The Preliminary Research On Callus Subculture Of Paeonia Suffruticosa

The callus of Paeonia Suffruticosa'Feng Dan Bai','Wu Long Peng Sheng','Shan Hu Tai'that induced from plantlets petiole as materials were used in this research to study the effect of different plant growth regulator,different light situation on the proliferation of callus subculture, and the effect of different medium contents on the browning of callus in subculture was studied. The total RNA extraction method was also explored to establish the basis for the molecular biology study of Paeonia Suffruticosa. The results were as follows:1. The callus that induced from the petiole of Paeonia Suffruticosa was subcultured . the weight gain rate of'Shan Hu Tai'with the 1/2MS+PVP+LH +NAA1.0 mg·L-1+KT0.3 mg·L-1 was 5.15 which was the highest. Within the various light quality treatment of CCFL, ant the the fluorescent lamp as the control group .The weight gain ratio of the fluorescent was highest, and it was 1.11. The weight gain effect of the 100%R,100%B was better than the various R/B light quality, and the white light was the poorest. The callus of Paeonia Suffruticosa'Feng Dan Bai'as explants, and the result was better with R:B=80%:20% and R:B=70%:30% light quality of LED ; The light intensity of 70μmol·s-1·m-2was best for the proliferation of callus of Paeonia Suffruticosa, the weight gain ratio was 0.57, and there was a big different with the other treatments.2. Within the treatment of the various contents of medium on the browning, the browning of'Shan Hu Tai'callus with the additives of NAA(0.5mg·L-1) +6-BA (0.5mg·L-1) or witrh the additives of NAA(0.5 mg·L-1)+ KT (0.3mg·L-1) was the lowest which was 25%. The browning of'Feng Dan Bai'callus was the lowest with the additives NAA(0.1mg·L-1)+TDZ(0.01mg·L-1) which was 7.1%. The concentration of the sugar, the kinds of the sugar, the content of the agar and the PH have a much effect on the browning ratio of callus. For the different sugar source, the browning rate with a sucrose concentration of 50 g·L-1 was the lowest which was 4.55%, the browning rate was lowest with maltose as sugar source, and was 4.76%, but, generally sucrose was the better choice for the subculture of callus, the browning rate of callus was only 5.56% after cultured for 30d. When with a little acid, the medium was better for the proliferation of the callus .With a PH of 6.0 or 6.5, after the callus was culture for 30d in the dark environment, the browned tissue was wrapped by the new proliferation callus, so it was seemed that the browning rate was 4%; With a agar concentration of 6.0 g·L-1, the proliferation effect of the callus was the best, the browning rate is only 5% after it was culture for 30d.3. The browning rate of the subculture callus was the lowest with a medium of 1/4MS+1/4Fe, and it is only 18.2%, and the most economic culture period was 20d. According to the species of the Paeonia Suffruticosa, the browning rate of callus subculture was least for the'Feng Dan Bai', as'Wu Long Peng Sheng'and'Shan Hu Tai'followed.4. According the experience about total RNA extraction of the Paeonia Suffruticosa callus, It seems that CTAB is more better than Trizol, for the Paeonia Suffruticosa was polyphenol and polysaccharide. Although CTAB was time and vigour, it is economic and practical, what is more, it is much cheaper than using test kit.