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ARS Home » Northeast Area » Beltsville, Maryland (BARC) » Beltsville Agricultural Research Center » Molecular Plant Pathology Laboratory » Research » Research Project #438568

Research Project: Developing Cas12a-Based Technologies for Early Detection of Exotic Phytoplasmas

Location: Molecular Plant Pathology Laboratory

Project Number: 8042-22000-306-39-S
Project Type: Non-Assistance Cooperative Agreement

Start Date: Sep 1, 2020
End Date: Aug 31, 2021

Objective:
The goal of this Agreement is to enhance research through collaborations that explore cutting-edge Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas genome editing technology as a tool for sensitive and specific detection of exotic phytoplasmas of agricultural importance. Two specific objectives will be pursued through this Agreement: 1. Identify molecular markers (target sequences) unique to two exotic phytoplasma species, 'Candidatus Phytoplasma solain' and 'Ca. Phytoplasma vitis'; and 2. Establish and optimize CRISPR-associated protein 12a (Cas12a)-based methods for ultra-sensitive detection of the two exotic phytoplasmas.

Approach:
Phytoplasma genome sequences will be retrieved from public databases. Additional sequences will be generated using nextGen Illumina and PacBio sequencing technologies. The obtained phytoplasma genome sequences will be subjected to comparative analysis using bioinformatics software. Exotic phytoplasma-specific genes and sequence blocks will be identified as detection targets. Detection of exotic phytoplasmas will be achieved by using Cas12a nuclease and in vitro transcribed guide RNAs that recognize the specific target sequences in respective phytoplasma genomes.