Abstract
Plant cell cultures constitute a potentially efficient and sustainable tool for the production of high added-value bioactive compounds. However, due to the inherent restrictions in the expression of secondary metabolism, to date the yields obtained have generally been low. Plant cell culture elicitation can boost production, sometimes leading to dramatic improvements in yield, as well as providing insight into the target biosynthetic pathways and the regulation of the genes involved. Among the secondary compounds successfully being produced in biotechnological platforms are taxanes and trans-resveratrol (t-R). In the current study, perfluorodecalins (PFDs) and hexenol (Hex) were tested for the first time with Taxus media and Vitis vinifera cell cultures to explore their effect on plant cell growth and secondary metabolite production, either alone or combined with other elicitors already established as highly effective, such as methyl jasmonate (MeJa), coronatine (Coro) or randomly methylated β-cyclodextrins (β-CDs). The total taxane content at the peak of production in T. media cell cultures treated with PFDs together with Coro plus β-CDs was 3.3-fold higher than in the control, whereas the t-R production in MeJa and β-CD-treated V. vinifera cell cultures increased 552.6-fold compared to the extremely low-yielding control. Hex was ineffective as an elicitor in V. vinifera cell cultures, and in T. media cell suspensions it blocked the taxol production but induced a clear enhancement of baccatin III. Regarding biosynthetic gene expression, a strong positive relationship was observed between the transcript level of targeted genes and taxol production in the T. media cell cultures, but not with t-R production in the elicited V. vinifera cell cultures.
Highlights
One of the most successful examples of biotechnological production of high added-value compounds is the use of Taxus spp. cell cultures to produce the well-known anti-cancer compound taxol as well as other taxanes used as semi-synthetic precursors of taxol and its analogs
We found that the addition of methyl jasmonate (MeJa) to Taxus spp. cell cultures at the production stage (Cusido et al, 2002) is an efficient method for enhancing taxane yield
Taking into account the aforementioned antecedents, the aim of this work was to study taxane and t-R production in elicited T. media and V. vinifera cell cultures, respectively, after supplementing optimum culture media (Cusido et al, 2002; Almagro et al, 2014) with the new elicitors, PFD (57.5% v/v), both gassed (PDFgas) and degassed (PDFdegas), or hexenol (40 μM Hex), in order to show if two different cell lines, both with a high capacity to produce bioactive compounds but with different biosynthetic pathways, are able to respond in a similar way to elicitation
Summary
One of the most successful examples of biotechnological production of high added-value compounds is the use of Taxus spp. cell cultures to produce the well-known anti-cancer compound taxol as well as other taxanes used as semi-synthetic precursors of taxol and its analogs. Many studies have shown the effectiveness of supplementing Taxus spp. and Vitis vinifera cell cultures with either biotic or abiotic elicitors to increase the accumulation of taxol and related taxanes (Vongpaseuth and Roberts, 2007; Exposito et al, 2009; Onrubia et al, 2013a; Ramirez-Estrada et al, 2016b), or t-R (Belchí-Navarro et al, 2012; Almagro et al, 2014). Coro, which is a phytotoxin produced by different pathovars of Pseudomonas syringae (Bender et al, 1999), is a natural analog of the active form of jasmonate, JA-Ile. In T. media cell cultures (Onrubia et al, 2013b), the yield of total taxanes increased remarkably after elicitation with Coro (1 μM), while taxol and baccatin III production was significantly enhanced by the combination of β-CDs and Coro (Ramirez-Estrada et al, 2015)
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