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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作者:
作者单位:

1.哈尔滨师范大学生命科学与技术学院,哈尔滨 150025;2.中国农业科学院作物科学研究所/作物基因资源与 育种全国重点实验室,北京100081

作者简介:

研究方向为植物遗传学,E-mail:59752006@qq.com

通讯作者:

郭长虹,研究方向为植物遗传学,E-mail:kaku3008@126.com
李新海,研究方向为玉米遗传改良与种质创新,E-mail:linxinhai@caas.cn

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基金项目:

国家自然科学基金(32001559);作物基因资源与育种全国重点实验室


Identification and Transcriptome Analysis of the High Methionine Mutant Zmts1 in Maize Kernels
Author:
Affiliation:

1.College of Life Science and Technology, Harbin Normal University,Harbin 150025;2.Institute of Crop Sciences, Chinese Academy of Agricultural Sciences/State Key Laboratory of Crop Gene Resources and Breeding,Beijing 100081

Fund Project:

Foundation projects: National Nature Science Foundation of China (32001559);State Key Laboratory of Crop Gene Resources and Breeding

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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ZmTS1Threonine 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,该蛋白含有一个苏氨酸合酶结构域,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个可能参与蛋氨酸代谢途径的关键候选基因,RNA-Seq与qRT-PCR的表达模式一致,表明7个基因能间接调控蛋氨酸代谢途径。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 primary limiting amino acid in maize-soybean meal diets for livestock and poultry. An imbalance in amino acids can inhibit protein synthesis in animals, thus affecting the quality of meat and milk. To analyze the regulatory mechanism governing methionine accumulation in maize grains, this study used homologous cloning strategy to identify a candidate gene, ZmTS1Threonine Synthase 1), which affects methionine content. Methionine content in Zmts1 mutant seeds increased by 60%. SDS-PAGE analysis revealed that the methionine residue-rich 10 kDa-δ alcohol-soluble protein in mature Zmts1 mutant kernel was significantly elevated compared to the wild type, confirming that this gene could significantly increase methionine content in maize kernels. Bioinformatics analysis indicated that the gene contained a threonine synthase structural domain and the encoded protein was hydrophilic. Transcriptome analysis revealed 1144 differentially expressed genes associated with methionine metabolism, of which 571 and 573 were up-regulated and down-regulated, respectively. GO and KEGG enrichment pathway analysis showed that the differentially expressed genes were mainly involved in the biosynthesis and metabolism of amino acids. qRT-PCR was used to further analyze seven key candidate genes that may be involved in methionine metabolism pathway. The transcriptional patterns revealed by both in RNA-Seq and qRT-PCR implied indirect functions of these genes in the methionine metabolic pathway. This study provides new germplasm resources for breeding methionine-rich maize, and offers a theoretical basis for understanding methionine regulation mechanisms in maize.

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引用本文

洪泽渊,鲁鑫,汤泽洋,等.玉米籽粒高蛋氨酸突变体Zmts1的鉴定与转录组分析[J].植物遗传资源学报,2025,26(2):331-341.

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  • 收稿日期:2024-05-15
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  • 在线发布日期: 2025-01-23
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