番茄<i>SlUDP</i>基因的克隆及其在镉、干旱和盐胁迫中的响应分析

doi:10.7668/hbnxb.20191459

摘要/Abstract

摘要： 非生物胁迫环境严重制约了番茄的生产，因此挖掘番茄耐非生物胁迫相关基因尤为重要。前期通过高通量测序筛选获得的UDP-糖基转移酶基因能够参与番茄镉胁迫应答反应，以番茄为试材，进一步研究该基因参与番茄抗逆的功能。首先，利用同源克隆法获得UDP全长的cDNA序列，对该序列进行生物信息学分析，再利用系统进化树分析该蛋白与其他植物的亲缘关系。其次，采用实时荧光定量PCR分析该基因的组织表达特性及其在3种不同非生物胁迫条件下（Cd、PEG-6000、NaCl）的表达模式。进一步利用转基因酵母分析胁迫表型。测序结果显示，该基因的cDNA全长为1 486 bp，包含一个1 452 bp的开放阅读框（ORF），编码483个氨基酸，将其命名为SlUDP。系统进化树分析结果显示该蛋白与马铃薯的亲缘关系最近。实时荧光定量PCR分析结果表明，该基因在番茄不同组织中存在表达差异，在叶中表达量最高，各部位的表达量为叶片 > 果实 > 根部 > 侧茎 > 主茎 > 花。转SlUDP基因酵母胁迫表型分析结果显示，该基因提高了酵母对Cd及干旱的耐受性。根据以上结果推测SlUDP可能在番茄响应重金属镉和干旱胁迫中起到了一定的作用。

关键词: 番茄, SlUDP基因, 非生物胁迫, 表达分析, 功能分析

Abstract: Abiotic stress environment severely restricted tomato production, so it is particularly important to find genes related to abiotic stress tolerance in tomato.In the early stage of this study, UDP-glycyltransferase gene screened by high-throughput sequencing was able to participate in the response to cadmium stress in tomatoes. Therefore, this study took tomatoes as the test material to further study the anti-resistance function of this gene.First, the full length UDP cDNA sequence was obtained by homologous cloning method, and the sequence was analyzed by bioinformatics.Phylogenetic tree was used to analyze the phylogenetic relationship between the protein and plants.Secondly, Real-time fluorescence quantitative PCR was used to analyze the tissue expression characteristics of the gene and its expression patterns under three different abiotic stress conditions (Cd, PEG-6000, NaCl).The stress phenotype of transgenic yeast was further analyzed.The sequencing results showed that the cDNA of this gene was 1 486 bp in length, containing a 1 452 bp open reading frame (ORF), encoding 483 amino acids, and was named SlUDP.Phylogenetic tree analysis showed that the protein was most closely related to potato.The results of Real-time fluorescence quantitative PCR showed that the expression of this gene was different in different tissues of tomato, and the expression level was the highest in leaves, and the expression level in each part was root > fruit > lateral stem > main stem > flower.The stress phenotype analysis results of transgenic yeast showed that SlUDP gene improved the yeast's tolerance to Cd and drought.According to the above results, SlUDP may play a certain role in tomato response to abiotic stresses such as heavy metals cadmium, high salt and drought.

Key words: Tomato, SlUDP gene, Abiotic stress, Expression analysis, Functional analysis

中图分类号:

Q78
S641.03

耿鑫鑫, 于丽杰, 陈超, 金晓霞. 番茄SlUDP基因的克隆及其在镉、干旱和盐胁迫中的响应分析[J]. 华北农学报, 2021, 36(2): 46-53. doi: 10.7668/hbnxb.20191459.

GENG Xinxin, YU Lijie, CHEN Chao, JIN Xiaoxia. Cloning of SlUDP Gene and Its Response to Cadmium,Drought and Salt Stress in Lycopersicon esculentum Mill.[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2021, 36(2): 46-53. doi: 10.7668/hbnxb.20191459.

http://www.hbnxb.net/CN/Y2021/V36/I2/46

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番茄SlUDP基因的克隆及其在镉、干旱和盐胁迫中的响应分析

Cloning of SlUDP Gene and Its Response to Cadmium,Drought and Salt Stress in Lycopersicon esculentum Mill.

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