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[摘要]
目的 研究铁皮石斛Dendrobiumcandidum原球茎液体悬浮培养的可行性以及接种量和培养液体积对原球茎生长的影响。方法 利用完全随机实验设计和正交试验设计研究不同基本培养基、接种量和培养液体积对原球茎生长的影响。结果 无论鲜重还是干重,铁皮石斛原球茎在液体培养基上均极显著好于固体培养基(P<0.001)。不同基本培养基对铁皮石斛原球茎生长影响的研究表明,培养天数为30d时,对于鲜重67-V极显著好于B5(P<0.01),B5极显著好于1/2MS(P<0.01),1/2MS显著好于MS(P<0.05);对于干重67-V与B5没有显著性差异(P>0.05),B5极显著好于1/2MS(P<0.01),1/2MS极显著好于MS(P<0.01)。接种量和培养液体积对原球茎生长影响的研究表明,接种量影响最大,体积其次,互作最小,若不考虑互作,对于鲜重和干重,最佳处理为接种量6.194g/瓶+培养液体积150mL/瓶或100mL/瓶;对于干重,若考虑互作,接种量为6.194g/瓶时,应选培养液体积150mL/瓶或100mL/瓶,接种量为3.102g/瓶时,应选培养液体积150mL/瓶或100mL/瓶,接种量为1.693g/瓶时,应选150mL/瓶或50mL/瓶。结论 液体悬浮培养对铁皮石斛原球茎的生长有利,获得了最佳基本培养基、接种量和培养液体积的最佳搭配方案,表明通过液体悬浮培养生产铁皮石斛原球茎具有较好的开发应用前景。
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[Abstract]
Object To study the feasibility of suspension culture of protocorm in Dendrobium cand idum Wall.ex Lindl.and effect of inoculum and medium volume on the grow th of protocorm.Methods Effect of four basic media MS,1/2 M S,67-V,and B5,inoculum and medium volume on the grow th of protocorm were studied by completely random experimental design and orthogonal test design.Results The grow th of D.candidum protocorms in liquid medium wasmarkedly better than that in solid medium(P<0.001).Among four basicmediaM S,1/2M S,67-V,and B5 for 30d,the 67-V medium was better than B5(P<0.01),B5better than 1/MS(P<0.01),1/2MS better than MS(P<0.05)for the grow th of protocorm in fresh weight; while there is no maskedly difference between 67-V and B5(P>0.05),B 5 was much better than 1/2M S(P < 0.01),and 1/2MS wasmuch better thanMS(P<0.01)in dry weight.Effect of inoculum on the grow th of protocorm was themost,and effect of medium volume the second,while the interaction between inoculum and medium volume was the least.The Results indicated that the treatment(inoculum 6.194g/flask and 150mL or 100mL liquid medium in 250mL flask)was the best for the grow th of protocorm,when the interaction between inoculum and medium volume was not considered.If the interaction between them was considered,treatments(inoculum 6.194g/flask and 150 mL or 100 mL liquid medium in 250 mL flask,inoculum 3.102 g/flask and 150 mL or 100 mL liquid medium in 250 mL flask,inoculum.1 693 g/flask and 150 mL or 50 mL liqid medium in 250 mL flask)were the best for the increase of dry weight of protocorm,respectively.Conclusion The grow th of D.cand idum protocorm in liquid medium is better than that in solid medium.Furthermore,the best basic medium and the best arrangement in pairs between inoculum and medium volume are obtained.D.cand idum is a raremedicinal plant,the probableway for producing protocorm s of D.cand idum is provided by the above study.
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[基金项目]
国家自然科学基金资助项目(30170023);国家杰出青年科学基金项目(30325047);全国高等学校优秀博士学位论文作者专项基金资助项目(199950);北京市科技新星计划资助项目(2003A61)