Construction of a BAC library for buckwheat genome research - an application to positional cloning of agriculturally valuable traits.
We have constructed a BAC library for common buckwheat Fagopyrum esculentum Moench. The library includes 142,005 clones with an average insert size of approximately 76 kb, equivalent to approximately a 7 to approximately 8-fold coverage of the genome. Polymerase chain reaction based screening of the library with AGAMOUS and FLORICAULA/LEAFY primers, has identified 7 and 9 BACs, respectively, which are consistent with the genome coverage. This library represents the first large insert genomic library for F. esculentum and it can be served as a genetic resource facilitating agricultural, pharmacological, physiological, and evolutionary studies of the species. To demonstrate the utilization of the library for characterizing agriculturally valuable traits, we developed a sequence tagged site marker tightly linked to the dwarf E locus as well as to the self-incompatibility complex locus and screened the library to initiate positional cloning of the causative genes.
Yasui
Yasuo
Y
Laboratory of Crop Evolution, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
Mori
Masashi
M
Matsumoto
Daiki
D
Ohnishi
Ohmi
O
Campbell
Clayton G
CG
Ota
Tatsuya
T
D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D018744:DNA, Plant / Q000032:analysis; D019612:Fagopyrum / Q000235:genetics*; D015723:Gene Library; D023061:Gene Order; D008040:Genetic Linkage; D018745:Genome, Plant / Q000235:genetics*
D018744:DNA, Plant