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
摘要冬季霜冻和强寒潮等极端天气严重制约核桃(Juglans regia)产业的健康发展。F-box基因作为E3泛素连接酶复合体的关键组分,已被证实参与植物逆境响应,但其在核桃抗寒中的作用尚不明确。为探究核桃的抗寒分子机制,本研究选取抗寒性较强的'麻核桃'和抗寒性较弱的'LL29'一年生枝作为实验材料,对这些枝条进行低温胁迫处理并对其韧皮部生理指标和转录组进行分析。结果表明,在低温胁迫下'麻核桃'的可溶性糖和可溶性蛋白含量高于'LL29'。此外,'麻核桃'的一年生枝在低温胁迫下大量基因上调表达。KEGG通路富集分析显示,差异表达基因主要涉及淀粉和蔗糖代谢、植物激素信号转导等通路。在此基础上,本研究进一步筛选出24个与抗寒性相关的候选基因,其中包括10个淀粉和蔗糖代谢途径基因以及14个转录因子。对其中4个F-box转录因子进行功能分析,结果显示,4个F-box转录因子分别属于FBL (F-box and leucine-rich repeat protein)、FBP (F-box and phloem protein 2)和FBU (F-box with unknown functional domain)亚族,转录因子基因的启动子区域含有8种不同类型的胁迫响应元件,其中JrFBU1基因含有低温响应的LTR元件。qPCR分析显示,JrFBU1和JrFBP在低温胁迫下表达量显著升高,且在'麻核桃'中的表达量始终高于'LL29',推测这2个基因可能在核桃抵御低温逆境中具有重要作用。本研究对于推动核桃抗寒性改良和栽培技术的进步具有重要的理论和实践意义。
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.
姜涛, 安孝莹, 梁曼曼, 申艳红. 核桃低温胁迫转录组解析及F-box基因抗寒功能初鉴[J]. 农业生物技术学报, 2025, 33(9): 1948-1960.
JIANG Tiao, AN Xiao-Ying, LIANG Man-Man, SHEN Yan-Hong. Transcriptome Analysis of Walnut (Juglans regia) Under Low Temperature Stress and Preliminary Identification of the Cold Resistance Function of F-box Genes. 农业生物技术学报, 2025, 33(9): 1948-1960.
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