摘要
蜡质诱导因子1(wax inducer 1/SHN1,WIN1/SHN1)与植物的抗逆性密切相关。为研究WIN1在耐旱经济灌木扁果枸杞(Lycium barbarum ssp.Bianguo)中的功能及其应用前景,本研究用反转录聚合酶链式反应(RT-PCR)技术从扁果枸杞中克隆了LbWIN1,构建了LbWIN1融合增强型绿色荧光蛋白(EGFP)的载体并表达在拟南芥(Arabidopsis thaliana)原生质体中进行亚细胞定位,用实时荧光定量PCR法(quantitative real-time PCR,qRT-PCR)分析LbWIN1的表达特征。结果表明,LbWIN1基因长1103 bp,含600 bp的开放阅读框,编码199个氨基酸,含有1个高度保守的AP2结构域。系统进化树分析发现,LbWIN1蛋白与茄科番茄(Solanum lycopersicum)SlWIN1和辣椒(Capsicum chinense)CcWIN1的同源性高达99%。亚细胞定位分析证明LbWIN1蛋白定位于细胞核。LbWIN1基因在叶、茎、根中均表达,其表达量受NaCl、渗透胁迫和外源脱落酸(ABA)诱导,表明LbWIN1参与旱生植物响应盐和干旱等非生物胁迫。
Wax inducer 1/SHN1(WIN1/SHN1)plays an important role in plant responses to stress resistance.To investigate the functions of WIN1 in the xerophytic economic shrub Lycium barbarum ssp.Bianguo and its application prospects,an AP2/ERF transcription factor-encoding gene,LbWIN1,was cloned using a reverse transcription-polymerase chain reaction.A pBI221::EGFP::LbWIN1 vector was constructed and expressed in the protoplasts of Arabidopsis thaliana for subcellular localization analysis.The expression patterns of LbWIN1 were analyzed using quantitative real-time PCR.The results showed that the LbWIN1 gene was 1103 bp in terms of its full length and contained a 600 bp open reading frame,encoding 199 amino acids.One highly conserved AP2 domain was found in LbWIN1.Phylogenetic analysis showed that LbWIN1 has 99%homology with WIN1 from Solanum lycopersicum and Capsicum chinense.LbWIN1 was located in the nucleus.Expression pattern analysis revealed that LbWIN1 is expressed in the leaves,stems,and roots,and its expression level was rapidly induced by NaCl,osmotic stress,and exogenous abscisic acid,indicating that LbWIN1 is involved in the stress response of L.barbarum ssp.Bianguo.
作者
袁惠君
苏照中
王春梅
张瑞艳
关玉晨
李学勇
鲍婧婷
YUAN Huijun;SU Zhaozhong;WANG Chunmei;ZHANG Ruiyan;GUAN Yuchen;LI Xueyong;BAO Jingting(School of Life Science and Engineering,Lanzhou University of Technology,Lanzhou 730050,Gansu,China;Lanzhou Institute of Husbandry and Pharmaceutical Sciences of CAAS,Lanzhou 730050,Gansu,China)
出处
《草业科学》
CAS
CSCD
北大核心
2023年第6期1449-1460,共12页
Pratacultural Science
基金
甘肃省自然科学基金(1606RJZA186)
国家自然科学基金(32060390、31460629)。
