报春石斛组培快繁技术体系的建立
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摘要
以报春石斛蒴果为外植体,通过对种子无菌萌发和原球茎诱导、原球茎增殖及丛生芽诱导、生根壮苗以及移栽驯化的研究,以期建立报春石斛的组培快繁技术体系。结果表明:培养基MS+6-BA 1.0mg·L~(-1)+NAA 0.1mg·L~(-1)+蔗糖20g·L~(-1)最适报春石斛种子无菌萌发和原球茎的诱导;原球茎增殖及丛生芽诱导的最佳培养基为MS+6-BA1.0mg·L~(-1)+NAA 0.1mg·L~(-1)+香蕉泥100g·L~(-1)+蔗糖20g·L~(-1),继代培养45d,增殖系数达8.2;最佳生根壮苗培养基为MS+6-BA 1.0mg·L~(-1)+KT 0.5mg·L~(-1)+NAA 0.2mg·L~(-1)+香蕉泥100g·L~(-1)+蔗糖30g·L~(-1)+活性碳1.0g·L~(-1),培养3个月,生根率达100%,根系较长,苗壮;3—4月是桂林地区移栽报春石斛组培苗的最佳季节,生根苗在栽培基质Ⅰ(树皮∶水苔∶珍珠岩∶蛭石=10∶2∶1∶1,体积比)的长势好于栽培基质Ⅱ(树皮∶珍珠岩∶蛭石=10∶1∶1,体积比),成活率为92.3%。
In order to establish a rapid propagation system of D.primulinum,immature seeds of Dendrobium primulinum Lindl.were used as explants for tissue culture,and mediums for aseptic seed germination,subculture of protocom,rooting and growth of strong plantlets were screened.The results showed that the medium MS+6-BA 1.0 mg·L~(-1)+NAA 0.1 mg·L~(-1)+sugar 20g·L~(-1) was optimum for seed germination.The optimum subculture medium that for proliferation of protocorm and shoot induction was MS+6-BA 1.0mg·L~(-1)+NAA 0.1mg·L~(-1)+mashed banana 100g·L~(-1)+sugar 20g·L~(-1),the propagation coefficient was 8.2after subcultured 45 days.The optimum rooting medium was MS+6-BA 1.0 mg·L~(-1)+KT 0.5 mg·L~(-1)+NAA 0.2 mg·L~(-1)+mashed banana100g·L~(-1)+sugar 30g·L~(-1)+activated carbon 1.0g·L~(-1),the rooting rate reached 100% and the roots had higher quality and more quantity after cultured 90 days.The best time for the transplantation of root regeneration plants was March to April in Guilin area,the rooted plants grew better in mediaⅠ(pine bark∶sphagna∶perlite∶vermiculite=10∶2∶1∶1)than that in mediaⅡ(pine bark∶perlite∶vermiculite=10∶1∶1),and the survival rate was 92.3%.
In order to establish a rapid propagation system of D.primulinum,immature seeds of Dendrobium primulinum Lindl.were used as explants for tissue culture,and mediums for aseptic seed germination,subculture of protocom,rooting and growth of strong plantlets were screened.The results showed that the medium MS+6-BA 1.0 mg·L~(-1)+NAA 0.1 mg·L~(-1)+sugar 20g·L~(-1) was optimum for seed germination.The optimum subculture medium that for proliferation of protocorm and shoot induction was MS+6-BA 1.0mg·L~(-1)+NAA 0.1mg·L~(-1)+mashed banana 100g·L~(-1)+sugar 20g·L~(-1),the propagation coefficient was 8.2after subcultured 45 days.The optimum rooting medium was MS+6-BA 1.0 mg·L~(-1)+KT 0.5 mg·L~(-1)+NAA 0.2 mg·L~(-1)+mashed banana100g·L~(-1)+sugar 30g·L~(-1)+activated carbon 1.0g·L~(-1),the rooting rate reached 100% and the roots had higher quality and more quantity after cultured 90 days.The best time for the transplantation of root regeneration plants was March to April in Guilin area,the rooted plants grew better in mediaⅠ(pine bark∶sphagna∶perlite∶vermiculite=10∶2∶1∶1)than that in mediaⅡ(pine bark∶perlite∶vermiculite=10∶1∶1),and the survival rate was 92.3%.
引文
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[15]仇硕,赵志国,赵健,等.翅萼石斛组织培养及快繁技术研究[J].广东农业科学,2018,45(9):48-52.
[16]HAO Z,QI G,QING F G,et al.First report of stem and root rot of the medicinal herb Dendrobium officinale(Orchidaceae)caused by Ceratobasidiumsp.AG-R in Guangxi,China[J].Plant Disease,2017,101(9):1679-1680.
[2]国家中医药管理局《中华本草》编委会.中华本草:傣药卷[M].上海:上海科学技术出版社,2005.
[3]梅媛,叶庆华,杨培明,等.报春石斛化学成分研究[J].中国医药工业杂志,2014,45(3):224-228.
[4]张莹,王雁,李振坚.报春石斛的组织培养与快速繁殖[J].植物生理学通讯,2007,43(4):749-750.
[5]张莹,王雁,李振坚.报春石斛再生体系的建立[J].浙江林学院学报,2009,26(4):603-606.
[6]李媛,朱丹红,潘会堂,等.3种石斛兰以茎段为外植体的离体快繁技术[J].东北林业大学学报,2013,41(8):77-81.
[7]孙永玉,李恒安,闫红,等.齿瓣石斛的无菌播种和组织培养[J].植物生理学通讯,2009,45(10):1017-1018.
[8]陈小强,新楠,孙宁,等.龙石斛种子萌发及原球茎增殖的研究[J].种子,2015,34(10):34-38.
[9]张华通,孙同高,林晓萍,等.铁皮石斛种子组织培养研究[J].林业与环境科学,2016,32(6):40-43.
[10]余乐.流苏、金钗、报春石斛种子非共生萌发以及流苏石斛离体保存的研究[D].重庆:西南大学,2009.
[11]杨柳平,刘畅庆,赵仁发,等.铁皮石斛种子诱导原球茎组培快繁体系的研究[J].广东农业科学,2012,48(7):54-57.
[12]郑晓君,叶静,管常东,等.兰科植物种子萌发研究进展[J].北方园艺,2010,34(19):206-209.
[13]付传明,赵志国,黄宁珍,等.铁皮石斛无菌播种产业化繁育技术研究[J].广西植物,2012,32(2):238-242.
[14]叶秀仙,黄敏玲,林榕燕,等.素心建兰无菌播种快繁技术研究[J].福建农业学报,2015,30(10):939-943.
[15]仇硕,赵志国,赵健,等.翅萼石斛组织培养及快繁技术研究[J].广东农业科学,2018,45(9):48-52.
[16]HAO Z,QI G,QING F G,et al.First report of stem and root rot of the medicinal herb Dendrobium officinale(Orchidaceae)caused by Ceratobasidiumsp.AG-R in Guangxi,China[J].Plant Disease,2017,101(9):1679-1680.