灰岩金线莲种子萌发与组培快繁技术研究

Seed germination and tissue culture propagation technique of Anoectochilus calcareus Aver.

  • 摘要: 【目的】利用灰岩金线莲种子无菌播种进行组培苗生产,为其工厂化组培育苗提供参考。【方法】采用灰岩金线莲种子为外植体,以MS或1/2MS为基本培养基,探讨不同激素组合对外植体进行无菌播种、原球茎增殖、丛生芽分化及生根壮苗的影响。【结果】采用0.1%升汞处理灰岩金线莲蒴果20 min,灭菌率可达100%,且种子萌发不受影响。在种子萌发培养基中添加香蕉泥有利于种子萌发,最适合种子萌发的培养基为1/2MS+0.5 mg/L 6-BA+0.2 mg/L NAA+60 g/L香蕉泥;在原球茎增殖培养中,低浓度的6-BA和NAA有利于灰岩金线莲原球茎增殖,最适合原球茎增殖的培养基为1/2MS+0.2 mg/L 6-BA+0.2 mg/L NAA+60 g/L香蕉泥;丛生芽分化的最适培养为MS+2.0 mg/L 6-BA+0.5 mg/L NAA+60 g/L香蕉泥+0.1%活性炭,原球茎分化丛生芽较多,生长势最佳;最适合灰岩金线莲生根壮苗的培养基为MS+0.5 mg/L 6-BA+2.0 mg/L NAA+60 g/L香蕉泥+0.1%活性炭。【结论】利用灰岩金线莲种子进行无菌播种,通过原球茎增殖和分化,可获得大量优质组培苗,是实现工厂化育苗的理想途径。

     

    Abstract: ObjectiveIn order to provide references for industrializing production of tissue-cultured seedlings of Anoectochilus calcareus Aver., tissue-cultured seedlings of A. calcareus were obtained by axenic sowing in vitro.MethodWith A. calcareus seeds as explants, MS or 1/2MS as basic culture medium, effects of different hormone combinations on aseptic sowing, protocorm proliferation, multiple shoot differentiation,promoting growth and rooting were studied. ResultResults showed that, 0.1%mercuric-chloride for 20 min showed better disinfection effect on A. calcareus capsule,with 100% of disinfection rate, and the seed germination was not affected. Adding banana puree to medium benefited seed germination. The optimal medium for seed germination was 1/2MS+6-0.5 mg/L 6-BA+0.2 mg/L NAA+60 g/L banana puree. In proliferation culture of protocorm, low concentration of 6-BA and NAA was beneficial to protocorm proliferation. The optimal medium for protocorm proliferation was 1/2MS+0.2 mg/L 6-BA+0.2 mg/L NAA+60 g/L banana puree. The optimal medium for multiple shoots differentiation was MS+2.0 mg/L 6-BA+0.5 mg/L NAA+60 g/L banana puree+0.1% activated carbon, protocorm could differentiate into more multiple shoots, showing growth vigor. The optimal medium for promoting growth and rooting of seedlings was MS+0.5 mg/L 6-BA+2.0 mg/L NAA+60 g/L banana puree+0.1%activated carbon. ConclusionA lot of high-quality tissue-cultured seedlings of A. calcareus can be obtained by aseptic sowing, proliferation and differentiation of protocorm. Therefore, aseptic sowing is ideal way to realize industrial production of A. calcareus seedlings.

     

/

返回文章
返回