Construction of a watermelon BAC library and identification of SSRs anchored to melon or Arabidopsis genomes.
A bacterial artificial chromosome (BAC) library was constructed for watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai var. lanatus) with an average insert-size of 106 kb, providing 21 haploid genome equivalents. The library was used to identify BAC clones that are anchored to probes evenly distributed on the genomes of melon or Arabidopsis. Twenty eight probes (representing 66% of the tested probes) from melon and 30 probes (65%) from Arabidopsis identified positive BAC clones. Two methods were implemented to identify SSRs from the positively hybridizing BAC clones. First, analysis of BAC end sequences revealed 37 SSRs. For the second method, pooled DNA of BACs identified by the melon probes was used to develop a shotgun library. The library was then screened with synthetic SSR oligonucleotides by hybridization. Sequence analysis of positively hybridizing shotgun clones revealed 142 different SSRs. Thirty eight SSRs were characterized using three watermelon cultivars, five plant introduction (PI) accessions of C. lanatus var lanatus and four PIs of C. lanatus var citroides. Of these, 36 (95%) were found to be polymorphic with up to six alleles per marker. Polymorphism information content values for polymorphic markers varied between 0.22 and 0.79 with an average of 0.53. The methods described herein will be valuable for the construction of a watermelon linkage map with SSRs evenly distributed on its genome that is anchored to the genomes of melon and Arabidopsis.
Joobeur
T
T
Department of Plant Pathology, Fungal Genomics Laboratory, North Carolina State University, Raleigh, NC 27965, USA.
Gusmini
G
G
Zhang
X
X
Levi
A
A
Xu
Y
Y
Wehner
T C
TC
Oliver
M
M
Dean
R A
RA
D000483:Alleles; D017360:Arabidopsis / Q000235:genetics*; D022202:Chromosomes, Artificial, Bacterial; D029773:Citrullus / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics; D015723:Gene Library; D005819:Genetic Markers; D018745:Genome, Plant; D006824:Hybridization, Genetic; D011110:Polymorphism, Genetic; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics*; D017422:Sequence Analysis, DNA
D018744:DNA, Plant; D005819:Genetic Markers