Research On Tissue Culture Technology And Vitrification Prevention Measure For Paeonia Suffruticosa

In this experiment, peony seeds and soil buds are taken as explants and used in vitro culture and vitrification study. Include established peony aseptic culture system; studding the physiological and biochemical changes during the formation of peony cotyledon callus; Peony test-tube vitrification seeding morphological and anatomical, physiological and biochemical, the control method. The results are as follows:1. Primary culture: The optimal culture medium for peony seed germination: MS+6BA1.0 mg·L^-1+TDZ0.25 mg·L^-1. The optimal culture medium for peony shoot germ-ination: MS+6BA0.5 mg·L^-1+GA30.25 mg·L^-1.2. The optimal medium for callus differentiation: MS+6-BA0.5 mg·L^-1+TDZ0.25 mg·L^-1+2,4-D1.5 mg·L^-1. The optimal medium for inducing indefinite root of regener-ation peony: 1/2MS+6-BA1.0 mg·L^-1+IBA4.0 mg·L^-1, in this condition. The'Tai Yang'and 'Feng Dan Bai' physiological and biochemical indexes of the morphogenetic callus had wider change range than those of the non-morphogenetic callus showing a clear upward trend. While the changes of physiological and bioch-emical indexes of non-morphogene-tic callus changed mildly.3. The establishment of peony propagation system: Making the single-factor test at first, selecting the test factors and levels, Making response surface analysis. Based on the analysis of the signi-ficance and interaction among the factors, the optimum medium is: BA1.05 mg·L^-1 +KT0.41 mg·L^-1+NAA0.77 mg·L^-1+TDZ0.05 mg·L^-1, the proliferation rate of the test results,can raise 4.36. We can obtain the best rooting medium: 1/2MS+6-BA2.0 mg·L^-1+NAA0.2 mg·L^-1+IBA3.0 mg·L^-1, in which the main effect factor is IBA. In this condition the rooting rate of 'painting' can reach 57.98 %.4. There are evident differences betwee the test tube vitrification peony seeds and the normal peony seeding in morphological and anatomical structures.The differences of morphological are: The normal seedling leaves are thin and flat, the plant are robust; the vitrification seeding leaves are small, longitudinal and curling shrinkage.The whole plat stems are transparent sweelling and small. Cytological difference are: The cell gap of normal is smaller and less closeld arranged, the normal peony hare larger number of chloroplast. The cell gap of vitrification seeding is bigger ,the is a larger hole in the cell and less chloroplasts.5. The physiological and biochemical changes of test tube vitrification peony and normal peony are as follows: Compared with normal seedings, vitrification seeding are obviously higher than the normal seedlings;water increased seeding, chlorophyll content, soluble protein, soluble sugar, sugar content, CAT, PAL activity are graclually the decreasing degree of the vitrific-ation., PODactivity and water content are the increase degree of the vitrification.6. The main methods to reduce the peony vitrification:Using good ventilation tampon can reduce the peony vitrification to 6.8 %; Increasing PVA to 3 g·L^-1 can reduce the vitrification rate to 10.7 %; Adjusting pH to 6.6, the vitrification can be controlled to a rate of 13.7 %; Drawing in Februany, we can control the vitrification rate to 3.3 %; When the active carbon is 4⁶ g·L^-1; Through a new medium 10 d, the three treatments have no vitrification phenomenon. 2 buds per bottle the vitrification can be controlled to a rate of 3.3 %; Using different carbon sources, B₅ medium and managing by double KH₂PO₄ can not reduce the rate of vitrification and have no significant effect on controlling the vitrification of the peony.