K.A. Timiryazev Institute of Plant Physiology of Russian Academy of Sciences, Botanicheskaya 35, 127276 Moscow, Russia.
Biology Faculty, M.V. Lomonosov Moscow State University, 119234 Moscow, Russia.
Biomolecules. 2023 Jun 9;13(6):969. doi: 10.3390/biom13060969.
The effects of methyl jasmonate (MeJ) on growth and taxoid formation in the cell culture of were investigated to elucidate the specifics of phytohormone action in dedifferentiated plant cells in vitro. The characteristics of the same suspension cell culture were compared in 2017 (the «young» culture) and in 2022 (the «old» culture)-1.5 or 6 years after culture induction, respectively. MeJ (100 µM) is added to the cell suspension at the end of the exponential growth phase. Cell culture demonstrated good growth (dry weight accumulation 10-18 g/L, specific growth rate µ = 0.15-0.35 day) regardless of its «age», cultivation system, and MeJ addition. UPLC-ESI-MS analysis revealed the presence of C14-hydroxylated taxoids (yunnanxane, taxuyunnanine C, sinenxane C, and sinenxane B) in the cell biomass. The content of C14-OH taxoids increased from 0.2-1.6 mg/gDW in «young» culture to 0.6-10.1 mg/gDW in «old» culture. Yunnanxane was the main compound in «young» culture, while sinenxane C predominated in «old» culture. Without elicitation, small amounts of C13-OH taxoids (<0.05 mg/gDW) were found only in «young» cultures. MeJ addition to «young» culture had no effect on the content of C14-OH taxoids but caused a 10-fold increase in C13-OH taxoid production (up to 0.12-0.19 mg/gDW, comparable to the bark of yew trees). By contrast, MeJ added to «old» culture was not beneficial for the production of C13-OH taxoids but notably increased the content of C14-OH taxoids (1.5-2.0 times in flasks and 5-8 times in bioreactors). These findings suggest that hormonal signaling in dedifferentiated yew cells grown in vitro is different from that in plants and can be affected by the culture's age. This might be a result of the high level of culture heterogeneity and constant auto-selection for intensive proliferation, which leads to the predominant formation of C14-OH taxoids versus C13-OH taxoids and a modified cell response to exogenous MeJ treatment.
为了阐明植物细胞体外去分化过程中植物激素作用的特点，研究了茉莉酸甲酯（MeJ）对 细胞培养物生长和紫杉烷形成的影响。2017 年（“年轻”培养物）和 2022 年（“年老”培养物）对相同的悬浮细胞培养物的特征进行了比较，分别为培养物诱导后 1.5 年和 6 年。MeJ（100 μM）在指数生长期结束时添加到细胞悬浮液中。无论其“年龄”、培养系统和 MeJ 添加如何，细胞培养均表现出良好的生长（干重积累 10-18 g/L，比生长速率μ=0.15-0.35 天）。UPLC-ESI-MS 分析表明，细胞生物量中存在 C14-羟基化紫杉烷（云南烷、紫杉烷 C、新云南烷 C 和新云南烷 B）。在“年轻”培养物中，C14-OH 紫杉烷的含量从 0.2-1.6 mg/gDW 增加到“年老”培养物中的 0.6-10.1 mg/gDW。在“年轻”培养物中，云南烷是主要化合物，而在“年老”培养物中，新云南烷 C 占主导地位。在没有诱导的情况下，仅在“年轻”培养物中发现少量的 C13-OH 紫杉烷（<0.05 mg/gDW）。MeJ 添加到“年轻”培养物中对 C14-OH 紫杉烷的含量没有影响，但导致 C13-OH 紫杉烷产量增加了 10 倍（高达 0.12-0.19 mg/gDW，与紫杉树皮相当）。相比之下，MeJ 添加到“年老”培养物中不利于 C13-OH 紫杉烷的产生，但显著增加了 C14-OH 紫杉烷的含量（在摇瓶中增加 1.5-2.0 倍，在生物反应器中增加 5-8 倍）。这些发现表明，体外生长的去分化紫杉细胞中的激素信号不同于植物中的激素信号，并且可能受到培养物年龄的影响。这可能是由于培养物高度异质性和不断的自我选择以进行强烈增殖，导致主要形成 C14-OH 紫杉烷而不是 C13-OH 紫杉烷，以及细胞对外源 MeJ 处理的反应发生改变所致。
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