Preliminary Study On In Vitro Regeneration Of Tea Plant(Camellia Sinensis L.)

Tea plant(Camellia sinensis(L.)O.Kuntze)is a perennial woody economic crops and its buds and one or two leaves are normally harvested as the best material.It is rich in a variety of substances and beneficial to human health,so is deeply loved by consumer around the world.It is particularly important to breed tea varieties with high yield and excellent quality,and plant regeneration in vitro technology is an important means of selection and breeding.Therefore,the construction of tea plant regeneration system through plant in vitro culture technology is of great significance to the subsequent genetic transformation system of tea plant and other research work.In this study,several tea varieties were used as the initial materials to study the construction of in vitro regeneration system of tea tree by inducing tea seed embryo,stem segment,leaf and so on.The main results are as follows:1.Establishment of aseptic system of tea tree.The most suitable disinfection method for tea tree seeds was found.According to the specific situation of tea seeds:(1)When the tea seeds are relatively fresh,the best disinfection method is to remove the peel outside the ultra-clean table → remove the seed shell in the ultra-clean table,and 75% alcohol for 20 s→ wash 3 times with sterile water → 100% 84 disinfectant solution for 10 min → wash 4times with sterile water.(2)When the tea seeds are stored for a long time and cause serious dehydration of the tea seeds,it is better to remove the seed shells outside the ultra-clean platform and use 75% alcohol for 3 minutes → 20%Na Cl O 10 min → wash 5 times with sterile water.2.Establishment of in vitro rapid propagation of stem segments of tea plant.(1)The stem segments which were cut axillary bud from tea seeds were induced in medium of MS+ 30 g / L sucrose + 3 g/L physical + 0.5 mg/L 6-BA + 0.5 mg/L KT + 0.1 mg/L NAA,and a large number of callus were induced,but only a few of them sprouted buds.(2)Callus was induced from stem segments containing axillary buds and differentiated into buds.The number of buds reached 4.3,and some buds had taken root.3.In vitro regeneration of tea tree embryos.(1)In vitro culture of mature embryo of tea plant induced a small amount of callus.(2)In vitro culture of immature embryo of tea plant induced callus,and the callus had already sprouted.4.Induction of tea leaves.The callus induction rate of tea leaves was 95.56%.For the construction of in vitro regeneration system of tea plant,only the stage of callus induction has been carried out at present.The varieties and culture conditions of callus induction from stem segments have been found,and the suitable medium for callus induction from leaves has been found.A lot of experimental materials were provided for the next stage of callus induction and rooting,which laid a foundation for the further construction of tea plant regeneration system.