A new identification system of sugi (Cryptomeria japonica) cultivars based on DNA typing

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This research subject was carried out for three years. The traditional local cutting cultivars and the plus-tree clones in sugi (Cryptomeria japonica) were reclassified based on DNA type by RAPD (random amplified polymorphic DNA) analysis. Furthermore, the new DNA markers called microsatellite DNA that possessed high variability and repeatability were developed.1.Reclassification of cutting cultivars : RAPD was used to discriminate the cutting cultivars in Kyushu region. About 550 individuals from 84 cultivars were analyzed. It was clarified that 53 cultivars were monoclonal and remaining 31 cultivar were clone-complex from the DNA type of RAPD.About 620 plus-tree clones in Kyushu region were also investigated to reveal the relation to the traditional cutting cultivars.2.Development of the new identification system : The microsatellite DNA regions, (GT) n, (CT) n, (TAT) n, and (AAT) n, were screened from genomic DNA using RAMP (random amplified microsatellite polymorphism) method. Thirty-one reliable markers were selected from the first screening pool. High heterozygosity (>0.5) was observed in twelve markers, and it was confirmed that microsatellite DNAs were very variable. The DNA sequences of 8 highly variable markers were determined, and the primers were designed for PCR (polymerase chain reaction). These markers are available to a new identification system using microsatellite DNA.