Optimization Of Tissue Culture And Rapid Propagation Of Fragaria Ananassa‘Benihoppe’

In this experiment,the shoot tips,stem nodes and internal shoots of strawberry rhizome of Fragaria ananassa‘Benihoppe’in the greenhouse were used as test materials.After the five stages of sterilizing explants,adventitious buds,subculture,rooting and seedling transplanting,the tissue culture and rapid propagation system of Fragaria ananassa‘Benihoppe’was established.The results showed that plant growth regulators of different varieties and concentrations used in the experiment further improved the induction rate and proliferation rate,and good rooting index and transplantation survival rate were also obtained.The rapid propagation system of Fragaria ananassa‘Benihoppe’was initially optimized,which provided a theoretical and practical basis for tissue culture and seedling breeding in the future.The conclusions are as follows:1.Screening for different sterilization times,The best sterilization method was obtained.The best sterilization of all three explants was 75%alcohol disinfection for 30s.0.1%HgCl2disinfection for 7min.Under this program,it has achieved good anti-virus effect and will not disinfect excessively poisonous plants.The optimal survival rate of shoot tips was 91.1%,the optimal survival rate of internal shoots was 55.53%,and the survival rate of stem nodes was62.23%.2.On the four-factor three-level orthogonal test,the optimal basic medium and plant hormone concentration induced by adventitious buds of strawberry were screened.The tip of strawberry in three explants is the best explant.The optimal induction medium was MS+2.0 mg·L^-16-BA+0.3 mg·L^-1NAA+0.3 mg·L^-1 IBA,and the induction rate was 92%,followed by the stem section of strawberry,and the optimal induction medium was MS+6-BA 1.0 mg·L^-1+ NAA0.2 mg·L^-1+IBA0.2 mg·L^-1,and the induction rate was 74.67%.The worst induction effect was strawberry internal bud.The optimal induction medium was MS+2.0mg·L^-16-BA+0.3 mg·L^-1NAA+0.3mg·L^-1IBA,and the induction rate was 65.33%.In the induction test of the shoot tip and inner bud of strawberry,the influence of four factors on the induction of adventitious buds was basic medium>6-BA>IBA>NAA.In the induction of stem segments of strawberry,the influence of four factors was in the order of basic medium>6-BA>NAA>IBA.Among the four factors,the most influential effect on adventitious bud induction is the type of basal medium.Through the single factor verification test,it can be seen that in the adventitious bud induction test,6-BA plays a leading role,and NAA and IBA have a good promoting effect.The three can be used together to achieve the best induction effect.3.Subculture proliferation test found:that sucrose is the best carbon source for strawberry proliferation,and the optimal concentration was 30 g·L^-1.The leaves were green and grow strong.On single factor test,the optimum concentration of 6-BA was 0.50 mg·L^-1 and the proliferation coefficient was 5.1.The optimal concentration of GA₃ was 0.11 mg·L^-1 and the proliferation coefficient was 4.13.The optimal concentration of IBA was 0.08 mg·L^-1 and the proliferation coefficient was 3.47.The optimal test results in response to phytohormone concentration were MS+0.56 mg·L^-16-BA+0.11 mg·L^-1GA₃+0.08 mg·L^-1IBA,and the optimal proliferation coefficient was 14.06.Among them,6-BA has a very significant effect on the proliferation coefficient.The impact of GA₃ and IBA is significant,The interaction between6-BA and GA₃ has a significant effect on plant proliferation coefficient.In order to ensure the optimal growth state of the plants,the optimal subculture time was 40d,and the optimal number of subcultures was 4 times.4.The best rooting medium was:1/2MS+30 g·L^-1sucrose+0.4 mg·L^-1IBA.The influence of three factors on rooting of tissue culture seedlings was as follows:basic medium>IBA> sucrose.On the basis of the optimal medium,1.0 g·L^-1 activated carbon is added to achieve the best rooting effect.The rooting rate was 100%,the average number of roots was 6.93,the average root length was 6.57 cm,and the rooting index was 45.53.The plants were robust and the roots were developed.5.In the experiment of transplanting seedlings,the best cultivation substrate was:V（grass soil）:V（meteorite）:V（perlite）=2:1:1,and the survival rate of tissue culture seedlings can reach 100%,and the plants grow robustly.