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United States Department of Agriculture

Agricultural Research Service

Title: Construction and characterization of two bacterial artificial chromosome libraries of pea (Pisum sativum L.) for the isolation of economically important genes)

Author
item Coyne, Clarice - Clare
item Mcclendon, M
item Walling, J
item Timmerman-vaughn, G
item Murray, S
item Meksem, K
item Lightfoot, D
item Shultz, Jeffry
item Keller, Karen
item Martin, Robert - Bob
item Inglis, D
item Rajesh, P
item Mcphee, Kevin
item Weeden, N
item Grusak, Michael - Mike
item Storlie, E
item Li, C

Submitted to: Genome
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/12/2007
Publication Date: 9/6/2007
Publication URL: http://hdl.handle.net/10113/9994
Citation: Coyne, C.J., Mcclendon, M.T., Walling, J.G., Timmerman-Vaughn, G.M., Murray, S., Meksem, K., Lightfoot, D.A., Shultz, J.L., Keller, K.E., Martin, R.R., Inglis, D.A., Rajesh, P.N., Mcphee, K.E., Weeden, N.F., Grusak, M.A., Storlie, E.W. 2007. Construction and characterization of two bacterial artificial chromosome libraries of pea (Pisum sativum L.) for the isolation of economically important genes. Genome 50:871-875.

Interpretive Summary: Genomic tools are playing an increasingly important role in basic plant science discovery for crop improvement. The aim here was to develop a genomic tool, a pea bacterial artificial chromosome (BAC) libraries that would enable the isolation of genes involved in plant disease resistance or controlling economic traits. The BAC libraries encompassed about 3.2 haploid genome equivalents. BAC plate-pool DNA successfully PCR amplified using seven of nine published pea resistance gene analogs (RGAs) and several other low copy number pea sequences. Individual BAC clones encoding specific sequences were identified. This HindIII BAC libraries of the pea, based on germplasm accession PI 269818, will be useful for the isolation of genes underlying disease resistance and other economic traits.

Technical Abstract: Pea (Pisum sativum L.) has a genome of about 4 Gbp that appears to share conserved synteny with model legumes having genomes of 0.2-0.4 Gbp despite extensive intergenic expansion. Pea plant inventory (PI) 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 controlling economic traits. The BAC libraries encompassed about 3.2 haploid genome equivalents among partially HindIII-digested DNA fragments of 105 kb mean size, that were inserted in one of two vectors. The plant-transformation, T-DNA, low copy, oriT based vector pCLD04541 library contained 55,680 clones. The pIndigoBAC-5, single copy, oriS based library contained 65,280 clones. Colony hybridization of a universal chloroplast probe inferred the libraries contained about 1% from clones 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 two replicated methods. First, high-density filters were probed with low copy number sequences. Secondly, BAC plate-pool DNA successfully PCR amplified using seven of nine 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 the pea, based on germplasm accession PI 269818, will be useful for the isolation of genes underlying disease resistance and other economic traits.

Last Modified: 8/24/2016
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