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| Identification, Expression Analysis and Development of CRISPR/Cas9-Edited Materials for the BnphyA Gene Family in Rapeseed (Brassica napus) |
| LI Pu, LIU Han-Xuan, XIE Chang-Gen* |
| College of Life Sciences, Northwest A&F University, Yangling 712100, China |
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Abstract Brassica napus, an economically vital oilseed crop in China, exhibits significant commercial and ornamental value. Phytochrome A (phyA), a core photoreceptor responsible for far-red light perception, plays crucial roles in regulating plant growth and abiotic stress responses. In this study, BnphyA gene family members in B. napus were systematically identified through bioinformatics screening and their expression patterns across tissues and under diverse abiotic stress conditions were analyzed. Fifteen BnphyA members were identified and categorized into 3 subfamilies. Phylogenetic analysis showed that Subfamily Ⅱmembers, including BnaA06G0057900ZS and BnaA09G0661700ZS, exhibited the closest evolutionary relationship with Arabidopsis thaliana AtphyA (AT1G09570). Structural analysis revealed conserved PHY-GAF-PAS_2-HATPase domains across all members. Cis-regulatory element screening identified light-responsive, developmental, and stress-associated motifs in BnphyA promoters. Chromosomal mapping indicated that homologs were predominantly localized near chromosomal termini. Tissue-specific expression profiling revealed upregulated BnphyA expression in roots and seeds, with predominant root-specific responses to cold, heat, drought, and osmotic stresses. Subcellular localization assays indicated cytoplasmic distribution of inactive BnphyA proteins. Heterologous complementation of A. thaliana phyA-211 mutants failed to restore wild-type phenotypes. CRISPR/Cas9-mediated editing generated multiple mutant lines in '19YB437' cultivars, including base substitutions (TGC→CGC) and deletions at the BnaA06G0057900ZS target site. This study provides a theoretical foundation and essential genetic resources for further investigation of the BnphyA gene family's functions.
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Received: 21 April 2025
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Corresponding Authors:
*changen.xie@nwafu.edu.cn
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