Cryptolepis buchanani微繁殖法的研究

Micropropagation of Cryptolepis buchanani Roem. & Schult.

建立Cryptolepis buchanani在試管中的再生步驟繁殖法，是利用在試管中生長的幼苗莖頂、子葉的節以及節的外植體；其中有最好表現的是節的外植體。栽培的方法是將節的外植體放到只含有各種細胞分裂素或者還包含生長素及吉貝素（gibberillic acid, GA3）的Murashige and Skoog（MS）培養基上。單獨使用不同的細胞分裂素或者結合使用生長素BAP，對於莖的增生有最好的效果。放在添加2 mg/L BAP、0.1 mg/L Kinetin（KN）、0.05 mg/L Napthalene acetic acid（NAA）以及0.05 mg/L吉貝素的MS培養基中，有60%的反應會產生最多的莖（12.5~13.0個/4.5到5.0公分長）。單獨將莖放到含有一種或多種生長素，例如IAA、IBA以及NAA的MS培養基中進行生根。在IBA濃度為1 mg/L時，會得到最多的微小莖來形成根（大約6.5到7.0個根/莖，其根的長度為4.0到4.5公分）。此方法可成功地馴化植物幼苗至盆中含有等比例消毒滅菌過的泥煤苔及堆肥中生長。

In vitro regeneration protocol was developed for multiplication of Cryptolepis buchanani by using shoot tip, cotyledonary node and nodal explants derived from seedlings grown in vitro. The best response was achieved with nodal explants. Cultures were established placing the nodal explants on Murashige and Skoog’s (MS; 1962) medium supplemented with various cytokinins singly or in combination with auxin and gibberellin. Of the various cytokinins used singly or in combination with auxins, 6-Benzyl aminopurine (BAP) was found to be most effective for shoot proliferation. The maximum number of shoots (12.5 to 13.0 shoots / explant with a shoot length of 4.5 to 5.0 cm) were produced on MS medium fortified with BAP 2mg/L, Kinetin (KN) 0.1mg/L, Napthalene acetic acid (NAA) 0.05 mg/L and gibberellic acid (GA3) 0.05 mg/L with 60% response. Individual shoots (grown on shoot proliferation medium) were rooted on MS medium supplemented with various auxins－ Indole acetic acid (IAA), Indole butyric acid (IBA), NAA－singly or in combination. Of these IBA 1 mg/L resulted in higher number of microshoots (80%) to form roots (about 6.5 to 7.0 roots / shoot and the root length of 4.0 to 4.5 cm). The in vitro raised plantlets were acclimatized successfully to pots containing a mixture of autoclaved peatmoss and compost in 1:1 ratio.