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菊花类胡萝卜素裂解双加氧酶 4a-5 基因启动子驱动发育中花瓣共轭基因的特异性转录。

The promoter of the carotenoid cleavage dioxygenase 4a-5 gene of Chrysanthemum morifolium (CmCCD4a-5) drives petal-specific transcription of a conjugated gene in the developing flower.

机构信息

Department of Biomolecular Science, Faculty of Science, Toho University, 2-2-1 Miyama, Funabashi, Chiba 274-8510, Japan.

出版信息

J Plant Physiol. 2013 Sep 15;170(14):1295-9. doi: 10.1016/j.jplph.2013.04.001. Epub 2013 May 2.

Abstract

Carotenoids comprise one of the major groups of pigments in flowers. Because carotenoids are physiologically indispensable pigments for all photosynthetic plants, their catabolism must be discretely regulated in photosynthetic organs and non-photosynthetic organs such as petals or fruits. In the chrysanthemum, carotenoid cleavage dioxygenase 4a (CmCCD4a), which is dominantly expressed in petals, cleaves carotenoid, leading to a white flower. CmCCD4a-5 was recently identified as a new member of the CmCCD4a family, but its detailed expression profile in plant tissues has not yet been established. In this study, we sequenced a 1094-bp region upstream of CmCCD4a-5 and assessed its petal-specific promoter activity. To evaluate the activity of this gene, we constructed two types of transgenic Arabidopsis thaliana that possessed, respectively, a fusion gene of a 1090-bp or 505-bp segment of the upstream region plus the β-d-glucuronidase (GUS) gene (1090bUR::GUS and 505bUR::GUS). GUS activity in the 505bUR::GUS strain was observed mainly in the anthers/pollen in flower buds, whereas GUS activity of the 1090bUR::GUS strain was observed in immature petals of the flower buds. Among the cis-acting elements located between positions -505 and -1090, no elements that have previously been reported to enhance the expression in petals or to suppress it in anthers/pollen were detected by PLACE analysis, indicating the existence of unknown cis-element(s). A semiquantitative reverse transcription-polymerase chain reaction analysis revealed that CmCCD4a-5 transcription was prominent in petals but was undetectable in roots, stems and leaves.

摘要

类胡萝卜素是花中主要色素之一。由于类胡萝卜素是所有光合植物生理上不可缺少的色素,它们的分解代谢必须在光合器官和非光合器官(如花瓣或果实)中进行离散调节。在菊花中,主要在花瓣中表达的类胡萝卜素裂解双加氧酶 4a(CmCCD4a)裂解类胡萝卜素,导致白花。CmCCD4a-5 最近被鉴定为 CmCCD4a 家族的一个新成员,但它在植物组织中的详细表达谱尚未建立。在这项研究中,我们对 CmCCD4a-5 上游的 1094bp 区域进行了测序,并评估了其花瓣特异性启动子活性。为了评估该基因的活性,我们构建了两种类型的拟南芥转基因植株,分别含有上游 1090bp 或 505bp 片段与β-葡糖苷酸酶(GUS)基因的融合基因(1090bUR::GUS 和 505bUR::GUS)。在花蕾中,505bUR::GUS 株系的 GUS 活性主要在花药/花粉中观察到,而 1090bUR::GUS 株系的 GUS 活性在花蕾的未成熟花瓣中观察到。在位置-505 和-1090 之间的顺式作用元件中,通过 PLACE 分析没有检测到以前报道的增强花瓣表达或抑制花药/花粉表达的元件,表明存在未知的顺式元件。半定量逆转录聚合酶链反应分析显示,CmCCD4a-5 转录在花瓣中明显,但在根、茎和叶中检测不到。

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