Effect of cell-cycle inhibition on synthesis of steroidal alkaloids by Solanum aviculare plant cells

Mak, Yunxian; Doran, Pauline M.

doi:10.1007/bf00129932

Cited by 9 publications

(3 citation statements)

References 8 publications

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“…Total metabolic activity inferred from total cellular protein content is relatively uniform in MeJA-elicited cultures (Naill and Roberts, 2005b), indicating that non-cycling cells are still metabolically active, but potentially redirect carbon flux away from primary metabolism towards secondary metabolism. Alkaloid accumulation increased in cultures of Solanum aviculare where cell cycle progression was inhibited using a cell cycle arrest agent, again suggesting that the metabolic flux may be directed towards secondary pathways in non-proliferating cells (Mak and Doran, 1993). Research to date in plant systems has been able to identify a significant G0 population in culture and presents indirect evidence to suggest a function of this population, but has not been able to explicitly correlate the non-cycling cell population to other metabolic information.…”

Section: Discussionmentioning

confidence: 99%

Methyl jasmonate represses growth and affects cell cycle progression in cultured Taxus cells

et al. 2014

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Methyl jasmonate (MeJA) elicitation is an effective strategy to induce and enhance synthesis of the anticancer agent paclitaxel (Taxol®) in Taxus cell suspension cultures; however, concurrent decreases in growth are often observed, which is problematic for large scale bioprocessing. Here, increased accumulation of paclitaxel in Taxus cuspidata suspension cultures with MeJA elicitation was accompanied by a concomitant decrease in cell growth, evident within the first three days post-elicitation. Both MeJA-elicited and mock-elicited cultures exhibited similar viability with no apoptosis up to day 16 and day 24 of the cell culture period, respectively, suggesting that growth repression is not attributable to cell death. Flow cytometric analyses demonstrated that MeJA perturbed cell cycle progression of asynchronously dividing Taxus cells. MeJA slowed down cell cycle progression, impaired the G1/S transition as observed by an increase in G0/G1 phase cells, and decreased the number of actively dividing cells. Through a combination of deep sequencing and gene expression analyses, the expression status of Taxus cell cycle-associated genes correlated with observations at the culture level. Results from this study provide valuable insight into the mechanisms governing MeJA perception and subsequent events leading to repression of Taxus cell growth.

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Section: Discussionmentioning

confidence: 99%

Methyl jasmonate represses growth and affects cell cycle progression in cultured Taxus cells

et al. 2014

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“…Hydroxyurea, an inhibitor of DNA synthesis, has been shown to arrest plant cell cultures previously (Mak and Doran, 1993;Yanpaisan et al, 1998;Swiatek et al, 2002) and it was effective in arresting Taxus cells in G 0/ G 1 , indicating that flow cytometry could be used to detect changes in the cell cycle distribution of Taxus cultures (Fig. 5).…”

Section: Stathmokinetic Methods For the Determination Of Noncycling Cellsmentioning

confidence: 99%

“…The population of noncycling, or G 0 , cells can be relevant to product accumulation. For example, noncycling cells may be specialized for secondary metabolite production (Mak and Doran, 1993;Yanpaisan et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

Cell cycle analysis ofTaxus suspension cultures at the single cell level as an indicator of culture heterogeneity

Naill

Roberts

2005

Biotechnol. Bioeng.

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Single cell growth and division was measured via flow cytometry in order to characterize the metabolic variability of Taxus cuspidata suspension cultures, which produce the valuable secondary metabolite Taxol. Good agreement was observed between the cell cycle distribution and biomass accumulation over the batch culture period. Specific growth rates of 0.13 days(-1) by fresh weight and 0.15 days(-1) by dry weight were measured. Elicitation with methyl jasmonate (MJ) significantly decreased both cell cycle progression and biomass accumulation, as the specific growth rate decreased to 0.027 days(-1) by fresh and dry weight. Despite the decrease in biomass accumulation for MJ elicited cultures, sucrose utilization was not significantly different from control cultures. MJ elicitation also increased the accumulation of paclitaxel and other taxanes. The accumulation of upstream taxanes (baccatin III and 10-deactylbaccatin III) increased during exponential growth, reached a maximum around day 12, and then declined throughout the stationary phase. The paclitaxel concentration increased during both exponential growth and stationary phase, reaching a maximum around days 20-25. Throughout the culture period, greater than 70% of the cells were in G(0)/G(1) phase of the cell cycle. Studies using bromodeoxyuridine (BrdU) incorporation showed that approximately 65% of the Taxus cells are noncycling, even during exponential growth. Although the role of these cells is currently unknown, the presence of a large, noncycling subpopulation can have a significant impact on the utilization of plant cell culture technology for the large-scale production of paclitaxel. These results demonstrate that there is a high degree of metabolic heterogeneity in Taxus cuspidata suspension cultures. Understanding this heterogeneity is important for the optimization of plant cell cultures, particularly the reduction of production variability.

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Implications of Cellular Heterogeneity on Plant Cell Culture Performance

Patil

Roberts

2012

Biotechnology for Medicinal Plants

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Effect of cell-cycle inhibition on synthesis of steroidal alkaloids by Solanum aviculare plant cells

Cited by 9 publications

References 8 publications

Methyl jasmonate represses growth and affects cell cycle progression in cultured Taxus cells

Methyl jasmonate represses growth and affects cell cycle progression in cultured Taxus cells

Cell cycle analysis ofTaxus suspension cultures at the single cell level as an indicator of culture heterogeneity

Implications of Cellular Heterogeneity on Plant Cell Culture Performance

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