Identification of Heterodera glycines (soybean cyst nematode [SCN]) cDNA sequences with high identity to those of Caenorhabditis elegans having lethal mutant or RNAi phenotypes.
The soybean cyst nematode (SCN; Heterodera glycines) is a devastating obligate parasite of Glycine max (soybean) causing one billion dollars in losses to the US economy per year and over ten billion dollars in losses worldwide. While much is understood about the pathology of H. glycines, its genome sequence is not well characterized or fully sequenced. We sought to create bioinformatic tools to mine the H. glycines nucleotide database. One way is to use a comparative genomics approach by anchoring our analysis with an organism, like the free-living nematode Caenorhabditis elegans. Unlike H. glycines, the C. elegans genome is fully sequenced and is well characterized with a number of lethal genes identified through experimental methods. We compared an EST database of H. glycines with the C. elegans genome. Our goal was identifying genes that may be essential for H. glycines survival and would serve as an automated pipeline for RNAi studies to both study and control H. glycines. Our analysis yielded a total of nearly 8334 conserved genes between H. glycines and C. elegans. Of these, 1508 have lethal phenotypes/phenocopies in C. elegans. RNAi of a conserved ribosomal gene from H. glycines (Hg-rps-23) yielded dead and dying worms as shown by positive Sytox fluorescence. Endogenous Hg-rps-23 exhibited typical RNA silencing as shown by RT-PCR. However, an unrelated gene Hg-unc-87 did not exhibit RNA silencing in the Hg-rps-23 dsRNA-treated worms, demonstrating the specificity of the silencing.
Alkharouf
Nadim W
NW
United States Department of Agriculture, ARS, Soybean Genomics and Improvement Laboratory, Beltsville, MD 20705-2350, USA.
Klink
Vincent P
VP
Matthews
Benjamin F
BF
D000818:Animals; D001483:Base Sequence; D017173:Caenorhabditis elegans / Q000145:classification / Q000235:genetics*; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000737:chemistry; D018509:DNA, Helminth / Q000737:chemistry; D030561:Databases, Nucleic Acid; D020224:Expressed Sequence Tags / Q000737:chemistry; D005804:Genes, Lethal; D010641:Phenotype; D034622:RNA Interference; D018510:RNA, Helminth / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D013025:Glycine max / Q000469:parasitology*; D014431:Tylenchoidea / Q000145:classification / Q000235:genetics*
D018076:DNA, Complementary; D018509:DNA, Helminth; D018510:RNA, Helminth