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园艺学报 ›› 2020, Vol. 47 ›› Issue (7): 1312-1322.doi: 10.16420/j.issn.0513-353x.2019-0956

• 研究论文 • 上一篇    下一篇

番茄转录因子基因SlWRKY16的克隆及原核表达分析

周 涛1,2,王 娟2,王露露1,王柏柯2,胡佳蕙1,2,兰海燕1,*,余庆辉2,*   

  1. 1新疆生物资源基因工程重点实验室/新疆大学生命科学与技术学院,乌鲁木齐 830046;2新疆农业科学院园艺作物研究所,乌鲁木齐 830091
  • 出版日期:2020-07-25 发布日期:2020-07-25
  • 基金资助:
    国家重点研发计划项目(2017YFD0101906);新疆农业科学院重点项目前期预研专项(xjzdy-003);国家现代农业产业技术体系建设专项资金项目(CARS-23-G25)

Cloning and Prokaryotic Expression Analysis of a Transcription Factor Gene SlWRKY16 in Tomato

ZHOU Tao1,2,WANG Juan2,WANG Lulu1,WANG Baike2,HU Jiahui1,2,LAN Haiyan1,*,and YU Qinghui2,*   

  1. 1Xinjiang Key Laboratory of Biological Resources and Genetic Engineering,College of Life Science and Technology,Xinjiang University,Urumqi 830046,China;2Institute of Horticultural Crops,Xinjiang Academy of Agriculture Sciences,Urumqi 830091,China
  • Online:2020-07-25 Published:2020-07-25

摘要: 从栽培番茄(Solanum lycopersicum)‘M82’中克隆到1个WRKY家族基因SlWRKY16。研究结果显示:SlWRKY16包含1 497 bp的完整开放读码框,编码498个氨基酸,理论分子量为55.5 kD,含有1个WRKYGQK保守结构域和C2H2锌指结构域,属于Ⅱb类WRKY转录因子基因,且定位于细胞膜上;栽培番茄SlWRKY16与潘那利番茄SpWRKY6(XP_015064265.1)、马铃薯StWRKY6(XP_006350473.1)同源性最高。qRT-PCR结果显示SlWRKY16在番茄不同组织中均有表达,叶片中的表达量最高;短时间非生物胁迫下该基因的表达量显著降低。此外,重组菌pET-30a-SlWRKY16在NaCl、甘露醇胁迫下生长均明显低于对照菌pET-30a。综上,番茄SlWRKY16基因可能在响应非生物胁迫过程中具有负调控效应。

关键词: 番茄, SlWRKY16, 原核表达, qRT-PCR, 亚细胞定位

Abstract: A WRKY family gene SlWRKY16 was cloned from cultivated processing tomato(Solanum lycopersicum‘M82’). The results showed that the open reading frame of SlWRKY16 was obtained from S. lycopersicum,which is 1 497 bp long and encodes 498 amino acids with an estimated molecular weight of 55.5 kD. Alignment of the deduced amino acid sequence of SlWRKY16 with WRKYs from other plant species indicates that SlWRKY16 contains a WRKYGQK conservative domain and a C2H2 zinc finger structure domain,which belongs to class Ⅱb WRKY transcription factor and locates on the plasma membrane. Further phylogenetic tree analysis showed that SlWRKY16 shared a high degree of sequence similarity with SpWRKY6(XP_015064265.1)and StWRKY6(XP_006350473.1)from S. pennellii and S. tuberosum,respectively. Moreover,quantitative RT-PCR analysis indicated that the expression of SlWRKY16 was tissue-specific and significantly decreased and down-regulated under short-term salt and drought stress. In addition,the recombinant strains(pET-30a-SlWRKY16)significantly grew lower than that of the control strain(pET-30a)when being applied with salt or simulated drought(mannitol)stress. Taken together,the above results suggest that SlWRKY16 from S. lycopersicum may have a negative regulatory effect in response to abiotic stress.

Key words: Solanum lycopersicum, SlWRKY16, prokaryotic expression, qRT-PCR, subcellular localization

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