‘金小童’大白菜BrcGASA3基因在盐碱胁迫下的表达分析及抗性鉴定  

Expression Analysis and Resistance Identification of BrcGASA3 in Chinese Cabbage‘Jinxiaotong’Cultivar under Saline-alkali Stress

作　　者：马天意 许家佳 路文婧 吴艳 沙伟[1] 张梅娟[1] 彭疑芳 MA Tian-yi;XU Jia-jia;LU Wen-jing;WU Yan;SHA Wei;ZHANG Mei-juan;PENG Yi-fang(College of Life Sciences,Agriculture and Forestry,Qiqihar University,Heilongjiang Provincial Key Laboratory of Resistance Gene Engineering and Preservation of Biodiversity in Cold Areas,Qiqihar 161006)

机构地区：[1]齐齐哈尔大学生命科学与农林学院、黑龙江省抗性基因工程与寒地生物多样性保护重点实验室,齐齐哈尔161006

出　　处：《生物技术通报》2025年第2期127-138,共12页Biotechnology Bulletin

基　　金：黑龙江省属高等学校基本科研业务费青年创新人才项目(145109212)。

摘　　要：【目的】对不同盐碱胁迫处理下耐盐碱性不同的大白菜(Brassica rapa ssp.pekinensis)品种中BrcGASA3的表达响应模式进行分析,对耐盐碱性相对较强的‘金小童’品种大白菜中BrcGASA3进行克隆,并在模式植物拟南芥(Arabidopsis thaliana)中进行遗传转化以鉴定BrcGASA3的盐碱胁迫响应特性。【方法】利用实时荧光定量PCR对盐碱耐性不同的大白菜品种在不同浓度NaCl和Na_(2)CO_(3)处理过程中BrcGASA3的表达模式进行检测;对‘金小童’品种大白菜中BrcGASA3的编码序列进行克隆;构建BrcGASA3过表达转基因拟南芥并进行盐碱胁迫处理,观察植株表型并进行生理生化指标测定及分析。【结果】在NaCl处理中,较低浓度处理时耐盐性较高的大白菜中BrcGASA3表达量较高,较高浓度处理时耐盐性较低的大白菜中BrcGASA3表达量较高;在Na_(2)CO_(3)处理中,低浓度处理下两种大白菜中BrcGASA3表达量差异相对较小,高浓度处理下耐盐性较高的大白菜中BrcGASA3表达量显著高于耐盐性较低的大白菜;成功克隆获得‘金小童’大白菜中BrcGASA3的编码序列并构建了多个BrcGASA3过表达转基因拟南芥纯合体株系,发现在NaCl和Na_(2)CO_(3)处理下BrcGASA3过表达转基因拟南芥显示出低于野生型拟南芥的胁迫耐性;测定出在不同浓度NaCl和Na_(2)CO_(3)处理下BrcGASA3过表达转基因拟南芥和野生型拟南芥的渗透调节物质的含量、丙二醛含量、叶绿素含量等生理生化指标,初步阐释了过表达BrcGASA3降低拟南芥植株盐碱胁迫耐性的生理响应情况。【结论】在抗盐碱性不同的大白菜品种中BrcGASA3在不同程度盐碱胁迫处理下的表达响应方式不同,BrcGASA3在拟南芥中过表达降低了植株在盐碱胁迫处理下的生长能力。【Objective】The expression patterns of BrcGASA3 responding to different saline-alkali stress treatments in Chinese cabbage(Brassica rapa L.ssp.pekinensis)cultivars with different saline-alkali tolerance were determined,BrcGASA3 of Chinese cabbage cultivar‘Jinxiaotong’,which is with stronger saline-alkali tolerance,was cloned,and the genetic transformation in the model plant Arabidopsis thaliana was carried on to identify the saline-alkali stress response characteristics of BrcGASA3.【Method】Expression patterns of BrcGASA3 in Chinsese cabbage cultivars with different salinity tolerance during treatments with different concentrations of NaCl and Na_(2)CO_(3)were detected using quantitative real-time PCR.The coding sequence of BrcGASA3 from the Chinese cabbage cultivar‘Jinxiaotong’was cloned.BrcGASA3 overexpressed transgenic A.thaliana was constructed and subjected to saline and alkaline stresses,then the phenotypes of the plants were observed and physiological and biochemical indexes were measured and analyzed.【Result】Under NaCl treatments,the expression of BrcGASA3 in Chinese cabbage with stronger salt tolerance was high when the NaCl concentration was low;while the expression of BrcGASA3 in Chinese cabbage with weaker salt tolerance was high when the NaCl concentration was high.Under Na_(2)CO_(3)treatments,the expression of BrcGASA3 was similar in the two Chinese cabbage cultivars when the Na_(2)CO_(3)concentration was low,but the expression of BrcGASA3 was significantly high in Chinese cabbage with stronger salt tolerance when the Na_(2)CO_(3)concentration was high.The coding sequence of BrcGASA3 in the Chinese cabbage cultivar‘Jinxiaotong’was successfully cloned,and the overexpressed transgenic A.thaliana lines of BrcGASA3 were successfully constructed,and it was found that BrcGASA3-overexpressed transgenic A.thaliana showed lowered stress tolerance comparing with the wild-type A.thaliana under NaCl and Na_(2)CO_(3)treatments.Physiological and biochemical indices including the contents of

关 键 词：BrcGASA3 ‘金小童’大白菜 盐碱胁迫 NACL处理 Na_(2)CO_(3)处理 表达分析 抗性鉴定 

分 类 号：S63[农业科学—蔬菜学]