一个影响酪氨酸代谢藜麦突变体的鉴定

doi:10.13560/j.cnki.biotech.bull.1985.2024-0301

摘要/Abstract

摘要：

【目的】利用收集的藜麦植株颜色突变体，测定该突变体的代谢组成，分析代谢通路之间变化和转化规律，构建出该突变体代谢变化基本模型，为进一步鉴定和克隆影响藜麦重要代谢通路的关键遗传位点提供材料基础。【方法】利用正向遗传学手段，从藜麦常规品种‘藜红1号’（Red Quinoa 1, RQ1）后代中，筛选到一植株和穗部红色消退而绿色加深（green quinoa 1, gq1）的自然突变株作为材料，与其原始亲本RQ1相比较，利用非靶向代谢组学鉴定灌浆时期幼穗差异代谢成分，通过KEGG（Kyoto encyclopedia of genes and genomes）代谢途径分析和差异代谢物关联分析，揭示GQ1基因突变引发的关键代谢通路的变化。【结果】经过连续4个世代的遗传分析表明，gq1突变体植株颜色变异能够稳定遗传，且是由单个遗传位点控制。与其原始亲本RQ1相对比，在藜麦gq1突变体中检测到了409个差异代谢物，其中含量升高的代谢物有110个，299个代谢物含量降低。代谢组学分析发现，在藜麦gq1突变体中，对植物次级代谢有着重要影响的酪氨酸和以其为核心衍生出的其他代谢产物发生了整体性的降低。此外，包括6种人体必需氨基酸在内的多种氨基酸和TCA循环中的成分，在gq1突变体发生了显著的减少。【结论】通过对这些差异代谢进行KEGG代谢途径富集分析，表明GQ1基因突变造成了以酪氨酸为核心的初级代谢组分和次级代谢组分整体降低，意味着该基因可以成为协同优化藜麦初级代谢和次级代谢的关键遗传位点。

关键词: 藜麦, 突变体gq1, 代谢组学, 酪氨酸代谢, 氨基酸含量, 初级代谢, 次级代谢

Abstract:

【Objective】 Quinoa, known as the “golden grain” for its rich beneficial secondary metabolites and nutritional components for human benefit, is explored in this study through the analysis of a color mutant isolated from quinoa natural population. By examining the metabolic composition of this mutant, analyzing the changes and transformation routes between metabolic pathways, and constructing a basic model of metabolic changes in the mutant, this study is aimed to provide a material basis for further identification and cloning of key genetic loci affecting important metabolic pathways in quinoa.【Method】 Using forward genetics, a natural mutant exhibiting reduced red pigmentation and intensified green coloration（green quinoa 1, gq1）was screened from the progeny of the quinoa cultivar, ‘Red Quinoa 1’（RQ1）. This mutant was compared with its original parent, RQ1, to identify differential metabolic components in the young panicle of grain filling stage using untargeted metabolomics. KEGG metabolic pathway analysis and correlation analysis of differential metabolites were employed to reveal the overall differences in metabolic pathways and some key nutritional components in the mutant gq1.【Result】 Genetic analysis conducted over four successive generations indicates that the color variation in the gq1 mutant quinoa plants was stably inherited and was controlled by a single genetic locus. Compared to its original parent RQ1, 409 differential metabolites were detected in the quinoa gq1 mutant, with the concentration of 110 metabolites increased and that of 299 metabolites decreased. Metabolomics analysis revealed a comprehensive decrease in tyrosine and its derivative metabolites in the quinoa gq1 mutant, which are crucial for plant secondary metabolism. Moreover, a significant reduction was observed in various amino acids, including six essential amino acids for humans, and components of the TCA cycle in the gq1 mutant. 【Conclusion】 The enrichment analysis of these differential metabolites through KEGG metabolic pathways indicates that the gq1 gene mutation leads to a general reduction in both primary and secondary metabolic components centered around tyrosine. This suggests that the gene could serve as a key genetic locus for the synergistic optimization of primary and secondary metabolism in quinoa.

