Minority Postdoctoral Research Fellowship for FY2006

该项目是根据2006年少数民族博士后研究奖学金和支持活动计划授予的。内生真菌与芽孢杆菌的共生关系及营养获取策略的研究本研究将利用报告基因技术来可视化生活在植物组织内部的细菌和真菌（内生菌）之间的相互作用。因此，荧光蛋白将被插入到内生真菌石香兰和内生细菌欧文氏菌。然后将微生物接种到黍属植物的叶子中，并使用荧光显微镜观察微生物在植物组织中的位置，并确定它们是否相互关联。接下来，为了确定微生物之间的关联是否通过增加营养物质的可用性而对它们中的一个或两个有益，微生物将被用作黍属叶片细胞间隙中糖的微观生物传感器。通过将真菌糖响应基因与质粒上的绿色荧光蛋白（gfp）融合并插入到B中，将制成真菌糖生物传感器。通过农杆菌介导的转化将其转化到表生植物中。构建的质粒将导致响应糖消耗的gfp荧光的积累。荧光强度将用于量化可用糖的量。将真菌生物传感器和先前构建的细菌生物传感器一起和分别接种到黍属物种中，并比较每种情况下的糖可用性。另外一项研究，以确定是否生长素生产的Balansia epichardium增加糖的可用性在细胞间隙的黍属物种将进行。通过质粒插入诱变的方法制备生长素缺陷突变体，并与B的野生型生长素生产菌株进行比较。表皮生长因子通过比较在E.生长素缺失突变体和生长素产生菌B侵染的黍属植物中的黑曲霉生物传感器表皮生长因子为了成功地进行拟议的调查，熟练掌握一些[分子遗传学]实验技术将获得。在该奖学金的任期内，罗伯茨博士将在加州大学伯克利分校的史蒂文·林道博士的指导下精通分子生物学。特别是这项研究，她将学习并成为善于构建质粒与报告基因，诱变，和农杆菌介导的转化。这些技能的获得将进一步提高她以前的植物微生物研究经验，重点是一般的研究技术。作为一个全面的研究人员，她将更好地准备开始在植物微生物相互作用的积极研究计划，她计划鼓励本科生获得实验室经验。她还希望担任对追求科学事业感兴趣的学生的导师。此外，她渴望通过分享通过研究项目获得的知识，成为科学界的一名有成就的成员。

This project is awarded under the Minority Postdoctoral Research Fellowships and Supporting Activities Program for 2006. AN INVESTIGATION OF SYMBIOTIC RELATIONSHIPS AND NUTRIENT ACQUISITION STRATEGIES OF ENDOPHYTIC FUNGI AND BACTERIAThis investigation will utilize reporter gene technology to visualize interactions between bacteria and fungi that live inside of plant tissue (endophytes). Thus, fluorescent proteins will be inserted into the endophytic fungus Balansia epichloe and the endophytic bacterium Erwinia herbicola. The microbes will then be inoculated into leaves of Panicum sp. and fluorescent microscopy will be used to visualize where the microbes reside in the plant tissue and to determine whether they associate with each other. Next, to determine whether the association between the microbes is beneficial to one or both of them by increasing nutrient availability, the microbes will be used as microscopic biosensors of sugars in the intercellular spaces of Panicum leaves. A fungal biosensor of sugar will be made by fusing a fungal sugar-responsive gene to green fluorescent protein (gfp) on a plasmid and inserted into B. epichloe via Agrobacterium-mediated transformation. The constructed plasmid will bring about the accumulation of gfp fluorescence in response to sugar consumption. Intensity of fluorescence will be used to quantify the amount of available sugar. The fungal biosensor and previously constructed bacterial biosensors will be inoculated into Panicum sp. together and separately, and sugar availability in each situation will be compared. An additional study to determine whether auxin production by Balansia epichloe increases sugar availability in the intercellular spaces of Panicum sp. will be conducted. Auxin deficient mutants will be made by the process of plasmid insertion mutagenesis and compared to wild-type auxin producing strains of B. epichloe. Increased sugar availability as a result of auxin will be determined by comparing fluorescence intensity visualized in E. herbicola biosensors in Panicum infected with auxin deficient mutants and auxin producing B. epichloe. In order to successfully conduct the proposed investigation, proficiency in a number of [molecular genetic] experimental techniques will be acquired. During the tenure of this fellowship Dr. Roberts will get well versed in molecular biology, under the mentorship of Dr. Steven Lindow at University of California, Berkeley. Specifically for this study, she will learn and become adept at constructing plasmids with reporter genes, mutagenesis, and Agrobacterium tumefaciens-mediated transformation. Acquisition of these skills will further enhance her previous plant microbe research experience, which focused on general research techniques. As a well-rounded researcher she will be better equipped to start an active research program in plant-microbe interactions where she plans to encourage undergraduates to attain laboratory experience. She also wants to serve as a mentor to students interested in pursuing scientific careers. Moreover, she aspires to be an accomplished member of the scientific community by sharing knowledge gained through her research program.