In Vitro Rapid Propagation And Regeneration System Of Niaowang Tea NW32 Was Established

Tea trees is one of the important economic crops in Guizhou Province.With the improvement of the quality and efficiency of the tea industry and the upgrading of tea gardens,the lack of good seedlings has become one of the bottlenecks in the development of the tea industry in Guizhou.The Niaowang tea group species is one of the core germplasm of Duyun Maojian and Yunwu Gong tea.The research team selected the NW32 strain from the Niaowang tea group species in the early stage.Its buds are abundant and full,which is an ideal material for making famous tea.At present,NW32 has been planted and demonstrated in some areas of Duyun.In order to further accelerate the promotion and shorten the breeding cycle,this study uses the axillary buds of the NW32 of Niaowang tea in the tea tree resource nursery as explants to establish an in vitro rapid propagation system,including disinfection conditions,axillary bud germination induction,multiplication induction,strong seedlings and rooting,seedling tempering and transplanting,and the establishment of in vitro regeneration systems of sterile seedling hypocotyls,stem segments and leaf explants,including healing explore the optimal conditions for wound tissue and bud differentiation induction,strong seedling induction,and rooting induction.The results of the study are as follows:（1）Establishment of in vitro rapid propagation system of the NW32 of Niaowang tea1.A two-factor orthogonal experiment was designed for the disinfection time of 75%ethanol（40 s,50 s,60 s）and the disinfection time of 10%sodium hypochlorite（10 min,13min,15 min）.Soaking in 75%ethanol for 60 s and 10%sodium hypochlorite for 13 min has the best disinfection effect.The survival rate is 66.67%,the pollution rate is 23.64%,and the mortality rate is 9.70%.2.Add different concentrations of 6-BA,IBA and GA₃to the MS medium for axillary bud germination induction culture.When the culture medium was MS+2.00 mg/L6-BA+0.80 mg/L IBA+1.00 mg/L GA₃,the germination rate of axillary buds was 95.56%,and the growth was good.3.Add three different combinations of different concentrations of 6-BA,NAA or 6-BA,IBA or 6-BA,IBA and GA₃to the MS medium to induce propagation and culture of axillary buds.It was finally obtained that when the culture medium was MS+3.00 mg/L 6-BA+0.20mg/L IBA+1.00 mg/L GA₃,the multiplication coefficient is 6.59,and the plants were growing well.When subcultured,the number of buds contained in a single explant was 2～3,multiplication coefficient can reach 7.49.4.Add different concentrations of 6-BA and IBA to MS medium to induce vigorous seedlings.When the culture medium is MS+2.00 mg/L 6-BA+0.80 mg/L IBA,the plant height difference is 2.38 cm,the sprouts are strong,and the growth is good.5.Before inoculation,soak in 50.00 mg/L IBA sterile solution for 5 minutes,and then inoculate into 1/2MS medium with different concentrations of IBA for rooting induction culture.It is concluded that when the culture medium is 1/2 MS+1.50 mg/L IBA,the rooting rate reaches 100.00%,the root length and lateral roots are more,and the growth vigor is better.6.Combine the cultivation time（3 d,5 d,7 d）with the transplanting substrate(V_{nutrient soil}=1,V_{nutrient soil}:V_perlite=2:1,V_{nutrient soil}:V_perlite:V_vermiculite=2:1:1)design a two-factor orthogonal experiment.The seedlings are obtained for 5 days,and the transplanting substrate is V_{nutrient soil}:V_perlite:V_vermiculite=2:1:1.The treatment is better,and the survival rate is 90.00%.（2）Establishment of the regeneration system of the NW32 of Niaowang tea1.The disinfection time of 75%ethanol（1 min,2 min,3 min）and the disinfection time of 20%sodium hypochlorite（10 min,15 min,20 min）were designed with a two-factor orthogonal test to disinfect the tea seeds.Soaking in 75%ethanol for 3 minutes and 20%sodium hypochlorite for 15 minutes has the best disinfection effect.The embryo survival rate is 85.00%,the contamination rate is 15.00%,and the mortality rate is 1.33%.2.Add different concentrations of GA₃to MS medium for seed embryo germination culture.It is best when the culture medium is MS+1.50 mg/L GA₃,and the germination rate of seed embryo is 86.67%.3.Add different concentrations of 6-BA,IBA or NAA to MS medium to induce callus and bud differentiation on the hypocotyls,stems and leaves of sterile seedlings.Hypocotyls are better when the medium is MS+4.00 mg/L 6-BA+1.00 mg/L IBA.The callus induction rate is 95.28%,and the adventitious bud differentiation rate is 23.33%.The stems and leaves are the best for callus induction in the medium of MS+9.00 mg/L 6-BA+6.00 mg/L IBA and MS+3.00 mg/L 6-BA+9.00 mg/L NAA.The induction rate is they were 99.33%and 96.00%.Respectively,growing callus with adventitious buds is beneficial to promote the survival and growth of strong seedlings.4.Add different concentrations of 6-BA and IBA to MS medium to cultivate adventitious shoots.It is best when the culture medium is MS+2.00 mg/L 6-BA+0.80 mg/L IBA,the plant height difference is 1.57 cm,and the growth is good.5.Soak the 50.00 mg/L IBA sterile solution for 5 minutes,and then inoculate it into 1/2MS medium supplemented with different concentrations of IBA for rooting culture.It is best when the medium is 1/2 MS+1.50 mg/L IBA for cultivation,the rooting rate is 87.78%,and the plant growth is good.After 5 days of cultivation,when the transplanting substrate is V_{nutrient soil}:V_perlite:V_vermiculite=2:1:1,the growth is good.