水茄组织培养技术体系的建立

Establishment of tissue culture technology system of solanum torvum

[目的]本文旨在建立高效的水茄下胚轴组织培养愈伤诱导和再生植株体系。[方法]以水茄下胚轴为外植体,采用组织培养技术,通过对25个激素组合的分析,研究了愈伤组织诱导与芽分化、继代和生根培养3个技术环节的激素配比,同时对愈伤诱导植株再生过程中的潮霉素筛选压进行了测定。[结果]结果表明,以MS为基础培养基,下胚轴愈伤组织诱导和芽分化以6-BA 2 mg·L^(-1)+IAA 0.1 mg·L^(-1)为宜,增殖系数达6.62;在继代培养中,使用MS+6-BA 2 mg·L^(-1)+IAA 0.1 mg·L^(-1)即可维持较高的增殖系数6.62;生根培养最佳培养基为1/2MS,培养2周后生根率达90%。8 mg·L^(-1)是水茄外植体再生较为合适的潮霉素筛选压浓度。[结论]本研究以优化激素配方6-BA 2 mg·L^(-1)+IAA 0.1 mg·L^(-1)作为基础培养基成分,为进一步建立农杆菌介导的遗传转化研究奠定了基础。

[Objective]The aim of present study was toestablish an efficient callus induction and plant regeneration system of Solanumtorvum. [Methods] Hypocotyl tissue of Solanumtorvum was used as explants, and tissue culture techniques were applied to analyze the effect of different phytohormone combinations at different concentrations on the callus induction, subculture, and rooting culture development. The screening pressure of hygromycin in callus induced during plant regeneration was also determined. [Results]The results showed that the optimal concentrations of plant growth regulators for the callus induction and the shoot regeneration were determined with MS 6-BA 2 mg·L-1 + IAA 0.1 mg·L-1 medium, in which the average shoot number was reached up to 6.62 per original explant, and it could be kept at the same level in the subsequent culture with the same medium. The medium with 1/2 MS was determined as the best one for rooting system development, in which root was observed on more than 90% of shoots in 2 weeks post treatment. The optimal concentration of hygromycin for regeneration of Solanumtorvum explants was measured as 8 mg·L-1. [Conclusion] This study provided a scientific support for further establishment of agrobacterium-mediated genetic transformation using optimized phytohormones formula of MS 6-BA 2 mg·L-1 + IAA 0.1 mg·L-1 as the basic medium.