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八氢番茄红素脱氢酶(PDS)基因是类胡萝卜素生物合成途径中关键基因,它参与叶绿体光保护、脱落酸合成等多种生物代谢,同时也是VIGS技术中常用的指示基因。本研究利用同源克隆技术克隆并分析了海南粗榧八氢番茄红素脱氢酶(PDS)基因,结果表明:海南粗榧PDS基因,c DNA全长1 758 bp,命名为Cm PDS(Gen Bank登录号:KX766035),其编码585个氨基酸的开放阅读框。预测该基因编码蛋白质分子量为64.99 k D,其等电位为7.11,是理化性质稳定的亲水蛋白。经过氨基酸序列比对,发现海南粗榧中的PDS和红掌、中国水仙的PDS同源性最高,为85%;与其他作物的PDS同源性也都在70%以上,并且包含一个明显的保守结构域。此外,通过施加茉莉酸甲酯来模拟逆境信号,发现Cm PDS基因表达量呈现了先快速下降,然后逐步恢复到正常表达水平的趋势。暗示茉莉酸信号启动的抗性机制会抑制植物的光保护机制。同时本研究也为利用VIGS技术鉴定和挖掘海南粗榧抗癌生物碱合成酶相关基因奠定基础。
The phytoene dehydrogenase (PDS) gene is a key gene in the carotenoid biosynthesis pathway. It is involved in various biological metabolism such as photoprotection of chloroplast and abscisic acid synthesis, and is also a commonly used indicator gene in VIGS technology. In this study, homologous cloning technique was used to clone and analyze the phytoene dehydrogenase (PDS) gene in Hainan. The results showed that the cDNA of PDS gene was 1 758 bp in length and named as Cm PDS Bank accession number: KX766035), which encodes an open reading frame of 585 amino acids. The molecular weight of this gene was predicted to be 64.99 kD and its equipotential was 7.11. It is a hydrophilic protein with stable physical and chemical properties. After amino acid sequence alignment, it was found that the PDS homologous to PDS and Anthurium in China, and Narcissus species in China had the highest homology (85%), and their PDS homology with other crops was over 70%, and included a significant Conserved domains. In addition, through the application of methyl jasmonate to simulate the signal of stress, it was found that the expression of Cm PDS gene showed a rapid decline first and then gradually returned to the normal expression level. The mechanism of resistance, which suggests that jasmonic acid signaling initiates, inhibits the photoprotection of plants. At the same time, this study also laid the foundation for the use of VIGS technology to identify and excavate the genes related to the anti-cancer alkaloid synthase in Hainan.