【作者】 苗兴芬；

【导师】 胡国华；

【作者基本信息】 东北农业大学 ， 作物遗传育种， 2010， 博士

【摘要】 大豆是重要的经济作物,即是人类重要的植物蛋白和油分来源之一,又是重要的饲料作物。目前我国生产上脂肪酸品种还不能满足生产生活需要,迫切需要利用现代分子育种技术改良现有大豆品种的五种脂肪酸含量。大豆五种脂肪酸含量都属于多基因控制的数量性状,对其数量性状位点进行分析和定位,在分子辅助育种上具有重要指导意义。本研究利用源自美国大豆Charleston和中国优质品系东农594杂交获得的154个株系组成的重组自交系群体,在2007-2009年和3个不同的地点对大豆的脂肪酸含量进行了分析。得到了控制大豆五种脂肪酸含量的QTL。1.改进了大豆肪肪酸快速气相色谱分析方法。直接将磨碎豆粉中的脂质皂化、甲脂化后,利用色谱柱HP519091J-413,进行气相色谱分析。结果表明:软脂酸、硬脂酸、油酸、亚油酸和亚麻酸的出峰时间依次为6.57、7.83、8.73、8.86、9.06 min。这种方法的优点是快速、操作方便、节省药品,即使在较低的杂交世代样品很少,用普通法不易提取脂质时,此法也能顺利地完成分析工作。2.在三年三点条件下用气相色谱法测得大豆各株系子粒5种脂肪酸的含量,利用Win QTL Cartographer 2.5复合区间作图法(CIM)进行了QTL分析。结果共检测到68个相关的QTL位点,分布在18个连锁群上。多年多点同时检测到的QTL共有20个,其中控制软脂酸性状的3个QTL,包括qPal-C2-2、qPal-A1-1和qPal-N-1;控制硬脂酸性状的5个QTL,包括qSt-C2-1、qSt-D1a-1、qSt-B1-1、qSt-B1-2和qSt-F-1;控制油酸性状有5个,包括qOle-B2-1、qOle-C2-3、qOle-G-1、qOle-H-1和qOle-M-1;控制亚油酸性状的有3个,包括qLin-B1-1、qLin-C2-1,qLinN-1和qLin-H-1;控制亚麻酸性状的有3个,包括qLino-B1-1、qLino-C2-1和qLino-J-1。这些QTL的一致性较高,为特异脂肪酸含量标记辅助育种奠定了基础。在三年三点7个环境下,共定位了68个与大豆脂肪酸含量含量相关的QTL,有48个QTL只在一个环境中检测到,有20个QTL被多次检测到。说明在不同的环境条件下,控制大豆脂肪酸含量QTL的表达不同,进一步说明了大豆脂肪酸含量QTL受环境影响。3.本研究在三年三点7个环境下,有19个QTL存在环境互作,共发现45对脂肪酸QTL上位互作,发生在18个连锁群上。在上位互作中,检测到8对影响大豆软脂酸含量的上位互作位点,8对影响大豆硬脂酸含量的上位互作位点,12对影响大豆油酸含量的上位互作位点,11对影响亚油酸含量的上位互作位点,7对影响亚麻酸含量的上位互作位点,几乎覆盖所有连锁群。在这些上位互作位点中,绝大部分发生在不同的两条连锁群上,也有在一条连锁群上的两个位点,也发现同一位点与多个QTL位点发生互作。证明了上位互作方式的广泛性和复杂性。检测到了主效QTL也参与了上位性互作,进一步证实了主效QTL和上位性互作是数量性状重要的遗传基础之一。4.东北大豆品种五种脂肪酸含量在不同地区品种间差异均十分显著,并有较明显的地区性分布趋势。其中,软脂酸和油酸分布趋势是黑龙江>吉林>辽宁。硬脂酸和亚油酸是辽宁>吉林>黑龙江,亚麻酸为吉林>辽宁>黑龙江。脂肪酸组分相关分析结果:软脂酸与硬脂酸(r=-0.57)、软脂酸与亚油酸(r=-0.56)呈显著负相关,软脂酸与油酸正相关,与亚麻酸负相关,但是都未达显著水平。硬脂酸与油酸负相关,与亚油酸和亚麻酸负相关,但是均未达显著水平。油酸与亚油酸负相关达到极显著水平(r=-0.853),与亚麻酸负相关未达到显著水平。亚油酸与亚麻酸负相关达到了显著水平(r=-0.47)。通过关联分析,筛选到与脂肪酸组分关联的SSR标记共有33个。其中,与软脂酸相关联的有12个SSR标记;与硬脂酸含量相关联的标记有13个,与油酸相关联的SSR标记有18个,与亚油酸相关联的SSR标记有18个,与亚麻酸相关联的SSR标记有16个。本研究筛选到的与脂肪酸性状QTL共分离或紧密连锁的分子标记可供大豆脂肪性状品质性状育种中使用标记辅助选择时参考。本论文在以下获得了创新:1.改进一种快速测定大豆脂肪酸方法,首次使用色谱柱HP519091J-413在大豆脂肪酸含量的分析,且操作简便,需样品量少,分析时间短。2.在多年多点条件下,定位了大豆脂肪酸含量QTL68个,其中被多次检测到的有20个,将环境影响因素降低到最小,多次被检测到的QTL和筛选出的与SSR标记紧密连锁;并发现45对脂肪酸QTL上位互作,19个与环境互作位点。对大豆特异脂肪酸品种选育分子辅助育种有重要的意义。更多还原

