Establishment Of Tissue Culture Rapid Propagation And Regeneration System Of Curcuma Chinensi

C.serrulata var.lannesiana‘Grandiflora’is a deciduous tree of Rosaceae.Among many varieties of C.serrulata var.lannesiana‘Grandiflora’,most of them are white,pink or red,and the green variety is especially rare.C.serrulata var.lannesiana‘Grandiflora’is such a unique variety that it has a high ornamental value in the full flowering and falling flowering period,and has a huge potential for development in gardens.The stock market is in short supply.In production,C.serrulata var.lannesiana‘Grandiflora’is mainly propagated by grafting,but this propagation method has many problems,such as occupying large space resources,long seedling cycle and high seedling price,which are susceptible to the influence of time,climate and environment.After multiple generations of propagation,it is easy to cause the accumulation of viruses,affecting the yield and quality of C.serrulata var.lannesiana‘Grandiflora’.Therefore,the application of plant tissue culture technology to establish asexual rapid propagation system of C.serrulata var.lannesiana‘Grandiflora’is the most effective way to produce high-quality seeds and seedlings in industrialization and standardization.It can also provide some reference for the tissue culture of other Cerasus plants.The main research results of this experiment are as follows:1.Establishment of tissue culture and rapid propagation system for stem segments of C.serrulata var.lannesiana‘Grandiflora’In order to establish a tissue culture and rapid propagation system for stem sections of C.serrulata var.lannesiana‘Grandiflora’,the optimal disinfection and sterilization scheme was as follows:75%ethanol solution was shaken for 30s,followed by 0.1%（m/v）Hg Cl₂ solution for shaken for 6min.The survival rate of explants could reach 46.67%.The optimal medium for induction of stem segments was MS+2.0 mg·L^-16-BA+0.2 mg·L^-1IBA,and the induction rate was 80.15%.The optimal medium for seedling growth was MS+2.0 mg·L^-16-BA+0 mg·L^-1NAA+0.05mg·L^-1 IBA,the proliferation coefficient reached 3.45,and the optimal medium for seedling growth was MS+2.0 mg·L^-16-BA+0.1 mg·L^-1GA+0.05 mg·L^-1 IBA,and the average growth rate of seedling height reached 2.53cm.Rooting culture of the best medium for 1/2 of the MS culture medium does not contain any hormone,rooting rate can reach 100%,the two-step rooting method can effectively increase seedling height and promote root growth,specifically for the 1/2 MS+1.0 mg·L^-1NAA cultivate grow adventitious roots primitive swelled after MS basal medium without any plant hormone,seedling height growth in an average of 4.23 cm,the maximum root number was 15.81;The optimal refining time was 5d.After refining,the seedlings were transplanted into the mixed matrix with the volume ratio of peat soil,river sand and vermiculite at 2:1:1,and the survival rate of transplantation could reach 96.37%.2.Establishment of callus regeneration system of C.serrulata var.lannesiana‘Grandiflora’In order to establish a callus regeneration system,the effects of different explants and plant hormone combinations on callus induction,adventitious bud differentiation,proliferation and rooting were studied by using perennial mother plant leaflets,annual grafted seedling leaflets,axillary bud induction leaflets and proliferative generation leaflets as explants.The results showed that the best disinfection scheme for the leaflets of the perennial mother plant was as follows:shake the leaflets with 75%ethanol solution for 30s,then turn to 0.1%sodium hypochlorite for 2min,and the survival rate of the explants could reach 67.74%.All of the four explants could induce callus,and all of them differentiated adventitious buds except the leaflets of the perennial mother plant.The higher the degree of explant larval,the greater the potential of later culture.The best medium for callus induction was MS+0.5 mg·L^-16-BA+1.0 mg·L^-12,4-D,and the induction rate was 96.22%.The optimal medium for differentiation was MS+1.0 mg·L^-16-BA+0.1mg·L^-12,4-D+0.1 mg·L^-1TDZ,and the differentiation rate was 78.14%.The optimal medium for proliferation was MS+1.0mg·L^-16-BA,and the proliferation coefficient reached 7.85.The optimal medium for rooting was 1/2MS without any hormone,and the regenerated plants with 100%rooting rate could be obtained.There were significant differences in the growth of regenerated plants from different explants after transplantation,and the regenerated plants induced by one generation of proliferative leaflets had the best growth.