<i>Napin</i>启动子特异性表达油菜<i>BnGPAT9</i>基因

doi:10.7668/hbnxb.20190708

摘要/Abstract

摘要： 甘油三磷酸酰基转移酶（GPAT）基因编码参与植物油脂生物合成的酶。为了提高油菜籽含油量，促进油菜增产增油，改善菜籽油脂肪酸组分，克隆甘蓝型油菜Napin启动子与三酰甘油合成相关基因BnGPAT9，构建pBI121-Napin-BnGPAT9植物过表达载体，利用农杆菌介导法转化拟南芥，采用气相色谱法分析种子中脂肪酸组分，通过索式抽提法测定种子中油脂含量，研究在种子中过表达BnGPAT9基因对种子油脂合成的影响。Napin启动子在线分析表明，克隆到的启动子具有大量TATA-box、CAAT-box以及ABRE、GCN4等特征元件，完全具备种子特异性表达功能；种子中脂肪酸组分分析显示，拟南芥种子中油酸（C18：1）含量提高0.70~1.01百分点，亚油酸（C18：2）和芥酸（C22：1）含量降低；拟南芥种子含油量测定结果显示，种子中含油量显著增加1.91~2.56百分点。综上，甘蓝型油菜BnGPAT9基因对植物油脂合成具有重要作用，利用Napin启动子在拟南芥种子中过表达BnGPAT9基因可提高种子含油量并改善脂肪酸组成成分，为BnGPAT9基因应用于油菜油脂改良方面打下一定基础。

关键词: 甘蓝型油菜, Napin启动子, BnGPAT9基因, 植物表达载体, 含油量

Abstract: Glycerol-3-phosphate acyltransferase(GPAT) genes encode enzymes involved in glycerolipid biosynthesis in plants. To investigate the increase of oil content in rapeseed,the yield of oilseed rape and improve the fatty acid composition of rapeseed oil,we cloned Napin promoter and triacylglycerol synthesisrelated gene BnGPAT9,constructed pBI121-Napin-BnGPAT9 seed-specific expression vector and transformed into Arabidopsis thaliana,analyzed fatty acid composition and oil content in seeds. Using soil Agrobacterium-mediated transformation of Arabidopsis thaliana,gas chromatography and soxhlet extraction method had been analyzed the fatty acid composition and oil content in Arabidopsis seeds. PlantCARE analysis of the Napin promoter showed that the cloned promoter could achieve seed-specific expression,as it had lots of characteristic elements,such as TATA-box,CAAT-box,ABRE and GCN4 motifs. The results showed that oleic acid (C18:1) content was increased by 0.70 to 1.01 percent point,linoleic acid (C18:2) and erucic acid(C22:1) content were decreased,oil content was increased by 1.91 to 2.56 percent point in Arabidopsis seeds,suggesting that overexpress BnGPAT9 gene may effectively increase seed oil content and fatty acid composition in rape. This study lays a foundation for the application of BnGPAT9 gene in the improvement of rapeseed oil.

Key words: Brassica napus L., Napin promoter, BnGPAT9 gene, Plant expression vector, Oil content

中图分类号:

S634.03
Q78

邢蔓, 洪波, 张泽荣, 张曦予, 陈拓, 吴宁柔, 官梅, 邬贤梦, 官春云, 熊兴华. Napin启动子特异性表达油菜BnGPAT9基因[J]. 华北农学报, 2020, 35(4): 1-7. doi: 10.7668/hbnxb.20190708.

XING Man, HONG Bo, ZHANG Zerong, ZHANG Xiyu, CHEN Tuo, WU Ningrou, GUAN Mei, WU Xianmeng, GUAN Chunyun, XIONG Xinghua. Seed-specific Expression of BnGPAT9 Gene Controlled by Napin Promoter in Arabidopsis thaliana[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(4): 1-7. doi: 10.7668/hbnxb.20190708.

http://www.hbnxb.net/CN/Y2020/V35/I4/1

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Napin启动子特异性表达油菜BnGPAT9基因

Seed-specific Expression of BnGPAT9 Gene Controlled by Napin Promoter in Arabidopsis thaliana

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Napin启动子特异性表达油菜BnGPAT9基因

Seed-specific Expression of BnGPAT9 Gene Controlled by Napin Promoter in Arabidopsis thaliana

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