細胞分裂素與透氣性容器封口對丹參組培苗增殖與玻璃質化之影響

Influence of Cytokinin and Ventilating Container Closure on Shoot Proliferation and Hyperhydricity of In Vitro Salvia miltiorriza Culture

陳威臣(Uei-Chern Chen) ； 蕭翌柱(Yih-Juh Shiau) ； 蔡新聲(Hsin-Sheng Tsay) ； 夏奇鈮(Chi-Ni Hsia)

《台灣農業研究》 54卷2期 (2005/06) Pp. 93-102

https://doi.org/10.6156/JTAR/2005.05402.03

摘要

本研究藉由改善細胞分裂素與培養容器透氣性，探討丹參(Salvia miltiorriza Bunge)組培苗大量繁殖及抑制玻璃質化苗形成之方法。將丹參莖節培養於含有1mg/l苄氨基嘌呤(benzyladenine, BA)與0.1mg/l奈乙酸(α-naphthaleneacetic acid, NAA)之全量MS (Murashige & Skoog)無基鹽類基本培養基中，可成功地誘導芽體形成及建立初代無菌丹參培養苗。無菌苗培養於添加0.25mg/l N-糠基腺嘌呤(N-furfuryladenine, kinetin)之全量MS基本培養基中，經6週培養後每個培植體可獲得6.5個芽體，優於0.5mg/l kinetin與0.5-1mg/l BA之處理，但所有處理之芽體均呈現玻璃質化。降低MS鹽類與kinetin濃度可部分改善玻璃質化苗的形成，其中以含有0.2mg/l kinetin之1/2MS基本培養基處理下，每個培植體可獲得5.6苗較多，但仍有高達46.6%玻璃質化苗形成。利用透氣性較高之藥包紙進行培養容器封口置換處理，可有效抑制丹參組培苗之玻璃質化，其中以雙層鋁箔紙封口培養2週，而後以三層藥包紙進行封口置換處理繼續培養4週，每個芽體可獲得4.8株正常苗之表現最佳，不僅能完全抑制玻璃質化苗形成，且苗長可達約4.78cm。綜合本研究結果，建議丹參組培苗可利用含有0.2mg/l kinetin之1/2MS基本培養基進行組培苗增殖，同時配合上述培養方式之藥包紙進行培養容器封口置換處理，不僅可建立丹參組培苗大量繁殖方式，更可完全抑制組培苗玻璃質化的產生。

關鍵字

丹參 ； 藥用植物 ； 微體繁殖 ； 玻璃質化

並列摘要

In vitro shoot multiplication of Salvia miltiorriza Bunge using nodal segment explants has been investigated in this study. Aseptic shoots were established on a full-strength MS medium containing 1 mg/l benzyladenine (BA) and 0.1 mg/l α-naphthaleneacetic acid (NAA) for shoot multiplication. In the cytokinin for shoot multiplication experiment, among the concentrations of BA (0.05, 0.1 mg/l) and kinetin (0.25, 0.5 mg/l) tested, the highest shoot multiplication (6.5 shoots per explant) was achieved in the medium containing 0.25 mg/l kinetin after 6 weeks of culture. Nevertheless, hyperhydric shoots were found for all cytokinin treatments. In order to improve the hyperhydricity disorder for in vitro shoot proliferation of S. mitrorriza, a medium with half-strength MS basal salts and lower concentration of kinetin was used for subsequent experiment. The highest normal shoots (3.0 per explant) were obtained from the 0.25 mg/l kinetin treatment along with a 46.6% hyperhydricity. Using the dispense paper instead of aluminum foil as container closure improved ventilation and reduced hyperhydricity in shoot cultures. Both the highest normal shoots (4.8 per explant) and the longest shoot length (4.78 cm) were obtained in the treatment using aluminum foil as container closure for the first 2 weeks of culture, then exchanging with dispense paper for the rest 4 weeks of culture. In conclusion, an in vitro shoot proliferation system for S. miltiorriza was established by culturing shoots in the medium containing half-strength MS basal salts and 0.2 mg/l kinetin. However, using ventilating dispense paper as container closure for certain culturing period was beneficial to overcome the hyperhydric disorder in shoot cultures.

並列關鍵字

Salvia miltiorrhiza ； Medicinal herbs ； Micropropagation ； Hyperhydricity ； Ventilating container closure