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 Posted: Wed 19:29, 06 Apr 2011 Post subject: Agrobacterium-mediated phytoene synthase gene into |
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Agrobacterium-mediated phytoene synthase gene into the callus of ginseng factors
To 10d, infection time to 10min, a total culture time was 1d, the infection, infection bacterial concentration will be best defined as 0.6. Bacterial concentration and the relationship between infection rate shown in Figure 2. 3025 volume 2015105O00.20.40.60.81.01.21.4 bacterial concentration / OD Figure 2 bacterial concentration and the relationship between infection rate 8OO clever 88,450 Changchun University of Technology (Natural Science) Vol.28 2. age 3 to determine the best cell infection bacterial concentration OD. . . 0.6, infection time was 10min, a total culture time was 1d, the infection. 15d callus cells cultured with high infection rates. Ginseng callus cells the relationship between age and infection rate shown in Figure 3. G oak 510152O2530 ginseng callus Age / d Figure 3 ginseng callus cells the relationship between age and infection rate of 2.4 to determine the best time of infection bacterial concentration OD. . . 0.6 cell age of ginseng cells 15d, a total culture time was 1d, the infection. Infection rates of infection between time and shown in Figure 4. Infection shows that the optimum time for 15min. 010152O253O35 infection time / min infection time and the relationship between infection rate of 2.5 to determine the best time of co-culture bacterial concentration OD. . . 0.6 cell age of ginseng cells 15d, infection time was 15rain, the best co-culture experiments of the time. Will be identified as the best co-culture time after infection 2d. Total culture time and the relationship between infection rate shown in Figure 5. Total incubation time / d Figure 5 co-culture time and the relationship between infection rate of 2.6 Identification of transgenic ginseng 2.6.1 Results from the external morphological features can be clearly seen that non-transgenic ginseng callus cells were white, while the transgenic ginseng callus cells showed orange color were significantly different. Normal ginseng and ginseng callus cells of transgenic contrast shown in Figure 6. (A) normal ginseng (b) transgenic ginseng ginseng Figure 6 normal and transgenic ginseng callus cells of transgenic compared 2.6.2PCR results of the PCR test results Ginseng shown in Figure 7. M123456M: XDNA / EcoRI ten HindIII; 1: positive control; 2: untransformed plants; 3 ~ 6: Figure 7 has been genetically transformed plants ginseng ginseng PCR results of transgenic cell lines and plasmids were amplified in the vicinity of specific 1.3kb band, while the negative control strains failed to amplify the specific bands corresponding, which initially confirmed that the gene has been transferred PSY ginseng callus cells. 3 Discussion of the concentration of bacterium foreign genes leaders, the bacteria in the logarithmic growth phase dynamic stronger, higher conversion efficiency. The results showed that infection bacterial concentration of 0.6, the bacteria in the logarithmic phase, with high conversion efficiency. The paper used in ginseng callus cell growth cycle is about 25 ~ 30d, in the course of the study found that cell growth 15d or so, has the highest efficiency of Agrobacterium transformation. Speculate at this time in the best ginseng callus cell growth state and differentiation rate is high, which will help the leaders of foreign genes. Ginseng callus cells as Agrobacterium-mediated transformation of the receptor, surface area, thick cell wall, leading to easy access to a wound Agrobacterium cells. Agrobacterium transformation of plants and when must such as adding O 4 pour Yao Liu Wei, et al: Agrobacterium-mediated phytoene synthase gene in callus of ginseng into the wound site 451, the survival factors 16h or more, to the T-DNA transfer. This period involves the attachment of Agrobacterium to the injury site _4], Vir region genes induced activation of T-DNA transfer and integration. Therefore, the infection takes a long time and a total culture time. But if the infection time is too long, post-cultured callus cells mostly appeared the phenomenon of soft rot,[link widoczny dla zalogowanych], and in the subculture, the pollution prone to the phenomenon of re-living bacteria, resulting in ginseng callus cells wither and die. The application of transgenic technology can not only expand the natural source of carotenoids, but also can significantly increase the accumulation of the original carotenoid content. Ginseng can be used as natural carotenoids not only the production of raw materials, while itself is a food or food processing, raw materials, thus increasing carotenoid content in ginseng can directly improve the quality of its appearance and make ginseng has a higher nutritional value.
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