Construction and characterization of a half million clone BAC library of durum wheat (Triticum turgidum ssp. durum)

• Published in: Theoretical and applied genetics Vol. 107; no. 5; pp. 931 - 939
• Main Authors: CENCI, A, CHANTRET, N, KONG, X, GU, Y, ANDERSON, O. D, FAHIMA, T, DISTELFELD, A, DUBCOVSKY, J
• Format: Journal Article
• Language: English
• Published: Heidelberg Springer 01.09.2003; Berlin Springer Nature B.V
• Subjects: Agronomy. Soil science and plant productions; Biological and medical sciences; Chromosomes, Artificial, Bacterial; Chromosomes, Plant - genetics; Classical and quantitative genetics. Population genetics. Molecular genetics; Classical genetics, quantitative genetics, hybrids; Cloning, Molecular; Construction; Fundamental and applied biological sciences. Psychology; Gene Library; Generalities. Genetics. Plant material; Genes; Genetics; Genetics and breeding of economic plants; Genetics of eukaryotes. Biological and molecular evolution; Genome, Plant; Libraries; Molecular genetics; Ploidies; Pteridophyta, spermatophyta; Triticum - genetics; Vegetals; Wheat; Monocotyledones; Construction; Polyploidy; Production quality; Triticum durum; BAC; Biological evolution; Cereal crop; Gene bank; Characterization; Genomic library; BAC chromosome; Gramineae; Angiospermae; Genetic improvement; Spermatophyta; Wheat; Protein content; Clone; Polyploid
• DOI: 10.1007/s00122-003-1331-z
• ISSN: 0040-5752

Durum wheat ( Triticum turgidum ssp. durum, 2 n = 4 x = 28, genomes AB) is an economically important cereal used as the raw material to make pasta and semolina. In this paper we present the construction and characterization of a bacterial artificial chromosome (BAC) library of tetraploid durum wheat cv. Langdon. This variety was selected because of the availability of substitution lines that facilitate the assignment of BACs to the A and B genome. The selected Langdon line has a 30-cM segment of chromosome 6BS from T. turgidum ssp. dicoccoides carrying a gene for high grain protein content, the target of a positional cloning effort in our laboratory. A total of 516,096 clones were organized in 1,344 384-well plates and blotted on 28 high-density filters. Ninety-eight percent of these clones had wheat DNA inserts (0.3% chloroplast DNA, 1.4% empty clones and 0.3% empty wells). The average insert size of 500 randomly selected BAC clones was 131 kb, resulting in a coverage of 5.1-fold genome equivalents for each of the two genomes, and a 99.4% probability of recovering any gene from each of the two genomes of durum wheat. Six known copy-number probes were used to validate this theoretical coverage and gave an estimated coverage of 5.8-fold genome equivalents. Screening of the library with 11 probes related to grain storage proteins and starch biosynthesis showed that the library contains several clones for each of these genes, confirming the value of the library in characterizing the organization of these important gene families. In addition, characterization of fingerprints from colinear BACs from the A and B genomes showed a large differentiation between the A and B genomes. This library will be a useful tool for evolutionary studies in one of the best characterized polyploid systems and a source of valuable genes for wheat. Clones and high-density filters can be requested at http://agronomy.ucdavis.edu/Dubcovsky/BAC-library/BAC_Langdon.htm