Rapid Identification of Rice Cultivar "Koshihikari" Using Loop-mediated Isothermal Amplification (LAMP)

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  • ＬＡＭＰ法を利用したコシヒカリの高精度・迅速識別
  • LAMPホウ オ リヨウ シタ コシヒカリ ノ コウセイド ・ ジンソク シキベツ

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Abstract

A simple and rapid method for identification of rice cultivar “Koshihikari” has been developed using loop-mediated isothermal amplification (LAMP). Primers for detection of Koshihikari (positive primer) and cultivars other than Koshihikari (negative primer) were designed for blast resistant gene Pi5-1and its susceptible allele. DNA amplification analysis of the top 48 cultivars using positive and negative primers showed that both primers had approximately 90% accuracy in detecting Koshihikari and cultivars other than Koshihikari. For rapid detection from rice grains, a simple DNA extraction method using sodium hydroxide was developed. LAMP amplification using DNA extracted by this method was almost equivalent to that using column-purified DNA. Using these methods, we tested LAMP detection of Koshihikari and Hitomebore (representative of cultivars excluding Koshihikari) grain contamination, and showed that both primers could detect 1% contamination.

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