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β-半乳糖苷酶(β-galactosidase,EC3.2.1.23)由植物中广泛分布的一类糖基水解酶组成,被认为与细胞壁多糖的代谢相关.棉花(Gossypiumhirsutum)β-半乳糖苷酶基因已被成功分离,被命名为GhGal1.RNA杂交实验显示该基因在棉花纤维发育的伸长期优势表达.为了分析GhGal1基因的时空表达调控,本研究构建了GhGal1启动子区域(1770bp)与β-葡糖醛酸糖苷酶(glucuronidase,GUS)基因融合的双元载体,通过农杆菌转化烟草植株.对转基因植株分析的结果表明:此转基因果实中的GUS活性比阴性和阳性对照的活性高.GUS组织定位分析表明:β-半乳糖苷酶基因能在根组织的分生区、子叶、维管束组织、果实和表皮毛中表达.此外,调控区域的序列分析揭示该序列含有一些果实/种子特异表达以及与表皮毛表达相关的保守元件.这些结果显示了GhGal1启动子在转基因烟草植株中的时空表达特征,并提供了GhGal1基因参与棉花纤维发育的一些重要线索.
β-galactosidase (EC 3.2.1.23) is composed of a wide range of glycosyl hydrolases distributed in plants and is believed to be involved in the metabolism of cell wall polysaccharides. Gossypium hirsutum beta-galactosidase The gene was successfully isolated and named as GhGal1.RNA hybridization assay showed that the gene was predominantly expressed in the elongation stage of cotton fiber.In order to analyze the temporal and spatial expression of GhGal1 gene, we constructed a GhGal1 promoter region (1770bp) Glucuronidase (GUS) fusion gene was used to transform tobacco plants via Agrobacterium tumefaciens.The results of transgenic plants showed that GUS activity in this transgenic fruit was higher than that of negative and positive controls.GUS Tissue localization analysis showed that the β-galactosidase gene could be expressed in the meristem, cotyledon, vascular bundle, fruit and epidermal hairs of the root tissues.In addition, sequence analysis of the regulatory regions revealed that the sequence contained some fruit / seed-specific Expression and the conserved components related to the expression of epidermal hair.These results show the spatial and temporal expression characteristics of GhGal1 promoter in transgenic tobacco plants and provide the reference gene of GhGal1 Some Important Clues to Cotton Fiber Development.