无患子优选树茎段离体培养体系的建立

Establishment for in vitro culture system of selective Sapindus mukorossi Gaertn stems

【目的】筛选无患子茎段离体快繁最佳培养基，建立无患子优良种苗快速繁育体系。【方法】以野外选优获得的无患子树带腋芽茎段为外植体，采用L9（33）正交试验对影响外植体灭菌的HgCl2浓度、HgCl2与酒精的作用时间，不定芽诱导萌发中的6-BA、NAA及蔗糖用量，继代增殖中6-BA与KT用量和芽苗生根中的MS培养基无机盐水平、6-BA和2,4-D用量进行优化。【结果】75%酒精浸泡1 min，再用0.2% HgCl2处理7 min是无患子茎段灭菌的最佳方法，其外植体成活率高达83.33%；MS＋2.0 mg/L 6-BA＋0.1 mg/L NAA＋20.0 g/L蔗糖是无患子茎段腋芽萌发的适宜培养基，诱导率高达96.67%；MS＋0.7 mg/L 6-BA＋0.1 mg/L KT是无患子茎段腋芽增殖的适宜培养基，30 d可增殖4.13倍；1/3 MS＋0.5 mg/L 6-BA＋0.7 mg/L 2,4-D是诱导芽苗生根的适宜培养基，平均生根率为41.50%。【结论】建立的无患子离体培养体系为：外植体茎段用75%酒精浸泡1 min，再用0.2% HgCl2灭菌7 min，在MS＋2.0 mg/L 6-BA＋0.1 mg/L NAA＋20.0 g/L蔗糖培养基中诱导腋芽萌发，在MS＋0.7 mg/L 6-BA＋0.1 mg/L KT培养基中进行增殖培养，在1/3MS＋0.5 mg/L 6-BA＋0.7 mg/L 2,4-D培养基中进行生根培养。

Objective]In this paper, the optimal medium of in vitro rapid propagation was selected in order to estab-lish rapid culture system for Sapindus mukorossi Gaertn seedlings . [Method]Taking S. mukorossi selective stems with axillary buds as explants,the concentration of HgCl2, action time of HgCl2 and alcohol in explants sterilization, the con-tents of 6-BA, NAA and sucrose in adventitious bud induction, the dosages of 6-BA and KT in shoot proliferation, and inorganic salt levels in MS, dosages of 6-BA and 2,4-D in seedling rooting was optimized by orthogonal experiment design L9（33）. [Result]The optimum stem sterilization method for S. mukorossi was as follows：immersing the stem in 75%alcohol for 1 min, and sterilized with 0.2% HgCl2 for 7 min. After this treatment, survival rate of explants was 83.33%. MS＋2.0 mg/L 6-BA＋0.1 mg/L NAA＋20.0 g/L sucrose was the most appropriate culture medium for axillary bud induction, and the induction rate was 96.67%. MS＋0.7 mg/L 6-BA＋0.1 mg/L KT was the best culture medium for axillary bud proliferation, and the proliferation multiple was 4.13 times after 30 days. 1/3MS＋0.5 mg/L 6-BA＋0.7 mg/L 2,4-D was the best culture medium for rooting,and the average rooting rate was 41.50%. [Conclusion]The in vitro culture system of S. mukorossi es-tablished preliminarily is as follows： immersing the stem explants in 75% alcohol for 1 min, sterilizing them with 0.2%HgCl2 7 min, using medium of MS＋2.0 mg/L 6-BA＋0.1 mg/L NAA＋20.0 g/L sucrose to induce axillary bud germination, putting them in MS＋0.7 mg/L 6-BA＋0.1 mg/L KT medium for multiplication culture, and using 1/3MS＋0.5 mg/L 6-BA＋0.7 mg/L 2,4-D medium for rooting.