摘要
半胱氨酸蛋白酶在植物调节盐胁迫应答方面起重要作用,克隆CaCP1,研究其抗盐性功能,为辣椒抗盐品种的培育提供参考。克隆CaCP1的启动子,根据CaCP1启动子中盐胁迫相关的作用元件,构建不同长度启动子片段的重组载体,进行烟草瞬时转化。通过对CaCP1超表达烟草转基因T3代株系进行盐处理,观察其表观症状,测定生理指标,并进行胁迫相关基因的表达分析。结果表明,盐胁迫下,CaCP1启动子-410--1 bp区段的GUS活性最强;CaCP1转基因植株叶片提前萎蔫和黄化,其中叶绿素含量和相对含水量均显著低于野生型,MDA和电解质渗漏率显著高于野生型,ROS清除酶CAT和SOD活性降低,POD活性升高;与野生型相比,转基因植株中ROS清除基因(NtSOD、NtPOD、NtCAT和NtAPX)和盐胁迫相关基因(NtLEA5、NtNHX1、NtP5CS1和NtSOS1)的表达量显著降低。CaCP1作为负调控因子介导植物对盐胁迫的防御反应。
Cysteine proteinases(CPs)play significant roles in salt stress responses of plants,thus the function of CaCP1 in salt resistance was studied by cloning CaCP1,which may provide reference for the breeding of pepper salt resistant varieties.First,the CaCP1 promoter of pepper(Capsicum annuum L.)was cloned,then recombinant vector with different length promoter fragments was constructed based on cis-regulatory elements associated with salt stress,and transient transformation of tobacco was performed.Salt treatment was carried out on T3generation tobacco transgenic lines overexpressing CaCP1,phenotype symptoms were observed,physiological indexes were measured,and the expressions of stress-related genes were analyzed.The results revealed that in response to salt treatment,the GUS expression in-410 to-1 bp region was the strongest.The leaves of CaCP1 transgenic plants wilted and yellowed earlier.The chlorophyll and relative water content were significantly lower than those of wild type,while MDA and electrolyte leakage rates were significantly higher than those of wild-type.ROS scavenger enzymes CAT and SOD activities decreased,while POD activity increased.Compared to wild-type,the expressions of ROS-scavenging genes(NtSOD,NtPOD,NtCAT,and NtAPX)and salt stress-related genes(NtLEA5,NtNHX1,NtP5CS1,and NtSOS1)significantly decreased.CaCP1 acts as a negative regulator to mediate the plant defense response to salt stress.
作者
杜清洁
周璐瑶
杨思震
张嘉欣
陈春林
李娟起
李猛
赵士文
肖怀娟
王吉庆
DU Qing-jie;ZHOU Lu-yao;YANG Si-zhen;ZHANG Jia-xin;CHEN Chun-lin;LI Juan-qi;LI Meng;ZHAO Shi-wen;XIAO Huai-juan;WANG Ji-qing(Department of Horticulture,Henan Agricultural University,Zhengzhou 450002;Henan Chuhe Garden Grass Service Limited Company,Zhengzhou 450000)
出处
《生物技术通报》
CAS
CSCD
北大核心
2023年第2期172-182,共11页
Biotechnology Bulletin
基金
国家自然科学基金项目(31801878)
河南省大宗蔬菜产业技术体系岗位专家项目(HARS-22-07-G4)。
