Journal: Synthetic and Systems Biotechnology
Article Title: Metabolic reprogramming of abscisic acid-producing strain Botrytis cinerea TB-31 toward terpenoid biosynthesis using a CRISPR/Cas9 ribonucleoprotein system
doi: 10.1016/j.synbio.2025.12.002
Figure Lengend Snippet: Transcriptomic analysis of B. cinerea strains TB-31, △ bcaba3 , and △ bcaba1234 . (A)∼(B) Regression analysis between transcriptomic data (RNA-seq) and RT‒qPCR validation. △ bcaba3 vs. TB-31: R 2 = 0.94, P < 0.001 (A); △ bcaba1234 vs. TB-31: R 2 = 0.97, P < 0.001 (B). (C) Heatmap of the relative transcriptional levels of the genes in the MVA pathway in △ bcaba1234 compared with those in TB-31. Gene transcription levels were normalized using Z score transformation, calculated as (X-mean)/SD, to represent the number of standard deviations above or below the mean transcription level. Upregulated genes are highlighted in red, while downregulated genes are shown in blue. The data are derived from three independent biological replicates. (D) Heatmap of the relative transcriptional levels of DEGs involved in other secondary metabolism pathways in △ bcaba1234 compared with those in TB-31. (E) Heatmap of the relative transcriptional levels of genes involved in intracellular primary metabolic pathways in the △ bcaba1234 strain compared with those in the TB-31 strain. Metabolic pathways are marked with gray boxes. PPP: pentose phosphate pathway; TCA: tricarboxylic acid cycle; GYC: glyoxylate cycle.
Article Snippet: The sgRNAs were transcribed using the T7 High Yield RNA Transcription Kit (Nanjing Vazyme Biotech Co., Ltd., Nanjing, China), and then purified using the RNA Clean & Concentrator Kit (Zymo Research Corp., Irvine, California, USA).
Techniques: RNA Sequencing, Biomarker Discovery, Transformation Assay, Derivative Assay