胰蛋白酶调控黄瓜丝裂原活化蛋白激酶途径中CsWRKY33诱导苯丙烷化合物的合成  

Trypsin regulates synthesis of phenylpropane compounds induced by CsWRKY33 in mitogen-activated protein kinase pathway of cucumber

作　　者：孙嘉炬 王贺敏 李欣[1] SUN Jiaju;WANG Hemin;LI Xin(College of Food and Bioengineering,Henan University of Science and Technology,Luoyang 471023,China)

机构地区：[1]河南科技大学食品与生物工程学院,河南洛阳471023

出　　处：《食品与发酵工业》2025年第5期95-102,共8页Food and Fermentation Industries

基　　金：国家重点研发计划项目(2017YFC1600802)。

摘　　要：丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)级联途径广泛参与植物的生理调控过程。胰蛋白酶作为诱抗剂可显著提高黄瓜的贮藏品质,但胰蛋白酶如何通过MAPK途径调控苯丙烷化合物的合成,从而影响果实抗性尚待阐明。为了研究胰蛋白酶诱导黄瓜抗性的分子机制,该研究进行了转录组学分析和广泛靶向代谢组学联合分析,并通过病毒诱导的基因沉默(virus-induced gene silencing,VIGS)进行验证。转录组结果显示,共有83个差异表达显著基因富集在MAPK途径上,其中48个基因显著下调,35个基因显著上调。基因集富集分析进一步从领头亚集中筛选出CsWRKY33转录因子。与拟南芥、烟草等植物的多序列比对结果显示,明确该转录因子属于WRKY家族,在N端有一个高度保守WRKY结构域。该研究将转录组与广泛靶向代谢组学联合分析发现,香兰素、二氢查尔酮-4′-O-葡萄糖苷、2-羟基-3-苯基丙酸等物质的合成显著上调。共表达网络分析结果显示,上述关键代谢物应受CsWRKY33的负调控。VIGS结果显示,CsWRKY33沉默后能够提高果实的抗性,延长保存时间。该研究发现,胰蛋白酶能够下调CsWRKY33的表达,促进香兰素、二氢查尔酮-4′-O-葡萄糖苷、2-羟基-3-苯基丙酸等具有较高抗氧化抗菌活性的化合物的合成,提高果实抗病性,延缓果实衰老腐败。The mitogen-activated protein kinase(MAPK)cascade pathway extensively participates in the physiological regulation processes of plants.Trypsin,as an inducer of resistance,significantly enhances cucumber storage quality.However,the mechanism by which trypsin regulates the synthesis of phenylpropanoid compounds through the MAPK pathway to influence fruit resistance remains to be elucidated.To investigate the molecular mechanism underlying trypsin-induced cucumber resistance,this study conducted a combined transcriptomic and widely targeted metabolomic analysis,validated through virus-induced gene silencing(VIGS).Transcriptomic results revealed that a total of 83 differentially expressed genes were enriched in the MAPK pathway,with 48 genes significantly downregulated and 35 genes significantly upregulated.Gene set enrichment analysis(GSEA)further identified the CsWRKY33 transcription factor from the leading edge subset.Multiple sequence alignment results with plants such as Arabidopsis thaliana and Nicotiana tabacum showed that the transcription factor belonged to the WRKY family and had a highly conserved WRKY domain at the N-terminus.The combined analysis of transcriptomics and widely targeted metabolomics demonstrated significant upregulation in the synthesis of compounds such as vanillin,dihydrochalcone-4′-O-glucoside,and 2-hydroxy-3-phenylpropanoic acid.Co-expression network analysis suggested that these key metabolites were negatively regulated by CsWRKY33.VIGS results showed that silencing CsWRKY33 enhanced fruit resistance and extended storage time.This study revealed that trypsin could downregulate the expression of CsWRKY33,promoting the synthesis of compounds with high antioxidant and antibacterial activity,such as vanillin,dihydrochalcone-4′-O-glucoside,and 2-hydroxy-3-phenylpropanoic acid.This enhancement leads to increased fruit disease resistance and delayed senescence and decay.

关 键 词：胰蛋白酶 转录组学 广泛靶向代谢组学 基因沉默 诱抗保鲜 

分 类 号：TS255.1[轻工技术与工程—农产品加工及贮藏工程]