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This article in CS

  1. Vol. 45 No. 1, p. 346-353
     
    Received: Sept 25, 2003
    Published: Jan, 2005


    * Corresponding author(s): bguo@tifton.usda.gov
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doi:10.2135/cropsci2005.0346

Generation of Expressed Sequence Tags (ESTs) for Gene Discovery and Marker Development in Cultivated Peanut

  1. M. Luoab,
  2. P. Dangc,
  3. B. Z. Guo *a,
  4. G. Hed,
  5. C. C. Holbrooke,
  6. M. G. Bausherc and
  7. R. D. Leeb
  1. a USDA-ARS, Crop Protection and Management Research Unit, Tifton, GA 31793
    b Univ. of Georgia Coastal Plain Experiment Station, Tifton, GA 31793
    c USDA-ARS, U.S. Horticultural Research Lab., Ft. Pierce, FL 34945
    d Center for Plant Biotechnology Research, Tuskegee Univ., Tuskegee, AL 36088
    e USDA-ARS, Crop Genetics and Breeding Research Unit, Tifton, GA 31793

Abstract

Expressed sequence tag (EST) libraries for cultivated peanut (Arachis hypogaea L.) were developed from two cDNA libraries constructed by means of mRNA prepared from leaves of peanut line C34-24 (resistant to leaf spots and Tomato spotted wilt virus) and immature pods of peanut line A13 (tolerant to drought stress and preharvest aflatoxin contamination). Randomly selected cDNA clones were partially sequenced to generate a total of 1825 ESTs, 769 from the C34-24 cDNA library and 1056 from the A13 cDNA library, in which 536 and 769 unique ESTs were identified, respectively. Results of BLASTx search showed that 52.8% of the ESTs from leaf tissue and 78.6% of the ESTs from the pod tissue have homology to genes of known function. Approximately 27.3 and 22.1% of ESTs matching homologous sequences in dbEST of GenBank on the basis of BLASTn algorithm have unknown functions. The ESTs were queried against MIPS functional catalog criteria and sorted according to putative function into 15 categories. A total of 1345 ESTs have been released to GenBank1

1 1345 ESTs have been submitted to GenBank database under accession numbers: CD037499 to CD038843
Four hundred unigenes have been selected from these ESTs and arrayed on glass slides for gene expression analysis, and 44 EST-derived simple sequence repeat (SSR) markers have been characterized for cultivated peanut, in which over 20% of the SSRs produced polymorphic markers among 24 cultivated peanut genotypes. This is the first report of ESTs in cultivated peanut, and further characterization of resistance and stress genes may explain mechanisms functioning in these two peanut lines.

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Copyright © 2005. Crop Science Society of AmericaCrop Science Society of America