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作物学报 ›› 2020, Vol. 46 ›› Issue (7): 1076-1086.doi: 10.3724/SP.J.1006.2020.94159

• 耕作栽培·生理生化 • 上一篇    下一篇

厚轴茶雄性不育株花药败育的生物学特性和细胞学研究

蒋会兵,杨盛美,刘玉飞,田易萍,孙云南,陈林波,唐一春()   

  1. 云南省农业科学院茶叶研究所 / 云南省茶学重点实验室, 云南勐海 666201
  • 收稿日期:2019-10-24 接受日期:2020-03-24 出版日期:2020-07-12 发布日期:2020-04-14
  • 通讯作者: 唐一春 E-mail:tyc188@126.com
  • 作者简介:E-mail: jianghb119@126.com
  • 基金资助:
    国家自然科学基金项目(31760224)

Biological characteristics and cytological studies on anther abortion of male sterile Camellia crassocolumna

JIANG Hui-Bing,YANG Sheng-Mei,LIU Yu-Fei,TIAN Yi-Ping,SUN Yun-Nan,CHEN Lin-Bo,TANG Yi-Chun()   

  1. Tea Research Institute, Yunnan Academy of Agricultural Sciences / Yunnan Provincial Key Laboratory of Tea Science, Menghai 666201, Yunnan, China
  • Received:2019-10-24 Accepted:2020-03-24 Online:2020-07-12 Published:2020-04-14
  • Contact: Yi-Chun TANG E-mail:tyc188@126.com
  • Supported by:
    National Natural Science Foundation of China(31760224)

摘要:

为明确厚轴茶(Camellia crassocolumna H. T. Chang)雄性不育株花器发育形态、花药和花粉败育时期及其细胞学特征, 利用体视显微镜、石蜡切片技术、染色体制片和DAPI染色法, 对厚轴茶雄性不育株和可育株开花进程、花器形态、花药发育过程、花粉母细胞减数分裂及小孢子发育过程比较观察。结果显示, 厚轴茶花属于完全花, 花药具四室、呈蝶形, 花药壁发育为基本型, 绒毡层细胞具双核, 于四分体时期形成分泌型细胞, 单核花粉期开始降解, 花粉母细胞经过减数分裂I、减数分裂II和胞质分裂后形成四面体型四分体, 小孢子呈三角形, 成熟花粉为二细胞型花粉。花蕾发育早期, 不育株雄蕊发育正常, 与可育株无明显差异。花蕾发育后期, 不育株花丝弯曲, 花药粘连、干瘪、褐化、坏死, 不裂药。不育株减数分裂期绒毡层细胞异常增生、排列混乱, 单核至双核花粉期绒毡层延迟降解。不育株花粉母细胞减数分裂过程中存在环状单价体、滞后染色体、染色体桥、染色体缺失、不均等分离、微核和多分体等异常现象。不育株小孢子胞质紊乱, 单核期花粉粒相互粘附, 花粉壁皱缩变形, 花粉细胞质和细胞核模糊不清, 成熟花粉细胞空瘪凹陷。研究结果表明, 厚轴茶雄性不育花器形态属雄蕊萎缩型和花药异常型, 花药发育受阻于减数分裂至单核花粉期, 存在花粉母细胞败育型和单核败育型。单核花粉期是其花药败育的主要时期。花药绒毡层异常发育和延迟降解, 花粉母细胞减数分裂染色体行为异常, 小孢子和花粉粒发育异常可能是其花药败育的主要原因。

关键词: 厚轴茶, 雄性不育, 花药, 形态学, 细胞学

Abstract:

The purpose of this study was to explore the flower morphology, anther and pollen abortion time and its cytological characteristics in male sterile plants of C. crassocolumna. The flowering process, floral morphology, anther structure, meiosis and microsporogenesis of male sterile (M350) and fertile plants (M352) of C. crassocolumna were observed by stereomicroscopy, paraffin section, chromosome preparation and DAPI staining. The results showed that the wild tea plant had a perfect flower, its anthers were tetrasporangiate shaped like a butterfly, with an anther wall development of basic type. It tapetal cells were binuclear, forming secretory cells at the tetrad stage, and degrading during mononuclear pollen stage. After the meiosis I, meiosis II and cytokinesis, pollen mother cells developed into tetrahedral tetrads. Microspores were triangular, and mature pollen was two-celled. The stamens of male sterile flowers developed normally in the early stage of flower bud development, and there was no difference between sterile and fertile flowers, while in the late stage of flower bud development, filaments were curved, anthers adhered and shriveled, anthers could not release pollen. The tapetal cells of male sterile flowers were proliferation abnormally and disordly at the meiosis of stage pollen mother cells, and the degradation of tapetum was delayed during the mononuclear and binucleate pollen periods. During meiosis of pollen mother cells of male sterile flowers, there were some abnormal appearances, such as chromosome ring, lagging chromosomes, chromosome bridges, chromosomal deletion, scattered chromosomes, unequal separation, micronucleus and abnormal tetrad. In sterile plants, the microspore cytoplasm was disorder, pollen grains adhered to each other at the mononuclear stage, pollen wall crumpled and deformed, cytoplasm and nucleus of pollen were blurred, the mature pollen cells were hollow and sunken. Taken together, the floral organ morphology of male sterile plants in C. crassocolumna belongs to stamen collapse type and anther abnormality type. The development of anthers is blocked from meiosis of pollen mother cells to uninucleate pollen stage, belonging to pollen mother cell abortion type and uninucleate pollen abortion type. Mononuclear period is the main period of anther abortion. Tapetum degradation delayed, chromosomal abnormalities of pollen mother cells in meiosis, and abnormal development of microspores and pollens, might be the main reasons for anther abortion in male sterile plants of C. crassocolumna.

