玉米籽粒高蛋氨酸突变体Zmts1的鉴定与转录组分析
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1.哈尔滨师范大学、中国农业科学院作物科学院作物科学研究所;2.中国农业科学院作物科学院作物科学研究所;3.哈尔滨师范大学;4.哈尔滨师范大学/中国农业科学院作物科学研究所

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国家自然科学基金青年科学基金项目(32001559)


Identification and Transcriptome Analysis of the High Methionine Mutant Zmts1 in Maize Kernels
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National Natural Science Foundation of China Youth Science Foundation Project(32001559)

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    摘要:

    蛋氨酸是畜禽类玉米-豆粕型日粮的第一限制性氨基酸,氨基酸不平衡会导致机体蛋白质合成受到抑制从而影响肉和奶的品质。为解析玉米籽粒蛋氨酸积累的调控机制,本研究利用同源克隆方法获得了影响蛋氨酸含量的候选基因ZmTS1(Threonine Synthase1),通过玉米突变体材料验证ZmTS1基因功能,发现与野生型相比,Zmts1突变体籽粒中蛋氨酸含量提高了60%,SDS-PAGE凝胶电泳结果表明Zmts1突变体成熟籽粒中富含蛋氨酸残基的10 KDa-δ 醇溶蛋白较野生型有了显著提高,验证了该基因可显著提高玉米籽粒中蛋氨酸含量。生物学信息分析结果表明,该基因含有一个苏氨酸合酶结构域,编码的蛋白属于亲水性蛋白。通过转录组分析挖掘与蛋氨酸代谢相关的差异表达基因,共筛选到1144个差异表达基因, 其中571个基因表达上调, 573个基因表达下调。GO和KEGG富集通路分析表明,差异表达基因主要参与氨基酸的生物合成和代谢途径。利用qRT-PCR进一步验证了7个参与蛋氨酸代谢途径关键候选基因,这7个基因能间接调控蛋氨酸代谢途径,RNA-Seq与qRT-PCR的表达模式相一致。本研究为高蛋氨酸玉米育种提供了新的种质资源,也为玉米蛋氨酸调控机制提供一定的理论依据。

    Abstract:

    Methionine is the first limiting amino acid in maize-soybean meal diets for livestock and poultry. An imbalance in amino acids leads to inhibition of protein synthesis in the organism, thus affecting the quality of meat and milk. To analyse the regulatory mechanism of methionine accumulation in maize grains, this study used homologous cloning to obtain a candidate gene, ZmTS1 (Threonine Synthase 1), which affects methionine content.Methionine content in Zmts1 mutant seeds increased by 60%, SDS-PAGE results showed that the methionine residue-rich 10 KDa-δ alcohol-soluble protein in the mature Zmts1 mutant kernel was significantly increased compared to the wild type, confirming that the gene could significantly increase the methionine content in maize kernels.The gene contained a threonine synthase structural domain and the encoded protein was hydrophilic, according to biological information analysis. The gene contained a threonine synthase structural domain and the encoded protein was hydrophilic, according to biological information analysis. Transcriptome analysis revealed 1,144 differentially expressed genes related to methionine metabolism, of which 571 were up-regulated and 573 down-regulated. Analysis of GO and KEGG enriched pathways showed that the differentially expressed genes were mainly involved in amino acid biosynthesis and metabolism pathways. Seven key candidate genes involved in the methionine pathway were further validated by qRT-PCR, which could indirectly regulate the methionine pathway, and the expression patterns of RNA-Seq and qRT-PCR were consistent. This study provides new germplasm resources for breeding methionine-rich maize, and also provides some theoretical basis for the mechanism of methionine regulation in maize.

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  • 收稿日期:2024-05-15
  • 最后修改日期:2024-06-20
  • 录用日期:2024-06-19
  • 在线发布日期: 2024-07-29
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