title: Caffeine production in tobacco plants by simultaneous expression of three coffee N-methyltrasferases and its potential as a pest repellant.
abstract: Caffeine (1,3,7-trimethylxanthine) is derived from xanthosine through three successive transfers of methyl groups and a single ribose removal in coffee plants. The methyl group transfer is catalyzed by N-zmethyltransferases, xanthosine methyltransferase (XMT), 7-methylxanthine methyltransferase (MXMT) and 3,7-dimethylxanthine methyltransferase (DXMT). We previously cloned three genes encoding each of these N-methyltransferases from coffee plants, and reconstituted the final sequence of the caffeine synthetic pathway in vitro. In the present study, we simultaneously expressed these coffee genes in tobacco plants (Nicotiana tabacum), using a multiple-gene transfer method, and confirmed successful caffeine production up to 5 microg g(-1) fresh weight in leaves of the resulting transgenic plants. Their effects on feeding behavior of tobacco cutworms (Spodoptera litura), which damage a wide range of crops, were then examined. Leaf disc choice test showed that caterpillars selectively fed on the wild-type control materials, or positively avoided the transgenic materials. The results suggest a novel approach to confer self-defense by producing caffeine in planta. A second generation of transgenic crops containing caffeine may save labor and agricultural costs and also mitigate the environmental load of pesticides in future.
authors.affiliation: Research and Education Center for Genetic Information, Nara Institute of Science and Technology, Nara, 630-0192, Japan.
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Uefuji
authors.forename: Yuko
authors.initials: Y
authors.lastname: Tatsumi
authors.forename: Masayuki
authors.initials: M
authors.lastname: Morimoto
authors.forename: Pulla
authors.initials: P
authors.lastname: Kaothien-Nakayama
authors.forename: Shinjiro
authors.initials: S
authors.lastname: Ogita
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Sano
mesh_terms: D000818:Animals; D002110:Caffeine / Q000096:biosynthesis* / Q000378:metabolism; D040503:Coffea / Q000201:enzymology* / Q000235:genetics; D005247:Feeding Behavior / Q000187:drug effects; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D018506:Gene Expression Regulation, Plant / Q000235:genetics; D007302:Insect Repellents; D007814:Larva / Q000187:drug effects; D008780:Methyltransferases / Q000235:genetics* / Q000378:metabolism*; D015394:Molecular Structure; D010572:Pest Control, Biological / Q000379:methods*; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D018411:Spodoptera / Q000187:drug effects; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism*
chemical_list: D007302:Insect Repellents; D002110:Caffeine; D008780:Methyltransferases