Effective Clonal Analysis of Trees in a Shakain Cutting Garden of <i>Cryptomeria japonica</i> Using Bulked DNA Samples

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  • Goto Susumu
    Education and Research Center, The University of Tokyo Forests, Graduate School of Agricultural and Life Sciences, The University of Tokyo
  • Matsui Yukari
    Kumamoto Prefecture Forestry Research Center

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  • バルク DNA を用いたスギ挿し木在来品種シャカイン採穂園の効率的なクローン分析法
  • バルク DNA オ モチイタ スギ サシ モク ザイライ ヒンシュ シャカインサイスイエン ノ コウリツテキ ナ クローン ブンセキホウ

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Contamination by different clones in a cutting garden is a serious problem for seedling quality. Clonal analysis of trees in a cutting garden can be performed using highly polymorphic microsatellite markers. However, clonal analysis of every tree is laborious and expensive. To establish an effective method for clonal analysis of trees in the Shakain cutting gardens of Kumamoto Prefecture, we conducted microsatellite analysis using bulked DNA. We bulked Shakain needle samples and those of other cultivars at proportions of 4:1 and 9:1. Then, we extracted DNA from the bulked samples and used it as a template for microsatellite analysis. We accurately identified the contaminated bulked samples by detecting peaks from other cultivars. We applied this method to two Syakain cutting gardens: cutting garden A consisting of 387 trees and cutting garden B consisting of 1,007 trees. Finally, we identified three contaminated trees by analyzing 103 samples (21.1%) in cutting garden A and 353 samples (35.1%) in cutting garden B, respectively. Thus, microsatellite analysis using bulked DNA is an effective method for clonal analysis of trees in a Shakain cutting garden of Cryptomeria japonica.

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