TRTech-PGR: PlantTransform: Boosting Agrobacterium-mediated transformation efficiency in the orphan crop tef (Eragrostis tef) for trait improvement

植物遗传转化在理解复杂性状的遗传基础方面起着关键作用。模式植物和一些主要作物（如水稻和玉米）的转化方法已经开发出来，这些作物通常缺乏必需营养素。相比之下，由于基因组信息有限，科学家和资助机构缺乏关注，对tef （Eragrostis tef）等未充分利用或孤立作物的遗传转化尚处于起步阶段。tef不含麸质，富含营养物质（矿物质、维生素、氨基酸）和有益的植物化学物质。这极大地推迟了这类作物的性状改进。因此，有必要开发一种有效的转化方法来改变tef基因型的理想性状，包括种子大小和颜色、矿物质含量、铁的生物利用度和成熟天数。此外，作为tef的野生祖先，印度lovegrass （E. pilosa）具有适应更广泛环境条件、早熟、耐倒伏等优良性状，是tef改良的潜在性状来源。该项目的主要目标是开发不同基因型的tef和E. pilosa转化工具。该项目还将为北卡罗来纳大学格林斯博罗分校的少数民族学生、一名博士后研究员和一名来自埃塞俄比亚的访问科学家提供培训机会，在埃塞俄比亚，大豆是最重要的主要作物。此外，该项目将支持在埃塞俄比亚农业研究所举办的tef遗传转化讲习班。近年来，利用玉米胚性调控基因Baby boom （Bbm）和wushel （Wus2）对包括tef在内的顽固性单子房的农杆菌介导转化显示出巨大的前景。在某些品种之间也观察到转化效率的差异。然而，这些方法仍有待优化。具体目标是：1)开发一种高效的tef遗传转化技术；2)开发毛囊草的转化方法。最近开发的几种农杆菌菌株，如胸腺营养不良菌株和工程毒力菌株，不同的tef基因型，在理想性状和目标组织（叶鞘和胚性愈伤组织）上存在差异，以及不同的组织培养条件，将进行测试，以开发一种有效的转化tef及其野生亲缘印度爱草的方法。该项目产生的资源将以出版物、协议、研讨会、演示和网站的形式与科学界免费共享。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Plant genetic transformation plays a key role in understanding the genetic basis of complex traits. Transformation methods have been developed for model plants and some major crops such as rice and corn, which are usually poor in essential nutrients. In contrast, genetic transformation for underutilized or orphan crops such as tef (Eragrostis tef), which are gluten-free and rich in nutrients (minerals, vitamins, amino acids) and beneficial phytochemicals, is at a very infant stage due to limited genome information and lack of attention from scientists and funding agencies. This has tremendously delayed trait improvement in such crops. Therefore, there is a need to develop an efficient transformation method for tef genotypes varying in desirable traits including seed size and color, mineral content, iron bioavailability, and days to maturity. Moreover, transformation method has not been developed for the Indian lovegrass (E. pilosa), which is a wild progenitor of E. tef, and has several desirable traits including adaptation to wider environmental conditions, early maturity, and lodging tolerance, and is a potential source of traits for tef improvement. The major goal of this project is to develop transformation tools for diverse genotypes of E. tef and E. pilosa. This project will also offer training opportunities for University of North Carolina–Greensboro minority students, a postdoctoral researcher, and a visiting scientist from Ethiopia where tef is the most important staple crop. Furthermore, this project will support a tef genetic transformation workshop to be organized at the Ethiopian Institute of Agricultural Research.Recently, the use of maize embryogenic regulator genes Baby boom (Bbm) and wushel (Wus2) has shown great promise for Agrobacterium-mediated transformation of recalcitrant monocots including tef. Differences in transformation efficiency have also been observed between some tef varieties. However, the methods remain to be optimized. The specific objectives are: 1) to develop an efficient genetic transformation technique for tef, and 2) to develop transformation method for E. pilosa. Several Agrobacterium strains recently developed for recalcitrant monocots such as the thymine auxotrophic strains and engineered virulent strains, different tef genotypes varying in desirable traits and target tissues (leaf sheath and embryogenic calli), and varying tissue culture conditions will be tested to develop an efficient transformation method for tef and its wild relative Indian love grass. Resources generated from this project will be freely shared with the scientific community in the form of publications, protocols, workshops, presentations and websites.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.