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转录组分析揭示了野生柴胡与内生真菌CHS3的关联对柴胡皂苷D产生的影响。

Transcriptome analysis revealing the effect of Willd association with endophytic fungi CHS3 on the production of saikosaponin D.

作者信息

Liu Guangjie, Liu Yuanzhen, Li Zhongmeng, Ren Yubin, Liu Bo, Gao Ning, Cheng Yupeng

机构信息

Key Laboratory of Basic and Application Research of Beiyao (Heilongjiang University of Chinese Medicine), Ministry of Education, China.

Heilongjiang Agricultural Reclamation Vocational College, China.

出版信息

Heliyon. 2024 Jun 22;10(12):e33453. doi: 10.1016/j.heliyon.2024.e33453. eCollection 2024 Jun 30.

Abstract

Saikosaponin D (SSd) is a naturally active product with strong pharmacological activity found in Willd. Studies have shown that endophytic fungi have great potential as sources of natural medicines. (CHS3), an SSd-producing endophytic fungus, was isolated from . To elucidate the effect of host plants on the production of SSd in CHS3, CHS3 was co-cultured with suspension cells of and SSd was detected using high-performance liquid chromatography (HPLC). Transcriptome sequencing (RNA-Seq) of CHS3 before and after co-culture was performed using an Illumina HiSeq 2500 platform. The results indicated that the content of SSd synthesised by CHS3 increased after co-culture with suspension cells of . Transcriptome analysis of CHS3 with differentially expressed genes (DEGs) showed that 1202 and 1049 genes were upregulated and downregulated, respectively, after co-culture. Thirty genes associated with SSd synthesis and 11 genes related to terpene backbone biosynthesis were annotated to the Kyoto Encyclopaedia of Genes and Genomes (KEGG). Combined with transcriptome data, it was speculated that the mevalonate (MVA) pathway is a possible pathway for SSd synthesis in CHS3, and the expression of key enzyme genes () was validated by qRT-PCR. In conclusion, the endophytic fungus CHS3 can form an interactive relationship with its host, thereby promoting SSd biosynthesis and accumulation by upregulating the expression of key enzyme genes in the biosynthesis pathway.

摘要

柴胡皂苷D(SSd)是一种在狭叶柴胡中发现的具有强大药理活性的天然活性产物。研究表明,内生真菌作为天然药物来源具有巨大潜力。从狭叶柴胡中分离出一种产SSd的内生真菌(CHS3)。为阐明宿主植物对CHS3中SSd产生的影响,将CHS3与狭叶柴胡悬浮细胞共培养,并使用高效液相色谱(HPLC)检测SSd。使用Illumina HiSeq 2500平台对共培养前后的CHS3进行转录组测序(RNA-Seq)。结果表明,CHS3与狭叶柴胡悬浮细胞共培养后合成的SSd含量增加。对CHS3进行差异表达基因(DEG)转录组分析表明,共培养后分别有1202个和1049个基因上调和下调。30个与SSd合成相关的基因和11个与萜类骨架生物合成相关的基因被注释到京都基因与基因组百科全书(KEGG)中。结合转录组数据推测,甲羟戊酸(MVA)途径可能是CHS3中SSd合成的途径,并通过qRT-PCR验证了关键酶基因的表达。总之,内生真菌CHS3可与其宿主形成相互作用关系,从而通过上调生物合成途径中关键酶基因的表达促进SSd的生物合成和积累。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5923/11250876/a5e5a0833bdf/gr1.jpg

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