青香蕉苹果MdAFS基因序列与表达模式分析

doi:10.13560/j.cnki.biotech.bull.1985.2017-0082

摘要/Abstract

摘要： 以青香蕉苹果叶片为材料，研究青香蕉苹果 ‘White Winter Pearmain’ α-法尼烯合酶（α-farnesene synthase，MdAFS）全长基因结构特征及编码蛋白质性质，分析MdAFS基因的诱导表达模式。采用生物信息学软件对 MdAFS 及其编码蛋白质进行详细分析，应用MEGA 6.0对不同物种来源的AFS构建系统进化树，利用 qRT-PCR技术研究MdAFS诱导表达模式。α-法尼烯合酶基因（GenBank No：AY563622）全长1 731 bp，由576氨基酸组成，是α螺旋为主要构成元件的混合型蛋白。不同物种的α-法尼烯合酶氨基酸序列相似度低，但α-法尼烯合酶N端RRx8W和C端的H-α1loop具有高度保守性。非生物胁迫和激素信号能诱导MdAFS基因上调表达，且MV、ABA和MeJA诱导该基因上调表达更加明显。α-法尼烯合酶氨基酸序列相似度低，但蛋白三维结构相似；α-法尼烯合酶在蛋白结构和功能上具有单萜合酶特征，初步揭示了非生物胁迫和激素信号能诱导青香蕉苹果α-法尼烯代谢途径的关键酶MdAFS基因上调表达。

关键词: 青香蕉苹果, α-法尼烯合酶, 倍半萜挥发物, 非生物胁迫, 表达分析

Abstract: This work is to study the full-length gene structural characteristics and encoded protein properties of α-farnesene synthase（MdAFS）from ‘White Winter Pearmain’ apple，and analyze the induced expression patterns of MdAFS gene. Bioinformatics software was used for analyzing the MdAFS gene and its encoded protein，secondary structure，and tertiary structure，then MEGA6.0 for constructing the phylogenetic trees with AFS from different species，and qRT-PCR for studying the expression pattern of MdAFS gene. The full-length of MdAFS gene（GenBank accession number：AY563622）was 1731 bp，contained the complete ORF，encoding 576 amino acids. The prediction analysis of secondary structure and tertiary structure showed that the protein was α-helix as the main parts of the mixed type proteins. The sequence similarity of α-farnesene synthase amino acid in different species was low，but the N-terminal RRx8W and C-terminal the H-α1loop of α-farnesene synthase was highly conservative. Abiotic stress and hormones induced MdAFS expression up-regulated，and more remarkably up-regulated expressions were observed in MV，ABA and MeJA treatments. In conclusion，α-farnesene synthase in ‘White Winter Pearmain’ apple has low similarity in amino acid sequence，but similar protein 3D structure. α-farnesene synthase presents monoterpene synthases characteristics in the protein structure and function. It is revealed that the abiotic stress and hormonal signal induce the up-regulation expression of key enzyme MdAFS gene in the α-farnesene metabolic pathway in ‘White Winter Pearmain’ apple.

Key words: ‘White Winter Pearmain’ apple, α-farnesene synthase, sequiterpene volatiles, abiotic stress, expression analysis

王茹茹,邓帅,丁瑞瑞,赵荣荣,张元湖. 青香蕉苹果MdAFS基因序列与表达模式分析[J]. 生物技术通报, 2017, 33(7): 75-82.

WANG Ru-ru ,DENG Shuai,DING Rui-rui ,ZHAO Rong-rong ,ZHANG Yuan-hu. The Sequence and Expression Analysis of MdAFS Gene in‘White Winter Pearmain’Apple[J]. Biotechnology Bulletin, 2017, 33(7): 75-82.

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