Construction and characterization of a BIBAC library of Jatropha curcas L. and identification of BIBAC clones containing genes associated with fatty acid metabolism

Jatropha curcas L. is a potentially significant bioenergy crop in the tropics and subtropics. Here we present a plant-transformation-competent binary bacterial artificial chromosome (BIBAC) library from Jatropha cultivar YN049-4. This library was constructed with BamH in the vector pCLD04541, consists of 30,720 clones and is arrayed in 80 384-well microtiter plates. Since 92.1% (28,293) of its clones were shown to contain Jatropha DNA inserts with an average size of 131.9 kb, the library is estimated to represent approximately 8.9 haploid genome equivalents of the species, thus providing a greater than 99% probability of discovering a particular single-copy sequence in the library. High-density clone filters were made from a subset of the library and hybridized with nine pairs of overgos designed from genes involved in fatty acid metabolism. Hybridization results showed that eight overgo pairs were able to identify positive clones from the subset of the library, with an average of 5.3 clones per probe, suggesting that it is suitable for Jatropha genomics and genetics research. Because this library, to our knowledge, represents the first large-insert, plant-transformation-competent BIBAC library for Jatropha, it will provide a vital resource for advanced genomics research, including isolation and characterization of genes and quantitative trait loci, integrative physical mapping and genome sequencing.