Construction and characterization of a plant transformation-competent BIBAC library of the black Sigatoka-resistant banana Musa acuminata cv. Tuu Gia (AA).
A plant transformation-competent binary bacterial artificial chromosome (BIBAC) library was constructed from Musa acuminata cv. Tuu Gia (AA), a black Sigatoka-resistant diploid banana. After digestion of high-molecular-weight banana DNA by HindIII, several methods of DNA size selection were tested, followed by ligation, using a vector/insert molar ratio of 4:1. The library consists of 30,700 clones stored in 80 384-well microtiter plates. The mean insert size was estimated to be 100 kb, and the frequency of inserts with internal NotI sites was 61%. The majority of insert sizes fell into the range of 100+/-20 kb, making them suitable for Agrobacterium-mediated transformation. Only 1% and 0.9% of the clones contain chloroplast and mitochondrial DNA, respectively. This is the first BIBAC library for banana, estimated to represent five times its haploid genome (600 Mbp). It was demonstrated by hybridization that the library contains typical members of resistance gene and defense gene families that can be used for transformation of disease susceptible banana cultivars for banana genetic improvement.
Ortiz-Vázquez
E
E
Centro de Investigación Científica de Yucatán, Mérida, Yucatán, México. eortiz@itmerida.mx
Kaemmer
D
D
Zhang
H-B
HB
Muth
J
J
Rodríguez-Mendiola
M
M
Arias-Castro
C
C
James
Andrew
A
D022202:Chromosomes, Artificial, Bacterial; D018744:DNA, Plant; D004926:Escherichia coli; D015723:Gene Library; D028521:Musa / Q000235:genetics*; D014170:Transformation, Genetic
D018744:DNA, Plant