Study On The Antibacterial Activity And The Active Ingredients Of Tinospora Capoillpes Gag-nep.

The antifungal activity of 104 species of plant in 64 families are collected from the north and southeast of Guangxi province.Based on the activity screening, the active ingredients of Tinospora capoillpes Gag-nep. showing high inhibitory activity against Colletotrichum gloeosporidides(Penz.)Sacc were isolated by activity-directed isolation method, and the chemical structures were identified. The antifungal mechanism of these Tinospora capoillpes Gag-nep. was also preliminarily studied.The antifungal activity of 116 species of plant extracts from 104 species of plant against C. gloeosporidides was determined by mycelium growth rate method. The results showed that 6 kinds of plant extracts had broad-spectrum antifungal activity, with the inhibitory rates of above 100% against C. gloeosporidides at 10mg/mL. These plants included T. capoillpes, Stephania kwangsiensis Lo., Stephania cepharantha Hayata, Acorus tatarinowii Schott., Coptis chinensis Franch.. 3 kinds of plant extracts with the inhibitory rates of above 80% included Tupistra wattii (C. B. Clarke), Eupatorium adenophorum Spreng and Saussurea costus (Falc.). The results showed that the inhibition rates of them were 88.33%,87.98%,81.24%.And most of the other plant extracts had significant antifungal activity against the C. gloeosporidide.The toxicity of the extracts of T. capoillpes., S. kwangsiensis, S. cepharantha, A. tatarinowii, C. chinensis. to C. gloeosporidide was further determined. The results showed that with the EC50 value of 0.7101mg/mL,2.2073mg/mL,0.8391mg/mL,0.1528mg/mL,0.8872mg/mL,3.0666mg/mL,0.1654mg/mL .the extract of T. capoillpes had the highest toxicity to C. gloeosporidide ; the extract of A. tatarinowi with had the least toxicity to C. gloeosporidide.and the rate of the highest toxicity is twenty times than the least.The extract of T. capoillpes. was isolated into 5 parts by the method of alkaloid isolation, and bioactivity measurement showed that the active ingredients mainly consisted in the chloroform fraction. Two active compounds were isolated from T. capoillpes by silica gel column chromatography using activity-directed isolation method, and identified for palmatine and jatrorrhizine by NMR and MS.The toxicity of palmatine to 11 kinds of plant pathogenic fungi was determined by mycelium growth rate method, and the results showed that the highest EC50 values of palmatine to C. gloeosporidide was 0.0348mg/mL; the results showed that the EC50 values of palmatine to Colletotrichum graminicola (Ces.) Wilson. and Gymnosporangium haraeanum Syd. were 0.1302mg/mL,0.1356mg/mL .higher than another kinds of plant pathogenic fungi. Palmatine also had strong inhibitory activity against animal pathogenic bacteria, the MIC values of palmatine to Bacillus subtilis (Ehrenberg) Cohn was 0.1 mg/mL. The MIC values of palmatine to Staphyloccocus aureus Rosenbach, Micrococcus lysodeikticus Fleming, Salmonella typhi (Schroeter) Warren & Scott were 0.2 mg/mL. The MIC values of palmatine to another 5kinds of animal pathogenic bacteria were in 0.3 mg/mL to 0.8mg/mL. The MIC values of palmatine to Micrococcus luteus(Schroeter) Cohn, Pseudomonas aeruginosa (Schroeter) Migula, Shigella dysenteriae (Shiga) Castellani & Chalmers were above 1mg/mL.The toxicity of jatrorrhizine to 11 kinds of plant pathogenic fungi was determined by mycelium growth rate method, and the results showed that the highest EC50 values of jatrorrhizine to C. gloeosporidide was 0.0240mg/mL; the results showed that the EC50 values of jatrorrhizine to C. graminicola and G. haraeanum were 0.1158mg/mL,0.1097mg/mL .higher than another kinds of plant pathogenic fungi. Jatrorrhizine also had strong inhibitory activity against animal pathogenic bacteria, the MIC values of jatrorrhizine to B. subtilis was 0.1 mg/mL. The MIC values of jatrorrhizine to B. cereus, M. lysodeikticus, P. vulgaris, S. typhi were 0.2 mg/mL. The MIC values of jatrorrhizine to another 4kinds of animal pathogenic bacteria were in 0.3 mg/mL to 0.6mg/mL. The MIC values of jatrorrhizine to M. luteu, P. aeruginosa, S. dysenteriae were above 1mg/mL.The mycelium shape of plant pathogenic fungi was changed by treating with the extracts of T. capoillpes and active compounds palmatine and jatrorrhizine, some of the mycelium were intumescent, monstrous, twisted, cracked and so on. Palmatine and jatrorrhizine led to different degree destruction of cell membranes of 4 kinds of pathogenic fungi, and also had different degree inhibition on the protein synthesis of them .