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| Transcriptome Analysis of Walnut (Juglans regia) Under Low Temperature Stress and Preliminary Identification of the Cold Resistance Function of F-box Genes |
| JIANG Tiao, AN Xiao-Ying, LIANG Man-Man, SHEN Yan-Hong* |
| College of Horticulture Science and Technology/Key Laboratory of Horticulture Germplasm Resource Discovery and Innovative Utilization, Hebei Science and Technology Normal University, Qinhuangdao 066000, China |
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Abstract Walnut (Juglans regia) is frequently subjected to extreme weather conditions such as winter frost and severe cold waves, which severely hinder the healthy development of the walnut industry. F-box genes, as critical components of the E3 ubiquitin ligase complex, have been demonstrated to participate in plant stress responses; however, their role in walnut cold resistance remains unclear. To explore the molecular cold resistance mechanism of walnuts, this study selected one-year-old branches of cold-resistant resources Juglans hopeiensis 'Mahetao' and cold-sensitive 'LL29' as experimental materials. Physiological indices and transcriptome analyses of the phloem were conducted following low-temperature stress treatment. The results revealed that 'Mahetao' exhibited higher soluble sugar and soluble protein contents under low-temperature stress compared with 'LL29'. Furthermore, a substantial number of genes were upregulated in one-year-old branches of 'Mahetao' under cold stress. KEGG pathway enrichment analysis indicated that the differentially expressed genes in 'Mahetao' under low-temperature stress were primarily associated with starch and sucrose metabolism, plant hormone signal transduction pathway, etc. Based on these findings, 24 candidate genes related to cold resistance were further screened, including 10 genes involved in starch and sucrose metabolism pathways and 14 transcription factors. Functional analysis of 4 F-box transcription factors revealed their classification into the FBL (F-box and Leucine-Rich Repeat Protein), FBP (F-box and phloem protein 2), and FBU (F-box with unknown functional domain) subfamilies. The promoter regions of the 4 F-box genes contained 8 distinct types of stress-responsive cis-acting elements, with the JrFBU1 gene harboring a low-temperature-responsive LTR element. qPCR analysis demonstrated that the expression levels of JrFBU1 and JrFBP were significantly upregulated under low-temperature stress and consistently higher in 'Mahetao' than in 'LL29', suggesting their potential critical roles in walnut cold resistance. This study provides theoretical and practical insights for advancing cold tolerance improvement and cultivation techniques in walnut.
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Received: 24 February 2025
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Corresponding Authors:
*papayacrazy@aliyun.com
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