香鳞毛蕨(Dryopteris fragrans)是一种多年生野生草本蕨类植物, 具有抗氧化、抑菌、抗银屑病和抗肿瘤等多种功效。香鳞毛蕨的野生资源匮乏, 通过组织培养建立其人工再生体系, 是保护香鳞毛蕨资源可持续利用的有效途径。通过对香鳞毛蕨孢子的无菌培养, 比较分析不同因素对原叶体增殖、孢子体诱导、愈伤组织诱导与增殖、丛生芽分化及生根的影响, 建立了快速繁殖体系, 为香鳞毛蕨规模化生产奠定基础。研究结果表明, 当培养基组分为1/2MS时, 原叶体生长状态较好, 颜色翠绿, 增殖倍数最高可达5.67±0.59, 此时有大量幼孢子体产生, 孢子体诱导率为(37.50±2.04)%; 愈伤组织诱导最佳培养基为1/2MS+2.0 mg·L^-1 6-BA+1.0 mg·L^-12,4-D, 诱导率可达(96.67%±5.77)%; 愈伤组织增殖最佳培养基为1/2MS+ 1.0 mg·L^-1 6-BA+0.5 mg·L^-1 2,4-D, 增殖倍数达13.30; 颗粒状愈伤组织在1/2MS培养基中生长出大量丛生芽, 诱导率为(53.33±3.33)%; 1/2MS+0.2 mg·L^-1 NAA培养基促进幼孢子体苗生根, 移栽成活率约为60%。

Dryopteris fragrans is a perennial herb fern with medical and economical values, such as anti-oxidation, bacteriostasis, anti-psoriasis, and anti-tumor. The wild resources of D. fragrans are scarce. Establishing a regeneration system for D. fragrans through tissue culture is needed to enable a sustainable use of this valuable resource. In this experiment, through sterile culture of spores of D. fragrans, the effects of different factors on prothallium proliferation, sporophyte induction, callus induction and proliferation, cluster bud differentiation, and rooting were compared and analyzed to establish a rapid propagation system, which laid the foundation for large-scale production of D. fragrans. The results showed that 1/2MS medium provided optimal growth with green color and a multiplication factor up to 5.67±0.59. The obtained plants have numerous young spores, and the spore induction rate was (37.50±2.04)%. The most efficient callus induction medium contains 1/2MS media supplied with 2.0 mg·L^-1 6-BA, and 1.0 mg·L^-1 2,4-D, which reached an induction rate up to (96.67±5.77)%. The optimal callus proliferation medium we obtained was 1/2MS media supplied with 1.0 mg·L^-1 6-BA, and 0.5 mg·L^-1 2,4-D, which reached a proliferation factor of 13.30. The obtained granular callus produced a large number of cluster buds (53.33±3.33)% in 1/2MS medium, and 1/2MS media supplied with 0.2 mg·L^-1 NAA medium promoted rooting, resulting in a transplanting survival rate of ~60%.