杠柳茎段组培快繁体系建立

Establishment of Rapid Propagation for Stem Segment of Periploca sepium Bunge by Tissue Culture

为建立杠柳茎段组培快繁体系,本试验以杠柳带芽茎段作为外植体,研究筛选外植体最佳的取材时间与消毒方法,带芽茎段诱导、增殖侧芽阶段和生根阶段最佳的培养基,以及组培苗驯化移栽炼苗时间和基质筛选。结果表明:外植体在5月份取材,消毒可采用75%乙醇预处理后,再通过0.1%HgCl2消毒5min,8%NaClO消毒10min,75%乙醇消毒1min组合消毒效果最好,污染率和死亡率均最低,且成活率最高达84.44%。在侧芽诱导培养阶段,最佳的培养基配方为MS+6-BA 1.5 mg/L+NAA 0.1 mg/L+蔗糖30 mg/L,光照强度为2500 lx,16 h/d,此培养条件下诱导率达93.33%。在侧芽增殖培养阶段,最佳的培养基配方为MS+6-BA 2mg/L+IBA 0.3mg/L+蔗糖30mg/L,适宜的光照强度为3000lx,此培养条件下增殖率达96.56%,增殖系数达6.28。杠柳组培苗最佳的生根培养基配方为1/2MS+IBA0.5mg/L+蔗糖25g/L,此培养条件下生根率达91.67%。杠柳组培苗最佳驯化时间为闭瓶炼苗5d+开瓶炼苗5d,最佳移栽基质组合为1/2泥炭+1/2蛭石的基质,此移栽基质下成活率达78.89%。本研究为杠柳快繁体系的建立和工厂化育苗提供了理论依据,为杠柳大规模推广应用奠定基础。

To establish a tissue culture and rapid propagation system of Periploca sepium,the study used stems bud as the explant and studied the optimumsampling time and disinfection method for explants,the optimum medium for lateral bud induction,proliferation,and rooting,respectively,as well as the selection of time and transplanting substrate for tissue culture seedling domestication.The results showed that the method of optimum disinfection was the combination of 0.1%HgCl2 for 5 min,8%NaClO for 10 min,and 75%alcohol for 1 min after pretreatment with 75%alcohol when sampling in the May.The contamination rate and mortality were the lowest,and the highest survival rate was 84.44%after disinfection by above method.The lateral bud induction rate reached 93.33%under the optimum culture ofMS+6-BA 1.5 mg/L+NAA 0.1 mg/L+sucrose 30 mg/L with 2500 lx and 16 h/d of the illumination intensity.While the lateral bud proliferation rate reached 96.56%under the optimum culture ofMS+6-BA 2 mg/L+IBA 0.3 mg/L+sucrose 30 mg/L with 3000 lx of the illumination intensity,and the proliferation coefficient reached 6.28.The optimal rooting medium was 1/2MS+IBA 0.5 mg/L+sucrose 25 g/L,and the rooting rate reached 91.67%under this condition.The optimal domestication time of seedlings was 5 d in closed bottle and then 5 d in open bottle.The optimal transplanting substrate was 1/2 peat and 1/2 vermiculite,in which the survival rate reached 78.89%.This study provided a theoretical basis for the establishment of rapid propagation system and factory seedling cultivation,and laid a foundation for large-scale popularization and application of Periploca sepium.