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ω-3脂肪酸脱氢酶(ω-3 FAD)是植物脂肪酸合成途径的关键限速酶,通过调节该酶的过量表达,可以使植物中的α-亚麻酸(ALA)含量增加。本研究采用RT-PCR的方法,以紫苏L1品系为材料,克隆得到一个ω-3脂肪酸脱氢酶基因PfFAD3。生物信息学分析结果显示PfFAD3开放阅读框为1 176 bp,编码319个氨基酸,推测其蛋白分子量为44.7 k D,等电点为9.19;PfFAD3较为保守,具有四个富含组氨酸的保守区域。q RT-PCR结果表明,PfFAD3基因具有组织表达特异性,在种子中的表达量远高于其它器官。同时,研究了外源Me JA和温度对紫苏茎和叶中PfFAD3基因表达的影响,结果显示外源Me JA能够快速诱导叶片中PfFAD3基因表达量的上调,而抑制PfFAD3基因在茎中的表达;低温能够诱导叶片中PfFAD3基因表达量上调,而高温则抑制叶与茎中PfFAD3基因的表达,表明PfFAD3基因可能参与紫苏的防御反应。
The omega-3 fatty acid dehydrogenase (ω-3 FAD) is the key rate-limiting enzyme in the plant fatty acid synthesis pathway. Increasing alpha-linolenic acid (ALA) levels in plants by regulating the overexpression of this enzyme can result in increased omega- In this study, RT-PCR method to perilla L1 material as a material, cloned a ω-3 fatty acid dehydrogenase gene PfFAD3. Bioinformatics analysis showed that the open reading frame of PfFAD3 was 1 176 bp, encoding 319 amino acids. The predicted molecular weight of PfFAD3 was 44.7 kD and the isoelectric point was 9.19. PfFAD3 was more conserved and had four conserved histidine residues . q RT-PCR results showed that the PfFAD3 gene has tissue-specific expression, and the expression level in the seed is much higher than other organs. At the same time, the effects of exogenous Me JA and temperature on PfFAD3 gene expression in stem and leaves of Perilla frutescens were studied. The results showed that exogenous Me JA could rapidly induce the up-regulation of PfFAD3 gene expression in leaves and inhibit the expression of PfFAD3 gene in stem ; Low temperature could induce the up-regulation of PfFAD3 gene expression in leaves, while high temperature inhibited the expression of PfFAD3 gene in leaves and stems, indicating that PfFAD3 may be involved in the defense response of Perilla frutescens.