利用甘蓝型油菜高密度SNP遗传图谱定位油酸、 亚麻酸及芥酸含量QTL位点

doi:10.3864/j.issn.0578-1752.2014.01.003

摘要/Abstract

摘要： 【目的】菜籽油包括多种脂肪酸组分，提高油酸（C18:1）含量，降低亚麻酸（C18:2）和芥酸（C22:1）含量是油菜育种改良和遗传研究的重要目标。本研究利用刚开发的油菜60K芯片构建的高世代重组自交系群体遗传连锁图谱，对3个不同环境中影响甘蓝型油菜品质的油酸、亚麻酸及芥酸含量进行QTL定位分析，研究结果可对脂肪酸组分QTL位点在不同的群体之间准确比较分析。【方法】以高芥酸亲本GH06为母本和低芥酸亲本P174为父本构建高世代重组自交系，分别于2008年在德国吉森、德国霍亨里特及2009年德国吉森3个不同的环境中设置田间试验，收获自交种子，采用近红外分析方法3次重复对种子的脂肪酸组分进行分析。利用油菜60K芯片对重组自交系群体进行基因型分析，DNA样品预处理及芯片处理严格按照Illumina Inc公司Infinium HD Assay Ultra操作说明进行。取最小阈值LOD 5.0利用MSTmap软件构建遗传图谱。QTL定位所用的遗传图谱包括2 756个SNP位点，覆盖甘蓝型油菜基因组1 832.4 cM。利用Windows QTL Cartographer复合区间作图法对油酸、亚麻酸及芥酸含量进行QTL定位。【结果】在3个环境中，油酸和芥酸含量均表现为极显著负相关，相关系数均达到-0.95，且表现为主基因控制的性状，芥酸和亚麻酸表现负相关，油酸与亚麻酸表现正相关。3个性状在3个环境中共检测到14个QTL，在A08和C03上都检测到油酸和芥酸含量重叠的主效QTL位点。在3个环境中，油酸主效QTL位点解释表型变异19%—31%，芥酸主效QTL位点解释表型变异19%—34%，两者表现加性效应相反。A08和C03染色体上的芥酸主效QTL位点加性效应在3个环境中为7.6到9.6，加性效应来自低油酸、高芥酸亲本GH06。亚麻酸属于典型的数量性状，受环境影响较大，在3个环境中检测到不同的微效QTL位点，解释表型变异3%—12%。遗传图谱与物理图谱比较分析发现，脂肪酸去饱和酶FAD2基因位于亚麻酸QTL qA05C18:3的置信区间，而脂肪酸延长酶FAE1基因位于芥酸QTL qA08C22:1的置信区间。【结论】利用该套油菜60K芯片准确定位了油酸、亚麻酸及芥酸QTL位点，位于A08和C03染色体上的芥酸主效QTL位点同时也是油酸的主效QTL位点，该研究结果有利于不同群体在使用该套SNP芯片分析及对脂肪酸组分定位后准确比较分析。

关键词: 甘蓝型油菜 , 单核苷酸多态性 , 脂肪酸 , QTL

Abstract: 【Objective】Rapeseed oil contains many fatty acid components and the major goal in breeding and genetic research is to increase of the oleic acid content and decreases of the linolenic acid and erucic acid content. QTL mapping of oil quality related oleic acid, linolenic acid and erucic acid content in Brassica napus was made by using the high density SNP genetic map which was constructed from the high generation RIL population with the newly developed rapeseed 60K chip array. 【Method】 The high generation RIL population was derived from the cross of high erucic acid female parent GH06 and low erucic acid male parent P174. Seeds harvested from Giessen and Hohenlieth in Germany in 2008 and again in Giessen in 2009 were measured by NIRS with three technical repeats. The RIL population genotype was analyzed with the newly developed rapeseed 60K chip array. The DNA preparation and the chip preparation were processed strictly according to Infinium HD Assay Ultra manual of Illumina Inc. The SNP linkage map was constructed by using MSTmap program with minimum LOD 5.0. The reference SNP genetic map contains 2756 SNP markers, covering 1 832.4 cM of B. napus genome. QTL mapping of oleic acid, linolenic acid and erucic acid content was conducted by composite interval mapping using software Windows QTL Cartographer. 【Result】 Trait analysis indicated that the oleic acid and erucic acid contents were controlled by major genes, and their correlation coefficient was higher than -0.95 in the three different environments at significant level. Linolenic content showed a negative and positive correlation with erucic acid and oleic acid content, respectively. Two major QTL located on A08 and C03 were detected both for oleic acid and erucic acid content at the same position. In the three different environments, the major oleic acid QTL accounted for the phenotype variation from 19% to 31%, and the major erucic acid QTL effects varied from 19% to 34%, and the two major QTL for oleic acid and erucic acid content gave opposite direction of additive effect. The additive effect of the major erucic acid QTL on A08 and C03 come from the low oleic acid and high erucic acid parent GH06 with value varied from 7.6 to 9.6 in three different environments. Different QTL for linolenic acid was found across three different environments with QTL effects varied from 3% to 12%, which was coincided with that the linolenic acid was a typical quantitative trait, and was easily affected by the environment factors. The genetic map and physical map comparison discovered that the FAD2 gene and FAE1 gene located on the QTL qA05C18:3 and qA08C22:1confidence interval, respectively. 【Conclusion】 The oleic acid, linoleic acid and erucic acid content QTL was mapped with a new set of rapeseed 60K chip array, and the major QTL on A08 and C03 for erucic acid content is also responsible for oleic acid content. The mapping results are helpful to the fatty acid composition QTL comparison in different populations based on the same rapeseed 60K chip array.

Key words: Brassica napus , SNP , fatty acid , QTL

刘列钊, 李加纳. 利用甘蓝型油菜高密度SNP遗传图谱定位油酸、亚麻酸及芥酸含量QTL位点[J]. 中国农业科学, 2014, 47(1): 24-32.

LIU Lie-Zhao, LI Jia-Na. QTL Mapping of Oleic Acid, Linolenic Acid and Erucic Acid Content in Brassica napus by Using the High Density SNP Genetic Map[J]. Scientia Agricultura Sinica, 2014, 47(1): 24-32.

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