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1、外植体接种取长寿花新发嫩枝梢约20厘米,用净水冲洗,在无菌操作台上用0.1%Hgcl_2(升汞)处理2分钟,无菌水冲洗4～5次,每次2～5分钟;然后用安福替民(次氯酸钠)配制的有效成分为2%的消毒液,浸泡消毒6分钟,无菌水洗3～5次,冲净,切下端梢部约1厘米茎尖,接入培养基中,深约为外植体的1/2长。2、瓶内培养初始培养基为 MS+BA2.0+NAA0.2+糖3%+琼脂0.7%,pH=5.8(一号基),培养约2～3周后,芽伸长并且部分愈伤组织膨大。不分切或轻量分切,转入MS+BA0.4+NAA0.5+糖3%+琼脂7%+NaH_2PO_4 (20mg),pH=5.8(二号基)中,2～3周后长成丛生苗,愈伤组织变得更大,上有大量芽丛。待芽长到约10厘米可以切分时,切成1个茎节1段,转入1/2MS+NAA0.1+糖20%+琼脂6.5%,pH=5.8(三号基)中生根。7～10天后,茎节与培养基接触处会有发霉状白毛,变粗后形成为大量根须,长达0.3厘米以上时即可出瓶移栽了。其分化形成不定根极易,几乎100%。
1, explants take longevity spent new shoot tip about 20 cm, rinsed with water, asepsis console with 0.1% Hgcl_2 (mercuric chloride) for 2 minutes, sterile water rinse 4 to 5 times, Each 2 to 5 minutes; then use An for the people (sodium hypochlorite) formulated active ingredients 2% disinfectant, soak disinfection 6 minutes, sterile washed 3 to 5 times, rinsed, cut the tip about 1 cm stem Sharp, access to culture medium, about 1/2 of the explant length. 2, the initial culture medium bottle for the MS + BA2.0 + NAA0.2 + sugar 3% + agar 0.7%, pH = 5.8 (one base), about 2 to 3 weeks after culture, bud elongation and part of the more Wound tissue swelling. Transferred into MS + BA0.4 + NAA0.5 + sugar 3% + agar 7% + NaH2PO4 (20 mg), pH = 5.8 Into clusters of seedlings, callus becomes larger, there are a lot of bud clusters. When the buds grow to about 10 cm in length, they can be cut into 1 stem section and transferred to 1 / 2MS + NAA 0.1 + sugar 20% + agar 6.5%, pH = 5.8 (base 3). After 7 to 10 days, there will be moldy white hair at the contact point between the stem section and the culture medium, and a large number of root bends after becoming thick. When the length reaches 0.3 cm or more, the bottle can be transplanted. Its differentiation into adventitious root extremely easy, almost 100%.