4-hydroxyphenylpyruvate dioxygenase (HPPD) gene, inhibitors of HPPD, method of conferring resistance to HPPD inhibitors and a method for detecting herbicide resistant HPPD genes

摘要

A method for the identification of herbicide resistant 4-hydroxyphenylpyruvate dioxygenase (HPPD) inhibitors comprising the steps of: (a) incubating a cell transformed with and expressing the nucleic acid which includes a sequence encoding HPPD proteins expressed in plants, the presence of a test compound to form a test culture, and in the absence of a test compound to form a control culture; (b) monitoring the level of homogentistic acid or oxidation products thereof, in the test culture and control; and (c) identifying as a compound that inhibits HPPD any compound that reduces the level of homogentistic acid or oxidation products thereof, in the test culture relative to the control culture. A variant HPPD protein is produced from Arabidopsis thaliana for use in the test culture is described. The amino acid encoding Arabidopsis thaliana HPPD being: MGHQNAAVSENQNHDDGAASSPGFKLVGFSKFV RKNPKSDKFKVKRFHHIEFWCGDATNVARRFSW GLGMRFSAKSDLSTGNMVHASYLLTSGD LRFLFT APYSPSLSAGEIKPTTASIPSFDHGSCRSFFSSHGL GVRAVAIEVEDAESAFSISVANGAIPSSPPIVLNEA VTIAEVKLYGDVVLRYVSYKAEDTEKSEFLPGFE RVEDASSFPLDYGIRRLDHAVGNVPELGPALTYV AGFTGFHQFAEFTADDVGTAESGLNSAVLASNDE MVLLPINEPVHGTKRKSQIQTYLEHNEGAGLQHL ALMSEDIFRTLREMRKRSSIGGFDFMPS PPPTYYQ NLKKRVGDVLSDDQIKECEELGILVDRDDQGTLL QIFTKPLGDDRRTIFIEIIQRVGCMMKDEEGKAYQ SGGCGGFGKGNFSELFKSIEEYEKTLEAKQLVG