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LP2 亮氨酸丰富重复受体激酶基因启动子指导转基因水稻中的器官特异性、光响应表达。

The LP2 leucine-rich repeat receptor kinase gene promoter directs organ-specific, light-responsive expression in transgenic rice.

机构信息

USDA-ARS, Western Regional Research Center, Crop Improvement and Utilization Research Unit, Albany, CA, USA.

出版信息

Plant Biotechnol J. 2009 Dec;7(9):867-82. doi: 10.1111/j.1467-7652.2009.00449.x. Epub 2009 Sep 28.

Abstract

Biotechnologists seeking to limit gene expression to nonseed tissues of genetically engineered cereal crops have only a few choices of well characterized organ-specific promoters. We have isolated and characterized the promoter of the rice Leaf Panicle 2 gene (LP2, Os02g40240). The LP2 gene encodes a leucine-rich repeat-receptor kinase-like protein that is strongly expressed in leaves and other photosynthetic tissues. Transgenic rice plants containing an LP2 promoter-GUS::GFP bifunctional reporter gene displayed an organ-specific pattern of expression. This expression corresponded to transcript levels observed on RNA blots of various rice organs and microarray gene expression data. The strongest beta-glucuronidase activity was observed in histochemically stained mesophyll cells, but other green tissues and leaf cell types including epidermal cells also exhibited expression. Low or undetectable levels of LP2 transcript and LP2-mediated reporter gene expression were observed in roots, mature seeds, and reproductive tissues. The LP2 promoter is highly responsive to light and only weak expression was detected in etiolated rice seedlings. The specificity and strength of the LP2 promoter suggests that this promoter will be a useful control element for green tissue-specific expression in rice and potentially other plants. Organ-specific promoters like LP2 will enable precise, localized expression of transgenes in biotechnology-derived crops and limit the potential of unintended impacts on plant physiology and the environment.

摘要

生物技术学家希望将基因表达限制在遗传工程谷类作物的非种子组织中,他们只有几种选择,即具有良好特征的器官特异性启动子。我们已经分离和表征了水稻叶穗 2 基因(LP2,Os02g40240)的启动子。LP2 基因编码一个富含亮氨酸的重复受体激酶样蛋白,在叶片和其他光合组织中强烈表达。含有 LP2 启动子-GUS::GFP 双功能报告基因的转基因水稻植物表现出器官特异性表达模式。这种表达与各种水稻器官的 RNA 印迹和微阵列基因表达数据中观察到的转录水平相对应。在组织化学染色的叶肉细胞中观察到最强的β-葡萄糖醛酸酶活性,但其他绿色组织和叶片细胞类型,包括表皮细胞,也表现出表达。在根、成熟种子和生殖组织中观察到 LP2 转录本和 LP2 介导的报告基因表达水平低或无法检测到。LP2 启动子对光高度敏感,在黄化的水稻幼苗中仅检测到微弱的表达。LP2 启动子的特异性和强度表明,该启动子将成为水稻和潜在其他植物中绿色组织特异性表达的有用控制元件。像 LP2 这样的器官特异性启动子将能够在生物技术衍生作物中精确、本地化地表达转基因,限制对植物生理学和环境的意外影响的潜力。

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