[go: up one dir, main page]

EP1151119A1 - Verfahren zur hemmung der expression von zielgenen in pflanzen - Google Patents

Verfahren zur hemmung der expression von zielgenen in pflanzen

Info

Publication number
EP1151119A1
EP1151119A1 EP00915151A EP00915151A EP1151119A1 EP 1151119 A1 EP1151119 A1 EP 1151119A1 EP 00915151 A EP00915151 A EP 00915151A EP 00915151 A EP00915151 A EP 00915151A EP 1151119 A1 EP1151119 A1 EP 1151119A1
Authority
EP
European Patent Office
Prior art keywords
plant
dna construct
sequence
chimeric
plants
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP00915151A
Other languages
English (en)
French (fr)
Inventor
Wolfgang Institute für WERR
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Biogemma SAS
Original Assignee
Rhobio SA
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Rhobio SA filed Critical Rhobio SA
Priority to EP00915151A priority Critical patent/EP1151119A1/de
Publication of EP1151119A1 publication Critical patent/EP1151119A1/de
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
    • C12N15/8237—Externally regulated expression systems
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
    • C12N15/8218—Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]

Definitions

  • the term "repression domain of the Drosophila engrailed gene (eng)” means a fragment of the Drosophila engrailed gene or of a derivative sequence thereof, said fragment comprising a nucleotide sequence that encodes a polypeptide interfering with the general transcription machinery and transcriptional activators. Said fragment advantageously comprises a minimal repression sequence coding for a polypeptide of 55 residues (Poole et al., 1985, Han K. et al., 1993). The whole engrailed gene may also be used but most preferably without its homeodomain.
  • Engrailed related domains which share the same repression activity, may also be used, such as those described in Smith et al. (1996).
  • the repressor sequence that is used may be a sequence that codes for at least the Kruppel-associated box-A (KRAB-A) domain of zinc finger proteins (Witzgall et al., 1994) for at least the RE-1- silencing transcription factor (REST) (Thiel et al., 1998), or for at least the BTB (for Broad-complex Tramtrac and Brie) domain, also known as POZ-domain (Ahmad et al., 1998 ; Huynh & Bardwell, 1998).
  • KRAB-A Kruppel-associated box-A domain of zinc finger proteins
  • REST RE-1- silencing transcription factor
  • BTB for Broad-complex Tramtrac and Brie domain
  • said transcription factor may preferably be selected from the group consisting of : STM ("Shootmeristemless"), member of the Knotted class of homeodomain proteins, which is an essential gene for development and function of the shoot apical meristem ;
  • ZmHox - member of the homeobox proteins more particularly ZmHox 1a/1 b and 2a/2b, which are expressed in maize meristems and proliferating cells from the early embryo to late reproductive organs. This expression pattern suggests a contribution to plant growth and morphogenesis;
  • Arabidopsis is a transcription factor belonging the family recently described as ARF1 family for Auxin Response Factor 1 (Ulmasov et al., 1997). Ms41-A analogues may also be used, such as those described in WO 97/23618 or the Arabidopsis gene Monopteros which encodes a transcription factor mediating embryo axis formation and vascular development (Hardtke et al., 1998), and the Arabidopsis gene ETTIN involved in floral development (Sessions, 1997).
  • the plants transformed with a chimeric DNA construct of the invention comprising such Ms-41-A factor are expected to be male sterile.
  • the promoter that is used is a tissue- specific promoter or a developmentally regulated promoter. This allows a conditional loss of functions that may be desired, for example to design new plant varieties.
  • 35S promoter or advantageously the double 35S constitutive promoter of CaMV as described in Kay et al. (1987) ; rice actin promoter followed by the rice actin intron (PAR- IAR) included in plasmid pAct1-F4 as described in Mc Elroy et al. (1991); the constitutive promoter EF-1 ⁇ of the gene encoding for plant elongation factor described in WO 90/02172 or in Axelos et al.
  • HMWG promoter High Molecular Weight Glutenin from barley (Anderson et al., 1989) ; the promoter of maize ⁇ zein (P ⁇ zein) included in p ⁇ 63 plasmid in Reina et al. (1990) allowing the expression in albumen of maize seeds ;
  • PGEA1 and PGEA6 promoters corresponding to the non coding 5' region of the genes GEA1 and GEA6, expressed in the grains in Arabidopsis thaliana (Gaubier et al., 1993) ; and ⁇ -phaseolin promoter (Riggs et al., 1989). • specific promoters which drive expression in particular plant tissues which are involved in the control of fertility :
  • parts of transgenic plants refer in particular to leaves, fruits, seeds, roots or cells that have been genetically transformed.
  • the preferred plants that are used for transformation may be for example selected from the group consisting of Arabidopsis thaliana, rice, tobacco, maize, Brassica, wheat, tomato and flowers (Petunia, rose, carnation).
  • Figure 5 represents a eng-STM construct.
  • Figure 6 represents a eng-AP3 construct.
  • the homeodomain of STM was removed in an independent experiment.
  • the plasmid pRT ⁇ eng-STM was first partially digested with Hindll, secondly to completion with BamHI and ends were ligated after fill in reaction with Klenow enzyme.
  • Plants are regenerated from these callus by modifying the hormonal and osmotic equilibrium of the cells according to the method described by Vain et al, 1989.
  • a method of genetic transformation leading to a stable integration of the modified genes in the genome of the plant involves the use of a gene gun.
  • the target cells are fragments of callus having a surface area of 10 to 20 mm 2 .
  • the fragments are laid in the center of a Petri dish containing a culture medium identical to the initiation medium, further containing 0.2 M of mannitol + 0.2 M of sorbitol (16 fragments per dish). Tissues are then bombarded as previously described.
  • the dishes are then sealed using Scellofrais® and cultivated in dark at 27°C.
  • the first planting is effected 24 hours afterwards, then every two-weeks during three months in a medium identical to the initiation medium but containing a selective agent.
  • the selective agents may be for example active ingredients of herbicidal agents (Basta®, Round up®) or antibiotics (hygromycin, kanamycin).
  • Example 5 Expression of the endogenous STM and AP3 genes
  • transgenic eng-STM or eng-AP3 phenocopy plants were subjected to RNA gel blot and RT-PCR analysis.
  • the chimeric transcripts derived from the CaMV 35S promoter were easily detectable in total or poly(A) + RNA from pooled seedlings or flowers (12 individual progeny) probed with the STM or AP3 coding regions.
  • the high abundance of chimeric transcripts interfered with the detection of the shorter native mRNA in these gel blot experiments.
  • Discrimination was achieved by probing with the natural 3'UTR sequences that are lacking in the transgenes. The result obtained for the STM 3'UTR confirms transcription of the native STM gene although at a significantly reduced level.
  • Example 6 Phenocopies rely on the incorporation of the chimeric eng-STM protein into the nuclear compartment Although the changes in size or identity of the expressing organs might explain the lower transcript levels observed in stm and ap3 phenocopy plants we aimed to elaborate further evidence against homology based gene silencing.
  • the chimeric eng-STM polypeptide therefore was expressed in C- terminal fusion with the hormone-binding domain of the glucocorticoid receptor in transgenic Arabidopsis plants. Due to this addition the resulting eng-STM-GR polypeptide should be cytoplasmatic and incorporated into the nuclear compartment only after hormone application. A linkage between the dexamethasone treatment and alterations in SAM activity thus should depend on the nuclear import of the chimeric eng-STM-GR protein and be incompatible with cosuppression.

