摘要
[目的]研究消毒处理方式、培养基成分和胚状体培养方法对油菜游离小孢子培养的影响。[方法]以B5培养基为基础培养基，添加不同浓度的蔗糖、琼脂及不同激素组合进行试验，对油莱DH系的组培技术进行优化。[结果]含2％Cl^-的NaCIO消毒15min与0．1％HgCl，消毒10min培养效果较好，都能达到良好的消毒和产胚效果；在游离小孢子提取的过程中，B5液体培养基中蔗糖浓度为2％时能达到较好的产胚效果；小孢子培养首先在32℃暗培养5～7d，然后25℃暗培养12～15d，最后25℃振荡暗培养3～7d胚状体就可完全成熟；1／2MS培养基（附加1．2％琼脂＋0．02％NAA＋2．0mg／L6-BA＋3．4mg／L AgNO3＋2％蔗糖）有利于胚的分化和苗的形成，其褐化程度小，玻璃化程度低。[结论]该研究结果有助于甘蓝型油菜DH系的规模化生产及高效转化体系的建立。
[Objective] The aim of this study is to reveal the disinfectants and disinfection methods, medium components, and embryoid culture method on dissociative microspore culture. [ Method] B5 as basic medium appended with different concentrations of sucrose, agar and different hormone combinations was used to optimize the culture technique for DH line in Brassica napus L. [Result] Both the 15 rain disinfection of Na-CIO containing 2% Cl^- and 10 min disinfection of 0.1% HgCl2 performed well in disinfection and subsequent embryo production; in the extraction process of dissociative microspores, B5 medium containing 2% sucrose could achieve a good embryo production effect; under dark condition microspores were firstly incubated at 32℃ 5 -7 d, then at 25℃ 12 - 15 d, and finally transferred to 25℃ oscillator（60 -65 r/min） for 3 - 7 d, when the embryoid would become full ripeness; 1/2 MS medium appended with 1.2% agar, 0.02% NAA, 2.0 mg,/L 6-BA, 3.4 mg/L AgNO3 and 2% sucrose was helpful for embryoid differentiation and plantlet generation, presenting low degree of browning and slight vitrification.[Conclusion ] The results may facilitate DH Line in rape production in large scale and high efficient transformation system.
出处
《安徽农业科学》
CAS
北大核心
2008年第32期13972-13974,14059,共4页
Journal of Anhui Agricultural Sciences
基金
贵州省科学技术基金项目(黔科合J字[2007]2081号)
贵州省农科院项目(院ZX[2007]013号)
贵州省优秀科技教育人才省长专项资金项目(黔省专合字(2005)40号)
贵州省科技攻关项目(黔科合NY(2006)3008号)
贵州省科技厅项目(院所创能[2008]-008)
