入侵杂草喜旱莲子草WRKY转录因子家族鉴定及其除草剂胁迫表达特征  

Identification of WRKY transcription factor family and herbicide stress expression characteristics of the exotic weed Alternanthera philoxeroides

作　　者：韩硕 韩晓文 胡义锋 杨静美 陈中义 朱永兴 尹军良 HAN Shuo;HAN Xiaowen;HU Yifeng;YANG Jingmei;CHEN Zhongyi;ZHU Yongxing;YIN Junliang(Engineering Research Center of Wetland Ecology and Agricultural Use,Ministry of Education/College of Agriculture/College of Horticulture and Gardening,Yangtze University,Jingzhou 434025,Hubei,China;College of Life Science,South-Central University for Nationalities,Wuhan 430074,Hubei,China;College of Agriculture and Biology,Zhongkai University of Agriculture and Engineering,Guangzhou 510225,Guangdong,China)

机构地区：[1]长江大学湿地生态与农业利用教育部工程研究中心/长江大学农学院/长江大学园艺园林学院,湖北荆州434025 [2]中南民族大学生命科学学院,湖北武汉430074 [3]仲恺农业工程学院农业与生物学院,广东广州510225

出　　处：《草业科学》2024年第11期2517-2534,共18页Pratacultural Science

基　　金：长江大学湿地生态与农业利用教育部工程研究中心开放基金项目(KFT202305、KF201909)。

摘　　要：喜旱莲子草(Alternanthera philoxeroides)是我国入侵范围广、入侵程度重的外来入侵杂草,给生态平衡造成严重危害。WRKY是一类植物特有转录因子,在响应除草剂胁迫中发挥重要功能。为系统鉴定并分析喜旱莲子草WRKY家族成员特征,探究其响应除草剂胁迫的诱导表达模式,本研究从该杂草中鉴定到66个WRKY基因,根据系统发育关系将其分为a、b、c 3个亚族,同一亚族成员具有相似的基因结构、保守基序以及蛋白质三级结构。蛋白质特征分析发现,所有ApWRKYs均为亲水性蛋白。表达模式分析发现,ApWRKYs表现时空表达特异性。防效测试表明,5种除草剂均能抑制植物的鲜重和叶绿素含量,其中氯氟比氧乙酸对鲜重抑制率最高,乙羧氟草醚对叶绿素含量抑制率最高;定量分析8个ApWRKYs的除草剂响应模式,发现2个ApWRKYs受草甘膦诱导后表达水平先上升后下降,ApWRKY38.c在施药后7 d内表达量相对稳定;异丙隆和乙羧氟草醚处理0.5~3 d时,3个ApWRKYs表达量呈下降趋势;ApWRKY49.c和ApWRKY53.c分别在氯氟吡氧乙酸和噁草酮处理7 d和3 d时被显著诱导上调表达。这说明ApWRKYs参与喜旱莲子草对除草剂胁迫的响应过程,针对不同的除草剂表现不同的时空诱导表达特征。本研究为进一步探究ApWRKYs响应除草剂胁迫中的生物学功能奠定了基础。Alternanthera philoxeroides is an exotic weed with a wide invasion region and heavy invasion level in China,which has seriously harmed the balance of ecosystem services.WRKY is a type of plant specific transcription factor that plays an important role in herbicide stress response processes.To identify WRKY family members in A.philoxeroides,their characteristics were analyzed and the expression patterns were explored in response to herbicide stress.Consequently,66 WRKY genes were identified in A.philoxeroides.These were divided into three subgroups(a,b,and c)according to their evolutionary relationships.The members in the same subgroup had similar gene structure,conservative motifs,and protein three-dimensional structure.Analysis of protein characteristics revealed that all ApWRKY proteins are hydrophilic.Expression patterns revealed that the expression of ApWRKYs was spatio-temporal specific.The herbicidal efficacy test showed that all five herbicides tested can decrease plant fresh weight and chlorophyll content.Among them,fluroxypyr exhibited the highest inhibition rate on fresh weight,whereas fluoroglycofen exhibited the highest inhibition rate on chlorophyll content.RT-qPCR analysis of herbicide response patterns of eight ApWRKYs showed that the expression level of two ApWRKYs increased first and then decreased after being induced by glyphosate,and the expression level of ApWRKY38.c was relatively stable within seven days after application.The expression levels of three ApWRKYs showed a decreasing trend after 0.5 to 3 days of treatment with isoproturon and fluoroglycofen.ApWRKY49.c and ApWRKY53.c were significantly induced to up-regulate their expression after 7 and 3 days of treatment with fluroxypyr and oxadiazon,respectively.These results indicate that ApWRKYs were involved in the response process of A.philoxeroides to herbicide stress and showed different spatio-temporal induction expression characteristics for different herbicides.This study lays the foundation for further exploration of the biological f

关 键 词：WRKY转录因子 进化 除草剂 胁迫应答 叶绿素 生物信息学 低钾胁迫 

分 类 号：S451[农业科学—植物保护]