摘要
为了快速鉴定目标性状遗传位点,开发与性状连锁的分子标记,用于剔除高世代选育株行中不利性状,从而加速育种进程。以大豆MS轮回群体中发生花色分离的叫株行为研究材料,利用其衍生的F6;7中20个紫花和17个白花纯合家系分别构建2个DNA混池,通过高通量重技术获得变异信息,明确SNP,并在SNP富集区内开发分子标记对目标性状进行连锁分析。结果显示,在2个DNA混池间发现329992个SNP位点,表明混池间的遗传背景已经非常相似,但未发现明显SNP富集区,表明可能存在较多假阳性位点;进一步对高质量(Quality〉100)的SNP变异位点进行筛选,并去除杂合SNP位点,最终获得3371个可信位点。其中,位于13号染色上的SNP变异有700个(占比20.77%),并在20-30 Mb的物理区间形成一个最大的SNP富集区,推测调控花色的叼位点可能位于此区间内。利用该区间内SNP信息开发出dCAPS-1.dCAPS-2分子标记,连锁分析结果显示其与叼位点紧密连锁(W1-(0.4 cM)-dCAPS-1-(2.3 cM)-dCAPS-2),表明叼位点位于SNP富集区内。综上所述,通过构建高世代株行的分离群体混池,利用高通量测序方法可以快速定位控制目标性状的遗传位点,且开发的dCAPS标记可以有效剔除不状,从而加速遗传育种程。
To rapidly determine the genetic loci for target traits and develop molecular markers linked to traits which could remove plants with disadvantage traits in high generation breeding line and accelerate the breeding process. In this study, a row of F6 generation plants performing flower color segregation were selected from soybean MS recurrent population and individually harvested to form F6 ∶7 population. Among the F6 ∶7 population, 20 and 17 homozygous family lines with purple and white flower color, respectively, were selected to construct two DNA pools. Following, genetic variation were derived from next-generation sequencing and SNP enriched region were determined, furthermore, molecular markers were also developed to confirm the linkage between markers and target traits. High throughput sequencing analysis showed a total of 329 992 SNP variations were identified between the DNA pools of purple and white color, indicating that the genetic background between the DNA pools have a high degree of similarity. However, no significant SNP rich region was found possible due to there exist lots of false positive SNP. After screening SNP variations with high standard(Quality>100) and removing heterozygous SNP variations, a total of 3 371 confidence SNP variation were obtained. Among them, 700(20.77%) distribute on chromosome 13, and most were clustered in the physical interval of 20 to 30 Mb, strongly indicating that W1 was localized in this region. Two new markers, dCAPS-1, dCAPS-2 in the region were developed and they were linked to W1 closely(W1-(0.4 cM)-dCAPS-1-(2.3 cM)-dCAPS-2). In conclusion, the locus underlying the target traits could be rapidly determined through constructing two segregation pools from high generation line and genotyping by next-generation sequencing. Moreover, the disadvantage traits could be effective removed by the linked dCAPS markers which will accelerate breeding process.
作者
林静
赵青松
张孟臣
杨春燕
赵团结
LIN Jing;ZHAO Qingsong;ZHANG Mengchen;YANG Chunyan;ZHAO Tuanjie(Institute of Cereal and Oil Crops,Hebei Academy of Agriculture and Forestry Sciences,National Center for Soybean Improvement,Shijiazhuang Sub-Center,Huang-huai-hai Key Laboratory of Biology and Genetic Improvement of Soybean,Ministry of Agriculture,The Key Laboratory of Crop Genetics and Breeding of Hebei,Shijiazhuang 050035,China1;National Center for Soybean Improvement,Key Laboratory of Biology and Genetics Improvement of Soybean,Ministry of Agriculture,National Key Laboratory for Crop Genetics and Germplasm,Nanjing Agricultural University,Nanjing 210095,China)
出处
《华北农学报》
CSCD
北大核心
2019年第6期47-53,共7页
Acta Agriculturae Boreali-Sinica
基金
国家“863”计划项目(2012AA101106)
国家产业技术体系(CARS-004-PS06)
现代农业科技创新工程项目(2019-4-3-1)
关键词
高世代混池
高通量测序
SNP变异
遗传定位
分子标记开发
Mixture pools of advance line
High-throughput sequencing
SNP variations
Genetic mapping
Molecular marker development