Key words: Camellia crassocolumna, male sterility, anther, morphology, cytology

图1

可育株M352和雄性不育株M350的花器官形态 A: 可育株M352; B: 雄性不育株M350。"

表1

可育株M352和雄性不育株M350开花期花器性状比较"

植株
Plant
花冠直径
Expansion
of corolla (cm)
花瓣长
Length of petal (cm)
花瓣宽
Width of petal (cm)
外轮花丝长
Length of out thrum (cm)
内轮花丝长
Length of inside thrum (cm)
花柱长
Length of style (cm)
雄蕊数量
Number of stamen (No.)
可育株M352
Fertile plant M352
7.28±0.57 a 3.53±0.30 a 2.80±0.24 a 1.66±0.18 a 0.94±0.18 a 1.61±0.18 a 190.62±16.37 a
不育株M350
Sterile plant M350
6.94±0.51 a 3.20±0.28 a 3.07±0.34 a 1.37±0.13 b 0.65±0.14 b 1.73±0.21 a 184.85±18.52 a

图2

可育株M352和雄性不育株M350花蕾发育过程 A~E: 可育株M352; F~J: 不育株M350。A和F: 花粉母细胞期; B和G: 减数分裂期; C和H: 单核期; D和I: 二核期; E和J: 展花期。"

图3

可育株M352花药发育过程 A: 造孢细胞期; B: 花粉母细胞期; C: 减数分裂期; D: 四分体时期; E: 单核期; F: 双核期; G和H: 成熟期。Ep: 表皮层; En: 内皮层; ML: 中层; T: 绒毡层; Sc: 造孢细胞; Pmc: 花粉母细胞; Tds: 四分体; Mpg: 单核花粉; Bpg: 双核花粉; Pg: 成熟花粉粒。"

图4

雄性不育株M350花粉败育过程 A: 造孢细胞期; B: 花粉母细胞期; C: 减数分裂期; D: 四分体时期; E: 单核期; F: 双核期; G和H: 成熟期。Ep: 表皮层; En: 内皮层; ML: 中层; T: 绒毡层; Sc: 造孢细胞; Pmc: 花粉母细胞; Mc: 减数分裂体; Tds: 四分体; Mpg: 单核花粉; Bpg: 双核花粉。"

图5

可育株M352花粉母细胞正常减数分裂过程 A: 间期I; B: 前期I细线期; C: 前期I偶线期; D: 前期I粗线期; E: 前期I双线期; F: 前期I终变期; G: 中期I; H: 后期I; I: 末期I; J: 前期II; K: 中期II; L: 后期II; M: 末期II; N: 四分体时期; O: 不同步分裂相。"

图6

雄性不育株M350花粉母细胞异常减数分裂过程 A~C: 环状、棒状单价体; D~G: 染色体单桥、多桥; H~K: 落后染色体; L: 染色体缺失; M~P: 染色体散乱排列; Q和R: 分裂不同步; S: 不对称排列; T~V: 微核; W: 三分体; X和Y: 多核四分体。"

表2

可育株M352和不育株M350花粉母细胞减数分裂中异常染色体的频率"

减数分裂时期
Meiosis stage
可育株M352 Fertile plant M352 雄性不育株M350 Male sterile plant M350
统计细胞数
Number of cells examined
异常细胞数量
Number of
abnormal cells
频率
Frequency (%)
统计细胞数
Number of cells examined
异常细胞数量
Number of abnormal cells
频率
Frequency
(%)
前期I Prophase I 177 20 11.30 205 47 22.93
中期I Metaphase I 90 16 17.78 97 36 37.11
后期I Anaphase I 87 13 14.94 92 44 47.83
末期I Telophase I 113 16 14.16 124 52 41.94
前期II Prophase II 98 12 12.24 108 48 44.44
中期II Metaphase II 72 11 15.28 93 33 35.48
后期II Anaphase II 93 12 12.90 105 40 38.10
末期II Telophase II 85 9 10.59 95 32 33.68
四分体时期 Tetrad 198 27 13.64 220 97 44.09

图7

可育株M352和不育株M350小孢子发育过程比较 A~E: 可育株M352; F~J: 不育株M350。A和F: 四分体时期; B和G: 单核早期; C和H: 单核期; D和I: 单核晚期; E和J: 双核期。"

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