紫穗槐WRKY42基因耐盐碱性的功能研究  

The Function of Salt and Alkaline Tolerance of WRKY42 Gene in Amorpha fruticosa

作　　者：孙宇[1] 张艺腾 成慧慧 SUN Yu;ZHANG Yiteng;CHENG Huihui(Heilongjiang Ecological Engineering Vocational College,Harbin 150080;Institute of Cultivation and Cultivation,Heilongjiang Academy of Agricultural Sciences,Harbin 150028;Key Laboratory of Northeast Saline-alkali Vegetation Restoration and Reconstruction,Ministry of Education,School of Life Sciences,Northeast Forestry University,Harbin 150040)

机构地区：[1]黑龙江生态工程职业学院,哈尔滨150080 [2]黑龙江省农业科学院耕作栽培研究所,哈尔滨150028 [3]东北盐碱植被恢复与重建教育部重点实验室,东北林业大学生命科学学院,哈尔滨150040

出　　处：《植物研究》2023年第4期612-621,共10页Bulletin of Botanical Research

基　　金：黑龙江省重点研发计划项目(JD22A008);黑龙江省教育科学规划重点课题项目(GZB1320205)。

摘　　要：为了研究植物WRKY转录因子感知盐碱胁迫信号并通过生理和生化调节途径来维持其耐性功能作用。克隆紫穗槐(Amorpha fruticosa)的AfWRKY42基因,分析盐(NaCl)和碱(NaHCO3)胁迫和组织器官的响应表达模式;通过超表达烟草(Nicotiana tabacum)研究其耐盐碱性功能。依据紫穗槐逆境胁迫下的转录组测序数据克隆AfWRKY42基因,生物信息分析其含有1个WRKY结构域,2个低复杂区域和1个螺旋区域。系统进化树分析氨基酸发现,AfWRKY42与木豆(Cajanus cajan)的WRKY47、藜豆(Mucuna pruriens)的WRKY42亲缘关系最近。通过拟南芥(Arabidopsis thaliana)叶肉原生质体的瞬时基因表达系统验证AfWRKY42蛋白定位在细胞核;AfWRKY42基因定量分析表明其在紫穗槐的嫩茎表皮中表达最高;检测经NaCl和NaHCO3处理的根和叶中的表达模式,结果表明总体受胁迫诱导AfWRKY42基因表达量增加,推测AfWRKY42基因与植物耐盐碱性调节相关。分析35S启动的过表达T3代转AfWRKY42基因烟草耐盐碱性表明,盐碱胁迫处理后,转基因烟草株系抗性提高,它的叶绿素和电导率比野生型的高,丙二醛含量显著降低,说明AfWRKY42在响应盐碱胁迫中扮演重要的调节角色。将为抗盐碱性育种提供一个WRKY转录因子候选基因,为改善紫穗槐和其他植物的抗逆性奠定基础。To investigate the role of WRKY transcription factors(TFs)in sensing saline-alkali stress signals and maintaining their tolerance function through physiological and biochemical regulatory pathways,and the WRKY42 gene of Amorpha fruticosa was cloned and the expression pattern in response to sal(t NaCl)and alkali(NaHCO3)stress and tissue organs was analyzed,and the salinity tolerance function was also studied its by overexpression in tobacco.In this study,the AfWRKY42 gene was cloned based on transcriptome sequencing data of A.fruticosa L.under stress.Bioinformatics analysis showed that AfWRKY42 contained a WRKY structural domain,two low-complexity regions and a helix region.Phylogenetic tree analysis of amino acids revealed that AfWRKY42 was most closely related to WRKY47 of Cajanus cajan and WRKY42 of Mucuna pruriens.The localization of AfWRKY42 protein in mesophyll protoplasts of Arabidopsis thaliana was confirmed in nucleus by a transient gene expression system.Quantitative analysis of AfWRKY42 gene showed the highest expression in the shoot epidermis of Sophora japonica.Detection of expression patterns in roots and leaves treated with NaCl and NaHCO3 showed an overall increasing trend induced by it,suggesting that overall stress induced an increase in AfWRKY42 gene expression,and AfWRKY42 gene was associated with the regulation of salinity tolerance in plants.Analysis of salinity tolerance in 35S-initiated overexpressing T3 generation of tobacco lines transgenic for AfWRKY42 gene showed that the transgenic tobacco lines showed increased resistance after salinity stress treatment,it had higher chlorophyll and electrical conductivity and significantly lower malondialdehyde content than the wild type,indicating that AfWRKY42 played an important regulatory role in response to salinity stress.It would provide a WRKY transcription factor candidate gene for salinity resistance breeding and lay the foundation for improving the resistance of A.fruticosa and other plants.

关 键 词：紫穗槐 WRKY转录因子 盐碱胁迫 烟草 转基因 

分 类 号：S793.2[农业科学—林木遗传育种]