摘要
为了建立优良同源四倍体桔梗的快速繁殖和离体保存技术,采用均匀设计和正交设计以添加不同激素的MS培养基为基本培养基进行筛选和优化。结果表明:同源四倍体桔梗最佳丛生芽培养基为MS+BA(6苄基嘌呤)0.6mg/L+NAA(萘乙酸)0.4mg/L+RTCA(病毒唑)3.0mg/L,丛生芽增殖系数可达到9.13。最佳壮苗培养基为:MS+BA0.2mg/L+IAA(吲哚乙酸)0.1mg/L+PP333(多效唑)0.2mg/L+RTCA3.0mg/L。最佳生根培养基为:1/2MS+IAA0.7mg/L+AC(活性炭)0.2mg/L+ABT(生根粉)0.2mg/L,生根率可达100%。最适室温离体保存条件为:1/4MS+蔗糖30mg/L+甘露醇30mg/L+PP3331.0mg/L+BA0.3mg/L+IAA0.1mg/L+RTCA3.0mg/L,保存150d时存活率仍可达到59.2%,同时恢复生长快,遗传稳定性好。本研究为优良同源四倍体桔梗的推广应用和优良种质离体保存、减少遗传变异提供了技术依据。
To establish a new technique for in vitro rapid propagation and conservation of germplasm of autotetraploid plant of Plalycodon grandiflorum, basal MS medium supplemented with, different hormones was filtrated and optimized by using uniform design and orthogonal design. The obtained results indicated that the optimized medium for crowd bud propagation was MS＋ BA 0. 60 mg/L＋ NAA 0. 40 mg/L＋ RTCA 3.0 mg/L and the crowd bud propagation coefficient was over 9.13. The most appropriate condition for plantlet growth was MS＋ BA 0.2 mg/L＋ IAA 0. 1 mg/L＋ PP3330. 2 mg/L ＋ RTCA 3.0 mg/L. The optimized medium for rooting was 1/2 MS＋ IAA 0. 7 mg/L＋ AC 0. 2 mg/L＋ ABT 0. 2 mg/L and the rhizogenesis rate reached to 100%. The most suitable medium for conservation in vitro at room temperature was 1/4 MS＋ sucrose 30 g/L＋ mannitol 30 g/L＋PP333 1.0 mg/L＋BA 0. 3 mg/L＋IAA 0. 1 mg/L＋ RTCA 3. 0 mg/L. The survival rate was as high as 59.2% after the materials being cultured in such medium for 150 days, and the materials could renew fleetly as well as maintained their genetic stability. This research provided techniques for the application, conservation in vitro of excellent germplasm and reduction of heredity variance of autotetraploids of Plalycodon grandiflorum.
出处
《药物生物技术》
CAS
CSCD
2007年第1期23-28,共6页
Pharmaceutical Biotechnology
关键词
桔梗
同源四倍体
均匀设计
正交设计
离体保存
Plalycodon grandiflorum, Autotetraploid, Uniform design, Orthogonal design, In vitro conservation
