白桫椤GGB途径快速繁殖与不完全组织培养  

Rapid proliferation via GGB and partial tissue culture of Sphaeropteris brunoniana

作　　者：李腾敏 刘雯雯 饶倩 陈彧 韩豫 吴二焕 饶丹丹 Li Tengmin;Liu Wenwen;Rao Qian;Chen Yu;Han Yu;Wu Erhuan;Rao Dandan(Hainan Academy of Forestry/Hainan Academy of Mangrove,,Haikou 571100,China;College of Tropical Agriculture and Forestry,Hainan University,Haikou 570228,China;Haikou Jinniuling Park Management Office/Haikou Municipal Institute of Landscape Architecture Science,Haikou 570206,China)

机构地区：[1]海南省林业科学研究院/海南省红树林研究院,海南海口571100 [2]海南大学热带农林学院,海南海口570228 [3]海口市金牛岭公园管理处/海口市园林科学研究所,海南海口570206

出　　处：《山西农业大学学报（自然科学版）》2024年第4期42-50,共9页Journal of Shanxi Agricultural University(Natural Science Edition)

基　　金：海南省科研院所技术创新专项(SQKY2022-0027)。

摘　　要：[目的]优化白桫椤的组织培养技术,可缩短植株繁育周期,进一步提高繁育效率。[方法]本研究以白桫椤的孢子为外植体,研究不同基本培养基对孢子萌发的影响;探讨不同植物生长调节剂对绿色球状体(GGB)诱导和增殖的影响;通过不完全组织培养法,筛选孢子体转化的最佳基质和培养方式。[结果]诱导孢子萌发最佳基质为1/10 MS,萌发率69.55%,10 d可见孢子萌发,在1/2 MS培养基下孢子萌发存在抑制情况;GGB诱导培养基以1/2 MS+NAA 0.2 mg·L^(–1)较适宜,诱导率为92.22%;GGB增殖培养基以1/2 MS+NAA 0.1~0.5 mg·L^(–1)+IBA 0~0.1 mg·L^(–1)较适宜,60 d增殖系数可达6.14;活性炭也有利于GGB增殖,其中1/2 MS+NAA 0.1 mg·L^(–1)+活性炭1.0 g·L^(–1)增殖效果最好,60 d增殖系数可达7.84;因孢子体难以在组培过程中直接获得,通过将GGB团移栽至瓶外基质中能成功诱导出孢子体,最适基质为泥炭∶珍珠岩∶椰糠(2∶1∶1),移栽90 d转化率为43.33%;Knauss不完全组织培养法在白桫椤离体繁殖中的具有可行性,在基质为泥炭∶珍珠岩(1∶1)时,32 d即能成功诱导出孢子体,孢子体转化率高达86.67%。[结论]通过GGB途径可以实现白桫椤的快速增殖,不完全组织培养法能有效提高幼孢子体的转化率,为白桫椤规模化生产育苗提供技术支撑。[Objective]The aim of current study was to optimize tissue culture techniques of Sphaeropteris brunoniana,and to shorten the plant breeding cycle and further improve breeding efficiency.[Method]In this study,the spores of S.brunoniana were used as explants to investigate the effects of different basic media on spore germination,to explore the effects of different plant growth regulators on the induction and proliferation of green globules(GGB),and to identify the optimal substrate and cultivation method for sporophyte transformation through the partial tissue culture method.[Result]The optimal substrate for inducing spore germination was 1/10 MS,with a germination rate of 69.55%and visible spore germination within 10 days.Spore germination was inhibited in 1/2 MS medium.The suitable medium for GGB induction was 1/2 MS+NAA 0.2 mg·L^(–1),with an induction rate of 92.22%.The optimal medium for GGB proliferation was 1/2 MS+NAA 0.1~0.5 mg·L^(–1)+IBA 0~0.1 mg·L^(–1),achieving a proliferation coefficient of 6.14 in 60 days.Activated charcoal also favored GGB proliferation,with the best proliferation effect seen in 1/2MS+NAA 0.1 mg·L^(–1)+activated charcoal 1.0 g·L^(–1),reaching a proliferation coefficient of 7.84 in 60 days.Due to the difficulty of directly obtaining sporophytes during tissue culture,transplanting GGB clusters to exvitro substrates successfully induced sporophytes.The optimal substrate was peat∶perlite∶coconut bran at 2∶1∶1,with a conversion rate of 43.33%after 90 days of transplanting.The Knauss partial tissue culture method proved feasible for the in vitro propagation of S.brunoniana.Using a matrix of peat∶perlite at 1∶1,sporophytes were successfully induced within 32 days,with a conversion rate of 86.67%.[Conclusion]The GGB pathway enabled rapid proliferation of S.brunoniana.The partial tissue culture method effectively improved the conversion rate of young sporophytes and provided technical support for the largescale production of S.brunoniana seedlings.

关 键 词：白桫椤 绿色球状体 孢子体 不完全组织培养 离体快繁 

分 类 号：S722.3[农业科学—林木遗传育种]