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一份玉米小籽粒发育突变体mn-like1的遗传分析与分子鉴定

袁鑫1，赵卓凡2，赵瑞清1，刘孝伟1，郑名敏3，刘育生4，董好胜4，邓丽娟4，曹墨菊1,*，黄强4,*   

  1. 1四川农业大学玉米研究所 / 农业农村部西南玉米生物学与遗传育种重点实验室, 四川成都611130; 2成都理工大学, 四川成都610051; 3成都师范学院化学与生命科学学院, 四川成都611130;4四川省原子能研究院 / 辐照保藏和效应四川省重点实验室, 四川成都610101
  • 收稿日期:2024-12-10 修回日期:2025-03-26 接受日期:2025-03-26 网络出版日期:2025-04-08
  • 基金资助:
    本研究由四川省科技计划项目(2022NSFSC0018, 2021YFYZ0011, 2024NSFSC1210, 2023YFH0020)资助。

Genetic analysis and molecular identification of a small kernel mutant mn-like1 in maize

YUAN Xin1,ZHAO Zhuo-Fan2,ZHAO Rui-Qing1,LIU Xiao-Wei1,ZHENG Ming-Min3,LIU Yu-Sheng4,DONG Hao-Sheng4,DENG Li-Juan4,CAO Mo-Ju1,*,HUANG Qiang4,*   

  1. 1 Maize Research Institute, Sichuan Agricultural University / Key Laboratory of Biology and Genetic of Maize in Southwest Region, Ministry of Agriculture and Rural Affairs,Chengdu 611130, Sichuan, China; 2 Chengdu University of Technology, Chengdu 610051, Sichuan, China; 3 College of Chemistry and Life Sciences, Chengdu Normal University, Chengdu 611130, Sichuan, China; 4 Sichuan Institute of Atomic Energy / Irradiation Preservation and Effect Key Laboratory of Sichuan Province, Chengdu 610101, Sichuan, China
  • Received:2024-12-10 Revised:2025-03-26 Accepted:2025-03-26 Published online:2025-04-08
  • Supported by:
    This study was supported by Sichuan Science and Technology Program (2022NSFSC0018, 2021YFYZ0011, 2024NSFSC1210, 2023YFH0020).

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摘要：

籽粒是玉米最重要的库器官，其正常发育需要充足的碳水化合物供应与顺畅的物质运输通道。本研究在选系中获得了1份籽粒发育缺陷突变体small kernel 18 (smk18)，经多年多点种植，突变体smk18性状遗传稳定，统计(B73×smk18) F2群体分离比，发现该性状受一对隐性单基因控制。将smk18突变性状回交5代于自交系RP125中构建近等基因系mn-like1 (RP125smk18 smk18)。表型鉴定显示，mn-like1植株相较于RP125株高、穗位高增加，百粒重、粒长、粒宽显著降低。通过分子标记将该突变基因定位于2号染色体分子标记Indel 4和Indel 5之间。该区间内有一报道的小籽粒基因Miniature1 (Mn1)，其编码细胞壁蔗糖转化酶，在籽粒发育早期控制糖类物质运输。通过扩增mn-like1中Mn1基因CDS，测序并比对发现突变体第5外显子上有9 bp缺失，使得Mn1蛋白第409~411的3个氨基酸缺失，蛋白结构变异。且授粉后13天mn-like1籽粒中Mn1表达量显著降低。通过将mn-like1与Mn1转座子插入突变体mn1-mu进行等位性测验，确认mn-like1为Mn1新的等位突变体。亚细胞定位、糖类物质测量以及糖原染色结果显示，Mn1在籽粒胚乳基底转移层特异表达，其突变后糖类物质运输通道受阻，mn-like1中蔗糖和淀粉含量显著降低，造成籽粒发育缺陷。本研究丰富了不同遗传背景下Mn1突变体的种质资源，为Mn1调控籽粒发育机制的解析和Mn1蛋白催化功能研究提供新的遗传材料。

关键词: 玉米, 基因定位, 籽粒发育, 细胞壁蔗糖转化酶, 物质运输

Abstract:

The kernel serves as the primary storage organ in maize, and its proper development requires on an adequate carbohydrate supply and efficient nutrient transport channels. In this study, we identified a natural mutant, small kernel 18 (smk18), exhibiting defects in kernel development. After multi-year and multi-location field trials, the smk18 mutant trait remained genetically stable. Segregation analysis of the (B73 × smk18) F2 population revealed that the mutant phenotype was controlled by a single recessive gene. The smk18 mutant was backcrossed to the inbred line RP125 for five generations to construct the near-isogenic mn-like1 (RP125smk18 smk18). Phenotypic evaluation showed that mn-like1 exhibited increased plant height and ear height compared to RP125, whereas hundred-kernel weight, kernel length, and kernel width were significantly reduced. Through molecular mapping, we localized the causal gene between Indel 4 and Indel 5 on chromosome 2. Within this interval, the Miniature1 (Mn1) gene had been previously reported to encode a cell wall invertase (INCW2) essential for carbohydrate transport during early kernel development. Sequencing of the Mn1 coding sequence (CDS) in mn-like1 revealed a 9-bp deletion in exon 5, leading to the loss of three amino acids (positions 409–411) in the Mn1 protein and alterations in its structure. Expression analysis showed that Mn1 transcript levels were significantly reduced in mn-like1 kernels at 13 days after pollination (DAP). An allelism test between mn-like1 and the transposon insertion mutant mn1-mu confirmed that mn-like1 is a novel allelic variant of Mn1. Further subcellular localization studies, carbohydrate quantification, and glycogen staining indicated that Mn1 is specifically expressed in the basal transfer layer of the endosperm. Mutation of Mn1 disrupted carbohydrate transport, leading to a significant reduction in sucrose and starch content in mn-like1 kernels, ultimately resulting in kernel developmental defects. In conclusion, this study expands the repertoire of Mn1 mutants in diverse genetic backgrounds and provides valuable genetic resources for elucidating the regulatory mechanisms of Mn1 in kernel development and the catalytic function of Mn1 protein.

Key words: maize, gene mapping, nutrient transport, cell wall invertase, kernel development

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