亚热带农业研究2025，Vol.21Issue(4)：261-270,10.DOI:10.13321/j.cnki.subtrop.agric.res.2025.04.006
甜菜蓝光受体基因BvCRY1和BvCRY2的克隆及亚细胞定位
Cloning and subcellular localization analysis of blue-light receptor genes BvCRY1 and BvCRY2 in Beta vulgaris L
摘要
Abstract
[Purpose]This study cloned the blue-light receptor genes BvCRY1 and BvCRY2in Beta vulgaris L.and employed bioin-formatics tools to analyze the characteristics of the proteins encoded by the two genes,aiming to provide theoretical reference for elu-cidating the function mechanisms of CRYs and the breeding high-quality varieties of B.vulgaris.[Method]BvCRY1 and BvCRY2 were cloned via RACE from the B.vulgaris variety"IMA1".Seamless cloning was employed to construct the recombinant expression plasmids 163-BvCRY1-hGFP and 163-BvCRY2-hGFP.These plasmids were transformed into Arabidopsis thaliana protoplasts via the PEG-mediated method,followed by subcellular localization observation.Subsequently,bioinformatics tools were utilized to analyze the physicochemical properties and structural characteristics of the proteins encoded by BvCRY1 and BvCRY2.[Result]The target genes were cloned and the recombinant plasmids 163-BvCRY1-hGFP and 163-BvCRY2-hGFP were constructed for transformation of A.thaliana protoplasts.Laser confocal microscopy revealed that the green fluorescent signal of BvCRY1 was distributed in both the cytoplasm and nucleus,whereas the fluorescent signal of BvCRY2 was specifically enriched in the nucleus.Bioinformatics analysis demonstrated that BvCRY1(2 058 bp)and BvCRY2(1 899 bp)encoded 685 and 632 amino acid residues,respectively.Both pro-teins were hydrophilic and unstable,with secondary structures predominantly consisting of random coils and α-helices.The proteins contained the conserved PHR and CCE domains of the CRY family,as well as multiple phosphorylation sites.Compared with the CRY proteins from other species,BvCRY1 and BvCRY2 were highly conserved in evolution and exhibited the highest similarity to the CRY protein from Chenopodium quinoa.[Conclusion]The subcellular localization differences suggest that BvCRY1 and BvCRY2 exhibit functional differentiation in the blue-light signaling pathway.The structural specificity and localization characteris-tics of BvCRYs may represent adaptive evolutionary outcomes in response to the harsh light conditions of northern arid environments,providing novel targets for breeding stress-resistant varieties of B.vulgaris.关键词
甜菜/隐花色素/生物信息/亚细胞定位Key words
Beta vulgaris L./cryptochrome/bioinformation/subcellular localization分类
农业科技引用本文复制引用
郑雯慧,高萍茹,何文锦..甜菜蓝光受体基因BvCRY1和BvCRY2的克隆及亚细胞定位[J].亚热带农业研究,2025,21(4):261-270,10.基金项目
国家糖料产业技术体系项目"糖料资源综合利用"(CARS-17) （CARS-17）
福建省自然科学基金项目"隐花色素介导蓝绿光调控机制"(2023J01509). （2023J01509）
