Quantitative proteomic and lipidomics analyses of high oil content GmDGAT1-2 transgenic soybean illustrate the regulatory mechanism of lipoxygenase and oleosin
Quantitative proteomic and lipidomics analyses of high oil content GmDGAT1-2 transgenic soybean illustrate the regulatory mechanism of lipoxygenase and oleosin
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DOI：
10.1007/s00299-021-02768-4
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发表时间：
2021-03
影响因子：
6.2
通讯作者：
Yang Xu;Fan Yan;Yajing Liu;Ying Wang;Han Gao;Shihui Zhao;Youcheng Zhu;Qingyu Wang;Jingwen Li
中科院分区：
文献类型：
--
作者：
Yang Xu;Fan Yan;Yajing Liu;Ying Wang;Han Gao;Shihui Zhao;Youcheng Zhu;Qingyu Wang;Jingwen Li
关键信息对转WT和GmDGAT 1 -2基因大豆的蛋白质组学和脂质组学分析表明，GmDGAT 1 -2的过量表达导致脂氧合酶表达下调，油酸含量上调，从而显著改变了大豆的脂肪组成和总脂肪酸含量。二酰基甘油酰基转移酶（DGAT）是脂肪酸代谢的关键限速酶，可调节油脂含量。本文中，从大豆中分离10 GmDGAT基因并将其转移到野生型（WT）拟南芥中。T3 GmDGAT 1 - 2转基因拟南芥种子中总脂肪酸含量是野生型的1.2倍。因此，将GmDGAT 1 - 2基因转入野生型大豆（JACK），获得了4个T3代转基因大豆株系。高效气相色谱分析和索氏提取结果表明，与野生型相比，转基因大豆中油酸（18：1）和总脂肪酸含量显著提高，而亚油酸（18：2）含量显著降低。棕榈酸（16：0）、硬脂酸（18：0）和亚麻酸（18：3）无显著差异。对于机理研究，使用蛋白质组学和脂质组学分析在WT（JACK）和转基因大豆豆荚之间鉴定了436个差异表达蛋白（DEPs）和180个差异表达代谢物（DEMs）。四种脂氧合酶蛋白在亚油酸代谢中下调，而四种油质蛋白在最终油脂形成中上调。结果表明，总脂肪酸和18：1脂肪酸组成增加，18：2脂肪酸组成减少。本研究为大豆的遗传转化和GmDGAT 1 - 2转基因大豆中总脂肪酸、18：1和18：2脂肪酸组成变化的分子机制的深入研究提供了新的思路。
Key messageProteomic and lipidomics analyses of WT and GmDGAT1-2 transgenic soybeans showed that GmDGAT1-2 over-expression induced lipoxygenase down-regulatation and oleoin up-regulatation, which significantly changed the compositions and total fatty acid.AbstractThe main goal of soybean breeding is to increase the oil content. Diacylglycerol acyltransferase (DGAT) is a key rate-limiting enzyme in fatty acid metabolism and may regulate oil content. Herein, 10GmDGATgenes were isolated from soybean and transferred into wild-type (WT)Arabidopsis. The total fatty acid was 1.2 times higher in T3GmDGAT1-2transgenicArabidopsisseeds than in WT. Therefore,GmDGAT1-2was transferred into WT soybean (JACK), and four T3transgenic soybean lines were obtained. The results of high-performance gas chromatography and Soxhlet extractor showed that, compared with those of JACK, oleic acid (18:1), and total fatty acid levels in transgenic soybean plants were much higher, but linoleic acid (18:2) was lower than WT. Palmitic acid (16:0), stearic acid (18:0), and linolenic acid (18:3) were not significantly different. For mechanistic studies, 436 differentially expressed proteins (DEPs) and 180 differentially expressed metabolites (DEMs) were identified between WT (JACK) and transgenic soybean pods using proteomic and lipidomics analyses. Four lipoxygenase proteins were down-regulated in linoleic acid metabolism while four oleosin proteins were up-regulated in the final oil formation. The results showed an increase in the total fatty acid and 18:1 composition, and a decrease in the 18:2 composition of fatty acid. Our study brings new insights into soybean genetic transformation and the deep study of molecular mechanism that changes the total fatty acid, 18:1, and 18:2 compositions inGmDGAT1-2transgenic soybean.