三个大豆ms3雄性不育突变体的等位突变分析
Allelic Mutation Analysis of Three Male Sterile ms3 Mutants in Soybean
朱玉雪 1黄圆圆 1徐研 1付爱根 1徐敏 1余君萍1
作者信息
- 1. 西北大学生命科学学院/西部资源与生物技术教育部重点实验室/陕西省生物技术与生化工程重点实验室,西安 710069
- 折叠
摘要
大豆雄性不育系对发挥大豆杂种优势具有重要价值,然而传统的三系杂交存在恢复系来源受限等问题,而环境敏感型细胞核雄性不育系在不同条件下育性可以改变.本文对ms3(Washington)、ms3(Flanagan)和ms3(Plainview)3个独立突变体的表型和突变位点展开了进一步研究,分别进行了不育表型鉴定、高通量测序、分子标记设计及突变位点验证、分析突变对MS3蛋白结构的影响、ms3(Plainview)花药半薄切片等实验.实验结果表明突变体花粉大量败育,只有零星被I2-KI染液染成黑色的不规则花粉粒.高通量测序结果显示,m3(Washington)和ms3(Flanagan)在MS3第3个外显子的PHD编码区域出现了大片段插入,导致MS3蛋白PHD结构域被破坏,这个等位基因命名为ms3-1;ms3(Plainview)在MS3第1个外显子缺失了一个A,导致移码突变,开放读码框仅编码40个氨基酸,蛋白功能完全丧失,这个等位基因命名为ms3-2.半薄切片结果显示,在花药发育中后期ms3(Plainview)的绒毡层和花粉发育出现异常.综上,本研究的结果为ms3的应用和MS3基因改造提供了材料和依据.
Abstract
Soybean male sterile lines play vital value for developing hybrid soybean varieties,while the traditional three-line hybridization system has problems due to a limited number of restorer lines.The environmental sensitive genic male sterile(EGMS)line can switch fertility under different conditions.In this research,the phenotypes and mutation sites of the three independent mutants ms 3(Washington),ms3(Flanagan)and ms 3(Plainview)were further studied.Phenotypic identification of sterility,high-throughput sequencing,molecular marker design and mutation site verification,analysis of the effect of mutation on MS3 protein structure,ms3(Plainview)anther semi-thin section experiments were carried out separately.These results showed that most of the pollens are aborted,only sporadic pollen grains with irregular shape were stained by I2-KI dye in the anthers of the three mutants.High-throughput sequencing results showed that ms3(Washington)and ms 3(Flanagan)had a large fragment insertion in the PHD coding region of the third exon of the gene MS3,eventually compromising the PHD domain of MS3 protein,which was named ms3-1.In ms3(Plainview),there is one base pair deletion in the first exon of MS3,resulting in frameshift mutation encoding only 40 amino acids.The loss-of-function allele was named ms3-2.Through semi-thin section analysis of ms 3(Plainview),its tapetum layer and pollen development were abnormal at the middle and late anther development stage.In summary,this study provided materials and basis for future application and genetic modification of ms 3 and MS3 in soybean breeding.
关键词
大豆/细胞核雄性不育/MS3/PHD结构域Key words
soybean/genic male sterile/MS3/PHD domain引用本文复制引用
基金项目
国家自然科学基金(32000598)
陕西省教育厅自然科学研究专项(19JK0858)
陕西省自然科学基础研究计划(2023-JC-YB-178)
陕西基础科学(化学、生物学)研究院科学研究计划(22JHZ007)
出版年
2024
国家科技期刊平台
NSTL
万方数据