Engineering herbicide metabolism in tobacco and Arabidopsis with CYP76B1, a cytochrome P450 enzyme from Jerusalem artichoke.
The Jerusalem artichoke (Helianthus tuberosus) xenobiotic inducible cytochrome P450, CYP76B1, catalyzes rapid oxidative dealkylation of various phenylurea herbicides to yield nonphytotoxic metabolites. We have found that increased herbicide metabolism and tolerance can be achieved by ectopic constitutive expression of CYP76B1 in tobacco (Nicotiana tabacum) and Arabidopsis. Transformation with CYP76B1 conferred on tobacco and Arabidopsis a 20-fold increase in tolerance to linuron, a compound detoxified by a single dealkylation, and a 10-fold increase in tolerance to isoproturon or chlortoluron, which need successive catalytic steps for detoxification. Two constructs for expression of translational fusions of CYP76B1 with P450 reductase were prepared to test if they would yield even greater herbicide tolerance. Plants expressing these constructs had lower herbicide tolerance than CYP76B1 alone, which is apparently a consequence of reduced stability of the fusion proteins. In all cases, increased herbicide tolerance results from more extensive metabolism, as demonstrated with exogenously fed phenylurea. Beside increased herbicide tolerance, expression of CYP76B1 has no other visible phenotype in the transgenic plants. Our data indicate that CYP76B1 can function as a selectable marker for plant transformation, allowing efficient selection in vitro and in soil-grown plants. Plants expressing CYP76B1 may also be a potential tool for phytoremediation of contaminated sites.
Didierjean
Luc
L
E.I. DuPont de Nemours and Company, Central Research and Development, DuPont Experimental Station, Wilmington, Delaware 19880-0328, USA.
Gondet
Laurence
L
Perkins
Roberta
R
Lau
Sze-Mei Cindy
SM
Schaller
Hubert
H
O'Keefe
Daniel P
DP
Werck-Reichhart
Danièle
D
D015660:7-Alkoxycoumarin O-Dealkylase / Q000235:genetics* / Q000378:metabolism; D000222:Adaptation, Physiological / Q000235:genetics / Q000502:physiology; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D002250:Carbon Radioisotopes; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D005818:Genetic Engineering / Q000379:methods; D006368:Helianthus / Q000201:enzymology; D006540:Herbicides / Q000378:metabolism* / Q000494:pharmacology; D008969:Molecular Sequence Data; D009251:NADPH-Ferrihemoprotein Reductase / Q000235:genetics / Q000378:metabolism; D010671:Phenylurea Compounds; D018515:Plant Leaves / Q000378:metabolism; D030821:Plants, Genetically Modified; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism*; D015003:Yeasts / Q000235:genetics / Q000378:metabolism
D002250:Carbon Radioisotopes; D018076:DNA, Complementary; D006540:Herbicides; D010671:Phenylurea Compounds; D011993:Recombinant Fusion Proteins; D003577:Cytochrome P-450 Enzyme System; D015660:7-Alkoxycoumarin O-Dealkylase; C113678:cytochrome P-450 CYP76B1 (Helianthus tuberosus); D009251:NADPH-Ferrihemoprotein Reductase