影响大白菜游离小孢子培养若干因素的研究

A Study on Affecting Factors of Isolated Microspore-derived Embryos of Chinese Cabbage

以太原地区大白菜主栽品种的F1“科萌夏秋王”、“科萌78”、“德阳01”等30个品种为试验材料,采用游离小孢子培养方法,研究了基因型、蔗糖、活性炭、细胞分裂素6-BA、生长素NAA、高温热激、氨基酸对小孢子胚胎发生的影响,培养基对子叶期胚分化的影响。研究结果表明：30个品种大白菜在植物组织培养基NLN-13液体培养基中有15个诱导出胚;添加不同浓度蔗糖以NLN-13培养效果最好;合适的活性炭浓度可明显提高小孢子的出胚率;NLN液体培养基中添加NAA和6-BA对小孢子出胚率影响不大,浓度超过一定剂量时出胚率反而降低;高温热激在12-48 h范围内小孢子出胚率差异不大,在此范围以外小孢子出胚率明显降低;添加一定量的氨基酸可提高小孢子出胚率;1 L B5＋0.2 mg 6-BA＋20 g蔗糖＋12 g琼脂培养基有利于小孢子子叶期胚长成再生植株。

Thirty cabbage varieties including Kemengxiaqiuwang, Kemeng78. Deyang01 ets, which were dominant varieties in Taiyuan, were used as the materials of this experimant. The aim is to study factors （genotype, sucrose, AC, 6-BA, NAA, high temperature treatment, amino acids） affecting the embryogenic rates in isolated microspore culture of Chinese cabbages and to study medium affecting the differentiation of cotyledonary embryos. The results showed that 15 varieties had produced microspore-derived embryos among 30 varieties of Chinese cabbage cultured in NLN-13 liquid medium. Sucrose in NLN medium were compared. NLN with 13% sucrose is the best in production of microspore-derived embryos. Contents suitable concentration of activated charcoal had improved on the frequency of microspore-induced embryos. The addition of auxins （NAA） and the cytokinins （6-BA） in NLN medium had no significant effect on the frequencies of microspore-induced embryos but when the concentration was excessive, the frequencies decreased. There as no significant difference at high temperature when the incubation time was within 12-48 h, but the frequencies of microspore-induced embryos were much lower over this duration. Suitable concentration of amino acids can enhance the embryogenic rates in isolated microspore cultue of Chinese cabbages. The B5 medium with 6-BA 0. 2 mg · L^-1 , 2 % sucrose and 1.2 % agar is good for microspore-derived cotyledonary embryos to regenerate plants.