山红柿组培快繁技术体系的建立

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。

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

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 com-pared,a system of primary culture,subculture proliferation,rooting and leaf regeneration was established.Pri-mary culture:dormant buds of Diospyros morrisiana Hance treated with 75%alcohol disinfection and 10%hy-drogen 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 sub-culture,the average plant height was up to 2.28cm.Rooting cultivation:After 3 d early darkness,in the MS me-dium 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%and 3.74,respectively.The in vitro rapid propagation system for Diospyros morrisiana Hance was successfully estab-lished.