南方农业学报2025，Vol.56Issue(4)：1224-1234,11.DOI:10.3969/j.issn.2095-1191.2025.04.020
海金沙孢子无菌萌发与原叶体生长条件的优化
Optimization of aseptic spore germination and prothallus growth conditions for Lygodium japonicum
摘要
Abstract
[Objective]The objectives of this study were to optimize the aseptic spore germination and the prothallus growth conditions of Lygodium japonicum,which could provide theoretical basis for the establishment of the rapid propa-gation system and large-scale planting of Lygodium japonicum.[Method]Using Lygodium japonicum spores as experimen-tal materials,the effects of different spore maturity levels,NaClO solution concentrations and sterilization durations,me-dium inorganic salt content,agar concentration,pH value of medium,inoculation concentration of spore suspension,cul-ture temperature,dark treatment duration,and gibberellin(GA3)concentrations were tested.Spore germination percen-tage and prothallus area were measured to identify optimal culture conditions.[Result]Spores from the same sporophyll exhibited varying maturity levels,with earlier mature and dispersed spores having higher germination percentage.Spores dispersed within 1-3 d achieved a final germination percentage of 38.4%after 15 d of culture,significantly higher than those dispersed later(P<0.05,the same below).The best reproductive material was mature spores naturally dispersed within 1-3 d after sporophyll harvest.The spores treated with different NaClO solution concentrations and disinfection durations were not contaminated within 15 d of culture,but 3%and 5%high-concentration NaClO inhibited spore germi-nation and prothallus growth.The optimal sterilization method was 1%NaClO solution soaking for 4 min.Knop's me-dium with lower inorganic salt content promoted spore germination and prothallus growth.The final germination percenta-ge and prothallus area after 15 d cultured at 7.5 g/L agar concentration were significantly higher than those at 8.5 and 9.5 g/L.The final germination percentage and prothallus area spore in the medium with pH 5.8 were significantly higher than those in the media with pH 5.4 and pH 6.2.The optimal medium formula was Knop's medium with 7.5 g/L agar,20.0 g/L sucrose,and pH value adjusted to 5.8.The optimal spore inoculation procedure was to inoculate 0.5 mL of spore suspen-sion at a concentration of 1.0 mg/mL.The optimal culture temperature was 30℃constant temperature,initiating germina-tion on 3 d of culture and achieving a final germination percentage of 52.5%on 15 d of culture,which was higher than constant 25℃and 20℃/30℃alternating temperatures,prothallus area was also the largest.The final germination per-centage of spores under dark treatment for 15 d was significantly higher than that without dark treatment(0 d),but it might not meet the efficiency requirements for large-scale reproduction.The final germination percentage of spores treated with GA3 concentration of 0.01 mg/L was significantly higher than that without GA3 treatment(0 mg/L),but when the GA3 concentration reached 100.00 mg/L,the final germination percentage of spores was significantly lower than that without GA3 treatment.[Conclusion]Optimal aseptic germination conditions for Lygodium japonicum spores are selected,which also favor early prothallus growth.关键词
海金沙/孢子萌发/原叶体生长/无菌培养Key words
Lygodium japonicum/spore germination/prothallus growth/aseptic culture分类
农业科技引用本文复制引用
黄满霖,刘薇,童家赟,詹若挺,刘赛..海金沙孢子无菌萌发与原叶体生长条件的优化[J].南方农业学报,2025,56(4):1224-1234,11.基金项目
国家自然科学基金项目(32170400) （32170400）
广东省乡村振兴战略专项资金种业振兴项目(2022-NJS-00-002) （2022-NJS-00-002）
东莞市科技特派员项目(2022441920000014) National Natural Science Foundation of China(32170400) （2022441920000014）
Seed Industry Revitalization Project of Guangdong Rural Revitalization Strategy Special Fund(2022-NJS-00-002) （2022-NJS-00-002）
Dongguan Science and Technology Com-missioners Project(2022441920000014) （2022441920000014）
