Construction of a BAC Library for a Defoliating Insect-Resistant Soybean and Identification of Candidate Clones Using a Novel Approach

TitleConstruction of a BAC Library for a Defoliating Insect-Resistant Soybean and Identification of Candidate Clones Using a Novel Approach
Publication TypeJournal Article
Year of Publication2009
AuthorsZhu, S., C. A. Saski, H. R. Boerma, J. P. Tomkins, J. N. All, and W. A. Parrott
JournalPlant Molecular Biology Reporter
Volume27
Pagination229-235
Date PublishedJun
ISBN Number0735-9640
Accession NumberISI:000266090300003
Keywordssoybean insect-resistance qtl genome walking bac library gene genome corn-earworm antibiosis resistance mexican bean beetle quantitative trait loci cyst-nematode resistance cutworm spodoptera-litura artificial chromosome library qtl
Abstract

Positional cloning of an insect-resistance quantitative trait locus (QTL) requires the construction of a large-insert genomic DNA library from insect-resistant genotypes. To facilitate cloning of a major defoliating insect-resistance QTL on linkage group M of the soybean genetic map, a bacterial artificial chromosome (BAC) library for PI 229358 was constructed and characterized. The HindIII BAC library contains 55,296 clones with an average insert size 131 kb. This library represents a 6-fold soybean haploid genome equivalents, allowing a 99.8% probability of recovering any specific sequence of interest in soybean. BAC filters were screened with a genomic DNA probe Sat_258sc2 obtained through genome walking from flanking sequences of a simple sequence repeat (SSR) marker, Sat_258, which links to the insect-resistance QTL. Thirteen BAC clones were identified positive for Sat_258sc2, and two of them were confirmed to carry Sat_258. The results suggest that this library is useful in positional cloning of the major insect-resistance QTL, and the approach presented here can be used to screen a BAC library for a SSR marker without requiring the creation of BAC pools.

URL<Go to ISI>://000266090300003
Alternate JournalPlant Mol Biol Rep