与T-BC4群体世代平衡的甘蓝型油菜双向导入系构建。另外分别得到来源于111个DH系对应239个单株BC2F1,360个单株BC2F2自交种；107个DH系对应501个单株BC3F,,105个DH系对应418个单株BC3F2自交种。以Ningyou7为父本的T-BC4群体,通过自交,得到来源于131个DH系对应BC4F2单株。经过回交,得到85个株系、133个单株的BC5F1回交种。对双向导入系进行了初步评估。结合开花时间,利用A10和C6 QTL区间内特异性分子标记对T-BC4群体和N-BC2群体进行了评估。调查两个群体约2000个单株开花时间,表明花期持续时间为30天,但即使来源于同一DH系的单株开花时间也有较大差异。同时,通过14对C6和3对A10连锁群开花QTL区间内分子标记进行评估,结果表明在双向导入系中同一QTL的作用刚好相反。开花时间早晚是由于导入相应染色体片段引起的,初步证明导入系群体构建成功。加密A9连锁群硫甙性状相关目标QTL所对应的遗传图谱置信区间。同时,利用T-BC4群体,通过A9连锁群硫甙QTL区间内分子标记进行筛选,为构建分离群体做准备。基于早期研究对对叶片、籽粒中各个硫苷分量和总量进行QTL定位分析,共有2个mQTL,表型方差均在20%以上。本研究利用白菜A9连锁群的BAC和Scaffold序列信息开发分子标记,加密QTL所对应的遗传图谱置信区间,完成目标QTL所对应的遗传图谱置信区间与白菜遗传图谱的比对。通过BAC和BAC-end共设计57对引物,其中3对定位到A9连锁群QTL区间内。经过QTL重新整合分析,目标QTL区间缩短为2cM。对来源于7个DH系810个BC4F2单株以及来源于11个DH系2000个BC4F3单株套袋自交,得到自交种,测定硫甙含量。同时利用目标QTL所对应遗传图谱置信区间内分子标记,对基因型进行筛选,结合基因型和表型,为构建分离群体奠定基础。更多还原

【Abstract】 Based on TN DH population with 202 lines, which is constructed by a winter and a semi-winter type of B. napus. In order to construct reciprocal introgression lines, Tapidor is used as the male parent to get the introgression lines called N-BC4 population. At the same time, a preliminary evaluation is made with the traits of flowering time. Meanwhile, using T-BC4 population is a good way to study the trait of glucosolinate in B. napus. Results stated as below:The work of construction of reciprocal introgression lines is successfully accomplished and the seeds of N-BC1, N-BC2, N-BC3, and N-BC4 populations are harvested. Four-thousand self-pollinated and hybrided seeds had been harvested from the N-BC4 populations. The seeds number of N-BC2F1, N-BC2F2,N-BC3F1, N-BC3F2 is 239、360、501 and 418.The seeds gained from the T-BC4 population is divided into two parts:self pollinated seeds of BC4F2 inherited from 133 DH liens and hybrided seeds of BC5F1 from 133 DH lines.In order to evaluate the introgression lines, markers from QTL region on C6 and A10 has been used to detect the trait of flowering time. In hypothesis, if the segements of QTL region within C6 is introgressed, the flowering time would be earlier than Ningyou7. On the contrary if the segements of A10 QTL region is introgressed, the flowerting time would be later than Ningyou7 in the T-BC4 populations. However, in the N-BC2 populations, there is a different situation. The introgression segments of A10 QTL region will flower earlier than Tapidor, vice verse. The results goes as what we wished.Confirm target QTL and construct NIL. Two in sixty-four mQTL, q.mcG-A9b and q.mcG-A9c, were two major QTL that controlled the content of total glucosinolate, each explaining more than 20% of phenotypic variation and controlled different glucosinolates in seeds, so the two mQTL, qmcG-A9b and q.mcG-A9c were the target QTL. Some polymorphic markers responding to BACs and Scaffold of B. rapa were mapped in the confidence intervals of the two target QTL, and the candidate genes for q.mcG-A9b were achieved through comparing A9 genomics between B. rapa and B. napus. By the tool of comaparative genomics analysis, the QTL region of A9 has been denstified with BAC-based and BAC end -based SSR markers. Three of fifty seven markers were mappped to the QTL region the QTL region was narrowed 2 cM. The materials of backcross populations derived from lines of TN DH population, and in BC4F2 population,11 NILs with the number of 2000, and in BC4F3 population,7 NILs with the number of 810 included target chromosome segments of varying length is developed through the foreground selection for target QTL.更多还原