【Abstract】 Soybean is an important economical crop, which is the major plant protein and oil for people consumption, and material of feeding stuff. Nowadays, the varieties of high fatty acids soybean, can not afford the need of people’s life and production. Therefore, it is imperious demand to use modern breeding technology to modify the content of fatty acids in soybean. Fatty acids are quantitative traits which were controlled by many genes. It has important meaning for breeding application to map these quantitative traits in soybean linkage groups.The population of 154 recombination inbred lines derived from the cross of America cultivar Charleston and Chinese elite line Dongnong 594. Fatty acids content of the population, planted in three different sites, were analyzed during 2007 to 2009. QTL that underlying Fatty Acids content of soybean were found.1. This paper improved the analysis method, the rapid determination on fatty acids content by Gas Chromatography(GC) in Soybean. The grinded soybean meal was used as material for saponification and methyl esterification directly, then measured by chromatography column HP519091J-413. The results showed that: the peak time of palmitic acid, stearic acid, oleic acid, linoleic acid, and linolenic acid were 6.57, 7.83, 8.73, 8.86, and 9.06 min. The advantages of this method fast, operating easily, and medicine-saving. Furthermore, it could obtain good results with few samples, which is difficult to recover the lipids from common ways.2. Five kinds of fatty acid contents of 154 F14 and F15 lines crossed by Charleston(♀)×Dongnong594(♂) were measured by GC under the condition of three years and three sites. The QTL of fatty acids contents were analyzed with method of composite interval mapping (CIM) by Win QTL Cartographer 2.5. In total, 68 related QTL distributed on the 18 linkage groups were obtained. 20 QTLs were detected in different years or different sites, including 3 QTLs related with palmitic acid contents, qPal-C2-2,qPal-A1-1 and qPal-N-1; 5 QTLs related with stearic acid contents, qSt-B1-1, qSt-B1-2, qSt-D1a-1, qSt-C2-1 and qSt-F-1; 5 QTLs related with oleic acid contents, qOle-B2-1、qOle-C2-3、qOle-G-1、qOle-H-1 andqOle-M-1; 4 QTLs related with Linoleic acid contents, qLin-B1-1、qLin-C2-1,qLinN-1 and qLin-H-1, and 3 QTLs related with Linolenic acid contents, qLino-B1-1, qLino-C2-1, and qLino-J-1. These stable QTLs laid the foundation for specific fatty acid contents breeding by marker-assisted selection.68 underlying fatty acids content QTLs were mapped under the 3 years and 3 sites. Among them, 48 QTLs were located in one environment, 20 were detected repeatedly. The results implied that different QTL controlled the expression of soybean fatty acid content and the fatty acid content of soybean QTLs underdifferent environmental conditions.3. 19 were environmental interaction QTLs and 45 pairs epistatic QTLs, related with soybean under 7 environments of 3 years and 3 sites, were located on eighteen linkage groups. 8 pairs epistatic QTLs were related with soybean palmitic acid contents, and 8 pairs epistatic QTL were related with soybean stearic acid contents. 12 pairs epistatic QTLs were related with soybean oleic acid contents. 11 pairs epistatic QTLs were related with soybean linoleic acid contents in all linkage groups. In these epistatic loci, the vast majority occured in two different linkage groups, or in a linkage group on two sites, it also found that with a locus were interacted with many QTL loci. It confirmed the extension and complexity of epistatic way. Major effect QTLs, involved in the epistatic, confirmed the main effect and epistatic QTLs is the genetic basis of quantitative traits. 4. It reached very significant of five fatty acids content among varieties from different regions in Northeast China, and a more visible trend was found in the regional distribution. The trends of palmitic acid and oleic acid distribution were Heilongjiang > Jilin > Liaoning. Stearic acid and linoleic acid distribution were Liaoning >Jilin>Heilongjiang. And linolenic acid distribution were Liaoning >Jilin>Heilongjiang.Correlation analysis results of fatty acid composition: palmitic acid and stearic acid was significantly negatively correlated (r = - 0.57, r = - 0.56) with linoleic acid content. Palmitic acid was positively correlated with oleic acid and negatively correlated with linolenic acid, but they are not significant negative correlation. Stearic acid was negatively correlated with oleic acid (r = -0.853), but none were significant negative correlation. Oleic acid was negatively correlated with linoleic acid (r = - 0.47).33 SSR markers were associated with the fatty acids composition. 12 SSR markers were associated with palmitic acid; 13 markers were associated with stearic acid, 18 SSR markers were associated with oleic acid, 18 SSR markers were associated with linoleic acid, and 16 SSR markers were associated with linolenic acid QTLs. In this study, the QTLs isolated or tightly linked molecular markers can be used for marker-assisted selection of soybean quality breeding.The paper obtained the Innovation in the following:1. Improved methods for papid Determination of soybean fatty acid. For the first time using the column HP519091J-413 in the analysis of soybean fatty acids, and it was easy to operate, less sample, and short analysis times.2. In multiple years and and sites, the mapping 68 QTLs of the fatty acid content of soybean, of which were repeatedly detected, 20 QTLs, will minimize the environmental impact factors, many QTLs were detected and screened with SSR markers closely linked; and detected 45 pairs of epistatic QTL fatty acids, 19 QTLs of interaction with the environment. These results were important in soybean molecular breeding with specific fatty acids.更多还原