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

了解微生物-植物相互作用对于防治农业上重要的植物病害和提高植物产量至关重要。农杆菌属物种在超过300种植物属上引起冠瘿病，包括许多经济上重要的作物。目前冠瘿病的控制方法不是很有效，导致高达80%的感染和苗圃植物的损失。或者，农杆菌介导的转化是产生具有增加的农业生产力的转基因植物的有力工具。但许多作物都不愿意转化。该奖项支持的研究，将采用新的方法开发的教授桑杰Swarup？他在新加坡国立大学（NUS）的实验室研究了转基因和未处理的A.利用16 S rRNA宏基因组学分析，了解农杆菌毒力蛋白如何操纵植物反应以增加感染，可以帮助改善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-微生物相互作用1。我建议调查根分泌物代谢物的变化，转基因拟南芥表达发根农杆菌效应蛋白，GALLS-CT，从β-雌二醇诱导启动子2，并测定这种蛋白质的表达是否改变植物根际微生物组。GALLS-CT蛋白从农杆菌转移到植物中3，并改变宿主防御基因的表达以增强感染（未发表的数据）。我将使用桑杰·斯瓦鲁普教授发明的新方法？他在新加坡国立大学（NUS）的实验室研究了转基因和未处理的A.利用16 S rRNA宏基因组学分析，目的是观察可能由GALLS-CT表达引起的微生物群落的变化。我还将收集诱导GALL-CT表达植物和未处理植物的根系分泌物，并使用超高压液相色谱-质谱法进行代谢组学分析（UHPLC-MS），以寻找可能由GALLS引起的代谢产物谱差异-该奖项在东亚和太平洋夏季研究所计划下支持美国研究生的夏季研究，由NSF和国家科学基金会共同资助。新加坡研究基金会。

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.