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作物学报 ›› 2021, Vol. 47 ›› Issue (5): 860-868.doi: 10.3724/SP.J.1006.2021.04153

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

绿豆雄性不育突变体msm2015-1的遗传学与细胞学分析

吴然然(), 林云, 陈景斌, 薛晨晨, 袁星星, 闫强, 高营, 李灵慧, 张勤雪, 陈新*()   

  1. 江苏省农业科学院经济作物研究所, 江苏南京 210014
  • 收稿日期:2020-07-11 接受日期:2020-10-14 出版日期:2021-05-12 网络出版日期:2020-10-30
  • 通讯作者: 陈新
  • 作者简介:E-mail: rrwu@jaas.ac.cn
  • 基金资助:
    国家重点研发计划项目(2019YFD1001301);国家重点研发计划项目(2019YFD1001300);江苏省自然科学基金项目(BK20190257);国家自然科学基金项目(31871696);江苏特粮特经产业技术体系集成创新中心(JATS[2019]399)

Genetic and cytological analysis of male sterile mutant msm2015-1 in mungbean

WU Ran-Ran(), LIN Yun, CHEN Jing-Bin, XUE Chen-Chen, YUAN Xing-Xing, YAN Qiang, GAO Ying, LI Ling-Hui, ZHANG Qin-Xue, CHEN Xin*()   

  1. Institute of Industrial Crops, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, Jiangsu, China
  • Received:2020-07-11 Accepted:2020-10-14 Published:2021-05-12 Published online:2020-10-30
  • Contact: CHEN Xin
  • Supported by:
    National Key Research and Development Program of China(2019YFD1001301);National Key Research and Development Program of China(2019YFD1001300);Natural Science Foundation of Jiangsu Province(BK20190257);National Natural Science Foundation of China(31871696);Jiangsu Agricultural Industry Technology System(JATS[2019]399)

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摘要：

单产水平低一直是我国绿豆存在的产业问题。绿豆杂种优势显著是提高产量的有效途径, 而雄性不育材料是实现杂种利用的宝贵资源。本研究利用60Co-γ辐射诱变技术首次在绿豆中获得雄性不育突变体, 并命名为‘不育绿豆2015-1’ (male sterile mungbean2015-1, 简写为msm2015-1)。msm2015-1营养生长期表型与‘苏绿1号’没有明显差异。花器官形态正常, 但花药发白且不能正常开裂散粉, 成熟期座荚率几乎为0。遗传分析显示, 育性分离群体中可育与不育分离比符合3∶1, 表明不育性状由单个隐性核基因控制。花粉发育的细胞学分析发现, msm2015-1的盛开花朵中花粉粒几乎未在柱头附着, 体外亦未萌发。花粉细胞质未被Alexander染液正常着色, I2-KI染色出现典败、圆败和染败3种类型。DAPI染色显示, msm2015-1花粉细胞核发育异常。醋酸洋红染色表明, 败育发生在花粉发育早期, 花粉母细胞在四分体时期减数分裂的不对称和异常多分裂是导致花粉败育的主要原因。

关键词: 绿豆, 雄性不育, 遗传分析, 花粉发育, 细胞学

Abstract:

In China, the low yield per unit has always been a problem in the mungbean industry. The heterosis of mungbean is an effective way to increase yield. The male sterile line is a valuable resource to realize the utilization of hybrid. In this study, 60Co-γ radiation mutagenesis technology was used to induce male sterile mutants in mungbean, and we gained one mutant named male sterile mungbean 2015-1 (msm2015-1) for the first time. There was no significant phenotype difference between msm2015-1 and Sulyu 1 during the vegetative growth stage. The floral organ of msm2015-1 was normal, but the anthers of msm2015-1 were white and could not crack and disperse powder normally in the flowering stage. In the mature stage, hardly any normal pods grew up. Genetic analysis showed the separation ratio of fertility and sterility in the fertile segregation population was in line with 3:1, indicating that the sterility trait was controlled by a single recessive nuclear gene. Cytological analysis of pollen development revealed that there were few pollen grains of msm2015-1 attached to mature stigmas. The pollen grains of msm2015-1 did not germinate in vitro. The cytoplasm of msm2015-1’s pollen could not be stained by Alexander dye. Three abortion types showed up when stained by I2-KI dye: typical abortion, spherical abortion and stained abortion. DAPI staining showed the nuclei of msm2015-1’s pollen developed abnormally. Magenta acetate staining indicated that abortion occurred in the early stage of pollen development, while the asymmetry and more anomalous fission of meiosis during the tetrad stage were the main causes of pollen abortion.

