Method to identify herbicides, fungicides or plant growth regulators 机翻标题: 暂无翻译,请尝试点击翻译按钮。

公开号/公开日
WO200120020 A2 2001-03-22 [WO200120020]WO200120020 A3 2001-09-27 [WO200120020] / 2001-03-222001-09-27
申请号/申请日
2000WO-EP08797 / 2000-09-08
发明人
BOUNAGA SAKINA;INZE DIRK;DE VEYLDER LIEVEN;MIRONOV VLADIMIR;
申请人
CROPDESIGN;
主分类号
IPC分类号
C07K-014/415C12N-015/81C12Q-001/02C12Q-001/18
摘要
(WO200120020) The present invention relates to a method for screening and identification of compounds or compositions useful as herbicides, growth regulators or fungicides involving at least the following steps of addition of the compound or composition to be screened or identified to a culture or culture area of a yeast strain transformed with and expressing one or more plant cell cycle control genes or mutants thereof (phytoyeast) as well as to a control yeast strain; and, determining the effect on the phenotype (growth and/or cell division and/or cell size/shape) of said phytoyeast compared to said control yeast.  The invention further relates to a bilogical screening assay (high throughput system) comprising the use of said phytoyeast expressing plant cell cycle control proteins to identify novel compounds or compositions that affect control yeast or phytoyeast phenotype.  The present invention also relates to methods for producing a pesticide, herbicide, plant growth regulator or fungicide, comprising the steps of (a) identifying a compound or composition as defined above or a derivative or homologue thereof, and, (b) mixing said compound, composition, derivative or homologue thereof with an acceptable carrier.  The invention also relates to the use of said compounds or compositions for inhibiting or stimulating plant growth and/or for increasing crop yield and/or for preventive or curative protection of the plant against fungal infection.
机翻摘要
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地址
代理人
代理机构
;
优先权号
1999EP-0870183 1999-09-10 2000US-60196240 2000-04-10
主权利要求
(WO200120020) CLAIMS 1. A method for screening and identification of compounds or compositions useful as herbicides, growth regulators or fungicides involving at least the following steps: (a) addition of the compound or composition to be screened or identified to a culture or culture area of a yeast strain transformed with and expressing one or more plant cell cycle control genes or mutants thereof (phytoyeast) as well as to a control yeast strain; and, (b) determining the effect on the phenotype (growth and/or cell division and/or cell size/shape) of said phytoyeast compared to said control yeast. 2. The method of claim 1 , wherein said plant polynucleic acids are expressed in yeast under the control of a repressible or inducible promoter, any other controlling element and/or culture conditions. 3. The method of claim 1 or 2 wherein said yeast is any strain of Saccharomyces cerevisiae, Candida albicans or more preferably Schizosaccharomyces pombe. 4. The method of any of claims 1 to 3, wherein said transformed yeast strain (phytoyeast) contains a plant cell cycle control gene or mutant thereof which is functionally complementing an essential endogenous cell cycle control gene or mutant thereof, involving at least the steps of: (a) addition of the compound or composition to be screened or identified to a culture or culture area of said phytoyeast expressing said plant cell cycle control protein or a mutant thereof, as well as to a control yeast strain; (b) determining the effect on the phenotype (growth and/or cell division and/or cell size/shape) of said phytoyeast compared to said control yeast; (c) identifying a compound or composition which affects said phenotype of said phytoyeast but not of said control yeast; and, (d) confirming whether said compound or composition also inhibits or stimulates the growth of plants. 5. The method of any of claims 1 to 3, wherein said transformed yeast strain (phytoyeast) expresses a plant cell cycle control gene or dominant mutant thereof, wherein said method involves at least the following steps: (a) addition of the compound or composition to be screened or identified to a culture or culture area of said phytoyeast expressing said plant cell cycle control gene or dominant mutant thereof as well as to a control yeast strain; (b) determining the effect on the phenotype (growth and/or cell division and/or cell size/shape) of said phytoyeast compared to said control yeast; (c) identifying a compound or composition which affects said phenotype of said phytoyeast but not of said control yeast; and (d) confirming whether said compound or composition also inhibits or stimulates the growth of plants. 6. The method of claim 5, wherein said transformed yeast (phytoyeast) expresses a plant cell cycle control gene which does not complement endogenous yeast cell cycle control genes but which affects the phenotype (growth and/or cell division and/or cell size/shape) of said phytoyeast. 7. The method of claim 5, wherein said transformed yeast (phytoyeast) expresses a plant cell cycle control gene with a dominant negative mutation resulting in growth arrest. 8. The method of claim 5, wherein said transformed yeast (phytoyeast) expresses a plant cell cycle control gene with a dominant positive mutation resulting in growth acceleration. 