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

C.J Coyne; M.T McClendon; J.G Walling; G.M Timmerman-Vaughan; S Murray; K Meksem; D.A Lightfoot; J.L Shultz; K.E Keller; R.R Martin; D.A Inglis; P.N Rajesh; K.E McPhee; N.F Weeden; M.A Grusak; C.-M Li; E.W Storlie

doi:10.1139/G07-063

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

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

C.J Coyne, M.T McClendon, J.G Walling, G.M Timmerman-Vaughan, S Murray, K Meksem, D.A Lightfoot, J.L Shultz, K.E Keller, R.R Martin, …

Genome, Vol.50(9), pp.871-875

09/2007

DOI: https://doi.org/10.1139/G07-063

Handle:

https://hdl.handle.net/2376/110078

PMID: 17893728

Abstract

fusarium wilt

PI 269818

banque de clones BAC

virus de la mosaïque du pois transmis par les graines

PSbMV

Fusarium oxysporum f. sp. pisi

pea seed-borne mosaic virus

BAC library

flétrissure fusarienne

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.

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Title: Construction and characterization of two bacterial artificial chromosome libraries of pea (Pisum sativum L.) for the isolation of economically important genes
Creators: C.J Coyne - USDA-ARS Plant Germplasm Introduction and Testing Unit, Washington State University, Pullman, WA 99164-6402, USA
M.T McClendon - USDA-ARS Plant Germplasm Introduction and Testing Unit, Washington State University, Pullman, WA 99164-6402, USA
J.G Walling - Department of Plant Science and Plant Pathology, Montana State University, Bozeman, MT 59717, USA
G.M Timmerman-Vaughan - New Zealand Institute for Crop and Food Research, P.O. Box 4704, Christchurch, New Zealand
S Murray - New Zealand Institute for Crop and Food Research, P.O. Box 4704, Christchurch, New Zealand
K Meksem - Department of Plant, Soil, and General Agriculture, Southern Illinois University, Carbondale, IL 62901-4415, USA
D.A Lightfoot - Department of Plant, Soil, and General Agriculture, Southern Illinois University, Carbondale, IL 62901-4415, USA
J.L Shultz - Department of Plant, Soil, and General Agriculture, Southern Illinois University, Carbondale, IL 62901-4415, USA
K.E Keller - USDA-ARS Horticulture Crops Research Laboratory, Corvallis, OR 97330, USA
R.R Martin - USDA-ARS Horticulture Crops Research Laboratory, Corvallis, OR 97330, USA
D.A Inglis - Northwestern Washington Research and Extension Center, Washington State University, Mount Vernon, WA 98273, USA
P.N Rajesh - USDA-ARS Grain Legume Genetics and Physiology Unit, Washington State University, Pullman, WA 99164-6434, USA
K.E McPhee - USDA-ARS Grain Legume Genetics and Physiology Unit, Washington State University, Pullman, WA 99164-6434, USA
N.F Weeden - Department of Plant Science and Plant Pathology, Montana State University, Bozeman, MT 59717, USA
M.A Grusak - USDA-ARS Children's Nutrition Research Center, Department of Pediatrics, Baylor College of Medicine, 1100 Bates Street, Houston, TX 77030-2600, USA
C.-M Li - USDA-ARS Children's Nutrition Research Center, Department of Pediatrics, Baylor College of Medicine, 1100 Bates Street, Houston, TX 77030-2600, USA
E.W Storlie - Augustana College, Department of Biology, 2001 South Summit Avenue, Sioux Falls, SD 57197, USA
Publication Details: Genome, Vol.50(9), pp.871-875
Academic Unit: Crop and Soil Sciences, Department of; Plant Pathology, Department of
Identifiers: 99900547013201842
Language: English
Resource Type: Journal article

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

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