Studies On Tissue Culture And Effects Of Different Antibiotics At Various Concentrations On Culture Of Ligusticum Chuanxiong Hort.

Ligusticum chuanxiong Hort. of Umbelliferae is truebom Chinese traditional medicine in Sichuan province in China. Ligusticum chuanxiong Hort. is planted in many regions in our country, and it is mainly planted in Du Jiangyan in Sichuan province. We make use of its dry rhizome clinically and it can cure a good deal of disease. The officinal effective components of Ligusticum chuanxiong Hort. are mainly in their rhizomes. Studies proved that the contents of naphtha and effective components in their rhizomes were generally the same as in their roots. Therefore, calli induced from roots probably have many kinds of officinal effective components. It is very important for production and medicine studies.The tissue culture and plant regeneration of Ligusticum chuanxiong Hort. were studied when the explants were roots, and a large number of regenerated plantlets were obtained. When the explants were roots, the optimal compounding of medium that induced calli, formed shoots and roots were the MS medium supplemented with 6-BA 0.8 mg/L and NAA 1.2 mg/L, the MS medium supplemented with 6-BA 2.2 mg/L and IAA 0.3 mg/L and the 1/2 MS medium supplemented with IBA 1.0 mg/L and NAA 0.7 mg/L, respectively. We are going to do further experiments to test the components in calli induced from Ligusticum chuanxiong Hort.'s roots.Comparing with root explants in tissue culture, leaf explants and stem explants were also studied in plant regeneration.Vitrification plantlets and etioiation plantlets appeared easily in tissue culture of Ligusticum chuanxiong Hort., so we chose and obtained the optimal compounding of medium for shoots growth: the MS medium supplemented with GA₃ 1.2 mg/L, 6-BA 1.8 mg/L and IAA 0.3 mg/L. The shoots growth medium could make the plantlets grow healthy and strong in a short time (about 14 days), and vitrification plantlets and etiolation plantlets rarely appeared.0.6%-0.7% agar and pH 5.8 were optimal for mediums above. The tube seedling could be successfully transplanted.In Agrobacterium mediated transformation of plant genetic transformation, how to choose an antibiotic and the concentration is very important. In Agrobacterium mediated transformation, antibiotics can be divided into two kinds according to their effects: one kind of antibiotics is used to inhibit Agrobacterium and other bacteria; the other kind of antibiotics is used to select transformers and test their offspring.The stems, leaves, roots and the corresponding calli of Ligusticum chuanxiong Hort. were selected to be experimental materials to carry out research on effects of four antibiotics on their growth. Different concentrations of various antibiotics were added into MS culture media with hormone combinations and then their influences on the growth of the explants and calli were compared. After analysis of variance and overall observation of tissue culture, it was concluded that when we chose kanamycin to select transformers, the best resistance selection concentration was between 20～40mg/L; when we chose rifampicin to select transformers, the best resistance selection concentration was about 200mg/L.Ampicillin and tementin were two ablastins. The concentrations of them varied with different bacteria and we should take the both effects of inhibiting bacteria and negative influence of inducing calli by antibiotics into consideration. We should choose the lowest concentration that inhibited bacteria and had the least negative influence of explant and callus culture by antibiotics. As a result, tementin was very harmful for explant and callus culture ofLigusticum chuanxiong Hort., so we were not able to choose tementin to be the ablastin; ampicillin had less influence on explant and callus culture of Ligusticum chuanxiong Hort., so we could choose ampicillin to be an ablastin, and its inhibition concentration was 200mg/L.