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光谱质量作为诱导龙葵悬浮细胞生物活性化合物产生的因素。

Spectral quality as an elicitor of bioactive compound production in Solanum aculeatissimum JACQ cell suspension.

机构信息

Plant Biotechnology, Program in Biotechnology and Biodiversity, Pro-Centro Oeste Network, Federal Institute of Education, Science and Technology Goiano (IF Goiano), Rio Verde, GO, Brazil.

Plant Biotechnology, Program in Biotechnology and Biodiversity, Pro-Centro Oeste Network, Federal Institute of Education, Science and Technology Goiano (IF Goiano), Rio Verde, GO, Brazil.

出版信息

J Photochem Photobiol B. 2020 Mar;204:111819. doi: 10.1016/j.jphotobiol.2020.111819. Epub 2020 Feb 8.

Abstract

Solanum aculeatissimum Jacq. is a common plant in much of Brazil. Despite containing metabolites with a wide range of pharmacological applications, there are few tissue culture reports for this plant. The possibility of large-scale in vitro production of this material has significant biotechnological potential. Therefore, the objective of this study was to investigate the effect of light conditions on the growth of cells in suspension, observing the production and yield of biomass and bioactive compounds and the enzymatic behavior. Calli obtained from leaf segments were cultured in solid medium supplemented with 1 mg L of 2,4-D, 2.5 mg L kinetin, pH 5.7, in the dark. After 110 days of subculture, the calli were transferred to liquid medium. Cells were kept in the dark under agitation at 110 rpm and 25 °C and subcultured every 30 days. After 90 days of culture, 20 mL aliquots of cell suspension were added to flasks containing approximately 20 mL of medium (1:1) and cultured at different wavelengths (white, green, blue, red, and blue/red) under a photoperiod of 16 h with irradiance of 50 μmol m s) and in the absence of light. The experiment was performed in a 6 × 6 factorial design (light condition × culture time). The cell cultures showed viability throughout the entire cycle, and chlorogenic and ferulic acids, orientin, quercitrin and, in higher amounts, quercetin, were detected in the first 7 days of culture. There was an increase in superoxide dismutase and catalase and a decrease in ascorbate peroxidase after exposure to different light conditions; for phenylalanine ammonia lyase, no differences were observed. The different light conditions were not sufficient to trigger responses in the concentrations of bioactive compounds, despite the detection of increased levels of the enzymes involved in cellular homeostasis.

摘要

刺茄是巴西大部分地区常见的植物。尽管其代谢物具有广泛的药理应用,但关于这种植物的组织培养报道很少。因此,大规模体外生产这种物质具有重要的生物技术潜力。本研究的目的是研究光照条件对悬浮细胞生长的影响,观察生物量和生物活性化合物的产生和产量以及酶的行为。从叶片段获得的愈伤组织在补充有 1 mg L 的 2,4-D、2.5 mg L 激动素、pH 5.7 的固体培养基中进行培养,在黑暗中。继代培养 110 天后,将愈伤组织转移到液体培养基中。细胞在黑暗中以 110 rpm 的速度搅拌,在 25°C 下保持,并每 30 天继代培养一次。培养 90 天后,向含有约 20 mL 培养基(1:1)的培养瓶中加入 20 mL 细胞悬浮液,在不同波长(白光、绿光、蓝光、红光和红蓝光)下培养,并在光周期为 16 h、光照强度为 50 μmol m s 的条件下培养在没有光的情况下。该实验采用 6×6 因子设计(光照条件×培养时间)进行。细胞培养在整个周期内均保持活力,在培养的前 7 天检测到绿原酸和阿魏酸、木犀草苷、槲皮苷,并且含量较高的还有槲皮素。暴露于不同光照条件下,超氧化物歧化酶和过氧化氢酶的含量增加,而过氧化氢酶的含量降低;苯丙氨酸解氨酶则没有差异。尽管检测到与细胞内稳态相关的酶的水平升高,但不同的光照条件不足以引发生物活性化合物浓度的变化。

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