摘要
目的建立金线莲工厂化高效育苗技术。方法以金线莲种子在培养基上萌发获得原球茎、不定芽,通过正交试验对愈伤组织的诱导和分化先行筛选,然后根据响应面分析法中心组合试验,以愈伤组织分化率为响应值,对6-BA、NAA、芋泥3因素进行优化。结果正交试验分析结果表明:影响愈伤组织诱导和分化的关键因子为外植体类型,不定芽类型最易诱导产生愈伤组织,诱导率较高,达到27.64%,筛选出的最优培养基为1/2MS+6-BA3.0mg/L+NAA0.5mg/L+芋泥200g/L,在该培养基上继代培养,愈伤组织月增殖系数达50倍。利用响应面优化得到的愈伤组织分化培养基为1/2MS+6-BA1.0mg/L+NAA0.46mg/L+芋泥200g/L,预测分化率为为83.16%,验证值为81.4%。生根壮苗培养1/2MS+NAA0.5 mg/L+80g/L香蕉+7g/L芋泥。结论因此利用响应面方法可有效优化金线莲愈伤组织分化培养条件,可应用于实际生产。
Objective To establish high-efficiency factory seeding technology on Anoectochilus roxburghii.Methods Using the protocorms and adventitious buds induced from its cultured seeds as explants,the medium for induction and differentiation of calli were pre-screened by orthogonal experiment,and then according to the Central Composite Design of response surface method,the 3 factors of 6-BA,NAA and mashed taro were evaluated using calli differentiation rate as a response value.Results The results of orthogonal design showed that explant type is the key factor affecting calli induction and differentiation,and aventitious buds were the most easy to induce calli,with a high induction rate of 27.64%.The best medium selected to induce calli was 1/2MS+6-BA 3.0 mg/L+NAA 0.5 mg/L+mashed taro 200g/L,and the monthly multiplication coefficient of calli was 50 times subcultured on it.The predicted value of calli differentiation rate reached to 83.16%,and the actual value of 81.4%was verified under the optimum medium of 1/2MS+6-BA1.0mg/L+NAA0.46mg/L+mashed taro 200g/L,optimized by Response Surface Methodology.Conclusion Therefore,the response surface method can effectively optimize the condition in the differentiation of calli from Anoectochilus roxburghii on subculture,and can be applied to practical production.
作者
林丛发
蒋元斌
郭慧慧
李志刚
林旭
LIN Cong-fa;JIANG Yuan-bin;GUO Hui-hui;LI Zhi-gang;LIN Xu(Ningde Agricultural Institute,Fu'an Fujian,355017,China)
出处
《时珍国医国药》
CAS
CSCD
北大核心
2020年第1期192-195,共4页
Lishizhen Medicine and Materia Medica Research
基金
福建省科技厅星火项目(2018S0053)。
关键词
金线莲
愈伤组织
诱导
分化
响应面分析法
Anoectochilus roxburghii
Callu
Induce
Differentiation
Response surface method
