蝴蝶兰的组织培养研究
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摘要
本试验以蝴蝶兰原优良品种满天红花梗为外植体,对休眠芽萌发诱导、叶片原球茎诱导、原球茎增殖、根诱导和组培苗的移栽等方面进行初步研究,得出以下结果:
     1、花梗休眠芽的诱导
     在夏季、秋季、春季三个季节选取含休眠芽的花梗节段进行培养,对污染率、休眠芽萌发诱导率和芽生长状况进行了比较。试验表明秋季选取的花梗外植体灭菌效果最好,休眠芽萌发诱导率最高。MS+BA 5.0培养基较适合休眠芽的诱导。
     2、无菌苗叶片的原球茎诱导
     本试验以无菌苗叶片为外植体诱导原球茎产生,结果表明,BA浓度5.0mg·L~(-1)最适合叶片原球茎诱导,添加15%(V/V)椰乳(CW)能提高原球茎的诱导率,并有利于原球茎的生长。MS+BA 5.0+CW 15%培养基较适合诱导叶片产生原球茎,诱导率最高达到38%。
     3、原球茎的增殖
     在MS培养基中添加一定浓度的BA和果汁能提高原球茎的增殖率。另外,添加聚乙烯吡咯烷酮(PVP)100~200mg·L~(-1)能有效控制培养基的酚污染、提高原球茎的增殖率。MS+BA 5.0+CW 15%培养基较适合原球茎的增殖。
    
     J‘西大学祝眨d卜学位论文
     4、根的诱导
     在1/2 Ms培养基中附加一定浓度NAA(0 .1一0.5mg’L一’)和多效哇
    MET(0 .1~0.5mg’L一‘)与BA配合,能诱导小苗生根。用多效哇诱导生
    根,幼苗矮壮,叶片较绿,有利提高幼苗的移栽成活率。添加0.39一’
    活性炭(AC)能大大提高生根苗的平均根数和平均根长。
     对根诱导较适合的培养基是:1/2 MS+B A 2.0+NAA 0.卜AC或
    1/ZMS+BA 2.0+MET 0.3。
     5、组培苗移栽
     土上!苦篇乐左牙湘于丁石,卜Z戈寸汗吞目J立伏禽己七心不破万县‘nl翻云六刁一谁才曰丢,卜Z人叹产1才曰日之习隆
     百冬观兀笠气1土刃尺水万万U圣a猫田联硒平葵啊权人。全从不分步,垦从进
    气性不好,易造成组培苗死亡。
In this experiments, we choosed to use the Phalaenopsis in vitro culture. These aspects of first step researching were proceed: inducenment flower stalks with dormancy buds prouting, inducing protocorm-like-body(PLB) from leaves, propagating PLB, rooting of PLB .The results are shown as below:
    1. Inducing dormancy buds from flower stalk
    We used the flower stalks with dormancy buds which selected in summer, autumn and spring to culture. Comparing their contamination rate, sprouting rate and growth condition of buds, proved that it is best to use the flower stalks which selected in autumn to culture. The effect of sterilization is the best and the induce rate of dormancy buds is the higliest. MS medium supplemented with 5.0 mg.L-1 BA were fit to induce sprouting.
    2. Inducing PLB from leaves
    5.0mg.L-1 BA is fit to induce PLB from tube-culture Phalaenopsis leaves. Appending 15% (v/v) coconut water (CW) can enhance the induce rate and advance the growth of the PLB. In MS+BA 5.0+CW 15% medium adapt to induce PLB from leaves, inducing rate is 38%.
    3. Multiplication of PLB
    
    
    
    BA and juice can prove the multiplication of PLB. Poly ( N-Viny 1-2-Pyrrolidinone) (PVP) can prevent the hydroxybenzene pollution in medium and prove the multiplication rate. MS+BA 5.0+CW 15% medium is suitable for PLB mutiplication.
    4. Rooting
    1/2 MS medium supplemented with 0.1-0.5 mg.L-1 NAA or MET can induce rooting of PLB. When rooting induced by paclobutrazol, seedlings were short and strong, leaves were greener than others. It promotes the survival rate of transplantation. Appending 0.3 g.L-1 active carbon (AC) can promote the average amount of roots and average root length.
    1/2 MS+BA 2.0+NAA 0.1+AC or 1/2 MS+BA 2.0+MET 0.3 medium are suitable for induce rooting.
    5. Transplantation
    Moisture and permeability affect the survival rate of tube-plantlet in transplantation. High degree of moisture and low permeability would lead to the death of plantlet.
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