TALE核酸酶（TALENs）有着设计简单、组装容易、特异性高、广适性强等优点，是基因定点改造中最具应用潜力的新工具。目前TALENs技术已在多个物种中成功应用，如用其定制基因功能缺失突变体，可加速基因功能研究。本实验室前期研究结果表明二穗短柄草BdHTAs基因过表达能提高农杆菌转化效率。为准确揭示BdHTAs基因功能，本申请拟首先建立TALENs的设计、组装、筛选和优化技术体系；再用TALENs技术创制BdHTAs13个成员基因功能缺失突变体，通过遗传学和分子生物学手段分析，明确BdHTAs基因功能；然后筛选出能显著提高农杆菌转化效率的成员BdHTA1，通过分子机制研究，揭示农杆菌转化机理。研究可望解析BdHTAs基因功能，为利用BdHTA1提高农杆菌转化效率提供理论依据；同时搭建二穗短柄草TALENs基因定点突变技术体系，为麦类作物基因功能解析和定向分子育种提供新的技术途径。

TALE Nucleases (TALENs) are recently discovered promising tools that can be used for targeted genome modifications. With remarkable targeting range, high target specificity,TALENs are easy to design and assemble to create DNA breaks at precise loci which can be reparied by NHEJ in any organism. Targeted mutagenesis induced by TALENs has been successfully estalished in several model organisms and TALENs hold much promise to understand gene function.Our previous investigation on BdHTAs, a histone gene family in Brachypodium distachyon, revealed that BdHTAs overexprsion could improve Agrobacterium transformation efficiency. In this study, we plan to create BdHTAs targeted mutagenesis mutants using TALENs, and investigate BdHTAs gene functions involved in Agrobacterium mediated transformation. So we will firstly develop methods including TALENs design, assembly, selection and optimization, and then create gene function lost mutants of 13 members of BdHTAs by TALENs and analyze gene function using different genetic and molecular methods. Finally, we will identify BdHTA1 and discover molecular mechanism of Agrobacterium mediated transformation in Brachypodium. Through the process of a creating BdHTA targeted mutagenesis mutants, we will develop routine targeted mutagenesis with TALENs in Brachypodium. TALENs technology is being deployed in DNA targeting for applications ranging from understand gene function in Brachypodium to improving traits in Triticeae crop plants.