Overexpression of BnaA08.NDPK Enhances Seed Oil Content and Modulates Fatty Acid Composition in Transgenic Arabidopsis: Insights from Comparative Transcriptomics and Co-Expression Network Analysis
- 1Oil Crops Research Institute, Chongqing Academy of Agricultural Sciences, Chongqing 401329, China
Abstract
Background/Objectives: Rapeseed (Brassica napus L.) is a major oilseed crop, and enhancing seed oil accumulation is a key breeding objective. However, the transcriptional regulation underlying natural variation in seed oil accumulation and fatty acid composition remains unclear. This study aimed to identify key lipid-associated modules and candidate genes and validate a selected gene. Methods: Twelve rapeseed lines with contrasting oil content were classified into high-, medium-, and low-oil groups. Phenotypic traits were evaluated across field seasons. Fatty acid composition was determined by gas chromatography with flame ionization detection (GC-FID). Transcriptome sequencing was performed on 30-day-after-pollination (DAP) developing seeds, followed by differential expression analysis and weighted gene co-expression network analysis (WGCNA). A candidate gene, BnaA08.NDPK (nucleoside diphosphate kinase), was screened and validated by qRT-PCR. For heterologous functional characterization, BnaA08.NDPK was overexpressed in Arabidopsis, and separate CRISPR/Cas9-edited Arabidopsis lines with disrupted endogenous AtNDPK were generated. Results: Five major fatty acids (C16:0, C18:0, C18:1n9c, C18:2n6c, and C18:3) accounted for >90% of total fatty acids. High-oil materials had higher C18:1n9c but lower C18:2n6c and C18:3, and lower desaturation ratios, than low-oil materials. Up-regulated differentially expressed genes (DEGs) in high-oil lines were enriched in fatty acid biosynthesis, elongation, and glycerolipid metabolism, while down-regulated DEGs were linked to fatty acid degradation and phenylpropanoid biosynthesis. WGCNA identified MEturquoise (positive) and MEgreen (negative) modules correlated with oil content. BnaA08.NDPK was highly expressed in high-oil lines and positively correlated with oil content. In transgenic Arabidopsis, overexpression of BnaA08.NDPK increased seed size, thousand-seed weight, and oil content, with higher C18:1n9c and lower C18:2n6c, whereas CRISPR/Cas9-edited Arabidopsis lines with disrupted AtNDPK showed the opposite trends. Conclusions: Heterologous functional characterization in Arabidopsis supports a positive role of BnaA08.NDPK in seed development and oil accumulation. These findings identify BnaA08.NDPK as a promising candidate gene for improving seed oil accumulation and fatty acid composition, and provide a valuable framework for gene discovery and molecular breeding in rapeseed.
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BibTeX
@article{yu2026overexpression,
title = {{Overexpression of BnaA08.NDPK Enhances Seed Oil Content and Modulates Fatty Acid Composition in Transgenic Arabidopsis: Insights from Comparative Transcriptomics and Co-Expression Network Analysis}},
author = {Shijie Yu and Zhi Zhao and Zhi Li and Cheng Zhu and Taocui Huang},
journal = {Metabolites},
year = {2026},
month = nov,
doi = {10.3390/metabo16110860},
url = {https://www.mdpi.com/2218-1989/16/11/860},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Overexpression of BnaA08.NDPK Enhances Seed Oil Content and Modulates Fatty Acid Composition in Transgenic Arabidopsis: Insights from Comparative Transcriptomics and Co-Expression Network Analysis
AU - Shijie Yu
AU - Zhi Zhao
AU - Zhi Li
AU - Cheng Zhu
AU - Taocui Huang
PY - 2026
DA - 2026/11/06
JO - Metabolites
DO - 10.3390/metabo16110860
UR - https://www.mdpi.com/2218-1989/16/11/860
AB - Background/Objectives: Rapeseed (Brassica napus L.) is a major oilseed crop, and enhancing seed oil accumulation is a key breeding objective. However, the transcriptional regulation underlying natural variation in seed oil accumulation and fatty acid composition remains unclear. This study aimed to identify key lipid-associated modules and candidate genes and validate a selected gene. Methods: Twelve rapeseed lines with contrasting oil content were classified into high-, medium-, and low-oil groups. Phenotypic traits were evaluated across field seasons. Fatty acid composition was determined by gas chromatography with flame ionization detection (GC-FID). Transcriptome sequencing was performed on 30-day-after-pollination (DAP) developing seeds, followed by differential expression analysis and weighted gene co-expression network analysis (WGCNA). A candidate gene, BnaA08.NDPK (nucleoside diphosphate kinase), was screened and validated by qRT-PCR. For heterologous functional characterization, BnaA08.NDPK was overexpressed in Arabidopsis, and separate CRISPR/Cas9-edited Arabidopsis lines with disrupted endogenous AtNDPK were generated. Results: Five major fatty acids (C16:0, C18:0, C18:1n9c, C18:2n6c, and C18:3) accounted for >90% of total fatty acids. High-oil materials had higher C18:1n9c but lower C18:2n6c and C18:3, and lower desaturation ratios, than low-oil materials. Up-regulated differentially expressed genes (DEGs) in high-oil lines were enriched in fatty acid biosynthesis, elongation, and glycerolipid metabolism, while down-regulated DEGs were linked to fatty acid degradation and phenylpropanoid biosynthesis. WGCNA identified MEturquoise (positive) and MEgreen (negative) modules correlated with oil content. BnaA08.NDPK was highly expressed in high-oil lines and positively correlated with oil content. In transgenic Arabidopsis, overexpression of BnaA08.NDPK increased seed size, thousand-seed weight, and oil content, with higher C18:1n9c and lower C18:2n6c, whereas CRISPR/Cas9-edited Arabidopsis lines with disrupted AtNDPK showed the opposite trends. Conclusions: Heterologous functional characterization in Arabidopsis supports a positive role of BnaA08.NDPK in seed development and oil accumulation. These findings identify BnaA08.NDPK as a promising candidate gene for improving seed oil accumulation and fatty acid composition, and provide a valuable framework for gene discovery and molecular breeding in rapeseed.
SN - 2218-1989
ER -