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<p>The <i>KNOTTED1-like homeobox</i> (<i>KNOX</i>) gene family represents a highly conserved group of genes in plants that play a crucial role in regulating leaf development. However, there has been a scarcity of studies focused on the identification and evolutionary analysis of the <i>KNOX</i> gene within the <i>Medicago</i> genus. In this study, we identified 71 <i>KNOX</i> genes across <i>Medicago truncatula</i>, <i>Medicago polymorpha</i>, and <i>Medicago sativa.</i> These genes were categorized into three primary subclasses: Class I, Class II, and NATM based on their phylogenetic relationships. Further sub-classification revealed five distinct evolutionary branches within Class I and II. Gene duplication events were observed in all three <i>Medicago</i> species, with annual alfalfa (<i>M. truncatula</i> and <i>M. polymorpha</i>) exhibiting segmental duplications, while <i>M. sativa</i> predominantly displayed tandem duplications. Promoter analysis indicated that the <i>KNOX</i> genes in <i>Medicago</i> contain elements associated with hormone regulation, light response, stress response, and tissue-specific expression<i>.</i> Analysis of tissue expression patterns demonstrated that the Class II-D/E subgroup was expressed in all examined tissues. Additionally, RT-qPCR data showed that <i>MsKNAT1, MsSTM2</i>, and <i>MsKNATM1/2</i> from Class I and NATM exhibited significantly higher expression levels (<i>P</i> &lt; 0.05) in alfalfa trifoliate leaves compared to multilobar leaves. Except for <i>MsKNAT8</i>, Class II genes were significantly higher (<i>P</i> &lt; 0.01) in seven leaflets compared to three leaves, and expression levels increased progressively with leaf age. Furthermore, the EST-SSR markers developed for the <i>MsSTM2</i> gene are proved to be effective in distinguishing between trifoliate and multifoliate alfalfa. The findings of this study provide valuable genetic resources for molecular marker-assisted breeding and functional research aimed at understanding the regulation of compound leaf development in alfalfa.</p>

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Characterization of the KNOX Gene Family in Regulating Alfalfa Compound Leaf Development

  • 期刊论文

  • 发表日期：

    2025年2月9日

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  • Bo Luo,
  • Jiaxiang Jiang,
  • Chenxi Liu,
  • Jia Wei,
  • Xia Wang,
  • Zhengfeng Cao,
  • Chuanjie Zhang,
  • Nana Liu,
  • Yanyan Lv,
  • Zhenwu Wei,
  • Xueyang Min

摘要

The KNOTTED1-like homeobox (KNOX) gene family represents a highly conserved group of genes in plants that play a crucial role in regulating leaf development. However, there has been a scarcity of studies focused on the identification and evolutionary analysis of the KNOX gene within the Medicago genus. In this study, we identified 71 KNOX genes across Medicago truncatula, Medicago polymorpha, and Medicago sativa. These genes were categorized into three primary subclasses: Class I, Class II, and NATM based on their phylogenetic relationships. Further sub-classification revealed five distinct evolutionary branches within Class I and II. Gene duplication events were observed in all three Medicago species, with annual alfalfa (M. truncatula and M. polymorpha) exhibiting segmental duplications, while M. sativa predominantly displayed tandem duplications. Promoter analysis indicated that the KNOX genes in Medicago contain elements associated with hormone regulation, light response, stress response, and tissue-specific expression. Analysis of tissue expression patterns demonstrated that the Class II-D/E subgroup was expressed in all examined tissues. Additionally, RT-qPCR data showed that MsKNAT1, MsSTM2, and MsKNATM1/2 from Class I and NATM exhibited significantly higher expression levels (P < 0.05) in alfalfa trifoliate leaves compared to multilobar leaves. Except for MsKNAT8, Class II genes were significantly higher (P < 0.01) in seven leaflets compared to three leaves, and expression levels increased progressively with leaf age. Furthermore, the EST-SSR markers developed for the MsSTM2 gene are proved to be effective in distinguishing between trifoliate and multifoliate alfalfa. The findings of this study provide valuable genetic resources for molecular marker-assisted breeding and functional research aimed at understanding the regulation of compound leaf development in alfalfa.