| The axillary buds of four breeds (P.suffruticosa'WuLongPengSheng', P.suffruticosa'HuHong', P.suffruticosa'LuoYangHong') were tested as explants in order to study the effects of hormone proportion, breeds variation on germination rate and vitrification rate in bud induction and that on multiplication coefficient and browning rate in subculture ; Using the test-tube plantlets P.suffruticosa'WuLongPengSheng'as tested materials to study the effects of hormone proportion on rooting induction , At the same time , changes of total soluble sugar, starch, free amino acids, soluble protein, total phenol contents, nucleic acid and chlorophy contents were determined in the process of rooting culture ,and the root system vigor in vitro was also determined. The rooting test-tube plantlets were domesticated and transplanted to explore the effects of different media on the survival rate of transplant. At last , studys on Anotomical and morphological of roots were investigated by paraffin sections, the results are as follows:1.The optimum hormone proportion for axillary bud germination was NAA:6-BA=0.3:0.5 (mg·L-1) , Differences between varieties manifested as P.suffruticosa'WuLongPengSheng'> P.suffruticosa'LuoYangHong'> P.suffruticosa'HuHong'. Effects of different concentrations of 6–BA on the vitrification rate of plantlets were significant, With the increasement of 6-BA concentration , the vitrification rate was gradually increased . The different concentration of NAA does not have the remarkable influence to the vitrification rate. When NAA×6-BA for 0.1×1.0 mg·L-1, the vitrification rate reaches 14.69% ,when NAA×6-BA for 0.3×0.5 mg·L-1 the vitrification rate reduced to4.05% only;differences between varieties manifested as, the'LuoYangHong'has the highest vitrification rate, nextis'HuHong', the'WuLongPengSheng'has the lowest,vitrification rate, it displays remarkable difference.between the three varieties.2. The optimum medium for subculture was'MS+NAA1.0mg·L-1+6-BA2.0mg·L-1', Effects of different concentrations of 6-BA on the multiplication coefficient were remarkable. When 6-BA is 2.0mg·L-1 the multiplication coefficient is biggest;differences between varieties on multiplication coefficient manifested as P.suffruticosa'LuoYangHong'>P.suffruticosa'WuLongPengSheng'>P.suffruticosa'HuHong',differences between three cultivars were remarkable.Effects of different concentrations of 6-BA on the browning rate in subculture were remarkable.with the increase of 6-BA concn. the browning rate was increasing.When 6-BA concn.was 3.0 mg·L-1, the browning rate is highest and when 6 - BA concn .was0.5 mg·L-1, the browning rate was the lowest ;differences between varieties on the browning rate manifested as P.suffruticosa'LuoYangHong'> P.suffruticosa'HuHong'> P.suffruticosa'WuLongPengSheng', differences between three cultivars were remarkable.3. In this work,tow method were used to induce root :Conventional culture and low temperature with no light,The result indicated that: Effect of basic medium on root-inducing of test-tube plantlet P.suffruticosa'WuLongPengSheng'was 1/2WPM>1/2MS+Ca2+,AC was inhibition to root-inducing ,The optimum medium for rooting culture was'1/2WPM+NAA3.0mg·L-1 +PVP1.0g·L-1 +GellanGum 2.0 g·L-1'with highest rooting rate 66.67%,the culture conditions was low temperature with no light, but the Conventional culture with the highest rooting rate 26.67%. With the increasement of NAA concn.the average root number was increasing, Effects of different hormone on average rooting number in the conventional culture condition manifested as:'ABA﹥NAA',however,when in the condition of low temperature with no light,it shows the opposite result:'NAA>ABA', the optimum hormone proportion for average root number was IBA/ABA=3.0/0.1 .Effects of different concentrations of IBA on average root length were not remarkable. ABA showed no positive promotion effects on the average root length, When NAA is 3.0 mg·L-1,ABA is 0 ,the average root length is longest; Effects of culture conditions on the average root length were conventional culture>low temperature with no light culture , Effects of the differences between the tow culture methods on root length were remarkable. the longest root reached 15cm long on the conventional culture condition.4. A complex physiological and biochemical changes were accompanied in test-tube plantlets rooting process. a significant increase of soluble sugar content, nucleic acid, phenol content, free amino acid content and chlorophyll content was presented in the adventitious root primordium induction period, at the same time , the soluble protein content and starch content dropped significantly, this may be due to that during the induction process of rooting primordium a large number of cell division, DNA reproduced quickly, various amino acids and nucleic acid content increased, a large amount of protein was consumed due to the decline of soluble protein content in vitro, some free amino acids into proteins, starch used to reduce the degradation of a soluble sugar rooting for the energy generated in the formation of adventitious root soluble sugar was transported to the base in vitro to ensure that the needs of adventitious root formation by the large number of nutrients, resulting in soluble sugar content also dropped significantly.5. The results of the transplantion of rooting shoots indicates that using three kind of matrix mixed with peat,pearlite and vermiculite , among the 20 shoots only 5 shoots were survived, the survival rate was low.6. The root system vigour increased with the increase of root number, but the quality of root system was not very good. Effects of culture conditions on the root system vigour manifested as: conventional culture > low temperature with no light culture.7. The result of paraffin sections indicated that the rhizome connection : the adventitious root primordium was induced from the callus,and the vascular bundle of rhizome was blocked by callus ,it results in the conducting tissue did not connect with each other. But the concrete position of root primordium needs further reseach. |
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