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目的研究白桦Betula platyphylla 3-羟基-3-甲基戊二酰辅酶A还原酶(3-hydroxy-3-methyl-glutaryl-Co A reductase,Bp HMGR)全长基因结构特征及编码蛋白性质,揭示其节律表达模式以及对外源NO信号分子的响应机制。方法通过RACE技术克隆白桦Bp HMGR基因,应用生物信息学软件对Bp HMGR及其编码蛋白进行详细分析,利用实时荧光定量PCR定量分析Bp HMGR基因在昼夜节律变化中的表达模式。根据Bp HMGR氨基酸序列构建系统进化树,分析昼夜节律以及外源NO对白桦叶片中Bp HMGR转录水平的影响。结果 Bp HMGR基因全长1 764 bp,包含完整开放阅读框,已上传至NCBI,并获得登录号KJ452334,其编码蛋白由587 aa组成。白桦Bp HMGR基因与大豆(XP_003519474.1)、葡萄(XP_002275827.1)、苜蓿(XP_003617066.1)亲缘关系较近。结论 Bp HMGR随昼夜周期变化呈现出节律表达特性,在夜晚21:00时达到峰值。初步揭示了外源NO能诱导白桦细胞内三萜代谢途径的关键酶Bp HMGR基因上调表达,进而促进三萜产物齐墩果酸的合成。
Aim To study the structural characteristics and the nature of the encoded protein of Betula platyphylla 3-hydroxy-3-methyl-glutaryl-CoA reductase (Bp HMGR) Rhythmic pattern of expression and response to exogenous NO signaling molecules. Methods The Bp HMGR gene was cloned by RACE technique. The bioinformatics software was used to analyze Bp HMGR and its encoded protein in detail. The expression pattern of BPR HMGR gene in circadian rhythm was quantitatively analyzed by real-time fluorescence quantitative PCR. Based on the amino acid sequence of Bp HMGR, a phylogenetic tree was constructed to analyze the circadian rhythm and the effect of exogenous NO on Bp HMGR transcription in birch leaves. Results The full-length Bg HMGR gene was 1 764 bp in length and contained a complete open reading frame. It was uploaded into NCBI and got accession number KJ452334. The encoded protein was composed of 587 aa. The Betula platyphylla Bp HMGR gene is closely related to soybean (XP_003519474.1), grape (XP_002275827.1) and alfalfa (XP_003617066.1). Conclusions Bovine HMGR exhibits a rhythmic expression pattern with diurnal cycle changes and peaks at 21:00 in the evening. The up-regulation of BGR HMGR, a key enzyme in the induction of triterpenoid metabolism in Betula platyphylla, was initially revealed, which further promoted the synthesis of oleanolic acid.