Experimental Study On Tissue Culture System Of Wild European Plum

Xinjiang wild European plum（Prunus domestica L.）,belongs to the Rosaceae Prunus.It is one of the rare wild fruit tree resources in Tianshan wild fruit forest in Xinjiang.At present,due to the influence of human activities and the natural environment,the living environment of this resource has been seriously damaged,and the population has been decreasing year by year and the current situation was precarious.Tissue culture is a rapid and effective breeding method,which plays an important role in protecting genetic resources and increasing the population of endangered plants.Therefore,in this study,the leaf,petiole,stem and seed explants of wild European plum were studied by tissue culture technology,and the optimum culture procedure was selected,and the problems in the culture results of these four explants were summarized.It provides reference for establishing the regeneration system and plants of wild European plum in the future.The main research results are as follows:（1）The stem segments of wild European plum were collected from May to mid-June;the best disinfection method was 75%alcohol disinfection for 60 s+0.1%Hg Cl₂ disinfection for 7 min,and the survival rate was 85.5%.The inducing ability of adventitious buds by three carbon sources was sucrose>glucose>D-fructose,and 30 g/L sucrose was the best sucrose concentration.Suitable for wild European plum stem segment primary culture formula:WPM+2.0 mg/L TDZ+0.4 mg/L IBA,adventitious bud induction rate was 91.67%;adventitious bud proliferation formula was WPM+0.7 mg/L TDZ+0.4 mg/L IBA,proliferation coefficient was 5.23;rooting culture formula was 1/2MS+0.4 mg/L IBA+0.3mg/L NAA,rooting rate was 36.67%.（2）In the leaf culture of wild European plum,the leaf reversion is beneficial to the induction of callus,and the callus induction rate was 100%.The induction ability of different parts of the leaf is in the order of leaf base>middle>upper.After 14 days of dark culture of wild European plum leaves and 21 days of dark culture of wild European plum tissue culture seedlings,In B5 medium,the formula supplemented with 0.2 mg/L 6-BA+2.0mg/L NAA was better than 7.5 mg/L TDZ+2.5mg/L IBA,the callus induction rate was96.67%and 93.33%respectively,and the induction rate of adventitious buds from leaves of tissue culture seedlings was 2.33%.A small number of adventitious buds could be obtained in B5+0.7 mg/L 6-BA+0.3 mg/L NAA adventitious bud proliferation culture.（3）The best disinfection method suitable for wild European plum field picking petiole is 75%alcohol disinfection for 45 s+0.1%Hg Cl₂ disinfection for 7 min,the lowest contamination rate is 3.33%;when dark treatment for 14 days,the best formula for callus induction and proliferation was B5+0.2 mg/L 6-BA+2 mg/L NAA.The petiole of wild Prunus humilis seedlings was cultured in dark for 21 days.The suitable formula for primary culture was B5+0.2 mg/L 6-BA+1.5 mg/L NAA,and the adventitious bud induction rate was 30.00%.The formula suitable for proliferation culture was B5+0.7 mg/L 6-BA+0.3 mg/L NAA,and the proliferation coefficient was 2.8.The formula suitable for rooting culture was B5+0.5mg/L IBA,and the rooting rate was 20%.（4）The more suitable disinfection method for wild European plum seeds was 75%alcohol disinfection for 60 s+0.1%Hg Cl₂ disinfecting for 5min.Direct light culture and peeling treatment（semi-peeling and full peeling）are beneficial to germination.Wild European plum seeds cultured on WPM and MS medium could induce seed germination,and the germination rate was 5.0%.Seed germination can be induced in WPM+0.2 mg/L NAA+0.5 mg/L 6-BA and MS+0.1mg/L IBA+2.0 mg/L 6-BA,and the germination rate was2.5%.The formula of proliferation culture was B5+0.2 mg/L NAA+2.0 mg/L 6-BA,the proliferation rate can reach 11.8,and rooting can be induced in the formula of 1/2MS+0.5mg/L IBA.