Study On Tissue Culture And Regeneration Of Viola Prionantha Bunge

Viola prionantha Bunge is an excellent wild ground-cover herb who is belong to Viola of Violaceae.This regeneration system of Viola prionantha was established in the study,and the cotyledon section,the leaf blade and petiole of Viola prionantha Bunge were used as explants for tissue culture.The system enriches the mode of reproduction of Viola prionantha Bunge,it provides technical assurance for its large-scale application in urban cover greening and makes full use of ornamental value of Viola prionantha Bunge.The main results of this paper were as follwing:1?The best sterilization methods of explants of Viola prionantha Bunge:(1)15 seconds is good to treat the cotyledon section by C2H5OH(70%),5-7minutes is good to treat the cotyledon section by HgCl2(0.1%).(2)The best time for sterilization the leaf blade is 30 seconds and 8 minutes respectively under C2H5OH(70%)and HgC12(0.1%).2?Primary culture:Under light conditions,the appropriate medium for the induction of normal buds from the cotyledon section and adventitious buds from the petiole of Viola prionantha Bunge was MS+6-BA 2.0mg·L-1+ NAA0.1 mg-L-1;the induction rate of the cotyledon section was 98.3%,the shoot forming index of the cotyledon section was 5.36.The leaf blade was not suitable as the explant to inducte adventitious bud directly.All three kinds of the explant(the cotyledon section?the leaf blade and the petiole)could induce callus.The cotyledon section could induced callus for the shortest time,followed by the petiole,the leaf blade the latest.The optimum plant growth regulator combination for the induction of callus from the cotyledon section and the petiole and the leaf blade of Viola prionantha Bunge was MS+6-BA 1.0 mg-L-1+2,4-D 0.5 mg-L-1,and the rates of the induction of callus respectively were 98.3%?96.7%?88.3%.3?Subculture:The best medium for the proliferation of normal buds and adventitious buds was MS+6-BA 1.5 mg·L-1 +NAA 0.075 mg-L-1,the proliferation rate was 6.70.The optimum combinations of plant growth regulators for the cotyledon section?the leaf blade and the petiole callus subculture of Viola prionantha Bunge is MS+6-B A 1.0 mg·L-1+2,4-D 0.75mg·L-1,with the callus growth and health,the multiplication rate of Viola prionantha Bunge callus was 4.97;The effect of multiple types of callus on the multiplication rate is not significant.The best medium for the differentiation of callus of Viola prionantha Bunge was MS+6-BA 1.5 mg-L-1 +NAA 0.1 mg·L-1,the differentiation rate was 100%and the average number of adventitious buds was 8.03.4?Seedling culture:The plants grown well and health in the MS medium without any plant growth regulators5?Rooting induction culture:The most suitable medium for the rooting of the renewable effective seedlings of Viola prionantha Bunge was 1/2MS+6-BA 1.0 mg·L-1,with the rooting rate was 92.3%,and the average number of rooting was 5.39,the length of root was 4.8(cm).6?Hardening seeding and transplanting:the optimized transplant medium for Viola prionantha Bunge was perlite,riversand and humus mixed by 1:2:2,which the rate of survival was highest,Up to 100%,and the growth is very well.