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园艺学报 ›› 2018, Vol. 45 ›› Issue (11): 2188-2198.doi: 10.16420/j.issn.0513-353x.2017-0737

• 研究论文 • 上一篇    下一篇

拟南芥异源表达紫斑牡丹PrLPAAT1对种子脂肪酸含量的影响

于 蕊,赵永青,张庆雨,白章振,孙道阳,胡佳媛,牛立新*,张延龙*   

  1. (西北农林科技大学风景园林艺术学院,陕西杨凌 712100)
  • 出版日期:2018-11-25 发布日期:2018-11-25

Effect of Ectopic Expression of Paeonia rockii PrLPAAT1 on Fatty Acids Content of Seed

YU Rui,ZHAO Yongqing,ZHANG Qingyu,BAI Zhangzhen,SUN Daoyang,HU Jiayuan,NIU Lixin*,and ZHANG Yanlong*   

  1. (College of Landscape Architecture and Arts,Northwest A & F University,Yangling,Shaanxi 712100,China)
  • Online:2018-11-25 Published:2018-11-25

摘要:

克隆得到的紫斑牡丹(Paeonia rockii)脂肪酸代谢相关基因PrLPAAT1,构建2S2::PrLPAAT1过表达载体并利用花序浸染法转化拟南芥,通过潮霉素抗性筛选以及PCR检测得到3个T3代纯合转基因拟南芥株系L1OX-11、L1OX-17和L1OX-19。半定量RT-PCR证明PrLPAAT1在3个转基因拟南芥株系中均显著表达。与野生型相比,过表达PrLPAAT1拟南芥的单株种子产量未产生明显变化。GC–MS测定结果表明,L1OX-11、L1OX-17和L1OX-19株系的总脂肪酸含量较野生型分别为无显著差异和提高了5.3%和5.2%;油酸(C18:1)含量分别提高了19.9%、24.6%和19.4%。实时荧光定量PCR分析表明,过表达PrLPAAT1拟南芥中油脂合成相关基因AtACP1、AtDGAT1和AtSUS3的表达水平均显著增加。由此推测PrLPAAT1在牡丹种子的脂肪酸合成过程中发挥重要的作用。

关键词: 紫斑牡丹, 溶血磷脂酸酰基转移酶, 脂肪酸, 三酰甘油

Abstract:

PrLPAAT1 gene was cloned from Paeonia rockii. The expression vector 2S2::PrLPAAT1 was constructed and transformed into wild-type(WT)Arabidposis plants by the floral dip method. Three positive T3 transformants L1OX-11,L1OX-17 and L1OX-19 were confirmed via hygromycin-resistant selection and PCR method. Semi-quantitative RT-PCR analysis indicated that PrLPAAT1 was highly expressed in transgenic Arabidopsis lines. Phenotypic analysis suggested that the seed yield had no significant changes in transgenic Arabidopsis lines,compared to WT plants. We analyzed the content and composition of FAs in mature seeds of the transgenic Arabidopsis using GC–MS. Compared with WT plants,seeds of L1OX-11,L1OX-17,and L1OX-19 line showed increased total fatty acids(FAs)content of 5.3%,and 5.2%,and the oleic acid(OA)content of 19.9%,24.6%,and 19.4%,respectively. Quantitative real-time PCR analysis indicated that a few endogenous genes(AtACP1,AtDGAT1,and AtSUS3)involved in lipid biosynthesis were up-regulated in PrLPAAT1-overexpressing lines. These results showed that PrLPAAT1 may play an important role in fatty acid biosynthesis of seed.

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