芦笋果聚糖:果聚糖6-果糖基转移酶基因Ao6G-FFT序列特征及表达模式研究

何娜1,2,张园1,2,林春1,2,3,毛自朝1,2,*,刘正杰1,2,3,*
1云南农业大学农学与生物技术学院, 昆明650201;2云南农业大学特色小宗作物研究中心, 昆明650201;3云南省高校作物种质创新及可持续利用重点实验室, 昆明650201

通信作者:毛自朝;E-mail: yhwynau@126.com;刘正杰;E-mail: 357315093@qq.com

摘 要:

为分析芦笋(Asparagus offcinalis)果聚糖合成关键基因功能特征, 本研究以二倍体芦笋为供试材料基于RT-PCR技术克隆果聚糖:果聚糖6-果糖基转移酶基因Ao6G-FFT, 并通过生物信息学方法分析基因、启动子和氨基酸序列特征, 通过qRT-PCR方法研究芦笋Ao6G-FFT基因在非生物胁迫处理下的表达情况。研究发现, Ao6G-FFT基因的cDNA及其基因组序列全长分别为1 812 bp17 351 bp, 基因组序列包含6个内含子和7个外显子, 编码含有603个氨基酸残基的亲水性蛋白, 并含有一个跨膜螺旋区和一个典型的信号肽结构。启动子区域(ATG上游约2 kb)含有TATA-box等保守元件外, 还存在大量与非生物胁迫和激素诱导响应的元件。组织特异表达分析表明, 该基因在不同组织器官中均有表达, 但贮藏根等地下部组织器官中基因表达量显著高于地上部分。 qRT-PCR分析表明, Ao6G-FFT基因受到冷胁迫、干旱胁迫、脱落酸和茉莉酸甲酯诱导表达, 推测该基因可能参与芦笋逆境胁迫应答。

关键词:芦笋; 6G-FFT基因; 克隆; 非生物胁迫; 表达分析

收稿:2020-07-04   修定:2021-01-13

资助:云南省农业基础研究联合专项[2018FG001(-019)和2017FG001(-002)]

Sequence characteristics and expression pattern of fructose 6G-fructose transferase gene Ao6G-FFT in Asparagus ofcinalis

HE Na1,2, ZHANG Yuan1,2, LIN Chun1,2,3, MAO Zichao1,2,*, LIU Zhengjie1,2,3,*
1College of Agriculture and Biotechnology, Yunnan Agricultural University, Kunming 650201, China; 2Institute of Improvement and Utilization of Characteristic Resource Plants, Yunnan Agricultural University, Kunming 650201, China; 3Key Laboratory of Crop Germplasm Innovation and Sustainable Utilization of Yunnan, Kunming 650201, China

Corresponding author: MAO Zichao; E-mail: yhwynau@126.com; LIU Zhengjie; E-mail: 357315093@qq.com

Abstract:

In order to analyze the functional characteristics of key genes of fructan synthesis in Asparagus
ofcinalis
, the fructose: fructose 6-fructosyltransferase gene Ao6G-FFT was cloned from diploid asparagus
by RT-PCR. The sequence characteristics of genes, promoters and amino acids were analyzed by bioinformatics. Expression pattern of
Ao6G-FFT gene in asparagus under abiotic stress was studied by qRT-PCR. The study found that the full length of cDNA and genome sequence of Ao6G-FFT gene were 1 812 bp and 17 351 bp, respectively. The genome sequence contained 6 introns and 7 exons, encoding hydrophilic proteins containing 603 amino acid residues, and contained a transmembrane helix region and a typical signal peptide conjunction. In addition to the conservative elements such as TATA-box, the promoter region (2 kb upstream of ATG) also contained many elements that respond to abiotic stress and hormone induction. Tissue-specifc expression analysis showed that the gene expressed in different tissues and organs, but the gene expressions in underground tissues and organs such as storage roots were signifcantly higher than those in aboveground parts. qRT-PCR analysis showed that Ao6G-FFT gene was induced by cold stress, drought stress, abscisic acid and methyl jasmonate, indicated that this gene might participate in asparagus stress response.

Key words: Asparagus ofcinalis; 6G-FFT gene; cloning; abiotic stress; expression analysis

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