Landscapes

  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Biomedical Technology (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Plant Pathology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Virology (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Peptides Or Proteins (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
EP00915151A 1999-02-09 2000-02-09 Verfahren zur hemmung der expression von zielgenen in pflanzen Withdrawn EP1151119A1 (de)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP00915151A EP1151119A1 (de) 1999-02-09 2000-02-09 Verfahren zur hemmung der expression von zielgenen in pflanzen

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
EP99420030 1999-02-09
EP99420030 1999-02-09
PCT/EP2000/001524 WO2000047754A1 (en) 1999-02-09 2000-02-09 A method for inhibiting the expression of target genes in plants
EP00915151A EP1151119A1 (de) 1999-02-09 2000-02-09 Verfahren zur hemmung der expression von zielgenen in pflanzen

Publications (1)

Publication Number Publication Date
EP1151119A1 true EP1151119A1 (de) 2001-11-07

Family

ID=8242281

Family Applications (1)

Application Number Title Priority Date Filing Date
EP00915151A Withdrawn EP1151119A1 (de) 1999-02-09 2000-02-09 Verfahren zur hemmung der expression von zielgenen in pflanzen

Country Status (4)

Country Link
EP (1) EP1151119A1 (de)
AU (1) AU770893B2 (de)
CA (1) CA2359749A1 (de)
WO (1) WO2000047754A1 (de)

