山红柿组培快繁技术体系的建立

Establishment of Rapid In-vitro Propagation System for Diospyros morrisiana Hance

以山红柿(Diospyros morrisiana Hance)当年生枝条上休眠芽为外植体材料,通过不同处理间对比研究对山红柿组培苗生长的影响,建立了初代培养、继代增殖、生根培养、叶片再生技术体系。初代培养:山红柿休眠芽经10%H2O2和75%乙醇消毒30 s消毒处理,有效解决了外植体褐化等问题。通过3种基本培养基的比较研究,发现(1/2N)MS培养基最适于豆柿组培苗的初代增殖生长培养。继代增殖:采用(1/2N)MS+6-BA1.0 mg/L+IAA 0.1 mg/L+TDZ 0.05 mg/L与(1/2N)MS+ZT 1.0 mg/L+IAA 0.1 mg/L培养基交替继代培养,平均株高可达2.28 cm。生根培养:在添加20 g/L蔗糖,活性炭0.05 g/L+IBA 0.1 mg/L的MS(1/2N)培养基中,前期暗培养3 d,平均根数最高可达2.08,生根率最高可达78.46%。叶片再生:在MS+TDZ 2.5 mg/L+NAA 0.2 mg/L的培养基中,先期暗培养14 d后转移至正常光照条件下培养,30 d后愈伤组织形成率、不定芽再生率和平均不定芽数分别达到93.3%,92%和3.74个。成功地建立了山红柿离体快繁技术体系。

In this study dormant buds on current-year shoots of Diospyros morrisiana Hance were used as explants,the influences of different treatments on seedling growth of Diospyros morrisiana Hance were compared,a system of primary culture,subculture proliferation,rooting and leaf regeneration was established. Primary culture： dormant buds of Diospyros morrisiana Hance treated with 75% alcohol disinfection and 10% hydrogen peroxide disinfection for 30 seconds,which effectively solved the problem of explant browning. Through a comparative study of three basic mediums,it was found that（ 1 / 2N） MS medium was the best for the first generation proliferation culture of Diospyros morrisiana Hance.Mulitiplication： using the medium of（ 1 / 2N） MS＋ 6-BA 1.0 mg / L＋IAA 0.1 mg / L＋TDZ 0.05 mg / L and（ 1 / 2N） MS＋ZT 1.0 IAA 0.1 mg / L for alternating subculture,the average plant height was up to 2.28 cm.Rooting cultivation： After 3 d early darkness,in the MS medium culture with 20 g / L sucrose and 0.05 g / L＋IBA 0.1 mg / L,the averange root number was up to 2.08 and the rooting rate was 78.46%.Regeneration of root： in the medium of MS＋TDZ 2.5 mg / L＋NAA 0.2 mg / L,after 14 days dark culture and then transferred to normal light illumination condition,30 days later,the callus formation rate,adventitious bud regeneration rate and the average number of adventitious buds were 93. 3%,92% and3.74,respectively.The in vitro rapid propagation system for Diospyros morrisiana Hance was successfully established.