Changes of alkaloids in Tibetan medicine Przewalskia tangutica during tissue culture and differentiation of its stem rapid propagation system

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1 386 1, 2, 3 1 1, 2, 3 1, 2, 3* Przewalskia tangutica 3 d 2 3 cm HPLC 4 MS 6-BA 2. mg/l NAA.5 mg/l 96.67% MS NAA.5 mg/l 71.67% HPLC μg/g R A (215) DOI: 1.751/j.issn Changes of alkaloids in Tibetan medicine Przewalskia tangutica during tissue culture and differentiation of its stem rapid propagation system LEI Tian-xiang 1, 2, 3, WANG Huan 1, CHEN Shi-long 1, 2, 3, ZHOU Dang-wei 1, 2, 3, CAI Xiao-jian 4, LI Song-ling 4, SHEN Jian-wei 1 1. Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 818, China 2. Key Laboratory of Adaptation and Evolution of Plateau Biota, Chinese Academy of Sciences, Xining 818, China 3. University of Chinese Academy of Sciences, Beijing 149, China 4. Institute of Soil and Fertilizer, Qinghai Academy of Agriculture and Forestry Sciences, Xining 8116, China Abstract: Objective To establish the stem rapid propagation culture system of Prezewalskia tangutica using aseptic seedling of P. tangutica as materials. Methods The stems from one month aseptic seedling were inoculated on Muragshige and Skoog media supplemented with different phytohormones to select the optimum medium for inducing differentiation of the adventitious buds and roots. Furthermore, the contents of four active alkaloids were measured with HPLC. Results The optimum medium was MS + 6-BA 2. mg/l + NAA.5 mg/l for inducing the propagation of the adventitious buds. The adventitious roots could be induced on MS + NAA.5 mg/l media. The inductivity was 71.67%. The HPLC data suggested that the content of total alkaloids is highest in root ( μg/g). The content of total alkaloids in callus was lower than those in roots, stems, and leaves. Conclusion The contents of total alkaloids in different fractions of P. tangutica seedlings are different, and the proportion of different alkaloids in total alkaloids is different, which indicates that the formation and dedifferentiation of organ can influence the contents of alkaloids in P. tangutica seedlings. Key words: Przewalskia tangutica Maxim.; tissue culture; differentiation; tropane alkaloids; inductivity Przewalskia tangutica Maxim. Solanaceae Przewalskia Maxim m Y Y KSCX2-EW-J m @163.com dangweizhou@sina.com

2 387 [1-2] 1984 [3] 54 [4] 1 [5-7] [8] [4,9-12] [4] m Przewalskia tangutica Maxim MS lx 12 h/d MS 8% 3 g/l ph Table 1 Effects of different phytohormone combination on inductivity and multiplication rates of P. tangutica adventitious buds 6-BA/(mg L 1 ) NAA/(mg L 1 ) /% I I I I I I I I I I I I The developmental state of plantlet of P. tangutica was showed by different sign: +++ is best, ++ express good, + express most of buds were not capable of generate new buds

3 HPLC 45 4 [11] MS 1 6-BA/NAA % 2.21 NAA 6-BA NAA MS 6-BA 2. mg/l NAA.5 mg/l A A C A- B- C- D- A-adventitious buds induced from axillary bud B-rooting differentiation C-regenerated plantlet D-callus Fig. 1 B D 1 Different phases of tissue culture of P. tangutica 2.2 NAA NAA 2 NAA NAA.5 mg/l 3 4 /% NAA /(mg L 1 ) 2 NAA Fig. 2 Effects of NAA with different concentration on rooting of P. tangutica stems 71.67% B 1-C 2.3 HPLC 4 HPLC μg/g [11] 4 /(μg g 1 ) b a c b a c c d c d b d (.5) Significant differences were expressed by differrent lowercase (P <.5) 3 4 Fig. 3 Contents of four alkaloids in different tissues of P. tangutica seedlings b c a a /

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