Comparative Analysis of Codon Usage Patterns in the Chloroplast Genomes of <i>Fagopyrum</i> Species

The non-random usage of synonymous codons encoding the same amino acid—referred to as codon usage bias (CUB)—varies substantially across genomes and significantly affects translational efficiency by modulating transcriptional and post-transcriptional processes. In chloroplast genomes, the optimizati...

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Main Authors: Qilin Liu, Shurui Li, Dinghong He, Jinyu Liu, Xiuzhi He, Chengruizhi Lin, Jinze Li, Zhixuan Huang, Linkai Huang, Gang Nie, Xinquan Zhang, Guangyan Feng
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Agronomy
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Online Access:https://www.mdpi.com/2073-4395/15/5/1190
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author Qilin Liu
Shurui Li
Dinghong He
Jinyu Liu
Xiuzhi He
Chengruizhi Lin
Jinze Li
Zhixuan Huang
Linkai Huang
Gang Nie
Xinquan Zhang
Guangyan Feng
author_facet Qilin Liu
Shurui Li
Dinghong He
Jinyu Liu
Xiuzhi He
Chengruizhi Lin
Jinze Li
Zhixuan Huang
Linkai Huang
Gang Nie
Xinquan Zhang
Guangyan Feng
author_sort Qilin Liu
collection DOAJ
description The non-random usage of synonymous codons encoding the same amino acid—referred to as codon usage bias (CUB)—varies substantially across genomes and significantly affects translational efficiency by modulating transcriptional and post-transcriptional processes. In chloroplast genomes, the optimization of CUB is critical for improving the efficacy of genetic engineering approaches. However, comprehensive analyses of CUB in <i>Fagopyrum</i> chloroplast genomes remain scarce. In this study, we performed an in-depth comparative analysis of codon usage patterns in the chloroplast genomes of nine <i>Fagopyrum</i> species. Our results revealed a marked AT-rich nucleotide composition, with base content in the order T > A > C > G. We identified 23 optimal codons and 29 high-frequency codons, most of which ended with A or U. Correlation analyses demonstrated that codon usage is strongly influenced by nucleotide skewness (GC and AT skews), protein properties (such as amino acid composition and the number of synonymous codons), and gene expression levels. Neutrality plot analysis (PR2 bias) and evaluations based on the effective number of codons (ENc) indicated that both mutational pressure and natural selection contribute to shaping CUB, with natural selection identified as the predominant evolutionary force. Comparative analyses with four model organisms indicated that <i>Arabidopsis thaliana</i> shares the highest codon usage compatibility with <i>Fagopyrum</i> chloroplast genomes, highlighting its suitability as a potential heterologous expression system. Phylogenetic reconstruction based on codon usage profiles yielded a fully resolved topology with 100% bootstrap support at all nodes, reinforcing the utility of codon usage data in evolutionary inference. This study elucidates the evolutionary determinants of codon usage variation in <i>Fagopyrum</i> plastomes and provides a robust methodological foundation for codon optimization in chloroplast-based synthetic biology. The validated codon adaptation metrics offer promising tools for improving heterologous protein expression and guiding transgene design in advanced breeding strategies.
