Wang Yinan, Wang Mengke, Li Min, Zhao Te, Zhou Lin
Henan Agricultural University, College of Plant Protection, Zhengzhou, Henan, China.
Henan Agricultural University, Henan Key Laboratory for Creation and Application of New Pesticides, Zhengzhou, Henan, China.
PeerJ. 2021 Apr 30;9:e11339. doi: 10.7717/peerj.11339. eCollection 2021.
Leonian () can cause wilting and roots rotting on pepper and other cash crops. The new fungicide cinnamaldehyde (CA) has high activity against this pathogen. However, its potential mechanism is still unknown.
In order to gain insights into the mechanism, isobaric tags for relative and absolute quantification (iTRAQ)-based quantitative proteomics was used to analyze treated with CA. The iTRAQ results were evaluated by parallel reaction monitoring (PRM) analysis and quantitative real-time PCR (qRT-PCR) analysis. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was used to speculate the biochemical pathways that the agent may act on.
The results showed that 1502 differentially expressed proteins were identified, annotated and classified into 209 different terms (like metabolic process, cellular process, single-organism process) based on Gene Ontology (GO) functional enrichment analysis and nine different pathways (glyoxylate and dicarboxylate metabolism, fatty acid metabolism and so on) based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. This study suggested that CA disordered fatty acid metabolism, polysaccharide metabolism and leucine metabolism. Based on PRM analysis, five proteins including CAMK/CAMK1 protein kinase, glucan 1,3-beta-glucosidase, 1,3-beta-glucanosyltransferase, methylcrotonoyl-CoA carboxylase subunit alpha and isovaleryl-CoA dehydrogenase were down-regulated in treated with CA. Furthermore, the qRT-PCR analysis showed that the gene expression level of the interested proteins was consistent with the protein expression level, except for , and .
CA destroyed the metabolic homoeostasisof , which led to cell death. This is the first proteomic analysis of treated with CA, which may provide an important information for exploring the mechanism of the fungicide CA against .
莱昂氏菌（Leonian）可导致辣椒和其他经济作物枯萎和根部腐烂。新型杀菌剂肉桂醛（CA）对这种病原菌具有高活性。然而，其潜在机制仍不清楚。
为深入了解其机制，采用基于相对和绝对定量等压标签（iTRAQ）的定量蛋白质组学分析经CA处理的莱昂氏菌。iTRAQ结果通过平行反应监测（PRM）分析和定量实时PCR（qRT-PCR）分析进行评估。利用京都基因与基因组百科全书（KEGG）富集分析推测该药剂可能作用的生化途径。
结果显示，基于基因本体论（GO）功能富集分析鉴定、注释并分类出1502个差异表达蛋白，分为209个不同类别（如代谢过程、细胞过程、单细胞过程），基于京都基因与基因组百科全书（KEGG）富集分析分为9条不同途径（乙醛酸和二羧酸代谢、脂肪酸代谢等）。本研究表明，CA扰乱了脂肪酸代谢、多糖代谢和亮氨酸代谢。基于PRM分析，经CA处理的莱昂氏菌中，钙/钙调蛋白依赖性蛋白激酶（CAMK/CAMK1蛋白激酶）、葡聚糖1,3-β-葡萄糖苷酶、1,3-β-葡聚糖基转移酶、甲基巴豆酰辅酶A羧化酶亚基α和异戊酰辅酶A脱氢酶这5种蛋白表达下调。此外，qRT-PCR分析表明，除了[未提及的某些情况]外，所关注蛋白的基因表达水平与蛋白表达水平一致。
CA破坏了莱昂氏菌的代谢稳态，导致细胞死亡。这是首次对经CA处理的莱昂氏菌进行蛋白质组学分析，可能为探索杀菌剂CA对莱昂氏菌的作用机制提供重要信息。
