为加强野生蕨类植物的保护和开发, 优化星蕨(Microsorum punctatum)孢子萌发方法及条件, 比较分析不同因素对原叶体增殖、孢子体诱导, 绿色球状小体(GGBs)诱导及其发育为幼孢子体的影响, 建立人工高效快速繁育技术体系。以成熟孢子为材料, 以MS、1/2 MS、1/3 MS和1/4 MS为基本培养基在无菌条件下进行萌发; 通过进行L₉(3⁴)正交试验, 研究无机盐浓度、植物生长调节剂及其质量浓度对原叶体发生和增殖的影响。当原叶体增殖到一定数量时, 以MS、1/2 MS、1/3 MS和1/4 MS为基本培养基筛选出适宜诱导孢子体的培养基; 随后, 以幼孢子体为材料诱导GGBs发育为新的幼孢子体并进行炼苗移栽。适宜孢子萌发的培养基为1/2 MS, 原叶体在MS+ 0.3 mg·L^–1 6-BA + 1.5 mg·L^–1 NAA中大量增殖, 60天后增殖系数约为9.6; 将原叶体切割后接入1/4 MS培养基中, 加无菌水培养90天后幼孢子体发生系数约为10.0; 幼孢子体在1/2 MS+1.5 mg·L^–1 6-BA + 0.1 mg·L^–1 NAA中可诱导出绿色球状小体GGBs, 诱导率达93.3%, GGBs在此培养基中增殖系数可达32.0; 1/2 MS培养基对GGBs的分化成苗率较高, 最高分化成苗率约为92%; 试管苗经炼苗后移栽成活率在90%以上。该研究分别建立了原叶体-受精-孢子体和幼孢子体-GGBs-幼孢子体2个体系, 尤其是GGBs的产生, 极大缩短了植株的再生周期, 从而创建星蕨完整的人工快繁技术体系。 研究结果可为保护星蕨野生资源、优质种苗繁育提供技术支撑, 也可为其它蕨类植物的人工繁育提供参考。

In order to strengthen the protection and development of wild ferns, optimize the methods and conditions of spore germination of Microsorum punctatum. The influences of different factors on prothallus proliferation, sporophyte induction, GGBs induction and its germination into young sporophyte were compared and analyzed. The technology system of artificial, efficient and rapid breeding was established. Mature spores were germinated under sterile conditions with MS, 1/2 MS, 1/3 MS and 1/4 MS as basic media. L₉ (3⁴) orthogonal test was carried out to study the effects of the concentration of inorganic salts, plant growth regulators and their mass concentration on the genesis and proliferation of prothallus. When the number of prothallus increased to a certain extent, MS, 1/2 MS, 1/3 MS and 1/4 MS were used as the basic medium to select the most suitable medium for sporophyte induction. At the same time, GGBs was induced to develop into a new young sporophyte by using the young sporophyte as the material and transplanting. The optimum spore germination medium was 1/2 MS, and the prothallus prolifed in MS+0.3 mg·L^–1 6-BA + 1.5 mg·L^–1 NAA, and the coefficient of increment was about 9.6 after 60 days. The prothallus was cut into 1/4 MS medium and cultured with sterile water for 90 days. The coefficient of young sporogenesis was about 10.0. Green globular GGBs was induced by young sporozoa in 1/2 MS+ 1.5 mg·L^–1 6-BA + 0.1 mg·L^–1 NAA, the induction rate was 93.3%, and the proliferation coefficient of GGBs in this medium reached 32.0. The palntlets differentiation rate of GGBs was higher on 1/2 MS medium, and the maximum seedling formation rate was about 92%. After transplanting, the survival rate of test tube palntlets was more than 90%. Two systems of prothalle-fertilization-sporophyte and neosporophyte GGBs-sporophyte were established, especially the generation of GGBs, which greatly shortened the regeneration cycle of plants, so as to establish a complete artificial rapid propagation technology system of Starfern. The research results can provide technical support for the protection of wild resources and the breeding of high-quality ferns, and can also provide reference for the artificial breeding of other ferns.