敬丹;骆翔;陈利娜;夏小丛;杨选文;李好先;王企;曹尚银;

• 1:中国农业科学院郑州果树研究所

摘要(Abstract)：

【目的】测定并分析不同核桃品种核仁脂肪酸成分,克隆核桃FAD2基因,探究其在核桃不饱和脂肪酸合成中的作用。【方法】采用气相色谱-质谱联用的方法对20份核桃种质资源核仁的脂肪酸组分进行测定并对各组分间的相对含量进行计算及分析;基于核桃全基因组注释信息,通过实时荧光定量PCR技术对核桃油酸脱氢酶FAD2基因在高、低亚油酸品种间的表达模式进行分析;通过RT-PCR技术扩增JrFAD2基因的CDS序列,对其进行结构分析,并利用农杆菌介导的瞬时表达系统选用带有GFP标签的pBI121载体构建35S::JrFAD2重组表达载体,利用激光共聚焦显微镜观察核桃FAD2在本氏烟草叶表皮细胞中的表达。【结果】20份核桃资源核仁脂肪酸主要组分为亚油酸、油酸、硬脂酸和棕榈酸,其中不饱和脂肪酸高达90%。实时荧光定量PCR结果显示,高亚油酸品种及低亚油酸品种的油酸脱氢酶FAD2基因的表达模式具有差异。其中WALNUT_00011902-RA在3个高亚油酸品种的表达量均显著高于低亚油酸品种。选择WALNUT_00011902-RA作为目标基因进行克隆,得到长度为1 152 bp的JrFAD2基因的CDS序列,编码383个氨基酸,含有ω-6脂肪酸去饱和酶的功能结构域,属于Membrane-FADS-like超家族。系统进化分析发现,核桃FAD2与大豆、油桐FAD2的亲缘关系较近。亚细胞定位结果显示,核桃FAD2定位于细胞质。【结论】JrFAD2基因可能在核桃不饱和脂肪酸合成过程中对亚油酸的合成及油酸和亚油酸的比例具有促进和调控作用。

关键词(KeyWords)： 核桃;脂肪酸;FAD2;基因克隆;亚细胞定位

Abstract:

Keywords:

基金项目(Foundation): 国家科技基础工作重点项目（2012FY110100）;; 中国农业科学院科技创新工程专项（CAAS-ASTIP-2019-ZFRI）

作者(Author): 敬丹;骆翔;陈利娜;夏小丛;杨选文;李好先;王企;曹尚银;

Email:

DOI: 10.13925/j.cnki.gsxb.20190390

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