Study On Tissue Culture And Plant Regeneration Of Persimmon(Diospyros Lotus L.)

Dormant buds taken from one year old persimmon branches as invitro explants materials,by studying the effect of different influence factors on the invitro rapid propagation system,Systematically studied the cultural technique system of early generations,subculture,rooting induction,leaf regeneration of Diospyros lotus L.The main research results are as follows:1.Primary culture of Diospyros lotus L.With 10%H₂O₂ disinfected dormant bud 10 min?20 min?30 min,survival of Diospyros lotus L.rate is 43.3%,63%and 43.3%respectively,the highest survival rate of disinfected time is 20 min dormant bud did not browning and have high survival rate,contamination rate and survival rate 10%H202 was better than using 75%ethanol surface disinfected.This method effectively solve the browning problems of sleeping bud early generation,obviously increase the survival rate of the initial culture.The bud germination vaccinates basic culture medium MS,1/2MS,MS?1/2N?,NN69 etc,which were adding ZT1.0 mg/L+NAA0.1 mg/L respectively,discovered the improvement?1/2N?the MS culture medium is suitable for multiplication growth of tissue culture seedling,the average height of the plant tissue culture is 2.4cm,growing well,the leaf blade is greatly paris green,basal callus small,better than other three kinds medium.2.Subculture of Diospyros lotus L.Studied the effects of different kinds of auxin and cytokinin on the growth and development of tissue culture seedling.The most suitable auxin combination for Diospyros lotus L.multiplication subculture is ZT2.0 mg/L and suitable concentration IAA.For the further study the effect of different hormone combination on Diospyros lotus L.multiplication subculture.Add 0.05 mg/L TDZ in the basis ZT2.0+IAA 0.1mg/l basic media,average plant height and multiplication is 2.53cm and 4.74 respectively,significantly improved transgenerational proliferation of multiplication,showes that appropriate concentration of TDZ has significant influence on transgenerational proliferation propagation of Diospyros lotus L.3.Leaf regeneration of Diospyros lotus L.Using L9?34?orthogonal experimental design,Optimize and filter out the different basic media?TDZ and NAA combination which effect the Leaf regeneration,Filter out the appropriate sugar concentration,hormone types,incubation time,the result showed that:in the MS media adding 40g/L sucrose?3.0 mg/L TDZ?0.1 mg/L NAA then culture in darkness 10 days,callus induction rate of Diospyros lotus L.were 100%,average number of adventitious buds were 4.26 PCS,regeneration rate were 94.28%,Effectively established leaf regeneration culture technology system of Diospyros lotus L.4.Rooting?acclimatization and transplants of Diospyros lotus L.:In MS,1/2MS,MS?1/2N?,NN69,etc taking rooting comparative test,Ms?1/2N?is suitable for rooting induction.NN69 medium doesn't suitable for rooting.Both IBA and IAA can induce tissue culturer rooting,IB A is better than IAA.For the further study the effects of cytokinin on tissue culturer rooting,When adding 0.5mg/L 6-BA?1.0mg/L IBA in?1/2N?Ms and dark culture three days,the average number of roots and rooting rates were 5.06?83.51%respectively,effect was the best.Meanwhile study the effects of mulching film on survival rate during transplanting,over filmed 30 d the Diospyros lotus L.seedling survival rate is 95%.