title: A non-destructive genotyping system from a single seed for marker-assisted selection in watermelon.
abstract: Genomic tools for watermelon breeding are becoming increasingly available. A high throughput genotyping system would facilitate the use of DNA markers in marker-assisted selection. DNA extraction from leaf material requires prior seed germination and is often time-consuming and cost prohibitive. In an effort to develop a more efficient system, watermelon seeds of several genotypes and various seed sizes were sampled by removing ⅓ or ½ sections from the distal ends for DNA extraction, while germinating the remaining proximal parts of the seed. Removing ⅓ of the seed from the distal end had no effect on seed germination percentage or seedling vigor. Different DNA extraction protocols were tested to identify a method that could yield DNA of sufficient quality for amplification by polymerase chain reaction. A sodium dodecyl sulfate extraction protocol with 1% polyvinylpyrrolidone yielded DNA that could be amplified with microsatellite primers and was free of pericarp contamination. In this study, an efficient, non-destructive genotyping protocol for watermelon seed was developed.
authors.affiliation: Department of Horticulture, University of Georgia, Athens, GA, USA. merujeff@uga.edu
authors.forename: G
authors.initials: G
authors.lastname: Meru
authors.forename: D
authors.initials: D
authors.lastname: McDowell
authors.forename: V
authors.initials: V
authors.lastname: Waters
authors.forename: A
authors.initials: A
authors.lastname: Seibel
authors.forename: J
authors.initials: J
authors.lastname: Davis
authors.forename: C
authors.initials: C
authors.lastname: McGregor
mesh_terms: D029773:Citrullus / Q000033:anatomy & histology / Q000235:genetics* / Q000254:growth & development; D018744:DNA, Plant / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D005838:Genotype; D060005:Genotyping Techniques / Q000379:methods*; D018525:Germination / Q000235:genetics*; D018895:Microsatellite Repeats / Q000235:genetics; D016133:Polymerase Chain Reaction; D011205:Povidone / Q000737:chemistry; D012639:Seeds / Q000033:anatomy & histology / Q000235:genetics* / Q000254:growth & development; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry
chemical_list: D018744:DNA, Plant; D012967:Sodium Dodecyl Sulfate; D011205:Povidone