EAPSI: Working to Increase Agricultural Productivity through Agrobacterium-Mediated Transformation in A. Thaliana

了解微生物与植物的相互作用对于防治重要的农业植物病害和提高植物产量至关重要。农杆菌在300多个植物属上引起冠瘿病，包括许多重要的经济作物。目前对冠瘿病的控制方法不是很有效，导致苗圃植物侵染和损失高达80%。另外，农杆菌介导的转化是创造具有更高农业生产力的转基因植物的有力工具。但许多作物难以转化。该奖项支持采用桑杰·斯瓦鲁普教授开发的新方法的研究。利用16S rRNA宏基因组分析研究了转基因和未经处理的拟南芥根际微生物组的群落组成。了解一种农杆菌毒力蛋白如何操纵植物对增加感染的反应，有助于改善对冠瘿病的预防和控制策略，以及创造更好的植物转化技术，以生产有用的转基因作物。细菌和植物代谢产物在根际环境中丰富，并调节植物与微生物的相互作用1。我建议研究从甜菜雌二醇诱导启动子中表达根源农杆菌效应蛋白（gals - ct）的转基因拟南芥根分泌物代谢物的变化，并测定该蛋白的表达是否改变了植物根际微生物群。GALLS-CT蛋白从农杆菌转移到植物中，改变宿主防御基因的表达以增强感染（未发表的数据）。我会采用桑杰·斯瓦鲁普教授开发的新方法？利用16S rRNA宏基因组分析研究了转基因和未经处理的拟南芥根际微生物组的群落组成。目的是观察可能由GALLS-CT表达引起的微生物群落的变化。我还将收集表达GALLS-CT的诱导植株和未处理植株的根分泌物，并使用超高压液相色谱-质谱（UHPLC-MS）进行代谢组学分析，寻找可能由GALLS-CT引起的代谢物谱差异。该奖项由美国国家科学基金会和新加坡国家研究基金会共同资助，隶属于东亚和太平洋暑期研究所项目，支持美国研究生进行暑期研究。

Understanding microbe-plant interactions is crucial to combating agriculturally important plant diseases and improving plant production. Agrobacterium species cause crown gall disease on over 300 plant genera, including many economically important crops. Current control methods for crown gall disease are not very effective, resulting in up to 80% infection and loss in nursery plants. Alternatively, Agrobacterium-mediated transformation is a powerful tool for creating transgenic plants with increased agricultural productivity. But many crops are recalcitrant to transformation. This award supports research that will employ novel methods developed by Professor Sanjay Swarup?s lab at the National University of Singapore (NUS) to look at the community composition of the rhizosphere microbiomes of transgenic and untreated A. thaliana using 16S rRNA metagenomic analysis. Understanding how an Agrobacterium virulence protein manipulates plant response to increase infection can help improve prevention and control strategies for crown gall disease, as well as create better plant transformation technologies for making useful transgenic crops. Bacterial and plant metabolites are abundant in the rhizosphere environment and modulate plant-microbe interactions1. I propose to investigate the changes in root exudate metabolites of transgenic Arabidopsis thaliana expressing an Agrobacterium rhizogenes effector protein, GALLS-CT, from a betaestradiol inducible promoter2, and assay if expression of this protein alters the plant rhizosphere microbiome. The GALLS-CT protein is transferred from Agrobacterium into the plant3 and alters host defense gene expression to enhance infection (unpublished data). I will employ novel methods developed by Professor Sanjay Swarup?s lab at the National University of Singapore (NUS) to look at the community composition of the rhizosphere microbiomes of transgenic and untreated A. thaliana using 16S rRNA metagenomic analysis. The goal is to observe changes in the microbial community that might be caused by GALLS-CT expression. I will also collect root exudates from induced GALL-CT expressing plants and untreated plants and perform metabolomics analysis using ultra high-pressure liquid chromatography-mass spectrometry (UHPLC-MS) to look for differences in metabolite profiles that might be caused by GALLS-CT.This award under the East Asia and Pacific Summer Institutes program supports summer research by a U.S. graduate student and is jointly funded by NSF and the National Research Foundation of Singapore.