| 摘要: |
| 以6份耐寒结球甘蓝杂交F1为试验材料,研究其胚胎发生过程及胚胎发生的诱导条件,为耐寒甘蓝材料的小孢子培养,以及其他芸薹属小孢子胚胎体系的建立及优化提供理论依据。采用4',6-二脒基-2-苯基吲哚 (DAPI) 染色方法对小孢子体内和体外的发育及胚胎发生过程、特征进行观察;NLN-13培养基中添加阿拉伯半乳聚糖(Arabinogalactan,AG)和阿拉伯半乳糖蛋白(Arabinogalactan-protein,AGP)对小孢子培养胚胎发生率的影响进行统计分析;小孢子体内正常发育方向是单核小孢子经历2 次不对称核分裂后发育成三核成熟花粉,小孢子单核晚期对应的花蕾长度主要集中在3.5~4.5 mm;体外培养中,热激胁迫(32.5 ℃)诱导小孢子从配子体发育途径向胚胎发生途径转换,只有少数胚性细胞最终发育成为胚状体;供试材料中有5份材料获得胚状体,材料间出胚量差异较大,其中'寒玉37’出胚量最高,为每蕾22.5个;在NLN-13培养基中添加10 mg/L AG或10 mg/L AGP,可以促进耐寒甘蓝胚状体发生。32.5 ℃热激胁迫1 d可有效诱导耐寒甘蓝小孢子培养中的胚胎发生,10 mg/L AG或10 mg/L AGP促进胚胎诱导。 |
| 关键词: 耐寒甘蓝 小孢子培养 胚胎发生 发育途径 |
| DOI：10.11841/j.issn.1007-4333.2015.02.011 |
| 投稿时间：2014-07-07 |
| 基金项目:"十二五"农村领域国家科技计划项目(2012BAD02B00); 江苏省农业科技支撑计划重点项目(BE2013429) |
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| Studies on isolated microspore culture and embryo developmental pathway in cold-tolerant white cabbage(Brassica oleracea L.var. capitata) |
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ZENG Ai-song1, GAO Bing1, SONG Li-xiao1, ZHANG Yun-xia1, LI Jian-qi2, YAN Ji-yong1
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| (1.Institute of Vegetable Crops, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China;2.Qingdao Bioivy Crop Science Co.Ltd, Qingdao 266109, China) |
| Abstract: |
| Microspore embryogenesis and culture conditions which affected microspore-derived embryos were studied with six F1 hybrids of white cabbage with cold tolerance.The aim was to construct and optimize microspore culture system of cold-tolerant white cabbage and other Brassica species hybrids.The microspores development and embryogenesis of six F1 hybrids of the white cabbage in vivo and in vitro were investigated with 4', 6-diamidino-2-phenylindol staining technique.The effects of arabinogalactan (AG) and arabinogalactan-protein (AGP) on embryo yield were estimated by statistical analysis.The microspore developmental stages from uninucleate, bicellular to mature pollen were observed in vivo, which underwent twice asymmetrical nuclear division.Flower bud length corresponding to late uninucleate microspores stage from 3.5 to 4.5 mm.Under temperature stress (32.5 ℃), in vitro microspores could be induced to deviate from their gametophytic development and switch to embryogenesis.A small portion of responsive microspores could develop into embryoid during isolated microspore culture.The outputs with various materials were obviously different.One of the six materials had no embryos.But ‘Hanyu No.37' possessed 22.5 embryos per bud, the highest level in the rest five materials with embryos.Adding 10 mg/L AG or 10 mg/L AGP to the medium of NLN-13 could improve the ability of microspore embryogenesis.The heat shock stress by 32.5 ℃ for 1 day could induce embryogenesis of the white cabbage.Either 10 mg/L AG or 10 mg/L AGP had a positive effect on embryogenesis. |
| Key words: cold-tolerant white cabbage microspore culture embryogenesis developmental pathway |
