铁皮石斛同源四倍体工厂化快繁工艺研究  

Rapid Propagation Tissue Culture Process of Dendrobium officinale Kimura et Migo Homologous Tetraploid

作　　者：刘静雯 梁晖辉 卢富华 李瑞兰 党仪泉 向增旭[1] LIU Jingwen;LIANG Huihui;LU Fuhua;LI Ruilan;DANG Yiquan;XIANG Zengxu(College of Horticulture,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学园艺学院,南京210095

出　　处：《西北植物学报》2018年第10期1927-1935,共9页Acta Botanica Boreali-Occidentalia Sinica

基　　金：现代农业产业技术体系建设专项资金(CARS-21)

摘　　要：以实验室前期诱导的铁皮石斛四倍体为研究材料,利用气孔、叶绿体、染色体计数及流式细胞仪等方法,对铁皮石斛同源四倍体株系的纯合性进行鉴定,在此基础上利用茎段扩繁和原球茎诱导、增殖、分化两种方案比较增殖效率,并对原球茎的倍性稳定性进行再鉴定。结果表明:(1)实验室的‘花自-2’株系为纯合同源四倍体。(2)茎段扩繁最佳增殖培养基为MS+2.0mg/L 6-BA+0.5mg/L NAA+15%香蕉汁,茎段诱导原球茎最佳培养基为MS+1.0mg/L 6-BA+0.5mg/L NAA+0.2mg/L KT,诱导率达到76.66%;原球茎增殖最佳培养基为MS+1.0mg/L 6-BA+0.5mg/L NAA+15%马铃薯提取液,原球茎分化最佳培养基为1/2 MS+0.5mg/L 6-BA+20%马铃薯提取液,原球茎分化率达到85.70%;生根最佳培养基为1/2MS+0.5mg/L NAA+15%香蕉汁,生根率达到92.77%。(3)原球茎根尖鉴定结果表明,诱导的倍性可以稳定遗传。(4)瓶苗移栽的最佳炼苗时间为5d,移栽成活率达92.30%。该试验建立了铁皮石斛同源四倍体株系新的快繁技术体系,为四倍体试管苗的工厂化生产和推广奠定了技术基础。Dendrobium o f ficinale Kimura et Migo homologous tetraploid was identified by means of stomata chloroplast chromosome counting and flow cytometry.On the basis of this,the multiplication efficiency was compared between the two schemes of stem segment propagation and protocorm induction,multiplication and differentiation,and the ploidy stability of protocorm was re-identified.The results showed that:(1)the strain‘Huazi-2’in the laboratory was a pure contract tetraploid.(2)The best medium for propagation of stem segments was MS+2.0 mg/L 6-BA+0.5 mg/L NAA+15%banana juice.For stem segment protocorm induction,the best culture medium for corm is MS+1.0 mg/L 6-BA+0.5 mg/L NAA+0.2 mg/L KT,the inducing rate is 76.66%;The best medium for protocorm multiplication is MS+1.0 mg/L 6-BA+0.5 mg/L NAA+15%potato extract.The best medium for protocorm differentiation is 1/2 MS+0.5 mg/L 6-BA+20%potato extract,the protocorm differentiation rate was 85.70%;The best medium for rooting is 1/2 MS+0.5 mg/L NAA+15%banana juice,the rooting rate was 92.77%.(3)The result of root tip identification of protocorm showed that the ploidy induced could be inherited stably.(4)The best time of transplanting was 5 days,and the survival rate of transplanting was 92.30%.A new rapid propagation technique system of autotetraploid lines of Dendrobium of ficinale Kimura et Migo was established in this experiment,which established the foundation for the industrial production and popularization of tetraploid plantlets.

关 键 词：铁皮石斛 同源四倍体 茎段 原球茎 快繁技术 

分 类 号：Q343.244[生物学—遗传学] Q813.1