甘蓝型油菜温敏细胞核雄性不育系TE5A 花药发育的细胞学研究

doi:10.3864/j.issn.0578-1752.2016.12.016

摘要/Abstract

摘要： 【目的】明确甘蓝型油菜温敏细胞核雄性不育系TE5A花药败育的时期及细胞学特征，为进一步研究其雄性不育的机理奠定理论基础。【方法】用不同温度（16℃和22℃）处理试验材料不育系TE5A，分别观察植株的育性及花器形态；并采用常规半薄切片技术，通过甲苯胺蓝、苯胺蓝和苏丹黑B等染料进行染色，观察并比较该不育系TE5A不育植株和可育植株花药发育的显微结构、胼胝质以及脂质体等脂类物质的异同；进一步通过透射电镜对其花药发育的超微结构进行比较观察。【结果】当把可育株油菜从16℃光照培养箱移到22℃光照培养箱后7 d，观察到花朵的育性转变为雄性不育。把22℃光照培养箱的不育株油菜移到16℃光照培养箱后，观察到先开放的花朵表现为雄性不育，而随后开放的花朵表现为雄性可育。在不育环境下的不育株TE5A花朵的花瓣大小和形态与可育株花朵没有明显差异，但不育株花朵的花丝显著短于可育株的花丝，并且花药萎缩、干瘪，没有花粉粒附着在上面。在不育环境下，不育系TE5A花药败育发生在花粉母细胞减数分裂时期，花粉母细胞减数分裂发生异常，没有二分体及四分体的形成，形成“拟小孢子”。胼胝质在花粉母细胞时期可以正常合成，但后续的降解滞后，在四分体时期没有降解，直至花粉粒成熟期才开始降解。绒毡层没有观察到明显异常，并能分泌脂质体等脂类物质。“拟小孢子”的花粉粒外壁发育异常，无法形成具有特殊柱状体和顶盖结构的花粉粒外壁，因此，不能结合孢粉素和脂质体等物质。随着花药的继续发育，最后“拟小孢子”逐渐降解，只剩下花粉空壳。【结论】甘蓝型油菜温敏细胞核雄性不育系TE5A花朵的花瓣大小和形态与可育株花朵没有明显差异，但花丝显著缩短，花药萎缩、干瘪，没有花粉粒附着在上面。不育系TE5A花药败育发生在花粉母细胞减数分裂时期，减数分裂异常导致无法形成二分体及四分体，并且胼胝质的异常降解及花粉粒外壁的异常对花药败育也有重要影响。甘蓝型油菜温敏细胞核雄性不育系TE5A可以作为一新型甘蓝型油菜温敏细胞核雄性不育系材料。

关键词: 甘蓝型油菜, 温敏细胞核雄性不育, 花药败育, 减数分裂

Abstract: 【Objective】The aim of this paper is to detect the vital abortion periods and cytological characteristics of the anther development in a thermo-sensitive genic male sterile line TE5A in Brassica napus. The results will provide a theoretical basis for further study on the mechanism of male sterility.【Method】The sterile line TE5A was used as test material and was conducted with two different treatment temperatures (16℃ and 22℃). Then the fertility and characteristics of the floral organs of the plants were observed, respectively. The differences of anther development between the sterile and the normal plants of the sterile line TE5A were observed and compared by using toluidine blue, aniline blue and sudan black B to stain the semi-thin sectionings. Further observations and comparisons of cell ultrastructure of the anther development were carried out by transmission electron microscopy.【Result】The flowers of the plants were observed to be sterile, after seven days when the fertile rapeseed plants were moved to the light incubator of 22℃ from the one of 16℃. After the sterile rapeseed plants were moved to the light incubator of 16℃ from the one of 22℃, it was observed that the sterile flowers were opened first and the fertile flowers were opened after. Compared with the fertile flowers, the size and characteristic of petals of sterile plants in the sterile environment displayed no obvious difference. But the filaments of the sterile plants were significantly shorter than the fertile's and the sterile anthers withered without pollen grains in them. The pollen abortion of the sterile line TE5A in the sterile environment occurred at the pollen mother cell meiosis stage. The pollen mother cells could not pass the meiosis, with no dyads and tetrads formed in TE5A, insteading of some "pseudo microspores". The callose was normal deposited around the pollen mother cells, however, the callose hadn't been degraded until the mature pollen stage. The tapetum cells which could secret liposomes were observed to be normal in the fertile line. The pollen exines of the "pseudo microspores" were abnormal. The special bacula and tectum structure of the pollen exine couldn't formed, so the pollen exine couldn't combine with sporopollenins, no liposomes and other substances combined. With the development of the anthers, the "pseudo microspores" were decayed gradually and only empty shells remained at last.【Conclusion】Compared with the fertile flowers, the size and characteristic of petals of sterile plants in the sterile line TE5A displayed no obvious difference. But the filaments of the sterile plants were significantly shorter and the sterile anthers withered without pollen grains in them. The vital abortion of the sterile line TE5A occurred at the pollen mother cell meiosis stage. The pollen mother cells couldn’t pass the meiosis, and neither dyads nor tetrads were formed. The abnormal degradation of the callose also play an important role in the anther development. The cytological characteristics of the thermo-sensitive genic male sterile line TE5A in B. napus were different from the previous materials. So these indicated that TE5A is a novel thermo-sensitive genic male sterile line.

Key words: Brassica napus; thermo-sensitive genic male sterile, anther abortion, meiosis

李可琪，曾新华，袁荣，闫晓红，吴刚. 甘蓝型油菜温敏细胞核雄性不育系TE5A 花药发育的细胞学研究[J]. 中国农业科学, 2016, 49(12): 2408-2417.

LI Ke-qi, ZENG Xin-hua, YUAN Rong, YAN Xiao-hong, WU Gang. Cytological Researches on the Anther Development of a Thermo-Sensitive Genic Male Sterile Line TE5A in Brassica napus[J]. Scientia Agricultura Sinica, 2016, 49(12): 2408-2417.

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