| 摘要: |
| 【目的】研究秋水仙碱对甘蓝小孢子胚胎发生及发育的影响，为促进甘蓝小孢子培养技术的有效利用提供依据。【方法】以甘蓝一代杂种及不同自交代数的高代自交系、低代自交系为试材，在游离小孢子培养基中添加不同质量浓度秋水仙碱，处理时间为48 h，以不添加秋水仙碱为对照（CK），在同等条件下进行小孢子培养，分别比较产胚量和胚胎发育状况的差异。【结果】（1）培养基中添加10 mg/L秋水仙碱的40份甘蓝材料中，有19份产生了胚状体，产胚材料比率为47.5%，较对照提高25%；（2）以9份甘蓝一代杂种及不同自交代数的高代自交系、低代自交系为试材，在培养基中添加1，10，50 mg/L秋水仙碱后，每蕾产胚数、子叶形胚率均较对照有所增加，其中一代杂种及低代自交系材料在1或10 mg/L秋水仙碱处理时增加幅度较大，每蕾产胚数和子叶形胚率分别为1.30～2.20个和15.0%～69.1%，而高代自交系材料在10或50 mg/L秋水仙碱处理时增加幅度较大，每蕾产胚数和子叶形胚率分别为0.78～1.98个和14.1%～70.6%；（3）在培养48 h时，用倒置显微镜观察各试材的小孢子，发现培养基中分别添加1，10，50 mg/L秋水仙碱时，膨大小孢子数量均较对照明显增多；在静止培养期间用倒置显微镜观察各试材小孢子的胚胎发育情况，发现培养基中添加1，10，50 mg/L秋水仙碱处理的多细胞原胚、球形胚、心形胚、鱼雷形胚频率均较对照高。【结论】在甘蓝小孢子培养基中添加秋水仙碱，可使培养无反应材料的部分材料产胚，使培养有反应材料的产胚量提高，同时也对胚胎的良好发育有促进作用。 |
| 关键词: 甘蓝（Brasscia oleracea var.capitata） 小孢子培养 秋水仙碱 胚胎发生 胚胎发育 |
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| 基金项目:陕西省农业攻关计划项目（2010K01-09）；国家“十一五”科技支撑计划项目（2006BAD01A7-2-04，2008BADB1B02）;陕西省“13115”重大科技专项（ZDKG-126） |
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| Effects of colchicine on embryogenesis and development of isolated microspore in Brasscia oleracea var.capitata |
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| Abstract: |
| 【Objective】The study was done to explore the effects of colchicine on embryogenesis and development of isolated microspore in cabbage to provide basis for promoting effective utilization of isolated microspore culture technique.【Method】Using cabbage F1 hybrid and the high generation inbred lines and the low generation inbred lines as test materials,the treatment group (adding different doses of colchicine in the medium of isolated microspores culture and treating 48 hours ) and the CK group(without adding colchicine in the medium of isolated microspores culture) were cultured under the same conditions.No.of embryo per bud and the situation of embryo development were compared respectively.【Result】(1) Adding 10 mg/L colchicine in the medium,there were 19 materials obtaining embryo in the 40 materials,the rate of the material which can obtain embryo was 47.5%.There was an increase of 25% compared with the CK;(2) Using nine cabbages F1 hybrids and the high generation inbred lines and the low generation inbred lines as test materials,adding 1,10 and 50 mg/L colchicine respectively in medium,No.of embryo per bud and the rate of cotyledon shaped embryo both increased compared with those of the CK.Among them,the materials of F1 hybrid and the low generation inbred lines cultured in the medium which was added 1 or 10 mg/L colchicine had much more increase,the increasing extent being 1.30-2.20 of embryo per bud and 15.0%-69.1% of the rate of cotyledon shaped embryo respectively,but for the materials of the high generation inbred lines cultured in the medium which was added 10 or 50 mg/L colchicine had much more increase,the increasing extent being 0.78-1.98 per bud and 14.1%-70.6% respectively;(3) Observing microspore of all the materials by inverted microscope after culturing 48 hours,it was found that all the numbers of the swelling microspores increased significantly in the treatment group (adding 1,10 and 50 mg/L colchicine respectively in medium) compared with that of the CK;Observing microspore embryos development of all the materials by inverted microscope during the period of static culture,it was found that the frequencies of multicellular proembryo,globular embryo,heart embryo and torpedo embryo in the treatment group (adding 1,10 and 50 mg/L colchicine respectively in medium) were all higher than those of the CK.【Conclusion】Adding colchicine in the medium of isolated microspore culture in cabbage,for the nonresponse materials which were cultured under normal condition,can induce embryoid in part of them,for the response materials which were cultured under normal condition,can increase embryo yield.It can not only promote embryoid induction,but also promote the development of embryo. |
| Key words: cabbage（Brasscia oleracea var.capitata） isolated microspore culture colchicine embryogenesis embryo development |
