摘要
用苹果（Ｍａｌｕｓ ｐｕｍ ｉｌａ Ｍｉｌｌ）胚珠诱导愈伤组织并建立悬浮细胞系。用２％ 纤维素酶、０．５％ 果胶酶的混合酶液分离悬浮培养物，可得到５．４×１０６ 个／ｇ ｆｒ． ｗ ｔ有活性的原生质体。这些原生质体在改良的ＭＳ、Ｋ８ｐ、Ｄ２ 培养基中均可发育成细胞团，在含２．０ ｍ ｇ／ＬＩＡＡ、２．０ｍ ｇ／ＬＮＡＡ、０．１ ｍ ｇ／ＬＢＡ 的ＭＳ固体培养基上形成愈伤组织，更换几次不同的培养基后，在分化培养基上分化出不定芽，在生根培养基上生根形成完整植株。
Calli were induced and suspension cell lines were set up from ovule of Malus pumila Mill. Protoplasts (5 40×10 6/g fr.wt) were isolated from suspension cell lines in enzyme mixture solution containing 2 0% Onozuka R 10, 0 5% pectinase, 0 65 mol/L mannital, 0 01 mol/L CaCl 2, 0 7 mmol/L KH 2 PO 4,0 3% dextran sulfate potassium salt, at pH 5 8 for 6 h at 26℃.The cell clumps were formed from protoplasts cultured in modified MS, K8p, D 2 media. Calli were formed on MS solid medium containing 2 0 mg/L IAA, 2 0 mg/L NAA , 0 1 mg/L BA. Shoots were differentiated on differentiated medium after several changes of the medium. Eventually, shoots rooted and developed into whole plantlets on a rooting medium.
基金
国家自然科学基金
关键词
苹果
原生质体培养
植株再生
Malus pumila
Protoplasts culture
Plant regeneration
