甘蓝型油菜雌性不育突变体FS-M1生物学特性及雌性不育遗传研究
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摘要
雌性不育现象在植物界普遍存在，目前已经在大豆、玉米、茶、山羊草、小麦和水稻等十几种作物中发现了雌性不育现象。雌性不育突变体对研究植物花器官发育、有性生殖和无融合生殖机制以及在杂种优势利用等方面具有重要意义。1996年江苏省农业科学院首次在甘蓝型油菜(Brassica napus L)中发现雌性不育突变体，定名为FS-M1。通过对突变体FS-M1形态解剖、细胞学观察、雌性不育遗传研究，获得如下结果：
1、突变体FS-M1和野生型间的花器官形态差异较大。突变体FS-M1的雌蕊显著短于雄蕊，无蜜腺，但在两弱雄蕊基部比野生型多出两个月牙形的花丝状器官。突变体FS-M1柱头顶端开裂呈“V”型，花柱明显短于野生型，子房扁平。突变体FS-M1胚珠发育正常，每角胚珠数与野生型相近。
2、突变体FS-M1雄性育性正常，雌性不育性高。突变体FS-M1天然授粉、人工授粉、自交套袋隔离、兄妹互交、剥蕾自交等结角率均为3％左右，平均每角粒数1.30-2.07粒，雌性不育率99.3％-99.8％。突变体FS-M1花粉活力同野生型，用FS-M1花粉授予正常油菜品种，杂交结实正常，F1植株自交结角率为97％，平均每角粒数21粒。
3、突变体FS-M1柱头乳突细胞较大，数目较少，粘着花粉能力弱，在开花2-3天后逐渐干瘪。对突变体FS-M1授粉后，在花柱中没有观察到花粉管伸长。切除FS-M1柱头及部分花柱后授粉、受精和结实过程正常。研究结果表明突变体FS-M1雌性不育现象可能与柱头乳突细胞发育畸变，功能缺失有关。
4、突变体FS-M1体细胞染色体数为2n=38，花粉母细胞减数分裂染色体配对正常；对突变体FS-M1与甘蓝型油菜N70、1059杂交F_1植株花粉母细胞减数分裂观察结果，杂种染色体在减数分裂中期配对，形成19个二价体，减数分裂行为正常。
5、突变体FS-M1与野生型及其它品种正反交F1群体植株雌蕊育性正常，F2代雌性不育株与可育株比为1：15；回交一代雌性不育株与可育株比为1：3。测验结果表明突变体FS-M1雌性不育性状受两对隐性基因控制。
Female sterile mutant FS-M1(Brassica. napus L.) was segregated from the cultivated variety Ningyou 10. A comparison between FS-M1 and its wild type Ningyou 10 was carried out by means of microscopic histological dissection and cytological observation. At the same time, the inheritance of female sterile of FS-M1 was studied. The main results were as follows:
1、The floral morphology of FS-M1 differ in some aspects to that of wild type. Pistil length of FS-M1 was shorter than that of wild type. The 6 stamens of FS-M1 were almost equal to each other in length, while that of wild type composed of 4 longer one and 2 shorter one. There were two extra crescents-like organ developed at the ovary base of FS-M1 with missing nectars. Ovary shape and length of FS-M1 was more flat and shorter respectively than that of wild type. Stigma end of FS-M1 shaped a fork-like looking, was differ obviously from wild type.
2、Result from the observation of agronomic trait of FS-M1 showed that: Pod set ratio of open-pollinated, artificial-pollinated, self-pollinated, sibling-pollinated, and constrained self-pollinated was about 3％respectively. Seed number per pod was about 2. It indicates that: female sterility of FS-M1 is steady. Seed set of hybrid from the cross of normal variety×FS-M1 was normal, so does open-pollinated of F_1. Seed set ratio of self-pollinated of F_1 was 97％, and seed per pod reached 21. It indicates that: the sterile pollens didn't affect the fertility of F_1.
3、The numbers of papillae cells of FS-M1 was sparse and its size was larger than that of wild type. Number of pollen grains observed on stigma of FS-M1 was less than that on wild type. The papillae cells of FS-M1 was drying and shriveling gradually in 2-3 days after flowering. There were few germinated pollens observed on surface of stigma of FS-M1 after hand-pollination. But no pollen tube found inside the style. Nevertheless there were pollen tubes existing inside the style of FS-M1, resulting in a normal seed-set when hand-pollination was made after excision of the stigma or part of the style. Results from above indicate that losing function of the papillae cells of FS-M1 resulting from its abnormal development is possibly a key factor causing the female sterility.
4、Cytological observation of FS-M1 showed that chromosome number in somatic cells of FS-M1 was 2n=38 and its meiotic chromosome behavior in PMCs was normal. Chromosomes behaviour of PMs meiosis in cross of (FS-M1×1059) F_1 and (FS-M1×N70) F_1 were normal. Chromosomes were paired normally, and formed 19 bivalents at MI and divided evenly to the two poles at AI. It indicats that performance of the mutant is not results from chromosome variation.
5、Genetic studies showed that the female sterility of FS-M1 was controlled by two pairs of recessive genes.