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spelling doaj-art-7ed5e700ce0940d2a3b04f263da0a4092025-08-20T01:57:04ZengMDPI AGAgronomy2073-43952025-05-01155119010.3390/agronomy15051190Comparative Analysis of Codon Usage Patterns in the Chloroplast Genomes of <i>Fagopyrum</i> SpeciesQilin Liu0Shurui Li1Dinghong He2Jinyu Liu3Xiuzhi He4Chengruizhi Lin5Jinze Li6Zhixuan Huang7Linkai Huang8Gang Nie9Xinquan Zhang10Guangyan Feng11College of Grassland Science and Technology, Sichuan Agricultural University, Chengdu 611130, ChinaCollege of Grassland Science and Technology, Sichuan Agricultural University, Chengdu 611130, ChinaCollege of Grassland Science and Technology, Sichuan Agricultural University, Chengdu 611130, ChinaCollege of Forestry, Sichuan Agricultural University, Chengdu 611130, ChinaCollege of Grassland Science and Technology, Sichuan Agricultural University, Chengdu 611130, ChinaCollege of Grassland Science and Technology, Sichuan Agricultural University, Chengdu 611130, ChinaCollege of Grassland Science and Technology, Sichuan Agricultural University, Chengdu 611130, ChinaCollege of Grassland Science and Technology, Sichuan Agricultural University, Chengdu 611130, ChinaCollege of Grassland Science and Technology, Sichuan Agricultural University, Chengdu 611130, ChinaCollege of Grassland Science and Technology, Sichuan Agricultural University, Chengdu 611130, ChinaCollege of Grassland Science and Technology, Sichuan Agricultural University, Chengdu 611130, ChinaCollege of Grassland Science and Technology, Sichuan Agricultural University, Chengdu 611130, ChinaThe non-random usage of synonymous codons encoding the same amino acid—referred to as codon usage bias (CUB)—varies substantially across genomes and significantly affects translational efficiency by modulating transcriptional and post-transcriptional processes. In chloroplast genomes, the optimization of CUB is critical for improving the efficacy of genetic engineering approaches. However, comprehensive analyses of CUB in <i>Fagopyrum</i> chloroplast genomes remain scarce. In this study, we performed an in-depth comparative analysis of codon usage patterns in the chloroplast genomes of nine <i>Fagopyrum</i> species. Our results revealed a marked AT-rich nucleotide composition, with base content in the order T > A > C > G. We identified 23 optimal codons and 29 high-frequency codons, most of which ended with A or U. Correlation analyses demonstrated that codon usage is strongly influenced by nucleotide skewness (GC and AT skews), protein properties (such as amino acid composition and the number of synonymous codons), and gene expression levels. Neutrality plot analysis (PR2 bias) and evaluations based on the effective number of codons (ENc) indicated that both mutational pressure and natural selection contribute to shaping CUB, with natural selection identified as the predominant evolutionary force. Comparative analyses with four model organisms indicated that <i>Arabidopsis thaliana</i> shares the highest codon usage compatibility with <i>Fagopyrum</i> chloroplast genomes, highlighting its suitability as a potential heterologous expression system. Phylogenetic reconstruction based on codon usage profiles yielded a fully resolved topology with 100% bootstrap support at all nodes, reinforcing the utility of codon usage data in evolutionary inference. This study elucidates the evolutionary determinants of codon usage variation in <i>Fagopyrum</i> plastomes and provides a robust methodological foundation for codon optimization in chloroplast-based synthetic biology. The validated codon adaptation metrics offer promising tools for improving heterologous protein expression and guiding transgene design in advanced breeding strategies.https://www.mdpi.com/2073-4395/15/5/1190codon usage biaschloroplast genome<i>Fagopyrum</i>synonymous codon selectionphylogenetic analysis
spellingShingle Qilin Liu
Shurui Li
Dinghong He
Jinyu Liu
Xiuzhi He
Chengruizhi Lin
Jinze Li
Zhixuan Huang
Linkai Huang
Gang Nie
Xinquan Zhang
Guangyan Feng
Comparative Analysis of Codon Usage Patterns in the Chloroplast Genomes of <i>Fagopyrum</i> Species
Agronomy
codon usage bias
chloroplast genome
<i>Fagopyrum</i>
synonymous codon selection
phylogenetic analysis
title Comparative Analysis of Codon Usage Patterns in the Chloroplast Genomes of <i>Fagopyrum</i> Species
title_full Comparative Analysis of Codon Usage Patterns in the Chloroplast Genomes of <i>Fagopyrum</i> Species
title_fullStr Comparative Analysis of Codon Usage Patterns in the Chloroplast Genomes of <i>Fagopyrum</i> Species
title_full_unstemmed Comparative Analysis of Codon Usage Patterns in the Chloroplast Genomes of <i>Fagopyrum</i> Species
title_short Comparative Analysis of Codon Usage Patterns in the Chloroplast Genomes of <i>Fagopyrum</i> Species
title_sort comparative analysis of codon usage patterns in the chloroplast genomes of i fagopyrum i species
topic codon usage bias
chloroplast genome
<i>Fagopyrum</i>
synonymous codon selection
phylogenetic analysis
url https://www.mdpi.com/2073-4395/15/5/1190
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