泼墨石斛高位芽组培快繁体系的建立

Establishment of tissue culture and rapid propagation system of Dendrobium Enobi Purple‘Splashi'

【目的】建立泼墨石斛(Dendrobium Enobi Purple‘Splashi’)高位芽组培快繁体系,为减少其无菌播种引起的子代植株花色性状分离提供技术支持。【方法】以泼墨石斛侧芽、高位芽和花梗为外植体,分析不同消毒时间对其不定芽诱导率的影响;以高位芽为外植体,分析不同激素组合对其不定芽诱导、丛生芽增殖及生根壮苗效果的影响。【结果】泼墨石斛高位芽的不定芽诱导效果优于侧芽和花梗,其中以0.1%HgCl2消毒2.5 min时的不定芽诱导率最高;以高位芽进行不定芽诱导的最佳培养基为MS+3.00 mg/L6-苄氨基嘌呤(6-BA)+0.50 mg/Lα-萘乙酸(NAA)+30.0 g/L蔗糖+5.0 g/L琼脂+2.0 g/L蛋白胨+0.5 g/L活性炭+100.0 m L/L椰汁,诱导率达73.3%;最佳增殖培养基为MS+4.00 mg/L NAA+0.20 mg/L 6-BA+30.0 g/L蔗糖+5.0 g/L琼脂+2.0 g/L蛋白胨+0.5 g/L活性碳+100.0 g/L土豆泥,增殖倍数达5.0倍;最佳生根壮苗培养基为MS+0.50 mg/LNAA+0.10 mg/L6-BA+30.0 g/L蔗糖+5.0 g/L琼脂+2.0 g/L蛋白胨+0.5 g/L活性碳+50.0 g/L香蕉泥+30.0 g/L土豆泥,平均株高7.30 cm,茎粗0.990 cm,生根数16条,根长6.10 cm,根粗0.140 cm;生根苗用水苔包裹根部后移栽至穴盘中,成活率达100.0%;以高位芽进行诱导获得的泼墨石斛再生植株,可保持其品种的典型泼墨花色特征。【结论】以高位芽为外植体建立的泼墨石斛组培快繁体系能同时实现种苗快繁和保持品种优良花色性状,可用于其种苗工厂化育苗。

【Objective】Phanerophyte tissue culture system of Dendrobium Enobi Purple‘Splashi’was established,in order to provide referential basis for solving the problem of flower color character segregation in progeny plant caused by aseptic seeding of D. Enobi Purple‘Splashi’.【Method】The lateral bud,phanerophyte and pedicel of D. Enobi Purple‘Splashi’were used as explant,and effects of different disinfection times on the adventitious bud induction rate of D.Enobi Purple‘Splashi’were studied. Besides,phanerophyte was utilized as explant to analyze the effects of different hormone combinations on adventitious bud induction,clustered buds proliferation,strong seedling and rooting of D. Enobi Purple‘Splashi’.【Result】The results showed that the adventitious bud induction effect of D. Enobi Purple‘Splashi’phanerophyte was better than that of lateral bud or pedicel,and the adventitious bud induction rate reached the highest level at2.5 min with disinfection of 0.1% HgCl2. The optimal medium for adventitious bud induction of phanerophyte was MS+3.00 mg/L 6-benzyl aminopurine(6-BA)+0.50 mg/L α-naphthylacetic acid(NAA)+30.0 g/L sucrose+5.0 g/L agar+2.0 g/L peptone+0.5 g/L active carbon+100.0 mL/L coconut juice,and the induction rate reached 73.3%.The optimal proliferation medium was MS+4.00 mg/L NAA+0.20 mg/L 6-BA+30.0 g/L sucrose+5.0 g/L agar+2.0 g/L peptone+0.5 g/L active carbon+100.0 g/L mashed potato,and proliferation times reached 5.0 times. The optimal medium of strong seedling and rooting was MS+0.50 mg/L NAA+0.10 mg/L 6-BA+30.0 g/L sucrose+5.0 g/L agar+2.0 g/L peptone+0.5 g/L active carbon+50.0 g/L mashed banana+30.0 g/L mashed potato,and the average plant height reached 7.30 cm,stem diameter being 0.990 cm,rooting number being 16,root length being 6.10 cm and root diameter being 0.140 cm. When rooted seedlings were transplanted to aperture disk with their roots being encapsulated with water moor,the survival percentage reached 100.0%. If the regenerated plants of D. Enobi Purple‘Splashi’were inducted by phanerophyte,typical flower color character of D.Enobi Purple‘Splashi’could be maintained.【Conclusion】The establishment of tissue culture and rapid propagation system of D. Enobi Purple‘Splashi’with phanerophyte as explant can be applied to industrial seedlings of D. Enobi Purple‘Splashi’since it can realize seedling rapid propagation and maintain fine color characters.