| Autor: |
Coyne CJ; USDA-ARS Plant Germplasm Introduction and Testing Unit, Washington State University, Pullman, WA 99164-6402, USA. coynec@wsu.edu, McClendon MT, Walling JG, Timmerman-Vaughan GM, Murray S, Meksem K, Lightfoot DA, Shultz JL, Keller KE, Martin RR, Inglis DA, Rajesh PN, McPhee KE, Weeden NF, Grusak MA, Li CM, Storlie EW |
| Jazyk: |
angličtina |
| Zdroj: |
Genome [Genome] 2007 Sep; Vol. 50 (9), pp. 871-5. |
| DOI: |
10.1139/g07-063 |
| Abstrakt: |
Pea (Pisum sativum L.) has a genome of about 4 Gb that appears to share conserved synteny with model legumes having genomes of 0.2-0.4 Gb despite extensive intergenic expansion. Pea plant inventory (PI) accession 269818 has been used to introgress genetic diversity into the cultivated germplasm pool. The aim here was to develop pea bacterial artificial chromosome (BAC) libraries that would enable the isolation of genes involved in plant disease resistance or control of economically important traits. The BAC libraries encompassed about 3.2 haploid genome equivalents consisting of partially HindIII-digested DNA fragments with a mean size of 105 kb that were inserted in 1 of 2 vectors. The low-copy oriT-based T-DNA vector (pCLD04541) library contained 55 680 clones. The single-copy oriS-based vector (pIndigoBAC-5) library contained 65 280 clones. Colony hybridization of a universal chloroplast probe indicated that about 1% of clones in the libraries were of chloroplast origin. The presence of about 0.1% empty vectors was inferred by white/blue colony plate counts. The usefulness of the libraries was tested by 2 replicated methods. First, high-density filters were probed with low copy number sequences. Second, BAC plate-pool DNA was used successfully to PCR amplify 7 of 9 published pea resistance gene analogs (RGAs) and several other low copy number pea sequences. Individual BAC clones encoding specific sequences were identified. Therefore, the HindIII BAC libraries of pea, based on germplasm accession PI 269818, will be useful for the isolation of genes underlying disease resistance and other economically important traits. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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