河南农业大学学报2024，Vol.58Issue(5)：783-790,8.DOI:10.16445/j.cnki.1000-2340.20240617.001
磷脂酶Dα1在TuMV激活叶绿素降解相关基因表达中的功能分析
The role of phospholipase Dα1 in the expression of chlorophyll degradation related genes activated by TuMV
摘要
Abstract
[Objective]Investigating the role of phospholipase Dα1(PLDα1)and its product phosphatidic acid(PA)in the expression of turnip mosaic virus(TuMV)-induced chlorophyll degradation related genes in plants.[Method]The impact of TuMV on chlorophyll content in Nicotiana benthamiana was assessed using a UV-2800 UV spectrophotometer.Real-time qPCR was employed to analyze the effect of TuMV infection or transient expression of the 6K2 protein on the mRNA relative expression levels of chlorophyll degradation related genes Nicotiana benthamiana NON-YELLOWING1(NbNYE1),Nicotiana benthamiana NON-YELLOWING2(NbNYE2)and Nicotiana benthamiana pheide a oxygense(NbPAO)in N.benthamiana.Knockout of Nicotiana benthamiana phospholipase D alpha 1(NbPLDα1)or reduction of PA levels generated by PLD using 1-butanol was conducted to investigate the influence on TuMV-induced changes in mRNA relative expression levels of NbNYE1,NbNYE2,and NbPAO.Exogenous application of PA was used to evaluate its effect on the mRNA relative expression levels of NbNYE1,NbNYE2,and NbPAO.[Result]TuMV infection resulted in a 48%reduction of chlorophyll content in wild-type N.benthamiana leaves,with mRNA expression levels of NbNYE1,NbNYE2,and NbPAO increasing by 7.61,15.80 and 5.19-fold of those in healthy wild-type N.benthamiana,respectively.The expression levels of NbNYE1,NbNYE2,and NbPAO in TuMV-infected NbPLDα1 knockout mutant N.benthamiana were 2.35,7.93 and 1.29-fold higher than those of healthy wild-type N.benthamiana,respectively.After 1-butanol treatment,there was no significant difference in expression levels of NbNYE1,NbNYE2,and NbPAO between TuMV-infected wild-type N.benthamiana and healthy plants.The expression levels of NbNYE1,NbNYE2,and NbPAO in wild-type N.benthamiana sprayed with PA were 12.39,3.64 and 8.26-fold higher than those of plants treated with water,respectively.In addition,the expression levels of NbNYE1,NbNYE2,and NbPAO by transient expression of the 6K2 protein were 5.64,4.67 and 3.45-fold higher than those of expressing GFP,respectively,in wild-type N.benthamiana.[Conclusion]TuMV-promoted expression of chlorophyll degradation related genes depends on NbPLDα1 and its product PA,with the virus-encoded 6K2 protein identified as a key pathogenic factor promoting the expression of these genes.关键词
芜菁花叶病毒/本氏烟/叶绿素降解/6K2蛋白/磷脂酸/磷脂酶DKey words
turnip mosaic virus/Nicotiana benthamiana/chlorophyll degradation/6K2 protein/phos-phatidic acid/phospholipase D分类
农业科技引用本文复制引用
林佳宇,张世杰,徐传涛,杜林林,孙炳剑,孙航军..磷脂酶Dα1在TuMV激活叶绿素降解相关基因表达中的功能分析[J].河南农业大学学报,2024,58(5):783-790,8.基金项目
国家自然科学基金项目(31901855) （31901855）
中国烟草总公司陕西省公司科技项目(KJ-2023-05) （KJ-2023-05）
