华富苹果离体叶片再生的主要影响因素研究

Studies on main influence factors of leaves regeneration of ‘HuaFu' Fuji apple in vitro

以花药培养选育的富士品种"华富"组培苗的叶片为试材,研究了离体叶片诱导再生不定芽过程中不同激素种类与组合、暗处理时间等因素对不定芽再生率的影响。结果表明:试管苗继代培养3～4代,取顶部嫩叶3～4片,剪除叶片边缘,叶片以近轴面接种到最佳诱导再生培养基MS+TDZ 2.0mg/L+IAA 0.75mg/L+蔗糖30g/L,pH值5.8,黑暗培养10～15d后,转入光照培养条件下,再生率达83.3%～86.7%。再生不定芽分化15～20d后转接到增殖培养基MS+BA 0.5mg/L+IBA 0.1mg/L上,当不定芽长度约2cm时再转接到生根培养基1/2MS+IAA 1.0mg/L+蔗糖2%,15d后,生根率可达95.0%以上。

In order to establish a high efficiency regeneration system of adventitious shoot,the effects of hormones and dark treament time,ect were studied by using leaves of＂HuaFu＂Fuji apple cultivar（Malus domestica） as a explant in vitro. The results showed that the optimal explant was the No 3～4 leaves from top of 25 d - old cultures.The leaves were cut to three parts along main vein without edge,then the obverse surface of middle parts was placed on the optimal inducing medium, which was MS ＋ TDZ 2.0 mg/L＋ IAA 0.75 mg/L＋ Sucrose 30 g/L,pH 5.8.After 10～15 days of dark treatment,all cultures were moved to the light- culture room,the regeneration rate reached 83.3%～86.7%.15 to 20 days later from differentiation,the adventitious buds were transferred to subculture medium,namely MS ＋ BA 0.5 mg/L ＋ IBA 0.1 mg/ L.When growing to 2 cm,they were transferred to rooting medium,that was 1/2MS ＋ IAA 1.0 mg/L＋ Sucrose 2%.After 15 days light - culture,rooting rate reached 95.0%.