摘要
杨树叶锈病是杨树上的一种严重病害,对“三北”防护林体系造成极大威胁。ERF亚家族是植物所特有的AP2/ERF转录因子家族的一个主要亚家族,在植物抗病等生物胁迫响应中发挥重要作用。对E4锈菌具有耐病性的杂交杨‘2114’在接种E4锈菌12 h后,Ptd-ERF71持续上调表达。为探究Ptd-ERF71表达在‘2114’与E4锈菌互作过程中的调控作用,对Ptd-ERF71进行基因克隆及生物信息学分析,并采用Gateway转基因技术构建过表达载体,在拟南芥中进行异源表达并验证其功能。结果表明:Ptd-ERF71基因具有AP2保守结构域,属于ERF亚家族。将Ptd-ERF71在拟南芥中进行遗传转化,筛选并获得Ptd-ERF71基因过表达植株Ptd-ERF71-OE#1和Ptd-ERF71-OE#2。拟南芥Ptd-ERF71-OE#1、Ptd-ERF71-OE#2和At-RAP2.6L-OE(拟南芥Ptd-ERF71同源基因过表达株系)对透明壳孢菌感病性均显著高于拟南芥Col-0(WT),但At-RAP2.6L的拟南芥突变体株系At-rap2.6l与WT的感病性无显著差异。进一步分析表明,Ptd-ERF71基因过表达可能影响XGA-β1,3、ATL79、ZFCCCH67等植物防御相关基因表达,抑制‘2114’对E4侵染的过度防御,从而降低免疫反应对生长的影响。这种抑制作用在拟南芥与病原物的亲和型互作关系中促进拟南芥的感病性。
Populus leaf rust is a serious disease affecting Populus species and poses a significant threat to the“Three-North”shelter forest system.The ethylene-responsive factor subfamily(ERF)is a major subfamily within the plant-specific AP2/ERF transcription factor family,playing an important role in plant responses to biotic stresses,including disease resistance.In the disease-resistant hybrid poplar‘2114’,Ptd-ERF71 was continuously upregulated 12 h after inoculation with E4 rust fungus.To explore the regulatory role of Ptd-ERF71 expression in the interaction between‘2114’and E4 rust fungus,gene cloning and bioinformatics analysis of Ptd-ERF71 were conducted.Additionally,a Gateway cloning system was employed to construct an overexpression vector,and heterologous expression was performed in Arabidopsis thaliana to validate its function.The results showed that the gene contains an AP2 conserved domain and belongs to the ERF subfamily.The Ptd-ERF71 gene was genetically transformed into A.thaliana,resulting in the selection of overexpression lines Ptd-ERF71-OE#1 and Ptd-ERF71-OE#2.The Ptd-ERF71-OE#1 and Ptd-ERF71-OE#2 lines,as well as the At-RAP2.6L-OE(overexpression line of the homologous gene of Ptd-ERF71),showed significantly higher susceptibility to Hyaloperonospora parasitica Noco2 compared to the wild type A.thaliana Col-0.However,there was no significant difference in susceptibility between the At-rap2.6l mutant line and the wild type.Further analysis showed that overexpression of Ptd-ERF71 may influence the expression of plant defense-related genes such as XGA-β1,3,ATL79,and ZFCCCH67,potentially inhibiting excessive defense responses in‘2114’against E4 infection,thereby reducing the impact of immune responses on growth.This inhibitory effect may enhance the susceptibility of A.thaliana in the context of compatible interactions with the pathogen.
作者
杨波
刘念
王倩文
陈俏丽
Yang Bo;Liu Nian;Wang Qianwen;Chen Qiaoli(Northeast Forestry University,Harbin 150040,P.R.China)
出处
《东北林业大学学报》
CAS
CSCD
北大核心
2024年第12期1-9,共9页
Journal of Northeast Forestry University
基金
中央高校基本科研业务费专项资金资助博士自主创新项目(2572018AA04)。
关键词
杨树叶锈病
ERF
落叶松-杨栅锈菌
转基因拟南芥
Populus leaf rust
ERF
Melampsora larici-populina
Transgenic Arabidopsis thaliana
