摘要
WRKY转录因子是植物特有的一类调控蛋白,在抵御生物和非生物逆境中起着重要作用。为初步明确该基因在抗病反应中的功能,本研究在分离青花菜BoiWRKY8基因的基础上,利用生物信息学方法进行序列分析,采用实时荧光定量PCR(qRT-PCR)明确其在核盘菌、灰霉菌侵染下的表达模式,并对过量表达植株进行抗病性鉴定。结果表明,BoiWRKY8的基因组全长为3170 bp,具2个内含子,长度分别为776和1422 bp。编码区全长为972 bp,编码323个氨基酸,WRKY结构域由60个氨基酸组成,包括一个WRKYGQK序列和一个C_(2)H_(2)锌指结构C-X_(4)-C-X_(23)-H-X_(1)-H。系统发育分析结果表明,BoiWRKY8与芸薹属植物的同源序列聚为一组,支持率达100%,与野甘蓝的关系最近。qRT-PCR结果显示,BoiWRKY8受核盘菌和灰霉菌的诱导,在6和12 h时的表达量最大,为对照的3.18/2.68和3.22/2.90倍;BoiWRKY8过量表达植株对核盘菌、灰霉菌的抗性显著增强,JA/ET通路标志基因BoPDF1.2的表达量显著提高。本研究结果为后续开展青花菜抗病机理研究和分子育种奠定了基础。
WRKY transcription factors are a distinct class of regulatory proteins found exclusively in plants,and they play an important role in combating various biotic and abiotic stresses.In this study,based on the isolation of BoiWRKY8 gene from broccoli,bioinformatic methods for sequence analysis were utilized,and qRT-PCR was applied to confirm the expression pattern of this gene under the infection of Sclerotinia sclerotiorum and Botrytis cinerea.Furthermore,overexpressed plants were subjected to disease resistance identification,aiming to preliminarily elucidate the function of BoiWRKY8 in disease responses.The results indicated that the genomic sequence of BoiWRKY8 was 3170 bp in length,containing two introns of 776 bp and 1422 bp.The complete coding sequence was 972 bp,encoding 323 amino acids.The WRKY domain of BoiWRKY8 was composed of 60 amino acid residues,including a WRKYGQK sequence followed by a C_(2)H_(2) zinc finger structure of C-X_(4)-C-X_(23)-H-X_(1)-H.Phylogenetic analysis results revealed that BoiWRKY8 clustered together with its homologous sequences from Brassica genus with a support value of 100%,and exhibited the closest relationship with wild cabbage.qRT-PCR results showed that the expression of BoiWRKY8 was significantly induced by the necrotrophic fungal pathogens S.sclerotiorum and B.cinerea,and the highest transcript levels were observed at 6 h and 12 h after inoculation,with fold changes of 3.18/2.68 and 3.22/2.90,respectively.BoiWRKY8 overexpressing broccoli plants exhibited increased resistance to both S.sclerotiorum and B.cinerea,concomitant with elevated expression of BoPDF1.2,one marker gene of the JA/ET pathway.Isolation and expression analysis of BoiWRKY8 gene provides a foundational framework for further investigation into the underlying mechanisms of broccoli’s resistance to diseases and molecular breeding.
作者
蒋明
何佳薇
方宇洁
尹龙飞
张慧娟
JIANG Ming;HE Jiawei;FANG Yujie;YIN Longfei;ZHANG Huijuan(College of Life Science,Taizhou University,Jiaojiang,Zhejiang 318000)
出处
《核农学报》
CAS
CSCD
北大核心
2024年第4期613-621,共9页
Journal of Nuclear Agricultural Sciences
基金
台州市科技计划项目(22nya01)
浙江省自然科学基金项目(LY19C150004)。
关键词
青花菜
表达分析
核盘菌
灰霉菌
过量表达
Brassica oleracea var.italica
expression analysis
Sclerotinia sclerotiorum
Botrytis cinerea
overexpression
作者简介
蒋明,男,教授,主要从事植物逆境生物学及分子调控研究。E-mail:jiangming1973@139.com;通讯作者:张慧娟,女,副教授,主要从事分子植物病理学研究。E-mail:zhanghj82@126.comxk。
