The Research On Tissue Culture Of Peony (Paeonia Suffruticosa)

The research took buds, leaves and leafstalks of different species of peony ( Paeonia Suffruticosa ) as explants to probe into the influence of different disinfectors and sterilized time on explants by Orthogonal design; Different basic medias, regulators, accessories and environmental factors were studied for their effects on buds of peony in the course of inducement, proliferation and inducing root, the main influence factors and effective concentration scale were analyzed. Further more to study the training and transplant of rooting peony; At the same time, the leaves and leafstalks of peony were used as explants in order to get the main influence factors and effective concentration scale in the callus inducement and differentiation, then adventitious buds from callus of leafstalk were induced root, to optimize the technique of callus inducement and differentiation. The results of experiments as follows:1. The buds of peony were used as explants, to study the effect of different disinfectors and sterilized time on explants. The results showed that: the optimal disinfector and sterilized time for bud are 0.1%HgCl2 and 8min, the suitable time for leaf and leafstalk is 3min and 5min.2. The buds of cv.'Da Hu Hong', cv.'Wu Long Peng Sheng'and cv.'Jin Ge'of peony were used as explants, to study the effect of different basic medias and species on bud inducing. The results showed that: the best medium is 1/2MS; the best percent of bud inducing is cv.'Jin Ge'.3. Different regulators and AC were studied in the course of bud inducing of cv.'Jin Ge'. The results showed that: the impact of different factor on bud inducing is different, 6-BA > NAA > AC > IAA; the best percent of bud inducing was obtained with medium 1/2MS + 6-BA 1.0mg/L+ NAA 0.2mg/L+IAA 0.3mg/L+AC 3.0g/L.4. The buds of cv.'Jin Ge'were used as explants, to study the effect of different basic medias and regulators on bud proliferation. The results showed that: the effect of factor on the proliferation of bud: NAA>basic medium>6-BA>IAA; the best percent of bud proliferation was obtained with medium 1/2MS+6-BA 1.0mg/L+NAA 0.5mg/L+GA3 0.2mg/L. In addition, different cycle of subculture had different impact on bud proliferation; the optimal cycle of subculture is thirty days.5. The leaves and leafstalks of different cv.'Da Hu Hong', cv.'Wu Long Peng Sheng'and cv.'Jin Ge'were used as explants, to study the effect of different species and types on percent of callus inducing. The results showed that: cv.'Jin Ge'had the best percent of callus inducing in the different species; leafstalk was better than leaf in the callus inducing.6. Different basic medias were studied in the percent of callus inducing of cv.'Jin Ge'. The results showed that: the optimal medium for leaf and leafstalk callus inducing are 1/2MS and WPM.7. The leaves and leafstalks of cv.'Jin Ge'were used as explants, to study the effect of different regulators and CH on the callus inducing. The results showed that: NAA, 2,4-D and 6-BA are the main effective factors in callus inducing by leaf, the optimal medium is 1/2MS+2,4-D 2.0mg/L+6-BA 1.0mg/L+NAA 1.0mg/L+CH 300mg/L; 6-BA and 2,4-D are the main effective factors in callus inducing by leafstalk, the optimal medium is: WPM+2,4-D 2.0mg/L+6-BA 1.0mg/L+NAA 1.0 mg/L+TDZ 0.5mg/L+CH 300mg/L.8. Different combination of regulators was studied in the callus differentiation of cv.'Jin Ge'. The results showed that: the leafstalk had higher percent of callus differentiation than leaf in the same cultural case; 2,4-D 0.25mg/L+TDZ 0.5mg/L is helpful to the callus differentiation; In addition, different cycles of subculture had different effect on callus differentiation, the optimal turning is 30d after inoculating.9. Different basic medias, regulators, sugar concentration and time of dark treatment were studied in the course of root inducing by cv.'Jin Ge'test-tube plantlet. The results showed that: the effect of factor on the percent of root inducing: basic medium>IAA>IBA>sugar concentration>time of dark treatment; the effect of factor on the average length of root: basic medium>time of dark treatment >IAA>sugar concentration >IBA; the best medium for root inducing is: WPM+IBA 1.0mg/L+IAA 0.5 mg/L +sugar 20 g/L + dark culture 5d; the best medium for root length is: WPM +IBA 1.0mg/L+IAA 0.5mg/L+ sugar 40g/L+dark culture 10d. In addition, they were used to study on the percent of root inducing, IBA 2.0mg/L is hopeful to the root inducing by the adventitious buds from leafstalk callus of cv.'Jin Ge'.10. The test-tube plantlets were used as explants, to study training and transplant. The results showed that: the optimal training time is 12d in the temperature of 15~20℃and the moisture of 85%~90%; the best percent of transplant survival is the combination of perlite and vermiculite (1:1).