Monday, November 13, 2006

Evaluation of a Peanut cDNA Library Enriched for Root-knot Nematode-Specific Transcripts by Suppression Subtractive Hybridization.

Sivananda Varma Tirumalaraju, Univ. of Florida-Agronomy Dept., 2183 Mccarty A Hall Po Box 110300, Gainesville, FL 32611 and Maria Gallo, Agronomy department, Plant Molecular and Cellular Biology Program, and The Genetics Institute, University of Florida, 303 Cancer/Genetics Research Complex, 1376 Mowry Road, Gainesville, FL 32610-3610.

Root-knot nematodes are the most damaging pests of peanut. Economic losses due to root-knot nematodes are significant in many peanut producing areas of Florida, Alabama, Georgia and Texas. Approximately, a 30% decrease in yield potential has been observed due to Meloidogyne arenaria (Neal) Chitwood race 1 alone. To identify possible candidate genes conferring resistance to this pest, we have constructed a cDNA library enriched with differentially expressed sequences, obtained by Mirror Orientation Selection PCR, from root tissues of the nematode resistant, cv. NemaTAM (Ne) and the nematode susceptible, cv. Florunner (FL) through suppression subtractive hybridization. Randomly picked clones (480) from the tester Ne-specific library and driver the FL-specific library were used for differential screening. From this screen, 140 Ne -specific clones and 123 FL -specific clones were obtained. Virtual northern blot analysis was performed to confirm the differential screening results. Further expression analysis by RT-PCR is currently in progress. The analysis of Ne- and FL- specific gene expression is an essential step in understanding the mechanism of peanut resistance to nematode infestation.

Handout (.pdf format, 84.0 kb)