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【目的】以高蛋白、高产、高配合力大豆品种冀豆12为遗传基础,创造高蛋白含量新种质,分析蛋白含量相关QTL及其连锁标记,挖掘QTL中包含的优异基因,为高蛋白育种提供新种质、新标记。【方法】以冀豆12为轮回亲本,来自东北地区的红丰11、茶秣食豆、绥农14等蛋白质含量不同的育成品种和地方品种为供体亲本,通过有限回交,创造高蛋白新种质。从3个组合的回交后代品系中,选择农艺性状一致、产量与冀豆12无显著差异的4个高蛋白(50%—53%)品系和3个中低蛋白(38%—41%)品系为试验材料,参照大豆公共遗传连锁图谱,分别在20个连锁群上,均匀选取并筛选在双亲间表现多态性的SSR标记,分析冀豆12及其后代品系的遗传相似性与遗传差异,确定与蛋白质含量相关的QTL及其连锁标记。通过对QTL候选区间内的基因功能注释,挖掘与蛋白质含量相关的优异基因。【结果】创制出一批蛋白质含量超过50%的高蛋白新种质。3个组合中分别筛选到209、201和199个双亲间多态性标记;亲本与后代间遗传相似性分析结果表明,同一组合的后代中高蛋白品系的轮回亲本遗传背景回复率高于低蛋白品系遗传背景回复率。3个组合中4个高蛋白品系与轮回亲本冀豆12之间的相似系数平均值为79.58%,显著高于3个低蛋白品系材料与轮回亲本冀豆12之间的相似系数平均值67.81%;轮回亲本冀豆12传递给高蛋白种质的SSR位点数平均为161个,传递给低蛋白材料的SSR位点平均数为135个,二者相差27%。遗传效应分析结果表明,共发掘出位于14个连锁群上的22个与蛋白质含量相关的QTL及其连锁SSR标记,其中,18个QTL的蛋白质含量增效基因来自轮回亲本冀豆12。进一步分析显示,4个共性高蛋白染色体区段对高蛋白含量形成具有关键作用,分别位于C1连锁群(第4染色体)的75.52—80.62cM、D2连锁群(第17染色体)的67.71—84.18 cM、G连锁群(第18染色体)的80.38—96.57 cM和I连锁群(第20染色体)的46.22—50.11 cM。通过基因功能注释和代谢途径分析,预测了4个区段内20个可能参与7-磷酸景天庚酮糖、半胱氨酸、谷氨酸、丝氨酸、甲硫氨酸、色氨酸等氨基酸的合成与代谢等蛋白质合成相关代谢途径的候选基因。【结论】冀豆12含有较多的高蛋白QTL位点,供体亲本的高蛋白遗传位点可以在以冀豆12为轮回亲本的后代中表现出来,并通过SSR分析检测到。以冀豆12为轮回亲本,通过有限回交,易于创造出高蛋白种质。
【Objective】 High-protein, high-yielding and high-combining soybean JIU 12 was used as a genetic basis to create new high-protein content germplasm. Protein content related QTLs and their linkage markers were analyzed. Excluding QTLs, Breeding provides new germplasm, new markers. 【Method】 With Jidou 12 as the recurrent parent, bred varieties and local cultivars with different protein content from Hongfeng 11, Siberian soybean, Suinong 14 in northeast China were used as donor parents, and the high protein New germplasm. Four high protein (50% -53%) lines and three low protein (38% -41%) lines with the same agronomic traits and no significant difference in yield and yield were selected from the three combinations of backcross progeny lines. Based on the common genetic linkage map of soybean, the SSR markers of polymorphism in parents were selected and screened on 20 linkage groups respectively, and the genetic similarity and genetic difference of Jidou 12 and its offspring lines were analyzed To determine the QTL associated with protein content and its linkage markers. Through the annotation of gene function in QTL candidate interval, the excellent genes related to protein content were excavated. 【Result】 A number of new high-protein germplasm with more than 50% protein content were created. 209, 201 and 199 polymorphic markers of parents were screened among the three combinations. The results of genetic similarity analysis between parents and offspring showed that the genetic background of the recurrent parents in the same combination was higher than that of the low-protein lines Genetic background response rate. The average similarity coefficient between the four high protein lines and the reincarnated Jidou 12 was 79.58%, which was significantly higher than the average of 67.81% similarity coefficient between the three low protein lines and the reincarnated Jidou 12, ; The average number of SSR loci transferred to high protein germplasm by reincarnation Jidou 12 was 161, and the average number of SSR loci transmitted to low protein material was 135, with a difference of 27%. Genetic analysis indicated that 22 QTLs associated with protein content and their linkage SSR markers were found in 14 linkage groups. Among them, the 18 QTLs were derived from the reincarnation parent Jidou12. Further analysis showed that four common high-protein chromosomal segments play a key role in the formation of high protein content, which are located at 75.52-80.62cM of C1 linkage group (chromosome 4), 67.71-84.18 cM of D2 linkage group (chromosome 17) , 80.38-96.57 cM for G linkage group (chromosome 18) and 46.22-50.11 cM for linkage group I (chromosome 20). Through gene function annotation and metabolic pathway analysis, it is predicted that 20 of the four segments may be involved in amino acids such as sedoheptulosin 7-phosphate, cysteine, glutamic acid, serine, methionine and tryptophan The synthesis and metabolism of protein synthesis related metabolic pathway candidate genes. 【Conclusion】 Jidou 12 contains more high-QTLs, and the high-protein genetic loci of donor parents can be detected in offspring of Jidou 12 as a recurrent parent and detected by SSR analysis. Jidou 12 as a reincarnated parent, through limited backcross, easy to create high-protein germplasm.