甜橙过氧化氢酶基因CsCAT2启动子克隆及活性分析  

Cloning and Activity Analysis of the CsCAT2 Promoter in Citrus sinensis

作　　者：陈烨[1] 单思杰 钟磊坚 曾卫 刘宇杭 李瑞民 CHEN Ye;SHAN Si-Jie;ZHONG Lei-Jian;ZENG Wei;LIU Yu-Hang;LI Rui-Min(College of Life Sciences,Gannan Normal University,Ganzhou 341000,China)

机构地区：[1]赣南师范大学生命科学学院,赣州341000

出　　处：《农业生物技术学报》2024年第12期2755-2763,共9页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(32260659);赣南师范大学2023年大学生创新创业训练计划项目(CX230074)。

摘　　要：柑橘黄龙病严重制约柑橘(Citrus spp.)产业的发展,黄龙病菌(Candidatus Liberibacter asiaticus)诱导柑橘韧皮部活性氧迸发导致细胞坏死,过氧化氢酶(catalase,CAT)是清除过氧化氢的关键酶。为明确甜橙(C.sinensis)CsCAT2基因(GenBankNo.XM_006473731)的表达模式,本研究克隆了CsCAT2的启动子,对CsCAT2启动子上顺式作用元件的分布进行分析,并利用截短的方法构建不同长度的CsCAT2启动子突变体,使用不同激素处理分析CsCAT2启动子活性的变化。结果表明,CsCAT2启动子序列中含有15个生物及非生物胁迫、激素信号响应相关的顺式作用元件;CsCAT2启动子全长具有最强的启动活性,截短突变体活性随长度减少而降低;脱落酸、茉莉酸、生长素、赤霉素和水杨酸处理显著提高CsCAT2启动子的活性,而乙烯利处理显著抑制了CsCAT2启动子的活性。本研究为深入解析甜橙过氧化氢酶基因CsCAT2的调控机制提供了数据支撑,丰富了柑橘与黄龙病菌互作过程中活性氧清除代谢的相关理论。Huanglongbing is a major impediment to the growth of the citrus industry.It is caused by Candidatus Liberibacter asiaticus which causes the production of reactive oxygen species in the phloem of citrus trees,resulting in cell death.The catalase(CAT)is essential for the elimination of hydrogen peroxide.In order to gain insights into the expression pattern of the catalase gene CsCAT2 in Citrus sinensis,the promoter of CsCAT2 was cloned,the cis-acting elements on the promoter were investigated,and truncations of the CsCAT2 promoter were constructed.Moreover,the response of the CsCAT2 promoter to various phytohormone treatments was evaluated.Analysis of the CsCAT2 promoter sequence revealed that it contained 15 cis-acting elements associated with biotic and abiotic stress and phytohormone signal response,predominantly for hormone signal response.The CsCAT2 promoter with its full-length had the highest promoter activity,and the activity of the truncated mutants decrease as their length decreasing.The activity of CsCAT2 promoter was significantly increased by abscisic acid,jasmonic acid,auxin,gibberellin and salicylic acid treatment,but treatment with ethephon significantly inhibited the activity of the CsCAT2 promoter.This study furnish data for further exploration of the regulatory process of the CsCAT2 gene from C.sinensis,and has broadened the theories concerning active oxygen scavenging metabolism during the interaction between Citrus spp.and Candidatus Liberibacter asiaticus.

关 键 词：甜橙 过氧化氢酶(CAT) 启动子 活性分析 植物激素 

分 类 号：S666[农业科学—果树学]