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

被引:17
作者
Coyne, C. J. [1 ]
McClendon, M. T.
Walling, J. G.
Timmerman-Vaughan, 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.
Li, C.-M.
Storlie, E. W.
机构
[1] Washington State Univ, USDA ARS, Plant Germplasm Intro & Testing Unit, Pullman, WA 99164 USA
[2] Montana State Univ, Dept Plant Sci & Plant Pathol, Bozeman, MT 59717 USA
[3] New Zealand Inst Crop & Food Res, Christchurch, New Zealand
[4] So Illinois Univ, Dept Plant Soil & Gen Agr, Carbondale, IL 62901 USA
[5] USDA ARS, Crop Genet & Prod Res Unit, Stoneville, MS 38776 USA
[6] USDA ARS, Hort Crops Res Lab, Corvallis, OR 97330 USA
[7] Washington State Univ, NW Washington Res & Extens Ctr, Mt Vernon, WA 98273 USA
[8] Washington State Univ, USDA ARS, Grain Legume Genet & Physiol Unit, Pullman, WA 99164 USA
[9] Baylor Coll Med, USDA ARS, Childrens Nutr Res Ctr, Dept Pediat, Houston, TX 77030 USA
[10] Augustana Coll, Dept Biol, Sioux Falls, SD 57197 USA
关键词
BAC library; pea seed-borne mosaic virus; PSbMV; fusarium wilt; Fusarium oxysporum f. sp pisi; PI; 269818;
D O I
10.1139/G07-063
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
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 I of 2 vectors. The low-copy oriT-based T-DNA vector (pCLD04541) library contained 55 680 clones. The single-copy oriS-based vector (pIndigo-BAC-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.
引用
收藏
页码:871 / 875
页数:5
相关论文
共 21 条
[1]  
[Anonymous], MOL CLONING
[2]  
Arumuganathan K, 1991, PLANT MOL BIOL REP, V9, P208, DOI [10.1007/BF02672069, DOI 10.1007/BF02672069]
[3]   A RAPID PROCEDURE FOR COLONY SCREENING USING NYLON FILTERS [J].
BULUWELA, L ;
FORSTER, A ;
BOEHM, T ;
RABBITTS, TH .
NUCLEIC ACIDS RESEARCH, 1989, 17 (01) :452-452
[4]   The role of iron-deficiency stress responses in stimulating heavy-metal transport in plants [J].
Cohen, CK ;
Fox, TC ;
Garvin, DF ;
Kochian, LV .
PLANT PHYSIOLOGY, 1998, 116 (03) :1063-1072
[5]  
Coyne C. J., 2000, Pisum Genetics, V32, P23
[6]   The potyvirus recessive resistance gene, sbm1, identifies a novel role for translation initiation factor eIF4E in cell-to-cell trafficking [J].
Gao, ZH ;
Johansen, E ;
Eyers, S ;
Thomas, CL ;
Noel Ellis, TH ;
Maule, AJ .
PLANT JOURNAL, 2004, 40 (03) :376-385
[7]   SYSTEMATIC SCREENING OF YEAST ARTIFICIAL-CHROMOSOME LIBRARIES BY USE OF THE POLYMERASE CHAIN-REACTION [J].
GREEN, ED ;
OLSON, MV .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (03) :1213-1217
[8]  
HARE W. W., 1949, JOUR AGRIC RES, V78, P239
[9]   INHERITANCE OF IMMUNITY TO MILDEW IN PERUVIAN FORMS OF PISUM-SATIVUM [J].
HARLAND, SC .
HEREDITY, 1948, 2 (02) :263-269
[10]  
Jones Jonathan D. G., 1992, Transgenic Research, V1, P285, DOI 10.1007/BF02525170