執行成果摘要
In previous studies, 5 eucalyptus clones (3 transgenic clones and 2 wild type clones) were selected and cultured in isolated green house. The integrations of transgenes of transgenic eucalyptus trees were confirmed by using PCR and transgene integration patterns were characterized by Southern analysis. Differences of expression level of transgenes were characterized by real-time PCR method. The lignin contents and total cellulose contents of wood samples were analyzed by using Klason method and CNS 3085 standard protocol. The syringyl and guaiacyl (S/G) ratio in lignin was measured by using pyrolysis-GC Mass method. In this year, the pulping characteristics of 5 selected eucalyptus clones were evaluated by micro-pulping experiments following the kraft pulping method. Meanwhile, the kappa number of pulp was evaluated according to the CNS 5470 (Method of test for KAPPA number of pulp) standard protocol. A new isolated net-room was supposed to be designed and constructed by the end of this year. The design of our second isolated net-room was finished and the plan of water and soil protection was confirmed. However, the construction of the isolated net-room would not be finished by the end of this year because the public tenders were failed. The second objective is to develop an efficient toxoids production system by using bioreactor culture techniques with elite transgenic cell lines of Taxus sumatrana. In previous studies, several elite transgenic cell lines with different combination of key toxoids biosynthesis genes were produced. High BC yield transgenic cell lines were selected and the cell growth rate and BC production were evaluated by using both disposable bioreactor and stir type bioreactor. The results indicated that production systems of taxoids by using transgenic cell lines performed well in 2L bioreactors. In this year, different combination of key gene constructs (BAPT and TS) related to taxoids biosynthesis were transformed into superior cell lines of Taxus sumatrana, and 54 new transgenic cell lines were induced and cultured. The transgenic cell line O1 harboring (sense TS , sense DBAT and antisense BAPT genes) was selected and the cell growth rate and BC production were evaluated by using both disposable bioreactor and stir type bioreactor. The results indicated that production systems of taxoids by using transgenic cell lines performed well in 2L bioreactors. Meanwhile, 2 culture experiments of transgenic cells (R1 and R2) were carried out by using 20L wave bioreactors, and the cell growth rate and taxoids production were evaluated.