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Evolutionary insight and characterization of WOX genes in callus development and differentiation of Peucedanum
Cheng Song1, Haoyu Pan1,2, Muhammad Arif3
1West Anhui University, Luan, 237012, China.
Main Conclusion:
The WOX genes essential for callus formation and development were successfully isolated from Peucedanum praeruptorum, confirming the involvement of PpWOX9, PpWOX14, and PpWOX15 in this process. WUSCHEL (WUS)-related homeobox (WOX) gene family is essential due to its pivotal role in orchestrating fundamental developmental processes, such as stem cell maintenance, organ morphogenesis, and embryonic patterning, which underpin plant growth and adaptability. They form intricate regulatory networks to ensure cellular homeostasis and enable indeterminate growth, revealing the evolutionary innovations and how plants achieve complex architectures. However, the specific role of WOX members in regulating callus development and differentiation in P. praeruptorum remains unclear. Using bioinformatics, genomics, and transcriptomics methods, we performed a systematic analysis of the structural composition, family classification, evolutionary relationships, and expression patterns of WOX genes in P. praeruptorum during callus formation. PpWOX proteins were categorized into three main subclades based on the phylogenetic tree. Cis-acting element analysis indicated that many PpWOX genes possess multiple copies of light-responsive elements (LREs), suggesting that light signals significantly influence the expression of these genes. Synteny analysis revealed eight whole-genome duplication (WGD) and segmental duplication events in their evolutionary history, which may have facilitated the expansion of PpWOX genes. The expression profile showed distinct expression levels for PpWOX genes during callus formation, with PpWOX9, PpWOX13, PpWOX14, PpWOX15, and PpWOX16 exhibiting higher expression levels compared to other PpWOX genes. qRT-PCR analysis demonstrated that the expression levels of PpWOXs varied across six different stages. During the somatic cell development stage, the expression levels of PpWOX9 and PpWOX15 were significantly elevated. Transcriptional co-expression analysis revealed that PpWOX9, PpWOX14, and PpWOX15 exhibited similar co-expression patterns. Subcellular localization studies using tobacco protoplasts showed that the PpWOX9 and PpWOX15 genes were specifically localized in the nucleus. Our findings serve as a scientific reference for the study of WOX genes in the conservation of P. praeruptorum resources.
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