Key words: mungbean, male sterile, genetic analysis, pollen development, cytology

图1

绿豆雄性不育突变体msm2015-1表型分析 A: 营养生长期; B: 花蕾; C: 盛开的花朵; D: 花蕾解剖图, 从左至右依次为旗瓣、翼瓣、龙骨瓣、雄蕊和雌蕊; E: 开花后第2天的小豆荚; F: 近成熟的豆荚。A图中标尺为10 cm, B~F图中标尺为1 cm。"

图2

绿豆雄性不育突变体msm2015-1花药及花粉粒形态观察 A: 花药, 白色箭头指示msm2015-1未正常开裂; B: ‘苏绿1号’柱头, 白色箭头指示大量花粉附着; C: msm2015-1柱头, 白色箭头指示几乎无花粉附着; D: ‘苏绿1号’花药的扫描电镜图; E: msm2015-1花药的扫描电镜图; F: ‘苏绿1号’成熟花粉粒的扫描电镜图; G: msm2015-1成熟花粉粒的扫描电镜图。A~C图中标尺为1 mm, D和E图中标尺为100 µm, F和G图中标尺为10 µm。"

表1

育性分离群体的遗传学分析"

表型
Phenotype		 观察值
Observed number		 理论值
Theoretical number		 χ2
(3:1)
可育株Fertile plants 187 193.5 0.873
不育株Sterile plants 71 64.5

图3

绿豆雄性不育突变体msm2015-1花粉体内及体外萌发 A: ‘苏绿1号’花粉柱头萌发, 白色箭头指示柱头上花粉粒及其萌发出的花粉管; B: msm2015-1花粉柱头萌发, 白色箭头指示柱头上无花粉粒附着; C: ‘苏绿1号’成熟花粉体外萌发; D: msm2015-1成熟花粉体外萌发。图中标尺为100 µm。"

图4

绿豆雄性不育突变体msm2015-1花粉活力的染色分析 A: ‘苏绿1号’成熟花粉的Alexander染色; B: msm2015-1成熟花粉的Alexander染色; C: Alexander染色花粉败育率统计; D: ‘苏绿1号’成熟花粉的I2-KI染色; E: msm2015-1成熟花粉的I2-KI染色; F: I2-KI染色花粉败育率统计; G: ‘苏绿1号’成熟花蕾的花药Alexander染色; H: msm2015-1成熟花蕾的花药Alexander染色。图中标尺为100 µm。"

图5

绿豆雄性不育突变体msm2015-1的花粉细胞核发育检测 A: ‘苏绿1号’成熟花粉DAPI染色, 白色和红色箭头分别指示生殖核和营养核; B: msm2015-1成熟花粉DAPI染色。图中标尺为20 µm。"

图6

绿豆雄性不育突变体msm2015-1花粉早期发育阶段分析 A: 花粉早期发育的各个时期, 白色箭头指示msm2015-1四分体时期的不对称和不均等分裂导致形成的大小不一的小孢子; B: 四分体时期出现的异常多分裂, 图中数字表示msm2015-1分裂出的小孢子个数。PMC: 花粉母细胞; Tetrad: 四分体; Msp: 小孢子; Pollen: 花粉。图中标尺为20 µm。"

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