摘要
为明确葡萄霜霉菌对枯草芽胞杆菌与嘧菌酯协同的反应,采用显微表型观察明确枯草芽胞杆菌与嘧菌酯协同对葡萄霜霉菌的抑制作用,RNA-seq分析葡萄霜霉菌基因对枯草芽胞杆菌与嘧菌酯的协同表达,并通过qRT-PCR验证显著差异表达基因,通过荧光强度和高效液相色谱测定对重要代谢成分活性氧和ATP酶的影响。结果显示,枯草芽胞杆菌与嘧菌酯协同对葡萄霜霉菌的抑制作用为98.87%,孢子囊明显干瘪,内含物大量释放,与细胞壁明显分离,明显优于单独处理;葡萄霜霉菌对枯草芽胞杆菌与嘧菌酯协同反应共有1116个(上调305个,下调811个)基因发生差异表达,明显高于单独处理;通过GO、KEGG功能注释和功能富集分析,显著差异变化基因有140个;显著差异变化基因对枯草芽胞杆菌与嘧菌酯协同反应主要导致活性氧含量增加,ATP下调,同时阻碍ABC的转运,从而影响RNA的转录水平,破坏蛋白质、氨基酸等的代谢;qRT-PCR验证与RNA-seq数据具有很好的相关性;葡萄霜霉菌协同反应后活性氧含量增加以及胞内ATP含量大大降低,显著抑制能量的合成。
In order to study the response of plant-pathogenic oomycete Plasmopara viticola to Bacillus subtilis and azoxystrobin,the synergistic inhibitiory effect of B.subtilis and azoxystrobin on P.viticola was determined by microscopic phenotype observation,RNA-seq and qRT-PCR to identify and analyze the differentially expressed genes.The important metabolic components of active oxygen and ATPase were determined by fluorescence intensity and high-performance liquid chromatography.The results showed that synergistic inhibitory effect of B.subtilis and azoxystrobin on downy mildew was 98.87%,as the sporangia were obviously shriveled,the contents were released in large quantities,and the cell wall was clearly separated,that was noticeably better than that of single treatment.We found that the expression of 1116 genes changed significantly after B.subtilis and azoxystrobin combined treatment,including 305 up-regulated genes and 811 down-regulated genes;it was also significantly higher than that of single treatment.According to the Gene Ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG)functional annotation and enrichment analysis,there were 140 significantly diffe-rent genes.The significantly different genes resulted in the increase of reactive oxygen,ATP down-regulation,the hindered transport of ABC,thus affecting the transcriptional level of RNA and resulting in destroyed metabolism of protein,amino acid,etc.There was a good correlation between qRT-PCR and RNA-seq data.The active oxygen increased and intracellular ATP content decreased significantly,so that energy synthesis of P.viticola was significantly inhibited.In summary,it will further clarify its potential synergy mechanism and provide theoretical basis for the development and application of new fungicide.
作者
毕秋艳
赵建江
陈丹
吴杰
路粉
王文桥
韩秀英
BI Qiu-yan;ZHAO Jian-jiang;CHEN Dan;WU Jie;LU Fen;WANG Wen-qiao;HAN Xiu-ying(Key Laboratory of Integrated Pest Management on Crops in Northern Region of North China,Ministry of Agriculture,Integrated Pest Management Center of Hebei Province,Plant Protection Institute,Hebei Academy of Agricultural and Forestry Sciences,Baoding 071000,China)
出处
《植物病理学报》
CAS
CSCD
北大核心
2021年第5期775-788,共14页
Acta Phytopathologica Sinica
基金
河北省自然科学基金资助项目(C2019301093)
国家重点研发计划(2016YFD0200505-6)。
