代谢组学揭示水稻—稻瘿蚊互作的潜在生化标识物

Metabolomics reveals potential biomarkers of rice gall midge(Orseolia oryzae) interactions

  • 摘要: 【目的】筛选稻瘿蚊(RGM)侵染下抗、感水稻材料差异代谢物,揭示水稻抗虫靶标化合物,为解析植物与昆虫互作关系及水稻害虫绿色防控提供理论依据。【方法】分别对抗RGM水稻材料GXM-001-2和易感RGM水稻材料TN1进行RGM接虫胁迫,接虫后48 h利用液相色谱—质谱联用仪(LC-MS)对RGM侵染及未侵染抗、感水稻材料的代谢物进行化学成分分析。【结果】RGM侵染下抗、感水稻材料的差异代谢物分析显示,LC-MS共检测到330种代谢物,其中155种为已知代谢物;被RGM侵染后,GXM-001-2中上调代谢物种类远少于其下调代谢物种类,也远少于TN1中的上调代谢物种类。RGM侵染下水稻潜在生化标识物种类分析表明,差异代谢物分为抗性特性(n=16)、易感特性(n=16)、侵染特性(n=7)和宿主特性(n=3)等4类。生化标识物及差异倍数分析表明,抗性特性物包括1-氨基环丙烷羧酸、3-磷酸甘油酸和二十烷酸等物质,其中6-甲基维生素D、谷氨酸、叶绿醇和酪氨酸等物质上调表达;易感特性物包括2-己酮酸、3-苯基乳酸和6-羟基己酸等物质,其中6-羟基己酸、葡萄糖和白皮杉醇等7种物质下调表达;侵染特性物包括γ-氨基丁酸、半乳糖酸和丙醇二酸等物质,其中腺嘌呤、尿苷、丙醇二酸和γ-氨基丁酸在抗、感材料中变化趋势一致;宿主特性物包括异亮氨酸、十六烷酸和环亮氨酸,前2种物质在GXM-001-2中显著高于TN1(P<0.05)。【结论】RGM侵染后从抗、感水稻材料中筛选到抗性特性物、易感特性物、侵染特性物和宿主特性物,其中脂肪酸及其衍生物(十六烷酸、二十烷酸、十四烷酸和β-羟基十四烷酸)、氨基酸(谷氨酸、酪氨酸、苯丙氨酸和γ-氨基丁酸)、酚类(阿魏酸)、萜类(叶绿醇)和胺类(乙酰苯胺、苄胺和乳酰胺)可能在植物抗虫防御反应中发挥重要作用,是RGM侵染水稻最重要的潜在生化标识物。

     

    Abstract: 【Objective】To detect differential levels of metabolites between rice gall midge(RGM) -resistant and -susceptible rice materials to provide a theoretical basis for analyzing rice-RGM interaction and assist in the development of green controls of RGM in rice.【Method】GXM-001-2(high resistance) and TN1(high susceptibility) were inoculated with RGM and liquid chromatography-mass spectrometry was employed to detect changes in metabolite levels after 48 h.【Result】330 metabolites were detected, of which 155 were known metabolites. TN1 displayed a much higher number of up-regulated metabolites after RGM infection than in GXM-001-2, where metabolites were largely down-regulated. The analysis of potential biochemical markers of rice infected by RGM showed that differentially regulated metabolites could be classified into four categories associated with resistance(n=16), susceptibility(n=16), infection(n=7) and host characteristics(n=3). Biochemical markers and fold change analysis showed that the resistance category included 1-aminocy-clopropanicarboxylic acid, 3-phosphoglyceric acid, eicosanoic acid, 6-methylvitamin D, glutamic acid, phyllol and tyrosine of which, the latter 4 were upregulated. Compounds associated with susceptibility included 2-hexketoic acid, 3-phenyllactic acid, 6-hydroxy hexanoic acid and other substances, of which 7 substances, including 6-hydroxy hexanoic acid, glucose and piceatannol, were downregulated. The characteristic compounds of infection included 4-aminobutyric acid, galactoic acid, propanedioic acid and adenine, uridine, propanedioic acid, and γ-aminobutyric acid showed similar up or down trends in GXM-001-2 and TN1. The host characteristic substances consisted of isoleucine, hexadecanoic acid and cycloleucine, with the former two substances at significantly higher levels in GXM-001-2 relative to TN1(P<0.05).【Conclusion】Under rice RGM condition, 16, 16, 7 and 3 metabolites associated with resistance, susceptibility, infection and host characteristics, respectively, were selected. Of these, fatty acids and their derivatives(hexadecanoic acid, eicosanoic acid, tetradecanoic acid and β-hydroxytetradecanoic acid), amino acids(glutamic acid, tyrosine, phenylalanine and g-aminobutyric acid), phenolic acids(ferulic acid), terpenoids(phyllol) and amines(acetanilide, benzylamine and lactamide) are the most important potential biomarkers of RGM infection, which could play important roles in plant insect defense responses.

     

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