植物合成生物学在探索和生产植物天然产物方面具有显著的理论优势。然而,由于缺乏有效的底盘系统和相关的启用技术,其对生物合成领域的贡献目前有限。合成生物学家经常避免将烟草作为底盘系统,因为它的运行周期长,遗传和代谢修饰的困难,复杂的代谢和纯化背景,尼古丁毒性,以及在准确控制农业生产方面的挑战。然而,烟草悬浮细胞底盘系统为这些挑战提供了可行的解决方案。这项研究的目的是开发具有高科学和工业潜力的烟草悬浮细胞底盘。这个底盘应该表现出快速增长,高生物量,优异的分散性,改造效率高,和最低的尼古丁含量。Nicotianabenthamiana,在分子技术中具有很高的适用性,用于诱导悬浮细胞。诱导的悬浮细胞,命名为NBS-1,表现出快速增长,优异的分散性,和高生物量,达到最大生物量476.39克/升(鲜重),显著高于BY-2。广泛使用的pEAQ-HT瞬时表达系统在NBS-1中的转化效率达到81%,与BY-2相比显著升高。使用转录组数据分析了BY-2和NBS-1的代谢特征和偏倚。研究发现,NBS-1中黄酮类化合物及其衍生物生物合成相关通路的基因表达显著增高,而与生物碱生物合成相关的途径明显低于BY-2。黄酮和生物碱的总含量进一步验证了这些发现。总之,我们的研究表明,NBS-1具有最低的尼古丁含量,并为为特定产品选择合适的底盘提供了有价值的指导。总之,本研究开发了具有优异生长和转化的烟草悬浮细胞底盘NBS-1,类黄酮含量高,尼古丁含量低,对烟草悬浮细胞底盘的发展具有重要的指导意义。
Plant synthetic biology has significant theoretical advantages in exploration and production of plant natural products. However, its contribution to the field of biosynthesis is currently limited due to the lack of efficient chassis systems and related enabling technologies. Synthetic biologists often avoid tobacco as a chassis system because of its long operation cycle, difficulties in genetic and metabolic modification, complex metabolism and purification background, nicotine toxicity, and challenges in accurately controlling for agricultural production. Nevertheless, the tobacco suspension cell chassis system offers a viable solution to these challenges. The objective of this research was to develop a tobacco suspension cell chassis with high scientific and industrial potential. This chassis should exhibit rapid growth, high biomass, excellent dispersion, high transformation efficiency, and minimal nicotine content. Nicotiana benthamiana, which has high applicability in molecular technology, was used to induce suspension cells. The induced suspension cells, named NBS-1, exhibited rapid growth, excellent dispersion, and high biomass, reaching a maximum biomass of 476.39 g/L (fresh weight), which was significantly higher than that of BY-2. The transformation efficiency of the widely utilized pEAQ-HT transient expression system in NBS-1 reached 81%, which was substantially elevated compared to BY-2. The metabolic characteristics and bias of BY-2 and NBS-1 were analyzed using transcriptome data. It was found that the gene expression of pathways related to biosynthesis of flavonoids and their derivatives in NBS-1 was significantly higher, while the pathways related to alkaloid biosynthesis were significantly lower compared to BY-2. These findings were further validated by the total content of flavonoid and alkaloid. In summary, our research demonstrates NBS-1 possesses minimal nicotine content and provides valuable guidance for selecting appropriate chassis for specific products. In conclusion, this study developed NBS-1, a tobacco suspension cell chassis with excellent growth and transformation, high flavonoid content and minimal nicotine content, which has important guiding significance for the development of tobacco suspension cell chassis.