title: A rapid and enhanced DNA detection method for crop cultivar discrimination.
abstract: In many crops species, the development of a rapid and precise cultivar discrimination system has been required for plant breeding and patent protection of plant cultivars and agricultural products. Here, we successfully evaluated strawberry cultivars via a novel method, namely, the single tag hybridization (STH) chromatographic printed array strip (PAS) using the PCR products of eight genomic regions. In a previous study, we showed that genotyping of eight genomic regions derived from FaRE1 retrotransposon insertion site enabled to discriminate 32 strawberry cultivars precisely, however, this method required agarose/acrylamide gel electrophoresis, thus has the difficulty for practical application. In contrast, novel DNA detection method in this study has some great advantages over standard DNA detection methods, including agarose/acrylamide gel electrophoresis, because it produces signals for DNA detection with dramatically higher sensitivity in a shorter time without any preparation or staining of a gel. Moreover, this method enables the visualization of multiplex signals simultaneously in a single reaction using several independent amplification products. We expect that this novel method will become a rapid and convenient cultivar screening assay for practical purposes, and will be widely applied to various situations, including laboratory research, and on-site inspection of plant cultivars and agricultural products.
authors: Graduate School of Environmental and Life Science, Okayama University, 1-1-1 Tsushimanaka Kitaku, Okayama, Okayama 700-8530, Japan. Electronic address: y_monden@cc.okayama-u.ac.jp.
authors: Yuki
authors: Y
authors: Monden
authors: FASMAC Co., Ltd., 5-1-3 Midorigaoka, Atsugi, Kanagawa 243-0041, Japan. Electronic address: ktakasaki@fasmac.co.jp.
authors: Kazuto
authors: K
authors: Takasaki
authors: FASMAC Co., Ltd., 5-1-3 Midorigaoka, Atsugi, Kanagawa 243-0041, Japan. Electronic address: sfuto@fasmac.co.jp.
authors: Satoshi
authors: S
authors: Futo
authors: Graduate School of Biomedical Engineering, Tohoku University, 6-6-05 Aramaki-aoba, Sendai, Miyagi 980-8579, Japan. Electronic address: kniwa@ecei.tohoku.ac.jp.
authors: Kousuke
authors: K
authors: Niwa
authors: Graduate School of Biomedical Engineering, Tohoku University, 6-6-05 Aramaki-aoba, Sendai, Miyagi 980-8579, Japan. Electronic address: m-kawase@ecei.tohoku.ac.jp.
authors: Mitsuo
authors: M
authors: Kawase
authors: Graduate School of Environmental and Life Science, Okayama University, 1-1-1 Tsushimanaka Kitaku, Okayama, Okayama 700-8530, Japan. Electronic address: ag20004@s.okayama-u.ac.jp.
authors: Hiroto
authors: H
authors: Akitake
authors: Graduate School of Environmental and Life Science, Okayama University, 1-1-1 Tsushimanaka Kitaku, Okayama, Okayama 700-8530, Japan. Electronic address: tahara@cc.okayama-u.ac.jp.
authors: Makoto
authors: M
authors: Tahara
mesh_terms: D000383:Agriculture / Q000379:methods*; D002845:Chromatography / Q000379:methods; D018556:Crops, Agricultural / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D018744:DNA, Plant / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D031985:Fragaria / Q000235:genetics*; D009693:Nucleic Acid Hybridization / Q000379:methods; D012680:Sensitivity and Specificity; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D018744:DNA, Plant
keywords: Cultivar discrimination; Multiplex PCR; Practical application; Retrotransposon; Strawberry