Identification, Classification of the MIKC-Type MADS-Box Gene Family, and Expression Analysis of Female and Male Flower Buds in Walnut (<i>Juglans regia,</i> Juglandaceae)

MIKC-type MADS-box transcription factors constitute one of the largest gene families in plants, playing pivotal roles in regulating plant growth and development, hormone signaling transduction, and responses to biotic and abiotic stresses. However, there have been no reports on the systematic identi...

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Main Authors: Caihua Guo, Olumide Phillip Fesobi, Zhongrong Zhang, Xing Yuan, Haochang Zhao, Shaowen Quan, Jianxin Niu
Format: Article
Language:English
Published: MDPI AG 2025-07-01
Series:Horticulturae
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Online Access:https://www.mdpi.com/2311-7524/11/7/787
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Summary:MIKC-type MADS-box transcription factors constitute one of the largest gene families in plants, playing pivotal roles in regulating plant growth and development, hormone signaling transduction, and responses to biotic and abiotic stresses. However, there have been no reports on the systematic identification and characterization of MIKC-type MADS-box proteins in walnuts. In this study, we identified 52 <i>JrMADS</i> genes in the walnut genome and transcriptome, and categorized them into 14 subfamilies through structural domain and phylogenetic tree analysis. It was found that these genes were unevenly distributed across 16 chromosomes. Within the MIKC-type MADS-box gene family, we identified three pairs of tandem-duplicated genes and 40 pairs of segmental duplicated genes, indicating that segmental duplication was the primary mechanism of gene amplification in walnut. Ka/Ks analysis showed that the family genes have undergone purifying selection during evolutionary processes. The promoter was predicted to contain cis-acting elements related to growth, development, plant hormones, and stress response. Expression profile analysis showed that <i>JrMADS</i> genes have different expression patterns in various tissues and developmental stages of male and female flower buds. Notably, an ancient clade of <i>TM8</i> (<i>JrMADS43</i>) genes was found, which is absent in Arabidopsis but present in other flowering plants. Another gene, <i>TM6</i> gene (<i>JrMADS4</i>), belongs to the AP3 subfamily and is a clade that has diverged from tomatoes. Through qPCR analysis, we verified the differential expression of <i>JrMADS</i> genes at different developmental stages (MB-1/2/3 and FB-1/2/3), with <i>JrMADS5</i>, <i>JrMADS8</i>, <i>JrMADS14</i>, <i>JrMADS24</i>, <i>JrMADS40</i>, <i>JrMADS46</i>, <i>JrMADS47</i>, <i>JrGA3ox1</i>, and <i>JrGA3ox3</i> showing significantly higher expression in male than in female flower buds. In summary, our results provide valuable information for further biological functions research on MIKC-type MADS-box genes in walnut, such as flower organ development, and lays a solid foundation for future studies.
ISSN:2311-7524