客家地区野生金线莲离体快速繁育的研究

A Study on the Rapid Propagation in vitro of wild Anoectochilus roxburghii from Hakka Region

以客家地区梅州野生金线莲(Anoectochilus roxburghii)为试材,研究了金线莲外植体消毒、原球茎诱导、增殖、生根过程中培养条件的优化,以期构建金线莲的高效繁育体系.结果表明,金线莲茎段经0.1%HgCl2溶液消毒8min,腋芽在MS+2.0mg/L 6-BA+0.1mg/L NAA+3%蔗糖+0.8%卡拉胶培养基上萌发伸长;在固体培养基MS+0.5mg/L TDZ+0.2mg/L NAA上原球茎诱导率达90.32%,在MS+0.6mg/L TDZ+0.1mg/L KT+0.5mg/L 2,4-D上液体培养时原球茎增殖系数为4.53;在1/2MS+1.0mg/L IBA+0.3mg/L NAA+0.2%活性炭的固体培养基上生根数可达3.4.金线莲原球茎在真菌污染下仍可正常生长,液体培养基添加棉花、玻璃珠、滤纸桥都有助于原球茎的增殖.

With wild Anoectochilus roxburghii from Meizhou Hakka region as explants,the optimum medium for the proto-corm induction,subculture proliferation in liquid medium and rooting were studied to establish the high efficiency system of rapid propagation in vitro.The results showed that,after being sterilized by immersing in 0.1%(w/v)HgCl2 solution for 8 min,the auxiliary buds of stem segments sprout on MS medium containing 2.0 mg/L 6-BA and 0.1 mg/L NAA with 3% sucrose and 0.8%carrageenan.The proto-corms were induced by 90.32%from the auxiliary buds on the solid medium MS+0.5 mg/L TDZ+0.2 mg/L NAA.Then proto-corms were propagated on liquid medium with 0.6 mg/L TDZ,0.1 mg/L KT and 0.5 mg/L 2,4-D with a 4.53-fold increase per subculture.1/2 MS solid medium supplemented with 1.0 mg/L IBA,0.3 mg/L NAA and 0.2% activated charcoal for rooting 3.4 and plantlet formation.The proto-corms could grow normally even under fungi contamination,and supplement with cotton,glass beads and filter paper bridge in the liquid MS medium promoted respectively the protocorm proliferation.