TAL effector nuclease (TALEN)-mediated generation of broad-spectrum powdery mildew resistant mlo plants

TAL 效应核酸酶 (TALEN) 介导的广谱白粉病抗性 mlo 植物的产生

• 批准号: 223610928
• 负责人: Professor Dr. Thomas Debener
• 金额: --
• 依托单位: Institut für Pflanzengenetik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2012
• 资助国家: 德国
• 起止时间: 2011-12-31 至 2015-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/223610928?language=en
• 关键词: TAL; effector; nuclease; TALEN; mediated

Durable broad-spectrum powdery mildew resistance conferred by loss-of-function mutant alleles of Mildew Locus O (MLO) genes is a conserved phenomenon in monocot and dicot plants. Some species that suffer notoriously from the powdery mildew disease, such as rose (Rosa spec.) or grapevine (Vitis vinifera), are for various reasons (e.g. polyploidy) not well accessible to forward and/or reverse genetic screens to obtain suitable mlo mutants. TAL effector nucleases (TALENs) provide a novel tool for site-specific mutagenesis of genes even within the context of complex eukaryotic genomes. In a collaborative effort we aim to establish TALENs that target MLO genes to generate tailor-made mlo loss-of-function alleles. Proof-of-principle will be provided in plant systems in which the analysis of mlo resistance is well established (barley and Arabidopsis thaliana). Next, TALENs will be used to generate powdery mildew resistant mlo mutants in rose, a plant species for which durable genetic resources to combat powdery mildew are lacking so far. The proposed research will not only open up new avenues for the targeted generation of powdery mildew resistant plant mutants but also provide valuable general insights for the deployment of TALENs in precision genome engineering.

在单子叶植物和双子叶植物中，由白粉菌O(MLO)基因功能缺失突变等位基因产生的持久的广谱白粉病抗性是一种保守的现象。有些种类的白粉病是臭名昭著的，如月季(Rosa Spec.)或葡萄(Vitis Vinifera)，由于各种原因(如多倍体)，正向和/或反向遗传筛选不能很好地获得合适的MLO突变体。TAL效应核酸酶(TALEN)为基因的定点突变提供了一种新的工具，即使在复杂的真核基因组中也是如此。在一项合作努力中，我们的目标是建立针对MLO基因的TALEN，以产生定制的MLO功能丧失等位基因。将在建立了MLO抗性分析的植物系统(大麦和拟南芥)中提供原则证明。接下来，TALEN将被用来在月季中产生抗白粉病的MLO突变体，这是一种到目前为止还缺乏持久的遗传资源来抗击白粉病的植物物种。这项研究不仅将为白粉病抗性植物突变体的定向产生开辟新的途径，还将为TALEN在精确基因组工程中的应用提供有价值的一般性见解。