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首页> 外文期刊>Transgenic Research >A sweetpotato SRD1 promoter confers strong root-, taproot-, and tuber-specific expression in Arabidopsis, carrot, and potato
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A sweetpotato SRD1 promoter confers strong root-, taproot-, and tuber-specific expression in Arabidopsis, carrot, and potato

机译:甘薯SRD1启动子在拟南芥,胡萝卜和马铃薯中赋予强烈的根,主根和块茎特异性表达

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摘要

Harvestable, starch-storing organs of plants, such as fleshy taproots and tubers, are important agronomic products that are also suitable target organs for use in the molecular farming of recombinant proteins due to their strong sink strength. To exploit a promoter directing strong expression restricted to these storage organs, we isolated the promoter region (3.0 kb) of SRD1 from sweetpotato (Ipomoea batatas cv. ‘White Star’) and characterized its activity in transgenic Arabidopsis, carrot, and potato using the β-glucuronidase (GUS) gene (uidA) as a reporter gene. The SRD1 promoter conferred root-specific expression in transgenic Arabidopsis, with SRD1 promoter activity increasing in response to exogenous IAA. A time-course study of the effect of IAA (50 μM) revealed a maximum increase in SRD1 promoter activity at 24 h post-treatment initiation. A serial 5′ deletion analysis of the SRD1 promoter identified regions related to IAA-inducible expression as well as regions containing positive and negative elements, respectively, controlling the expression level. In transgenic carrot, the SRD1 promoter mediated strong taproot-specific expression, as evidenced by GUS staining being strong in almost the entire taproot, including secondary phloem, secondary xylem and vascular cambium. The activity of the SRD1 promoter gradually increased with increasing diameter of the taproot in the transgenic carrot and was 10.71-fold higher than that of the CaMV35S promoter. The SRD1 promoter also directed strong tuber-specific expression in transgenic potato. Taken together, these results demonstrate that the SRD1 promoter directs strong expression restricted to the underground storage organs, such as fleshy taproots and tubers, as well as fibrous root tissues.
机译:植物的可收获的储存淀粉的器官,例如肉质的主根和块茎,是重要的农艺产品,由于其强大的沉陷强度,它们也适合用于重组蛋白的分子农作。为了利用启动子来指导仅限于这些存储器官的强表达,我们从甘薯(Ipomoea batatas cv。'White Star')中分离了SRD1的启动子区域(3.0 kb),并使用转基因植物鉴定了其在转基因拟南芥,胡萝卜和马铃薯中的活性。 β-葡萄糖醛酸酶(GUS)基因(uidA)作为报告基因。 SRD1启动子赋予转基因拟南芥根特异性表达,SRD1启动子活性响应于外源IAA而增加。对IAA(50μM)的影响的时程研究显示,在治疗后24小时内SRD1启动子活性最大增加。 SRD1启动子的连续5'缺失分析确定了与IAA诱导型表达相关的区域以及分别控制表达水平的包含正负元素的区域。在转基因胡萝卜中,SRD1启动子介导了强的主根特异表达,如GUS染色在几乎整个主根中都很强,包括次生韧皮部,次生木质部和血管形成层。 SRD1启动子的活性随着转基因胡萝卜中主根直径的增加而逐渐增加,并且比CaMV35S启动子的活性高10.71倍。 SRD1启动子还指导转基因马铃薯中的块茎特异性强表达。综上所述,这些结果表明SRD1启动子指导强表达局限于地下存储器官,例如肉质的主根和块茎以及纤维根组织。

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