甘蓝和青花菜杂种小孢子培养

Isolated-microspore Culture of F1 Hybrids between Brassica oleracea var. capitata and var. italica

摘要:

对甘蓝（Brassica oleracea var．capitata）×青花菜（Brassica oleracea var．itdica）的20个杂种及相应的父母本进行游离小孢子培养，并对影响甘青杂种小孢子胚胎发生的主要因子进行探讨，适于小孢子培养的培养基为l/2NLN，附加0.5 mg L^-1 NAA、0.05 mg L^-1 BA、5 mg L^-1 AgNO₃、0.2 mg L^-1 2,4-D和0.1 mg L^-1活性炭。结果有14个杂种能产生胚状体，诱导率70％；不同杂种间小孢子胚胎发生频率存在很大差异，最高的是绿洲808×夏宝，平均每蕾16.2个胚。诱导杂种胚状体发生的最佳时期是小孢子单核靠边期至双核期，34 ℃热激2 d有利于小孢子细胞对称分裂。在含糖170 g L^-1的液体培养基中培养3 d，添加低糖（含糖110 g L^-1）的培养液，可显著提高出胚率。

Abstract:

Microspore culture of twenty F1 hybrids between cabbage （Brassica oleracea var. capitata） and broccoli （Brassica oleracea var. italica） and their corresponding parents were carried out, and the major factors affecting microspore embryogenesis was investigated. Liquid medium 1/2NLN was used for microspore culture, which contained 0.5 mg L^-1 NAA, 0.05 mg L^-1 BA, 5 mg L^-1 AgNO₃, 0.2 mg L^-1 2,4-D and 0.1 mg L^-1 active carbon. Results showed that fourteen F1 hybrids could produce embryoids, the induced frequency being 70%. The highest embryo producing rate appeared in＇ Luzhou808×Xiabao＇ with averaging 16.2 embryos developed per flower bud. The optimum period of microspores for embryoid development was from late uninucleate （late anaphase I） stage to binucleate （anaphase II） stage. The heat-shock treatment at 34℃ for 2 days was beneficial for stimulating symmetric unclean division of microspore cells. The embryo production was significantly increased in liquid medium containing 170 g L^-1 sucrose for 3 days and then by adding low sucrose content （110 g L^-1）.