Australian Centre for Plant Functional Genomics, Hartley Grove, Urrbrae, SA, Australia.
Plant Biotechnol J. 2010 Jan;8(1):47-64. doi: 10.1111/j.1467-7652.2009.00465.x. Epub 2009 Nov 30.
Engineering of plant protection in cereals requires well characterized tissue-specific and wounding/pathogen-inducible promoters for targeted expression of pathogen responsive and resistance genes. We describe the isolation of seven wheat and rice defensin genes expressed in early developing grain and during grain germination, two developmental stages that are particularly vulnerable to pathogens and insects. Comparison of three-dimensional (3D) models of these rice and wheat PRPI defensins indicated variations in spatial architectures that could reflect their functional diversities. Wheat and rice were stably transformed with promoter-GUS fusion constructs and the spatial and temporal activities of four promoters were studied using whole-mount and histological assays. PRPI promoters were active before and at anthesis in both transgenic wheat and rice with activity mainly in the ovary. In rice, GUS activity was also observed in vascular tissue of the lemma, palea and anthers. After fertilization, GUS was strongly expressed in the outer cell layers of the pericarp and in the main vascular bundle of the grain. During, and a short time after, seed germination, wheat promoters were active in transgenic rice embryos, roots and/or coleoptiles. All wheat and rice promoters were strongly induced by wounding in leaf, stem and grain of transgenic rice plants. These results suggest that PRPI promoters will be useful for specific targeting and accumulation of proteins conferring resistance to pathogens in vulnerable tissues of developing and germinating grain.
在谷物中进行植物保护工程需要具有良好特征的组织特异性和创伤/病原体诱导启动子,以便靶向表达对病原体有反应和抗性的基因。我们描述了七个在早期发育谷物和谷物萌发过程中表达的小麦和水稻防御素基因的分离,这两个发育阶段特别容易受到病原体和昆虫的侵害。对这些水稻和小麦 PRPI 防御素的三维 (3D) 模型进行比较表明,空间结构存在差异,这可能反映了它们的功能多样性。使用启动子-GUS 融合构建体稳定转化小麦和水稻,并用整体和组织学分析研究了四个启动子的时空活性。PRPI 启动子在小麦和水稻的开花前和开花期均具有活性,活性主要在子房中。在水稻中,GUS 活性也在颖片、外稃和花药的维管束中观察到。受精后,GUS 在果皮的外层细胞和谷物的主维管束中强烈表达。在种子萌发期间和之后不久,小麦启动子在转基因水稻的胚胎、根和/或幼叶中活跃。所有小麦和水稻启动子在转基因水稻叶片、茎和谷物受伤时均被强烈诱导。这些结果表明,PRPI 启动子将有助于在发育和萌发谷物的脆弱组织中特异性靶向和积累赋予对病原体抗性的蛋白质。