• Journal of Plant Biotechnology
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• 제3권2호
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• pp.107-112
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• 2001

The kinetics of Ti-transformed Salvia miltiorrhiza cell cultures was studied in 250-$m\ell$ shake flasks by using B5 medium with addition of 30 gfL of sucrose. In the cell cultures, the maximum cell mass obtained was 11.5 g DW/L on day 15. The highest amount of phenolic compounds - rosmarinic acid (RA) and lithospermic acid B (LAB) reached 871.3 mg/L (day 15) and 121.3 mg/L (day 13), respectively. The total tanshinone production, i.e., intracellular plus extracellular cryptotanshinone, tanshinone 1, and tanshinone IIA, was 5.3 mg/L on day 13. For the cultivations in 2.4-L stirred bioreactors, the residual sugar level and medium conductivity were a little higher in a small turbine impeller reactor ($T_s$) than those in a large turbine impeller reactor ($T_L$), while a higher cell density was obtained in the $T_L$. For the production of tanshinones and phenolics, better results were obtained in the $T_L$ than in the $T_s$. In the $T_L$, similar or even a little higher production titers of tanshinones and phenolic compounds were achieved compared to those in the flasks. The results suggest that the shake flask results could be successfully scaled up to the $T_L$ reactor. Such a large impeller reactor like $T_L$ may be better than a small impeller one for the large-scale production of the valuable metabolites by the suspension cultures of Ti transformed S.miltiorrhiza cells. This is considered due to the beneficial culture environment in the $T_L$, such as low shear rates as estimated theoretically.

• Journal of Applied Biological Chemistry
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• 제49권1호
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• pp.1-7
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• 2006

Differential display reverse transcription PCR (DDRT-PCR) analysis was performed on lily 'Marcopolo' bulb scale for isolation of expressed genes during bulblet formation. Cu/Zn lily-superoxide dismutase (LSOD) of 872 bp gene, with ability to scavenge reactive oxygen in stress environment, was isolated. Northern blot analysis showed expression levels of LSOD maximized 12 days after bulblet formation. Ti plasmid vectors were constructed with sense and antisense expressions of LSOD gene and transformed into potato. Southern blot analysis of transgenic potatoes revealed different copies of T-DNA were incorporated into potato genome. In transgenic potatoes, lily SOD gene was overexpressed in sense lines and not in antisense lines. In native polyacrylamide gel electrophoresis analysis, additional engineered LSOD was detected in sense overexpressed transgenic line only. Transgenic potatoes were subjected to oxidative stress, such as herbicide methyl viologen (MV). Transgenic potato lines with sense orientation exhibited increased tolerance to MV, whereas in antisense lines exhibited decreased tolerance. In vitro tuberization of transgenic potato with sense orientation was promoted, but was inhibited in transgenic potato with antisense orientation.