Construction of two BAC libraries from cucumber (Cucumis sativus L.) and identification of clones linked to yield component quantitative trait loci

被引:20
作者
Nam, YW [1 ]
Lee, JR
Song, KH
Lee, MK
Robbins, MD
Chung, SM
Staub, JE
Zhang, HB
机构
[1] Sogang Univ, Dept Life Sci, Seoul 121742, South Korea
[2] Texas A&M Univ, Dept Soil & Crop Sci, Inst Plant Genom & Biotechnol, College Stn, TX 77843 USA
[3] Univ Wisconsin, USDA ARS, Madison, WI 53706 USA
[4] Univ Wisconsin, Dept Hort, Madison, WI 53706 USA
关键词
BAC libraries; cucumber; molecular markers; PCR screening; quantitative trait loci; yield components;
D O I
10.1007/s00122-005-2007-7
中图分类号
S3 [农学(农艺学)];
学科分类号
0901 ;
摘要
Two bacterial artificial chromosome (BAC) libraries were constructed from an inbred line derived from a cultivar of cucumber (Cucumis sativus L.). Intact nuclei were isolated and embedded in agarose plugs, and high-molecular-weight DNA was subsequently partially digested with BamHI or EcoRI. Ligation of double size-selected DNA fragments with the pECBAC1 vector yielded two libraries containing 23,040 BamHI and 18,432 EcoRI clones. The average BamHI and EcoRI insert sizes were estimated to be 107.0 kb and 100.8 kb, respectively, and BAC clones lacking inserts were 1.3% and 14.5% in the BamHI and EcoRI libraries, respectively. The two libraries together represent approximately 10.8 haploid cucumber genomes. Hybridization with a C(0)t-1 DNA probe revealed that approximately 36% of BAC clones likely carried repetitive sequence-enriched DNA. The frequencies of BAC clones that carry chloroplast or mitochondrial DNA range from 0.20% to 0.47%. Four sequence-characterized amplified region (SCAR), four simple sequence repeat, and an randomly amplified polymorphic DNA marker linked with yield component quantitative trait loci were used either as probes to hybridize high-density colony filters prepared from both libraries or as primers to screen an ordered array of pooled BAC DNA prepared from the BamHI library. Positive BAC clones were identified in predicted numbers, as screening by polymerase chain reaction amplification effectively overcame the problems associated with an overabundance of positives from hybridization with two SCAR markers. The BAC clones identified herein that are linked to the de (determinate habit) and F (gynoecy) locus will be useful for positional cloning of these economically important genes. These BAC libraries will also facilitate physical mapping of the cucumber genome and comparative genome analyses with other plant species.
引用
收藏
页码:150 / 161
页数:12
相关论文
共 54 条
[1]  
Arumuganathan K, 1991, PLANT MOL BIOL REP, V9, P208, DOI [DOI 10.1007/BF02672069, 10.1007/BF02672069]
[2]   Towards an expanded and integrated linkage map of cucumber (Cucumis sativus L.) [J].
Bradeen, JM ;
Staub, JE ;
Wye, C ;
Antonise, R ;
Peleman, J .
GENOME, 2001, 44 (01) :111-119
[3]  
Budiman MA, 2000, GENOME RES, V10, P129
[4]  
BURKE DT, 1987, SCIENCE, V236, P801
[5]  
CANTLIFFE DJ, 1981, J AM SOC HORTIC SCI, V106, P133
[6]   Molecular cloning and tissue-specific expression of an anionic peroxidase in zucchini [J].
Carpin, S ;
Crèvecoeur, M ;
Greppin, H ;
Penel, C .
PLANT PHYSIOLOGY, 1999, 120 (03) :799-810
[7]  
Chang YL, 2001, GENETICS, V159, P1231
[8]   Construction of two BAC libraries from the wild Mexican diploid potato, Solanum pinnatisectum, and the identification of clones near the late blight and Colorado potato beetle resistance loci [J].
Chen, Q ;
Sun, S ;
Ye, Q ;
McCuine, S ;
Huff, E ;
Zhang, HB .
THEORETICAL AND APPLIED GENETICS, 2004, 108 (06) :1002-1009
[9]   CONSTRUCTION AND CHARACTERIZATION OF A BACTERIAL ARTIFICIAL CHROMOSOME LIBRARY OF ARABIDOPSIS-THALIANA [J].
CHOI, S ;
CREELMAN, RA ;
MULLET, JE ;
WING, RA .
PLANT MOLECULAR BIOLOGY REPORTER, 1995, 13 (02) :124-128
[10]   Targeted isolation of simple sequence repeat markers through the use of bacterial artificial chromosomes [J].
Cregan, PB ;
Mudge, J ;
Fickus, EW ;
Marek, LF ;
Danesh, D ;
Denny, R ;
Shoemaker, RC ;
Matthews, BF ;
Jarvik, T ;
Young, ND .
THEORETICAL AND APPLIED GENETICS, 1999, 98 (6-7) :919-928