DEVELOPMENT AND VALIDATION OF A UNIVERSAL PLANT VIRUS MICROARRAY FOR DETECTION AND IDENTIFICATION OF PLANT VIRUSES
Floral and Nursery Plants Research Unit
2009 Annual Report
1a.Objectives (from AD-416)
To develop a universal plant virus micro array for detection and identification of plant viruses.
1b.Approach (from AD-416)
Develop microarrray containing probes representing all known plant viruses.
A project to develop a Universal Plant Virus Microarray has been initiated as a collaborative effort between the Floral and Nursery Plants Research Unit, ARS; the Donald Danforth Plant Science Center; Washington University; the University of Utah; Oklahoma State University, and Cornell University. The goals are to be able to detect any plant virus in extracts of infected plants, and to identify previously characterized viruses to the species level, or previously uncharacterized virus to at least the viral family or genus level. Prior to development of the full microarray, the system is being validated for approximately 50 characterized viruses represented by a total of 750 virus-specific oligonucleotides, plus control host genes, on a ‘MiniPlantViroChip’ array. To date all but one of the virus isolates tested have been successfully detected with the majority of oligonucleotides representing the virus in question, without amplification of the samples. The tested samples represent viruses with both RNA and DNA genomes, including members of the families Geminiviridae and Caulimoviridae (DNA), as well as species in the Bromoviridae, Flexiviridae, Potyviridae, and genus Tobamovirus (RNA), including multiple strains of some viruses. These results demonstrate the specificity and sensitivity of the array over a wide dynamic range of probe intensities, and form a strong background for development of the full scale Universal array. The Universal Plant Virus Microarray will be of value to diagnosticians in the National Plant Diagnostic Network, plant introduction and quarantine facilities, clean stock virus certification programs, and plant propagators introducing new varieties.
Research activities conducted under this agreement were monitored by regular email and phone communication and in person at scientific meetings.