9. The method of any of claims 1 to 3, wherein said transformed yeast (phytoyeast) expresses a first plant cell cycle control gene or mutant thereof with the activity of its encoded protein being modulated by the expression of a second plant cell cycle control gene or mutant thereof, wherein said method comprises the following steps: (a) addition of the compound or composition to be screened or identified to a culture or culture area of a strain of said double transformed yeast (phytoyeast) expressing at least said first and said second plant cell cycle control gene or mutants thereof as well as to said phytoyeast only expressing said first plant cell cycle control gene; (b) determining the effect on the phenotype (growth and/or cell division and/or cell size/shape) of said double transformed phytoyeast expressing at least said first and said second plant cell cycle control gene compared to said phytoyeast only expressing said first plant cell cycle control gene; (c) identifying a compound or composition which affects said phenotype of said double transformed phytoyeast expressing at least said first and said second plant cell cycle control gene compared to said phytoyeast only expressing said first plant cell cycle control gene; and (d) confirming whether said compound or composition also inhibits or stimulates the growth of plants. 10. The method of claim 9, wherein said first plant cell division cycle control protein is preferably encoded by the A. thaliana cdc2a gene or mutant thereof and said second plant cell cycle control protein or mutant thereof is encoded by another cell cycle control gene or mutant thereof, preferably an A. thaliana CKI or mutant thereof. 11 . A method for identifying plant specific compounds or compositions comprising the steps of a method of any of claims 1 to 10 wherein said compound or composition affects the phenotype (growth and/or cell division and/or cell shape/size) of a yeast strain transformed with one or more plant cell cycle control genes (phytoyeast) and not of a wild type control yeast and not of a control yeast expressing an animal or human homologue(s) of the plant cell cycle control gene(s) expressed in the phytoyeast. 12. A method for identifying fungicide compounds or compositions comprising the steps of a method of claim 1 to 10 wherein said compound or composition inhibits growth of the wild type control yeast but not of the mutant control yeast being either a phytoyeast and or a yeast expressing the corresponding animal or human cell cycle control gene. 13. The method for identifying a compound or composition of any of claims 1 to 12, comprising solubilising said test compound or composition in a solvent, diluting it and adding it to the cell culture medium of said yeast strains. 14. The method of any of claims 1 to 13, wherein said compound or composition is added to a disk or to a well on a culture plate, in a standard diffusion assay using solidified media, or is added directly into liquid media. 15. The method of any of claims 1 to 14, wherein said yeast are exposed to a range of concentrations of said test compound or composition. 16. A method of any of claims 1 to 15, wherein the determination of the effect on the phenotype (growth and/or cell division and/or cell size/shape) consists of measuring the decrease or increase of the degree of growth and/or cell division and/or analyzing the phenotype of said phytoyeast and said control yeast upon addition of said compound or composition. 17. Use of transformed yeast cells expressing at least one plant cell cycle control gene or a mutant thereof as defined in any of claims 1 to 3 for the screening or identification of compounds or compositions which abolish, retard or stimulate plant growth. 18. Use of a recombinant vector comprising at least one polynucleic acid encoding at least part of a plant cell cycle control protein or a mutant thereof as defined in any of claims 1 to 3 to transform yeast for the screening or identification of compounds or compositions which abolish, retard or stimulate plant growth. 19. The use of claim 18, wherein said recombinant vector is a plasmid, more particularly a vector comprising a selectable marker and transcriptional control elements for the expression of said plant or animal/human cell cycle control polynucleic acids in yeast. 20. The method or use of any of claims 1 to 19, wherein said plant or animal cell cycle control polynucleic acid is integrated into the yeast genome by random, non-homologous or homologous recombination. 21. A biological screening assay comprising the use of transformed yeast (phytoyeast) as defined in any of claims 1 to 20 expressing plant cell cycle control proteins to identify novel compounds or compositions that affect control yeast or phytoyeast phenotype. 22. The use of a screening assay according to claim 21 for identifying compounds or compositions which act as herbicides, plant growth regulators or fungicides. 23. A high throughput screening system to determine compounds or compositions which act as herbicides, plant growth regulators or fungicides involving a method, use or biological assay of any of claims 1 to 22, preferably a microtiter plate screening system. 24. A compound or composition identifiable by a method according to any of claims 1 to 23. 25. A method for producing a pesticide, herbicide, plant growth regulator or fungicide, comprising the steps of: (a)identifying a compound or composition as defined in any of claims 1 to 24 or a derivative or homologue thereof, and, (b)mixing said compound, composition, derivative or homologue thereof with an acceptable carrier. 26. Use of a compound or composition according to claim 24 in a method according to claim 25 for inhibiting or stimulating plant growth and/or for increasing crop yield and/or for preventive or curative protection of the plant against fungal infection. 27. A yeast strain expressing CDC2a.AF and a cyclin-dependent kinase inhibitor of A. thaliana resulting in an elongated cell type. 28. A compound screening assay comprising the use of a yeast strain of claim 27.
