Studies On Tissue Culture Of Paeonia Ostii And The Effects Of Cultured Conditions On The Content Of Paeonol In Callus

Paeonia ostii is a kind of Paeonia suffruticosa Andr which can be used for medicine, food and enjoyment. It is particularly worth of the clinical application and development of the paeonol in its root. Plant tissue culture is a fast way to acquire consistent materials. However, there are less researches on officinals than ornamentals in the studies of tissue culture of Paeonai surffruticosa, especially on the physiological activity. So, in order to offer the theoretical reference for tissue culture of officinal species of peony, the researches on finding the best explants material, safe and effective method of sterilization and the best culture medium for callus culture of Paeonia ostii, as well as the bud and root inducing had carried out. Then, the callus was used to be materials of the research, which is to discover the effects of external culture conditions such as Cu, different hormone combination, light qualities, the phenolic precursor on paeonol’s content, aimed to offer a basis for paeonol’s synthesis and production by tissue culture.1.Tissue culture of Paeonia ostiiThe callus was induced by different explants and used to dedifferentiate buds and roots. The results indicate that the tender stipe obtained by GA3solution treating is the best explant for callus inducing, and its effective sterilization method is that2.5%NaC10treat15-20minutes. The studies on brownings show that there is no remarkable difference in basal mediums such as MS,1/2MS and WPM, and adding in1000mg·L-1PVP as antioxidantand can remarkably decrease the rate of browning, indeed to0%. So, the optimum medium for callus inducing and multiplication is:MS+BA1.0mg·L-1+NAA0.2mg·L-1+LH500mg·L-1+PVP1000mg·L-1, inducing rate reach to97%, or more, and the weight gain multiple is2.86. The callus cultured under the light have much more weight than the one cultured in dark. Adding in DNP can markedly restrain the growth of callus, and Phe can apparently enhance the weight gain multiple, but the difference among the treatments of different concentration of Phe is not obvious. Modulating the ratio of BA and NAA in the medium all can’t make the bud inducing come true, but the callus can achieve root inducing though the rooting rate is low when the concentration of NAA is1.Omg·L-1or1.5mg·L-1, but not considering the IB A as a key factor, because the results of the studies show that IBA added in the basal medium solely can not carry out the rooting, although its concentration is high enough.2. The effect of external conditions on the content of paeonol.Cu:The research was carried out to study the effects of Cu on content of the paeonol in callus by modulating the concentration of Cu in the medium (0.00025,0.025,2.5,25mg·L-1) to culture callus for10d. The results indicate that the concentration and the increment of grosses of the paeonol increased with the increasing concentration of increased Cu, and the results gained from high Cu2+treatment have significant difference from the one gained from the low Cu2+treatment, except the treatments of two low concentration of Cu. Analyzing the relationship between the Cu and paeonol content, it can be seen that Cu2+concentration has a remarkably positive correlation with paeonol content, that is to say that adding Cu2+in moderate concentration can promote the synthesis of the paeonol effectively.Light quality:It can be found that the light quality for the concentration and the increment of grosses of the paeonol in callus are in the order of blue> red> dark> fluorescence, and there is remarkably difference from each other, except for the difference between blue and red. Moreover, the variation of the grosses of the paeonol in callus under the fluorescence lamp is negative, so it is obvious that the synthesis of the paeonol in callus had been restrained and the paeonol had decomposed under the fluorescence lamp.6-BA and NAA:Callus induced from the tender stipes were used as materials for investigating the effects of different concentration of6-BA and NAA on the content of paeonol. After10day’s culture, the results show that the content of the paeonol in callus can be increased remarkably by enhancing the concentration of6-BA or NAA in the trails of single factor, and both of the concentration of6-BA and NAA are1.0mg-L"1, the content of paeonol is at the highest level. In the trial of interaction of6-BA and NAA, the concentration and the increment of grosses of the paeonol in callus is the highest when both the concentration of6-BA and NAA are1.0mg·L-1.DNP and Phe:The growth of the callus of Paeonia ostii is obviously restrained by adding0.2、0.5、1.0mmol·L-1DNP into the medium for4d. The content of the paeonol in the treatments of0.5and1.0mmol·L-1DNP is seriously decreased, compared with the treatment of0.2mmol·L-1DNP, and the variation of the grosses of the paeonol is negative. While in the treatment of adding30、50.80mg·L-1Phe, the concentration and the increment of grosses of the paeonol in callus are remarkably increased, by comparing with the control. But there are not remarkable differences among different concentration treatments.