[1]郭新亚,章文华,林峰.大豆油体蛋白基因GmOLE2的功能分析[J].南京农业大学学报,2021,44(3):477-486.[doi:10.7685/jnau.202008027]
 GUO Xinya,ZHANG Wenhua,LIN Feng.Functional analysis of soybean oleosin gene GmOLE2[J].Journal of Nanjing Agricultural University,2021,44(3):477-486.[doi:10.7685/jnau.202008027]
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大豆油体蛋白基因GmOLE2的功能分析()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
44卷
期数:
2021年3期
页码:
477-486
栏目:
生物与环境
出版日期:
2021-05-10

文章信息/Info

Title:
Functional analysis of soybean oleosin gene GmOLE2
作者:
郭新亚 章文华 林峰
南京农业大学生命科学学院, 江苏 南京 210095
Author(s):
GUO Xinya ZHANG Wenhua LIN Feng
College of Life Sciences, Nanjing Agricultural University, Nanjing 210095, China
关键词:
油体蛋白脂肪酸非生物胁迫大豆
Keywords:
oleosinfatty acidabiotic stressGlycine max
分类号:
Q942.5
DOI:
10.7685/jnau.202008027
摘要:
[目的] 本文旨在对编码栽培大豆(Glycine max)油体蛋白(oleosin,OLE)的GmOLE2基因进行功能研究,分析其蛋白结构、在油脂合成中的调控作用以及对非生物胁迫的响应。[方法] 运用生物信息学方法,分析其蛋白保守结构、亲疏水性以及进化关系;利用农杆菌侵染获得转基因拟南芥纯合株系,并利用气相色谱检测转基因拟南芥种子脂肪酸组分;对转基因拟南芥进行多种非生物胁迫处理,观察并统计相关性状和指标。[结果] GmOLE2蛋白具有OLE蛋白家族高度保守的结构域——脯氨酸结,且蛋白中间部分是一个由约24个氨基酸组成的超长疏水性结构域。GmOLE2是种子特异性表达的基因,过表达GmOLE2导致拟南芥种子内总蛋白含量增加约25%,总可溶糖含量下降10%,油脂含量下降约10%,饱和脂肪酸和三不饱和脂肪酸含量减少,单不饱和脂肪酸含量增加。过表达GmOLE2拟南芥种子可以缓解冻害,-20℃处理后的种子萌发率仍达到75%,是野生型萌发率的2倍。GmOLE2 mRNA积累受MeJA和ABA调控:10 μmol·L-1 MeJA处理GmOLE2过表达拟南芥幼苗,其主根生长受到抑制的情况下,侧根丰度及长度显著大于未处理组,且在24 h内GmOLE2基因表达量显著增加,是未处理组的8~10倍。将GmOLE2过表达拟南芥的种子播种在含0.5 μmol·L-1 ABA的培养基上,萌发3 d后,经RT-qPCR检测发现GmOLE2基因表达量显著升高,是未处理组的8倍。[结论] GmOLE2基因具有典型油体蛋白结构特征。在拟南芥中过表达GmOLE2基因,虽然减少种子总油含量但可以调节脂肪酸构成,且其种子具有较好的抗冻性,可受MeJA和ABA诱导表达参与植物对非生物胁迫的响应。
Abstract:
[Objectives] The objectives of this article were to investigate the function of soybean oleosin gene GmOLE2 by analyzing its protein structure,the regulation of oil synthesis and its response to abiotic stress,and thus provide genetic sources for the improvement of soybean seed quality and seed oil content. [Methods] The conserved protein structure,hydrophilicity and hydrophobicity and evolutionary relationship were analyzed by using bioinformatics tools;the fatty acid composition of Arabidopsis thaliana seeds was detected by gas chromatography. Transgenic A.thaliana seeds were treated under various abiotic stress,and related traits were observed and quantified. [Results] The GmOLE2 protein had a conserved domain "proline knot",and the middle part was an ultra-long hydrophobic structure composed of about 24 amino acids. GmOLE2 was highly expressed in seeds. Overexpression of GmOLE2 resulted in an about 25% increase in total protein in A.thaliana seeds,an about 10% decrease in total soluble sugar,and an about 10% decrease in oil,in which monounsaturated fatty acids increased accompanied by reduction of saturated fatty acids and triunsaturated fatty acids. The overexpression of GmOLE2 in A.thaliana enhanced the tolerance to freezing,and the seed germination rate still reached 75% at -20℃,twice higher than those of wild-type. GmOLE2 was regulated by MeJA and ABA:when the primary roots were inhibited,the abundance and length of the GmOLE2 seedling lateral roots were significantly longer than those of the untreated group upon 10 μmol·L-1 MeJA treatment,and also the expression level of GmOLE2 increased by 8 to 10 times within 24 h. The germination rate of GmOLE2-OEs seeds on 0.5 μmol·L-1 ABA medium was significantly higher than that of wild type,and also the expression level of GmOLE2 in GmOLE2-OEs was 8 times higher than that of the control group. [Conclusions] GmOLE2 had typical OLE protein structural characteristics. The overexpression of the GmOLE2 in A.thaliana could regulate fatty acid composition,reduce the total oil content in the seed and confer resistance to freezing. GmOLE2 was regulated by MeJA and ABA.

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备注/Memo

备注/Memo:
收稿日期:2020-08-23。
基金项目:国家转基因生物新品种培育重大专项(2018ZX08004003);南京农业大学高层次人才启动基金项目(680804016);江苏省自然科学基金项目(BK20200555)
作者简介:郭新亚,硕士研究生。
通信作者:林峰,副教授,主要从事植物磷脂代谢与逆境生理研究,E-mail:fenglin123@njau.edu.cn。
更新日期/Last Update: 1900-01-01