Quantitative real-time PCR to determine allele number for the astringency locus by analysis of a linked marker in Diospyros kaki Thunb

Takashi Akagi, Shinya Kanzaki, Mai Gao, Ryutaro Tao, Dan E. Parfitt, Keizo Yonemori

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Persimmon (Diospyros kaki Thunb.) is a polyploidy fruit tree species of economic importance to East Asia. Natural astringency loss is an important trait in persimmon breeding programs. Quantitative real-time PCR was used to determine the number of AST/ast alleles for fruit astringency in persimmon (D. kaki Thunb.). To this end, the cultivar Jiro was transformed with one or two copies of a gene encoding NADP-dependent sorbitol-6-phosphate dehydrogenase (S6PDH), which was used as a standard for measuring the allele number of a sequenced marker tightly linked to the recessive ast locus for nonastringency. Primers for markers linked to the AST or ast allele were then used to measure the AST to ast ratio directly in the progeny of a full-sib cross. From determination of the AST to ast ratio and the results of the S6PDH copy number, the number of AST and ast alleles at the AST/ast locus was estimated. This research supported the hypothesis that D. kaki is a hexaploid with six AST and/or ast alleles. In addition to the determination of the allelic status of the AST locus, the application of real-time PCR for confirmation of the ploidy level and allelic composition of target genes in autopolyploids or allopolyploids was demonstrated.

Original languageEnglish
Pages (from-to)483-492
Number of pages10
JournalTree Genetics and Genomes
Volume5
Issue number3
DOIs
Publication statusPublished - Jul 1 2009
Externally publishedYes

Keywords

  • D. kaki
  • Genotype
  • Persimmon
  • Polyploid
  • Real-time PCR

ASJC Scopus subject areas

  • Forestry
  • Molecular Biology
  • Genetics
  • Horticulture

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