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Effects of editing DFR genes on flowers, leaves, and roots of tobacco

文献类型： 外文期刊

作者： Jiang, Jiarui 1 ; Huang, Haitao 1 ; Gao, Qian 1 ; Li, Yong 2 ; Xiang, Haiying 1 ; Zeng, Wanli 1 ; Xu, Li 1 ; Liu, Xin 1 ; Li, Jing 1 ; Mi, Qili 1 ; Deng, Lele 1 ; Yang, Wenwu 1 ; Zhang, Jianduo 1 ; Yang, Guangyu 1 ; Li, Xuemei 1 ;

作者机构： 1.China Tobacco Yunnan Ind Co LTD, Technol Ctr, 181 Hongjin Rd, Kunming 650000, Yunnan, Peoples R China

2.Jiangsu Acad Agr Sci, 50 Zhongling St, Nanjing 210014, Jiangsu, Peoples R China

关键词： DFR; Gene editing; CRISPR; Cas9; Flavonoid metabolism

期刊名称：BMC PLANT BIOLOGY （ 影响因子：5.3； 五年影响因子：5.9 ）

ISSN： 1471-2229

年卷期： 2023 年 23 卷 1 期

页码：

收录情况： SCI

摘要： BackgroundDFR is a crucial structural gene in plant flavonoid and polyphenol metabolism, and DFR knockout (DFR-KO) plants may have increased biomass accumulation. It is uncertain whether DFR-KO has comparable effects in tobacco and what the molecular mechanism is. We employed the CRISPR/Cas9 method to generate a knockout homozygous construct and collected samples from various developmental phases for transcriptome and metabolome detection and analysis.ResultsDFR-KO turned tobacco blossoms white on homozygous tobacco (Nicotiana tabacum) plants with both NtDFR1 and NtDFR2 knockout. RNA-seq investigation of anthesis leaf (LF), anthesis flower (FF), mature leaf (LM), and mature root (RM) variations in wild-type (CK) and DFR-KO lines revealed 2898, 276, 311, and 101 differentially expressed genes (DEGs), respectively. DFR-KO primarily affected leaves during anthesis. According to KEGG and GSEA studies, DFR-KO lines upregulated photosynthetic pathway carbon fixation and downregulated photosystem I and II genes. DFR-KO may diminish tobacco anthesis leaf photosynthetic light reaction but boost dark reaction carbon fixation. DFR-KO lowered the expression of pathway-related genes in LF, such as oxidative phosphorylation and proteasome, while boosting those in the plant-pathogen interaction and MAPK signaling pathways, indicating that it may increase biological stress resistance. DFR-KO greatly boosted the expression of other structural genes involved in phenylpropanoid production in FF, which may account for metabolite accumulation. The metabolome showed that LF overexpressed 8 flavonoid metabolites and FF downregulated 24 flavone metabolites. In DFR-KO LF, proteasome-related genes downregulated 16 amino acid metabolites and reduced free amino acids. Furthermore, the DEG analysis on LM revealed that the impact of DFR-KO on tobacco growth may progressively diminish with time.ConclusionThe broad impact of DFR-KO on different phases and organs of tobacco development was thoroughly and methodically investigated in this research. DFR-KO decreased catabolism and photosynthetic light reactions in leaves during the flowering stage while increasing carbon fixation and disease resistance pathways. However, the impact of DFR-KO on tobacco growth steadily declined as it grew and matured, and transcriptional and metabolic modifications were consistent. This work offers a fresh insight and theoretical foundation for tobacco breeding and the development of gene-edited strains.

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