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为了定位向日葵体细胞胚基因和筛选相关标记用于辅助育种,以31个重组自交系为材料,利用下胚轴表皮细胞诱导体细胞胚,获得了与体细胞胚发生率相关的,位于第5、10、13连锁群上的3个QTL位点,可分别解释表型变异的14.61%、10.04%和14.07%。同时,获得了与每块胚数相关的9个QTL位点,分别位于第2、8、10、11、12和19连锁群上,可解释表型变异的6.64%~16.96%,其中4个QTL位点位于第10连锁群上且位置相邻(在34.57~47.10cM之间),可解释表型变异的40.39%。为了验证所获结果,在31个重组自交系中,筛选出2个体细胞胚发生率高的株系和1个低的株系杂交,构建了2个F2分离群体。利用AFLP和SSR标记对F2群体进行扫描,在与体细胞胚发生率相关的第5、10、13条连锁群上分别增加了24、6和18个新标记,标间距从6.80~11.40cM缩短到5.10~5.60cM。通过对F2群体与亲本基因型进行QTL区域的对比分析,预选出7个体细胞胚发生率高和7个低的株系,利用F3家系诱导体细胞胚进行验证。结果表明,预选结果与实际获得的结果完全一致,控制向日葵每块胚数的基因主要位于第10连锁群上,控制体细胞胚发生率的基因主要位于第5、13连锁群上;4个AFLP标记(e32m50-1、e38m49-2、e40m49-1、e32m62-10)是最靠近控制体细胞胚发生基因的标记,可以作为该性状的辅助选择标记。
In order to locate the sunflower somatic embryo gene and the related markers for screening breeding, 31 recombinant inbred lines were used as materials to induce somatic embryos by hypocotyl epidermal cells, Three QTL loci on the 5, 10 and 13 linkage groups explained 14.61%, 10.04% and 14.07% of the phenotypic variation, respectively. At the same time, 9 QTL loci associated with each embryo were obtained, which were located on the 2nd, 8th, 10th, 10th, 11th, 12th and 19th linkage groups respectively, which explained 6.64% ~ 16.96% of the phenotypic variation, of which 4 QTL loci are located on the 10th linkage group and are adjacent to each other (between 34.57 and 47.10 cM), accounting for 40.39% of phenotypic variation. In order to verify the results obtained, two lines with high incidence of somatic embryos and one low line were selected from 31 recombinant inbred lines, and two F2 segregating populations were constructed. Using AFLP and SSR markers to screen F2 population, 24, 6 and 18 new markers were added to the 5, 10, and 13 linkage groups related to the somatic embryogenesis rate, and the standard spacing was shortened from 6.80 to 11.40 cM To 5.10 ~ 5.60cM. By comparing the QTLs between F2 population and their parents, seven lines with high incidence of somatic embryos and seven low lines were preselected and F3 lines were used to verify the somatic embryos. The results showed that the preselected results were completely consistent with the actual results. The genes controlling the number of embryos per sunflower were mainly located on the 10th linkage group, and the genes controlling the incidence of somatic embryos were mainly located on the 5th and 13th linkage groups. The 4 AFLP The markers (e32m50-1, e38m49-2, e40m49-1, e32m62-10) are the closest markers to the control of somatic embryogenesis genes and may serve as ancillary selection markers for this trait.