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United States Department of Agriculture

Agricultural Research Service

Research Project: Molecular Fingerprinting of Microbial Populations in Phytoplasma-Diseased Plants

Location: Molecular Plant Pathology

Project Number: 1245-22000-282-08
Project Type: Specific Cooperative Agreement

Start Date: Sep 23, 2009
End Date: Jul 31, 2014

Objective:
1. Characterize microbial communities associated with tissue necrosis during the terminal stage of lethal yellowing disease of palms. 2. Determine genetic variability/diversity of phytoplasmas affecting potato and other crops. The ultimate goal of the project is to gain new knowledge of ecology, niche adaptation and pathogenicity evolution of phytoplasmas, causal agents of economically important plant diseases worldwide.

Approach:
For Objective 1, necrotic tissues will be harvested from lethal yellows (LY) disease-affected palm trees and total DNA will be extracted from selected tissues. Microbial populations will be characterized using 454 pyrosequencing technology with primers designed to target V3 and V6 hypervariable regions of bacterial 16S rRNA genes. For Objective 2, total DNA will be extracted from symptomatic tissues of phytoplasma-infected plants, and phytoplasma DNA will be enriched through cesium chloride gradient centrifugation. Phytoplasma genome survey sequencing will be conducted using 454 pyrosequencing technology using coded primers. For both Objectives, the sequence data obtained will be analyzed and annotated using a suite of bioinformatic tools.

Last Modified: 9/1/2014
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