カキの品種同定のためのＲＡＰＤ分析

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タイトル別名
  • Evaluation of RAPD Analysis for Cultivar Identification of Persimmons.
  • カキ ノ ヒンシュ ドウテイ ノ タメ ノ RAPD ブンセキ

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The effectiveness of RAPD analysis for cultivar identification of persimmons (Diospyros kaki) was evaluated by using 10 base primers (Operon Technologies, CA). A protocol of reproducible RAPD analysis for persimmon was established by determining suitable buffer and optimum concentrations of DNA polymerase, MgCl2, primer, and dNTP. The 12.5 μl reaction mixture, found to be reliable for RAPD for persimmon, contains 50 mM Tris-HCI (pH 8.9), 20 mM NaCl, 1.0%Triton-X 100, 0.1% gelatin as a buffer solution (Levi et al., 1993), 0.06 units-μl-1 AmpliTaq DNA polymerase, 2.0 mM MgCl2, 0.28 μM primer, and 0.1 mM each of dNTP per reaction. Among these elements, the kinds of buffer and concentrations of ampliTaq DNA polymerase and MgCl2 were important for reproducibility of RAPD. Amplification was performed by 45 cycles of 1 min at 94°C, 1 min at 45 °C, 2 min at 72°C in the Perkin Elmer Cetus DNA Thermal Cycler. 5 ng•μl-1 DNA were used as a template. By this reproducible protocol, we selected 2 primers (OPA-06 and OPA-08) among 20 primers (OPA-01 to OPA-20) as effective ones for cultivar identification of persimmons. The fifteen cultivars tested were completely distinguishable from each other by RAPDs, using OPA-06 or OPA-08. Furthermore, two bud mutants of 'Hiratanenashi', i. e. 'Tonewase' and 'Sugitawase', showed different DNA patterns with a few additional minor bands by RAPD, using OPA-06 primer. It may be possible to identify persimmon sports by this method. In addition, polymorphisms among 11 Diospyros species were observed by RAPD, using OPA-10 primer. The same method revealed little polymorphisms among intraspecific levels of D. kaki or D. lotus. The potential of RAPD analysis for persimmon identification is discussed.

収録刊行物

  • 園芸学会雑誌

    園芸学会雑誌 64 (3), 535-541, 1995

    一般社団法人　園芸学会

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