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Background <p>Alfalfa (<i>Medicago sativa</i> L.), a key perennial leguminous forage, suffers severe yield and quality losses under diverse abiotic stresses. The plant-specific B3 superfamily transcription factors regulate plant growth, development and abiotic stress responses. However, their systematic identification and stress-responsive expression patterns in alfalfa remain uncharacterized, especially under salt, drought, cold, and low-phosphorus stresses.</p> Results <p>A total of 146 <i>MsB3</i> genes were identified from the genome of alfalfa cultivar Zhongmu No.1 and classified into four subfamilies with distinct member numbers, 89 belonging to the REM subfamily, 42 to the ARF subfamily, 8 to the LAV subfamily and 7 to the RAV subfamily. All <i>MsB3</i> genes were renamed based on their chromosomal distribution and subfamily classification. Gene structure and conserved motif analyses of <i>MsB3</i> genes showed variable exon counts, subfamily-specific motifs, and a conserved B3 DNA-binding domain in all MsB3 proteins. Collinearity analysis detected 18 tandem and 13 segmental duplication events. Transcriptomic data demonstrated tissue-specific expression for three <i>MsB3</i> genes, while 73 <i>MsB3</i> genes showed widespread expression across six alfalfa tissues including elongated stems, post-elongated stems, flowers, leaves, nodules and roots. Further analysis of transcriptomic data under salt, drought, cold and low-phosphorus stresses identified numerous stress-responsive <i>MsB3</i> members, with salt stress inducing the largest number of responsive genes at 25 and low-phosphorus stress eliciting the fewest at 12. Among these stress-responsive genes, 14 genes exhibited stress-specific expression patterns, seven responded to three different abiotic stresses, and only <i>MsARF16</i> was responsive to all four stresses, the expression pattern of which was further validated by qRT-PCR.</p> Conclusion <p>This study systematically identified and characterized the B3 superfamily genes in alfalfa. Abiotic stress-responsive candidate genes were identified via transcriptomic analysis under multiple stress conditions. The findings furnish essential insights into subsequent functional investigations of alfalfa <i>B3</i> genes and the genetic improvement of alfalfa abiotic stress resistance.</p>

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Genome-wide characterization and expression analysis of the B3 superfamily in response to abiotic stresses in alfalfa (Medicago sativa L.)

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年6月26日

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  • Yajing Wu,
  • He Zhu,
  • Siqi Wang,
  • Yuxuan Ding,
  • Tao Zhu,
  • Qingchuan Yang,
  • Xue Wang

摘要

Background

Alfalfa (Medicago sativa L.), a key perennial leguminous forage, suffers severe yield and quality losses under diverse abiotic stresses. The plant-specific B3 superfamily transcription factors regulate plant growth, development and abiotic stress responses. However, their systematic identification and stress-responsive expression patterns in alfalfa remain uncharacterized, especially under salt, drought, cold, and low-phosphorus stresses.

Results

A total of 146 MsB3 genes were identified from the genome of alfalfa cultivar Zhongmu No.1 and classified into four subfamilies with distinct member numbers, 89 belonging to the REM subfamily, 42 to the ARF subfamily, 8 to the LAV subfamily and 7 to the RAV subfamily. All MsB3 genes were renamed based on their chromosomal distribution and subfamily classification. Gene structure and conserved motif analyses of MsB3 genes showed variable exon counts, subfamily-specific motifs, and a conserved B3 DNA-binding domain in all MsB3 proteins. Collinearity analysis detected 18 tandem and 13 segmental duplication events. Transcriptomic data demonstrated tissue-specific expression for three MsB3 genes, while 73 MsB3 genes showed widespread expression across six alfalfa tissues including elongated stems, post-elongated stems, flowers, leaves, nodules and roots. Further analysis of transcriptomic data under salt, drought, cold and low-phosphorus stresses identified numerous stress-responsive MsB3 members, with salt stress inducing the largest number of responsive genes at 25 and low-phosphorus stress eliciting the fewest at 12. Among these stress-responsive genes, 14 genes exhibited stress-specific expression patterns, seven responded to three different abiotic stresses, and only MsARF16 was responsive to all four stresses, the expression pattern of which was further validated by qRT-PCR.

Conclusion

This study systematically identified and characterized the B3 superfamily genes in alfalfa. Abiotic stress-responsive candidate genes were identified via transcriptomic analysis under multiple stress conditions. The findings furnish essential insights into subsequent functional investigations of alfalfa B3 genes and the genetic improvement of alfalfa abiotic stress resistance.