Key words: quinoa, mutant gql, metabolomics, tyrosine metabolism, amino acid content, primary metabolism, secondary metabolism

姜宇舢, 兰倩, 王芳, 姜亮, 裴成成. 一个影响酪氨酸代谢藜麦突变体的鉴定[J]. 生物技术通报, 2024, 40(10): 253-261.

JIANG Yu-shan, LAN Qian, WANG Fang, JIANG Liang, PEI Cheng-cheng. Characterization of a Quinoa Mutant Affecting Tyrosine Metabolism[J]. Biotechnology Bulletin, 2024, 40(10): 253-261.

图/表 7

图1 ‘藜红1号’（RQ1）及其突变体（gq1）生长发育各阶段的表型比较 A：2周时期幼苗；B：2周时期幼苗侧面；C：灌浆期植株；D：灌浆期幼穗；E：种子；比例尺：A, B, D, E = 1 cm, C = 10 cm

Fig. 1 Phenotypic comparison between ‘Red Quinoa 1’（RQ1）and its mutant（gq1）during different development stages A: 2-week-old seedling; B: side view of 2-week-old seedling; C: plants during the filling stage; D: young panicle of grain filling stage; E: seed. Scale bar: 1 cm for A, B, D, and E, respectively, and 10 cm for C

表1 突变体gq1和‘藜红1号’（RQ1）杂交F2世代性状分离比的卡方测验

Table 1 Test of Chi-square on segregation rate of F2 population between gq1 and RQ1

杂交组合 Hybrid combination of F₂	总计 Total	红色单株个体数 Number of red individuals	绿色单株个体数 Number of green individuals	x²（3∶1）	P
RQ1/gq1	163	128	35	1.082	0.298

图2 ‘藜红1号’（RQ1）及其突变体（gq1）中差异代谢物分析概述 A：gq1对RQ1的OPLS-DA模型得分散点图；B：差异代谢物层次聚类分析热图；C：基于倍数变化的差异代谢物的火山图

Fig. 2 Overview of differential metabolite analysis in gq1 vs RQ1 A: Score plots of the OPLS-DA model for gq1 vs RQ1. B: Heatmap of hierarchical clustering analysis for differential metabolites. C: Volcano plot of differential metabolites based on fold change

图3 ‘藜红1号’（RQ1）及其突变体（gq1）中差异代谢物KEGG分类热图

Fig. 3 Heatmap of KEGG classification for differential metabolites between RQ1 and gq1

图4 ‘藜红1号’（RQ1）及其突变体（gq1）中差异代谢物KEGG通路分析 A：gq1组对RQ1组的代谢通路富集分析；B：gq1组对RQ1组的KEGG代谢互作分析

Fig. 4 KEGG pathway enrichment analysis of differential metabolites between RQ1 and gq1 A: KEGG pathway enrichment analysis of qg1 vs RQ1; B: KEGG category netplot of qg1 vs RQ1

图5 gq1突变体中氨基酸的变化 A：gq1和RQ1中鉴定出的14种差异表达氨基酸含量；B：酪氨酸代谢途径中代谢物变化概述，虚线箭头表示多个酶步骤。TAT：酪氨酸转氨酶；TAL：酪氨酸解氨酶；TYDC：L-酪氨酸脱羧酶；HPD：羟苯丙酮酸双加氧酶；AOC3：伯胺氧化酶；AOX：醛氧化酶；差异的显著性（* P ≤ 0.05，** P ≤ 0.01，*** P ≤ 0.001）

Fig. 5 Variation of amino acid content in mutant gq1 A: Contents of 14 differentially expressed amino acids in gq1 and RQ1. B: An overview of metabolites and their expressions in tyrosine metabolism pathway. Arrows with dashed lines designate multiple enzymes steps. TAT: Tyrosine aminotransferase; TAL: tyrosine ammonia-lyase; TYDC: L-tyrosine decarboxylase; HPD: hydroxyphenylpyruvate dioxygenase; AOC3: primary-amine oxidase; AOX: aldehyde oxidase. Variables of significance（* P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001）

图6 gq1突变体中代谢物和代谢通路之间的转变基本模型

Fig. 6 Basic model of transition between metabolites and metabolic pathways in the gq1

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