State Key Laboratory of Tree Genetics and Breeding, School of Forestry, Northeast Forestry University, Harbin 150040, China.
Institute of Biotechnology, Jilin Provincial Academy of Forestry Sciences, Changchun 130033, China.
Int J Mol Sci. 2022 Nov 24;23(23):14679. doi: 10.3390/ijms232314679.
In the large-scale breeding of conifers, cultivating embryogenic cells with good proliferative capacity is crucial in the process of somatic embryogenesis. In the same cultural environment, the proliferative capacity of different cell lines is significantly different. To reveal the regulatory mechanism of proliferation in woody plant cell lines with different proliferative potential, we used Korean pine cell lines with high proliferative potential 001#-001 (Fast) and low proliferative potential 001#-010 (Slow) for analysis. A total of 17 glutathione-related differentially expressed genes was identified between F and S cell lines. A total of 893 metabolites was obtained from the two cell lines in the metabolomic studies. A total of nine metabolites related to glutathione was significantly upregulated in the F cell line compared with the S cell line. The combined analyses revealed that intracellular glutathione might be the key positive regulator mediating the difference in proliferative capacity between F and S cell lines. The qRT-PCR assay validated 11 differentially expressed genes related to glutathione metabolism. Exogenous glutathione and its synthase inhibitor L-buthionine-sulfoximine treatment assay demonstrated the positive role of glutathione in the proliferation of Korean pine embryogenic cells.
在大规模培育针叶树的过程中,培养具有良好增殖能力的胚胎细胞对于体细胞胚胎发生至关重要。在相同的培养环境中,不同细胞系的增殖能力存在显著差异。为了揭示具有不同增殖潜力的木本植物细胞系增殖的调控机制,我们对具有高增殖潜力的 001#-001(快速)和低增殖潜力的 001#-010(缓慢)的红松细胞系进行了分析。在 F 和 S 细胞系之间共鉴定出 17 个与谷胱甘肽相关的差异表达基因。在代谢组学研究中,从两个细胞系中获得了 893 种代谢产物。与 S 细胞系相比,F 细胞系中共有 9 种与谷胱甘肽相关的代谢物显著上调。综合分析表明,细胞内谷胱甘肽可能是介导 F 和 S 细胞系增殖能力差异的关键正调控因子。qRT-PCR 检测验证了 11 个与谷胱甘肽代谢相关的差异表达基因。外源性谷胱甘肽及其合成酶抑制剂 L-丁硫氨酸亚砜胺处理实验证明了谷胱甘肽在红松胚胎细胞增殖中的积极作用。