文冠果懸浮體系的建立和原生質(zhì)體的分離.pdf

1、河北農(nóng)業(yè)大學(xué)碩士學(xué)位論文文冠果懸浮體系的建立和原生質(zhì)體的分離姓名：劉磊申請學(xué)位級別：碩士專業(yè)：植物學(xué)指導(dǎo)教師：顧玉紅2009-06-12Establishment of Suspension Culture and Preparation of Protoplasts in Xanthoceras sorbifolia Bunge Author: LIU Lei Major: botany Supervisor: GU Yu-hong

2、 Abstract Xanthoceras sorbifolia Bunge is a plant belonged to Sapindaceae, which is one of the peculiar woody fuel and medicinal plants in China. This study dealt with the induction of embryogenic callus from mature embr

3、yos, establishment of suspension cell lines. Moreover, the leaves of tissue culture seedlings from Xanthoceras sorbifolia Bunge were used as experiment materials.Through selecting multi-impact factors of the separation o

4、f protoplasts, a good system of protoplast isolation was established. The results obtained as follows. The study used mature embryo as a test material and screened the optimal culture conditions of callus induction, call

5、us subculture. In addition, the activities of antioxidant enzyme and contents of solubie protein in the callus from the seed of Xanthoceras Sorbifolia Bunge were investigated. The results showed that callus was induced f

6、rom stem on B5 supplemented with 2 mg/L 2,4-D + 20 g/L sucrose ＋ 5.5 g/L Agar(pH5.8)，at the temperature of (25±2)℃ with darkness. B5 + 1 mg/L 2,4-D + 1 mg/L 6-BA + 20 g/L Sucrose + 5.5 g/L Agar(pH5.8)could be the b

7、est culture conditions of callus subculture , with Low-light training, (25±2)℃. POD activity of white callus and yellow callus was significantly higher than transparent callus; CAT activity of white callus and tran

8、sparent callus was extremely obviously higher than yellow callus, CAT activity of white callus was significantly higher than transparent callus; SOD activity of white callus was significantly higher than transparent call

9、us and yellow callus; Soulble protein content of yellow callus was significantly higher than white callus and transparent callus, soulble protein content of white callus was significantly higher than transparent callus.T

10、he development of callus was related to the activities of antioxidant enzyme and contents of solubie protein. The experiment obtained callus from subculture culture for materials, established suspension cell line of Xan

11、thoceras Sorbifolia Bunge and studied its growing characteristic. The results showed that the optimum initial inoculum amount was 2g in 40ml liquid culture medium, and the optimum hormone concentration were 0.5 mg/L 2,4-

12、D and 0.5 mg/L 6-BA and 0.5 mg/L NAA. The highest cell mitotic index, cell activity and increased content of fresh weight appeared on cell suspension culture[0] 10th day, 3th day and 14th day each other. The leaves of ti

13、ssue culture seedlings from Xanthoceras Sorbifolia Bunge were used as experiment material and the influences of mannitol, cellulase, pectinase, and enzyme time on protoplast isolation from them were studied. The results