洋葱AcSCL4的克隆及其功能分析  

Cloning and function analysis of AcSCL4 in onion

作　　者：张旭[1,2] 吴小旭 张智慧 胡云捷[1,2] 秦蕾 王勇 ZHANG Xu;WU Xiaoxu;ZHANG Zhihui;HU Yunjie;QIN Lei;WANG Yong(Key Laboratory of Biology and Genetic Improvement of Horticultural Crops(Northeast Region),Ministry of Agriculture and Rural Affairs,Northeast Agricultural University,Harbin,Heilongjiang 150030,China;College of Horticulture and Landscape Architecture,Northeast Agricultural University,Harbin,Heilongjiang 150030,China)

机构地区：[1]东北农业大学农业农村部东北地区园艺作物生物学与种质创制重点实验室,黑龙江哈尔滨150030 [2]东北农业大学园艺园林学院,黑龙江哈尔滨150030

出　　处：《西北农林科技大学学报（自然科学版）》2024年第2期127-134,共8页Journal of Northwest A&F University(Natural Science Edition)

基　　金：黑龙江省重点研发计划项目(GA21B012);黑龙江省自然科学基金联合引导项目(LH2020C012)。

摘　　要：【目的】初步探讨AcSCL4在洋葱成花中的功能,为解析洋葱成花分子调控机制提供理论基础。【方法】以长日生态型洋葱品系SB2为试验材料,根据已有的洋葱转录组数据,通过qRT-PCR筛选目的基因AcSCL4后进行基因克隆,并对其进行生物信息学分析;构建AcSCL4基因的过表达载体pB221-3300-AcSCL4并转化农杆菌,采用花序浸染法浸染拟南芥,并对T_(3)代纯系转基因植株进行表型分析和开花相关基因表达量分析。【结果】洋葱AcSCL4基因CDS长1515 bp,编码504个氨基酸;AcSCL4蛋白N端高度变异,C端有GRAS结构域。聚类分析发现,AcSCL4与石刁柏和棉花的SCL4蛋白亲缘关系较近。与野生型拟南芥相比,AcSCL4过表达植株表现晚花表型。荧光定量PCR结果表明,与野生型拟南芥相比,AcSCL4过表达植株开花调控基因CO和FT的表达量极显著下降。【结论】AcSCL4通过调控CO及FT基因表达来抑制洋葱开花。【Objective】The function of AcSCL4 in flowering was preliminarily studied to provide basis for understanding molecular regulation mechanism of onion flowering regulation.【Method】AcSCL4 was identified by qRT-PCR and bioinformatics analysis based on the transcriptome data of a long-day onion inbred line SB2.The AcSCL4 overexpression vector pB221-3300-AcSCL4 was constructed and transformed to Arabidopsis by Agrobacterium tumefaciens.Homozygous Arabidopsis transgenic lines(T_(3))were used for further phenotypic characterization and genes expression identification.【Result】The cDNA of AcSCL4 gene was 1515 bp and encoded 504 amino acids.AcSCL4 had conserved GRAS domain at the C terminus and was highly variable at the N terminus.AcSCL4 was phylogenetically closer to the SCL4 of Asparagus officinalis and Gossypium hirsutum.Overexpression of AcSCL4 delayed the flowering time of wild-type Arabidopsis.The expressions of CO and FT were significantly decreased in AcSCL4 overexpression transgenic lines.【Conclusion】AcSCL4 suppressed flowering by regulating CO and FT expressions.

关 键 词：洋葱 AcSCL4 基因克隆 开花调控 

分 类 号：S633.203.3[农业科学—蔬菜学]