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PsEND1启动子:一种通过早期花药消融产生基因工程雄性不育植物的新工具。

The PsEND1 promoter: a novel tool to produce genetically engineered male-sterile plants by early anther ablation.

作者信息

Roque Edelín, Gómez María D, Ellul Philippe, Wallbraun Michael, Madueño Francisco, Beltrán José-Pío, Cañas Luis A

机构信息

Departamento de Biología del Desarrollo, Instituto de Biología Molecular y Celular de Plantas (C.S.I.C.-U.P.V.), Campus de la Universidad Politécnica de Valencia, Avda. de los Naranjos s/n., 46022 Valencia, Spain.

出版信息

Plant Cell Rep. 2007 Mar;26(3):313-25. doi: 10.1007/s00299-006-0237-z. Epub 2006 Oct 3.

Abstract

PsEND1 is a pea anther-specific gene that displays very early expression in the anther primordium cells. Later on, PsEND1 expression becomes restricted to the epidermis, connective, endothecium and middle layer, but it is never observed in tapetal cells or microsporocytes. We fused the PsEND1 promoter region to the cytotoxic barnase gene to induce specific ablation of the cell layers where the PsEND1 is expressed and consequently to produce male-sterile plants. Expression of the chimaeric PsEND1::barnase gene in two Solanaceae (Nicotiana tabacum and Solanum lycopersicon) and two Brassicaceae (Arabidopsis thaliana and Brassica napus) species, impairs anther development from very early stages and produces complete male-sterile plants. The PsEND1::barnase gene is quite different to other chimaeric genes previously used in similar approaches to obtain male-sterile plants. The novelty resides in the use of the PsEND1 promoter, instead of a tapetum-specific promoter, to produce the ablation of specific cell lines during the first steps of the anther development. This chimaeric construct arrests the microsporogenesis before differentiation of the microspore mother cells and no viable pollen grains are produced. This strategy represents an excellent alternative to generate genetically engineered male-sterile plants, which have proved useful in breeding programmes for the production of hybrid seeds. The PsEND1 promoter also has high potential to prevent undesirable horizontal gene flow in many plant species.

摘要

PsEND1是一种豌豆花药特异性基因,在花药原基细胞中表现出非常早期的表达。后来,PsEND1的表达局限于表皮、连接组织、药室内壁和中层,但在绒毡层细胞或小孢子母细胞中从未观察到。我们将PsEND1启动子区域与细胞毒性的芽孢杆菌RNA酶基因融合,以诱导PsEND1表达所在细胞层的特异性消融,从而产生雄性不育植株。嵌合的PsEND1::芽孢杆菌RNA酶基因在两种茄科植物(烟草和番茄)和两种十字花科植物(拟南芥和甘蓝型油菜)中的表达,从非常早期阶段就损害了花药发育,并产生了完全雄性不育的植株。PsEND1::芽孢杆菌RNA酶基因与之前在类似方法中用于获得雄性不育植株的其他嵌合基因有很大不同。其新颖之处在于使用PsEND1启动子,而不是绒毡层特异性启动子,在花药发育的最初阶段产生特定细胞系的消融。这种嵌合构建体在小孢子母细胞分化之前阻止了小孢子发生,并且没有产生可育的花粉粒。这种策略是产生基因工程雄性不育植株的一种极好的替代方法,已证明其在杂交种子生产的育种计划中很有用。PsEND1启动子在防止许多植物物种中不期望的水平基因流动方面也具有很高的潜力。

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