法律状态
(WO200120020) LEGAL DETAILS FOR WO200120020  Actual or expected expiration date=2003-03-10    Legal state=DEAD    Status=LAPSED     Event publication date=2000-09-08  Event code=WO/APP  Event indicator=Pos  Event type=Examination events  Application details  Application country=WO WOEP0008797  Application date=2000-09-08  Standardized application number=2000WO-EP08797     Event publication date=2001-03-22  Event code=WO/A2  Event type=Examination events  International application published without international search report  Publication country=WO  Publication number=WO200120020  Publication stage Code=A2  Publication date=2001-03-22  Standardized publication number=WO200120020     Event publication date=2001-03-22  Event code=WO/AK  Event indicator=Pos  Event type=Designated states  Designated states    Event publication date=2001-03-22  Event code=WO/AL  Event indicator=Pos  Event type=Designated states  Designated countries for regional patents    Event publication date=2001-06-28  Event code=WO/DFPE  Event type=Examination events  Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)    Event publication date=2001-09-27  Event code=WO/A3  Event indicator=Pos  Event type=Examination events  Later publication of ISR with revised front page  Publication country=WO  Publication number=WO200120020  Publication stage Code=A3  Publication date=2001-09-27  Standardized publication number=WO200120020     Event publication date=2001-09-27  Event code=WO/AL  Event indicator=Pos  Event type=Designated states  Designated countries for regional patents    Event publication date=2001-09-27  Event code=WO/AK  Event indicator=Pos  Event type=Designated states  Designated states    Event publication date=2003-03-10  Event code=WO/EETL  Event type=Event indicating Not In Force  PCT Application validity period expired. LEGAL DETAILS FOR DESIGNATED STATE AU7517200  Actual or expected expiration date=2002-05-16    Legal state=DEAD    Status=LAPSED   Corresponding cc:  Designated or member state=AU Corresponding appl: AU7517200  Application date in the designated or member state=2000-09-08   Application number in the designated or member state=2000AU-0075172 Corresponding cc:  Designated or member state=AU Corresponding pat: AU7517200  Publication stage code in the designated or member state=A  Publication date in the designated or member state=2001-04-17   Publication number in the designated or member state=AU200075172    Event publication date=2002-05-16  Event code=AU/STCHG  Patent Status changed by the national office Corresponding cc:  Designated or member state=AU  LEGAL DETAILS FOR DESIGNATED STATE DE  Actual or expected expiration date=2002-08-22    Legal state=DEAD    Status=LAPSED   Corresponding cc:  Designated or member state=DE     Event publication date=2002-08-22  Event code=WO/REG  Event code=DE/8642  Event indicator=Neg  Event type=Non-entry into national phase  Event type=Event indicating Not In Force  Reference to a national code Impact abolished for de - i.e. PCT appl. not ent. German phase Wirkung weggefallen fuer de Corresponding cc:  Designated or member state=DE  LEGAL DETAILS FOR DESIGNATED STATE JP  Actual or expected expiration date=2004-06-11    Legal state=DEAD    Status=LAPSED   Corresponding cc:  Designated or member state=JP     Event publication date=2004-06-11  Event code=WO/NENP  Event type=Event indicating Not In Force  Non-entry into the national phase in: Corresponding cc:  Designated or member state=JP
专利类型码
A2A3
国别省市代码
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