铁皮石斛种子液体静置培养及其对离体植株生长的影响

Static Liquid Culture of Seeds and Its Effect on Plantlets Growth of Dendrobium officinale in vitro

【目的】研究铁皮石斛种子液体静置培养过程中,基本培养基对种子萌发、原球茎发育的影响以及来自不同培养基的原球茎及其体积大小对离体植株生长的影响。【方法】采用l/2B5、l/2N6、l/2MS、l/2花宝1号、1/2花宝2号等液体培养基静置培养铁皮石斛种子,观察统计种子萌发和原球茎发育生长情况;原球茎转至固体培养基后,比较来自不同液体培养基和体积大小(30目、40目)的原球茎,生长形成离体植株的形态类型。【结果】1/2N6培养基培养获得的原球茎直径平均值最高,显著大于1/2MS培养基、1/2花宝1号培养基和1/2花宝2号培养基;原球茎在固体培养基上培养90 d,可发育形成单芽、多芽、愈伤组织等形态类型,来自不同液体培养基的原球茎,发育形成多芽、愈伤组织的比例均存在显著差异,原球茎大小对单芽、多芽、愈伤组织产生具有显著影响。【结论】铁皮石斛种子液体静置培养中,基本培养基是影响原球茎发育的重要因素,1/2N6是较适宜的培养基;来自不同液体培养基的原球茎及其大小显著影响离体植株的生长,大于30目的原球茎发育形成正常单芽植株比例较高。

[Objective]The effect of different basic mediums on seeds germination,protoconns development and the effect of liquid mediums,protocorm sizes on the plantlets growth in vitro during static liquid culture of Dendrobium officinale were studied.[Method]The seeds of Dendrobium officinale were cultured in different liquid mediums(1/2B5,1/2N6,1/2MS,l/2Hyponex No.1,l/2Hyponex No.2).The process of seeds germination and protocorms development were observed,and the diameters of protoconns were also measured.Protocorms from different liquid mediums were separated by sizes(30 and 40 mesh)and placed on solid mediums of the same composition,respectively.The effect of liquid mediums and protocorm sizes on plantlets growth in vitro were tested.[Result]The diameters of protocorms cultured in 1/2N6 medium were significantly larger than that in 1/2MS medium,l/2Hyponex No.1 medium and l/2Hyponex No.2 medium.Single bud,multiple bud and callus were found after protocorms cultured on solid medium for 90 days.The ratio of multiple bud and callus derived from protocorms cultured in 1/2B5,1/2N6 and 1/2MS liquid medium was significantly different.The ratio of single bud,multiple bud and callus derived from protocorms of two sizes was significantly different.[Conclusion]Seed germination and protocorm development were greatly influenced by mediums.1/2N6 was most suitable for static liquid culture of seeds in Dendrobium officinale.Protocorm size significantly affected the plantlet growth in vitro.As the sizes of protocorms were larger than 30 mesh,protocorms inclined to form normal plantlets of single bud.