1、突变体FS-M1和野生型间的花器官形态差异较大。突变体FS-M1的雌蕊显著短于雄蕊，无蜜腺，但在两弱雄蕊基部比野生型多出两个月牙形的花丝状器官。突变体FS-M1柱头顶端开裂呈“V”型，花柱明显短于野生型，子房扁平。突变体FS-M1胚珠发育正常，每角胚珠数与野生型相近。
2、突变体FS-M1雄性育性正常，雌性不育性高。突变体FS-M1天然授粉、人工授粉、自交套袋隔离、兄妹互交、剥蕾自交等结角率均为3％左右，平均每角粒数1.30-2.07粒，雌性不育率99.3％-99.8％。突变体FS-M1花粉活力同野生型，用FS-M1花粉授予正常油菜品种，杂交结实正常，F1植株自交结角率为97％，平均每角粒数21粒。
3、突变体FS-M1柱头乳突细胞较大，数目较少，粘着花粉能力弱，在开花2-3天后逐渐干瘪。对突变体FS-M1授粉后，在花柱中没有观察到花粉管伸长。切除FS-M1柱头及部分花柱后授粉、受精和结实过程正常。研究结果表明突变体FS-M1雌性不育现象可能与柱头乳突细胞发育畸变，功能缺失有关。
4、突变体FS-M1体细胞染色体数为2n=38，花粉母细胞减数分裂染色体配对正常；对突变体FS-M1与甘蓝型油菜N70、1059杂交F_1植株花粉母细胞减数分裂观察结果，杂种染色体在减数分裂中期配对，形成19个二价体，减数分裂行为正常。
5、突变体FS-M1与野生型及其它品种正反交F1群体植株雌蕊育性正常，F2代雌性不育株与可育株比为1：15；回交一代雌性不育株与可育株比为1：3。测验结果表明突变体FS-M1雌性不育性状受两对隐性基因控制。
Female sterile mutant FS-M1(Brassica. napus L.) was segregated from the cultivated variety Ningyou 10. A comparison between FS-M1 and its wild type Ningyou 10 was carried out by means of microscopic histological dissection and cytological observation. At the same time, the inheritance of female sterile of FS-M1 was studied. The main results were as follows:
1、The floral morphology of FS-M1 differ in some aspects to that of wild type. Pistil length of FS-M1 was shorter than that of wild type. The 6 stamens of FS-M1 were almost equal to each other in length, while that of wild type composed of 4 longer one and 2 shorter one. There were two extra crescents-like organ developed at the ovary base of FS-M1 with missing nectars. Ovary shape and length of FS-M1 was more flat and shorter respectively than that of wild type. Stigma end of FS-M1 shaped a fork-like looking, was differ obviously from wild type.
2、Result from the observation of agronomic trait of FS-M1 showed that: Pod set ratio of open-pollinated, artificial-pollinated, self-pollinated, sibling-pollinated, and constrained self-pollinated was about 3％respectively. Seed number per pod was about 2. It indicates that: female sterility of FS-M1 is steady. Seed set of hybrid from the cross of normal variety×FS-M1 was normal, so does open-pollinated of F_1. Seed set ratio of self-pollinated of F_1 was 97％, and seed per pod reached 21. It indicates that: the sterile pollens didn't affect the fertility of F_1.
3、The numbers of papillae cells of FS-M1 was sparse and its size was larger than that of wild type. Number of pollen grains observed on stigma of FS-M1 was less than that on wild type. The papillae cells of FS-M1 was drying and shriveling gradually in 2-3 days after flowering. There were few germinated pollens observed on surface of stigma of FS-M1 after hand-pollination. But no pollen tube found inside the style. Nevertheless there were pollen tubes existing inside the style of FS-M1, resulting in a normal seed-set when hand-pollination was made after excision of the stigma or part of the style. Results from above indicate that losing function of the papillae cells of FS-M1 resulting from its abnormal development is possibly a key factor causing the female sterility.
4、Cytological observation of FS-M1 showed that chromosome number in somatic cells of FS-M1 was 2n=38 and its meiotic chromosome behavior in PMCs was normal. Chromosomes behaviour of PMs meiosis in cross of (FS-M1×1059) F_1 and (FS-M1×N70) F_1 were normal. Chromosomes were paired normally, and formed 19 bivalents at MI and divided evenly to the two poles at AI. It indicats that performance of the mutant is not results from chromosome variation.
5、Genetic studies showed that the female sterility of FS-M1 was controlled by two pairs of recessive genes.
引文
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89、Yokoo. Female sterility in an.indica-japonica crosses of rice. Jap. J. Breed ,1984, 34:219-227
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94、Chih-Wei Tung, Kathlcen G. Dwyer, Mikhail E. Nasrallah, and June B. Nasrallah. Genome-Widc Identification of Genes Expressed in Arabidopsis Pistils Specifically along the Path of Pollen Tube Growth. Plant Physiology,(2005), 138: 977-989
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101、郑丽霞植物成花转变过程中的分子机理研究．中山大学研究生学刊：自然科学与医学版．2001．022(004)．21-27
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104、杨鹭生，李国平．被子植物胚囊结构的观察方法．实验室研究与探索，2004，23(2)：9-11