Biol. Pharm. Bull., 27(9),1333-1339, September 2004

Regular Articles

New Qualitative Detection Methods of Genetically Modified Potatoes

Takahiro WATANABE,^*,a Hideo KURIBARA,^b,i Takashi MISHIMA,^c Hiroyuki KIKUCHI,^a Misao KUBO,^b Takashi KODAMA,^b,i Satoshi FUTO,^d Kikuko KASAMA,^e Akie TOYOTA,^f Masanori NOUNO,^g Ayako SAITA,^g Kunihiko TAKAHASHI,^h Akihiro HINO,ⁱ Hiroshi AKIYAMA,^a and Tamio MAITANI^a

^a National Institute of Health Sciences; 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501, Japan: ^b Center for Food Quality, Labeling and Consumer Services; 2-1 Shintoshin, Chuo-ku, Saitama, Saitama 330-9731, Japan: ^c Japan Food Research Laboratories; 2-3 Bunkyo, Chitose, Hokkaido 066-0052, Japan: ^d FASMAC Co., Ltd.; 5-1-3 Midorigaoka, Atsugi, Kanagawa 243-0041, Japan: ^e Hatano Research Institute, Food and Drug Safety Center; 729-5 Ochiai, Hadano, Kanagawa 257-8523, Japan: ^f Hiroshima Prefectural Institute of Public Health and Environment; 1-6-29 Minami-cho, Minami-ku, Hiroshima 734-0007, Japan: ^g Center for Inspection of Imported Foods and Infectious Diseases Yokohama Quarantine Station; 107-8 Nagahama, Kanazawa-ku, Yokohama, Kanagawa 236-0011, Japan: ^h Saitama Prefectural Institute of Public Health; 639-1 Kamiokubo, Sakura-ku, Saitama 338-0824, Japan: and ⁱ National Food Research Institute; 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan. * To whom correspondence should be addressed. e-mail: tawata@nihs.go.jp

In Japan, 8 lines of genetically modified (GM) potato (2 lines of NewLeaf^® potato; NL, 3 lines of NewLeaf Plus^® potato; NLP, and 3 lines of NewLeaf Y^® potato; NLY) have already been authorized as safe for use in foods and feeds. We have developed polymerase chain reaction (PCR) methods for the qualitative detection of the GM potatoes for the screening and the identification of NL, NLP and NLY. The gene encoding uridine diphosphate (UDP)-glucose pyrophosphorylase (UGPase) was used as a taxon specific gene. We designed the primer pair to detect the cryIIIA genes as a screening method for GM potatoes because the gene should be inserted in all 8 lines of the GM potatoes. For identification of NL, NLP and NLY, we further designed three specific primer pairs for the different recombinant DNAs (r-DNA) specifically introduced into NL, NLP, or NLY. In addition, to identify the 3 lines of NLY that have been introduced with the same r-DNA, the three line-specific primer pairs for the border sequence between the r-DNA and genomic DNA of NLY 3 lines were designed. Six lines of GM potato used as the test material were specifically identified using the each primer pair under the same PCR condition. The detection limits of all the GM potatoes should be approximately 0.1%. Furthermore, the specificity and reproducibility of the methods were confirmed in a six-laboratory collaborative study.

Key words genetically modified potato; detection method; NewLeaf; NewLeaf Plus; NewLeaf Y; uridine diphosphate (UDP)-glucose pyrophosphorylase (UGPase)