Families Citing this family (26)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1714975A3 (de) 2001-12-26 2007-02-28 National Institute of Advanced Industrial Science and Technology Transkription-regulierendes Gen und Peptid
EP2278018A3 (de) 2002-08-07 2012-01-04 BASF Plant Science GmbH Nukleinsäuren die für Proteine kodieren, die an abiotischer Stressantwort beteiligt sind
CN103131673A (zh) 2003-04-15 2013-06-05 巴斯福植物科学有限公司 编码非生物胁迫反应相关蛋白质的核酸序列，以及提高对环境胁迫耐性的植物和植物细胞
JP2005295879A (ja) * 2004-04-09 2005-10-27 Japan Science & Technology Agency 花の形態が改変された植物体の生産方法およびこれを用いて得られる植物体、並びにその利用
JP2005295878A (ja) * 2004-04-09 2005-10-27 Japan Science & Technology Agency 花芽形成遅延植物体の生産方法、及びこれを用いて得られる植物体、並びにその利用
JP2006006248A (ja) * 2004-06-28 2006-01-12 Japan Science & Technology Agency 葉の形態形成が制御された植物体の生産方法およびこれを用いて得られる植物体、並びにその利用
JP2006020607A (ja) * 2004-07-09 2006-01-26 Japan Science & Technology Agency 葉の形態が改変された植物体の生産方法およびこれを用いて得られる植物体、並びにその利用
JP2006042730A (ja) * 2004-08-06 2006-02-16 Japan Science & Technology Agency 単子葉植物の雄性不稔体の生産方法およびこれを用いて得られる植物体、並びにその利用
CN103289961A (zh) 2004-09-24 2013-09-11 巴斯福植物科学有限公司 编码与非生物性胁迫反应相关的蛋白质的核酸序列和环境胁迫抗性增加的植物细胞和植物
JP2006101827A (ja) * 2004-10-08 2006-04-20 Japan Science & Technology Agency 雄性不稔形質転換植物体の生産方法およびこれを用いて得られる植物体、並びにその利用
JP2006280242A (ja) * 2005-03-31 2006-10-19 Japan Science & Technology Agency 完全不稔性植物体の生産方法およびこれを用いて得られる植物体、並びにその利用
EP2221382A3 (de) 2006-03-24 2010-12-01 BASF Plant Science GmbH Mit der abiotischen Stressreaktion assoziierte Proteine und Homologe
WO2008043849A2 (en) 2006-10-13 2008-04-17 Basf Plant Science Gmbh Plants with increased yield
EP2064330A2 (de) 2007-05-22 2009-06-03 BASF Plant Science GmbH Pflanzen mit erhöhter toleranz und/oder resistenz gegenüber umweltstress und erhöhter biomasseproduktion
CN103695459A (zh) 2007-09-18 2014-04-02 巴斯夫植物科学有限公司 产量提高的植物
US8809059B2 (en) 2007-09-21 2014-08-19 Basf Plant Science Gmbh Plants with increased yield
WO2009077611A2 (en) 2007-12-19 2009-06-25 Basf Plant Science Gmbh Plants with increased yield and/or increased tolerance to environmental stress (iy-bm)
US20110010800A1 (en) 2008-02-27 2011-01-13 Basf Plant Science Gmbh Plants with increased yield
MX2011001899A (es) 2008-08-19 2011-04-05 Basf Plant Science Gmbh Plantas con aumento de rendimiento al aumentar o generar una o mas actividades en una planta o una de sus partes.
EP2337853A1 (de) 2008-09-23 2011-06-29 BASF Plant Science Company GmbH Pflanzen mit erhöhtem ertrag (lt)
BRPI0924536A2 (pt) 2008-10-23 2015-08-11 Basf Plant Science Gmbh Método para produzir uma célula transgênica com teor de ácido gama-aminobutírico (gaba) aumentado, para aumentar o rendimento, molécula de ácido nucleico isolada, construção de ácido nucleico, vetor, célula hospedeira, processo para produzir um polipeptídeo, polipeptídeo, anticorpo, e, núcleo de célula, célula, núcleo de célula vegetal, célula vegetal, tecido vegetal, material de propagação, pólen, progênie, material coletado ou uma planta
EP2350289A1 (de) 2008-10-23 2011-08-03 BASF Plant Science GmbH Pflanzen mit erhöhtem ertrag (nue)
WO2011009801A1 (en) 2009-07-23 2011-01-27 Basf Plant Science Company Gmbh Plants with increased yield
AU2010320547B2 (en) 2009-11-17 2016-06-09 Basf Plant Science Company Gmbh Plants with increased yield
EP2961845A4 (de) 2013-03-01 2016-08-10 Univ California Verfahren und zusammensetzungen zum targeting von rna-polymerasen und zur nichtcodierung einer rna-biogenese an spezifischen loci
WO2016128470A1 (en) 2015-02-11 2016-08-18 Basf Se Herbicide-resistant hydroxyphenylpyruvate dioxygenases

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5866755A (en) * 1993-06-14 1999-02-02 Basf Aktiengellschaft Animals transgenic for a tetracycline-regulated transcriptional inhibitor

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO0047754A1 *

Also Published As

Publication number Publication date
CA2359749A1 (en) 2000-08-17
AU3656300A (en) 2000-08-29
AU770893B2 (en) 2004-03-04
WO2000047754A1 (en) 2000-08-17

Similar Documents

Publication Publication Date Title
AU770893B2 (en) A method for inhibiting the expression of target genes in plants
US6005167A (en) Male-sterile plants, method for obtaining male-sterile plants and recombinant DNA for use therein
US5977441A (en) Control of plant gene expression
US5723765A (en) Control of plant gene expression
Southerton et al. Eucalyptus has a functional equivalent of the Arabidopsis floral meristem identity gene LEAFY
Huang et al. The Arabidopsis ACT11 actin gene is strongly expressed in tissues of the emerging inflorescence, pollen, and developing ovules
US20030126642A1 (en) Compositions and methods for modulating plant development
HUT73336A (en) Methods and compositions for controlling plant development
US9587243B2 (en) Ubiquitin regulatory elements
Schoenbeck et al. The alfalfa (Medicago sativa) TDY1 gene encodes a mitogen-activated protein kinase homolog
WO2005122751A1 (en) Nucleic acid molecules and their use in plant male sterility
BRPI0011101B1 (pt) Vetor, e, métodos para produzir embriões derivados assexualmente, para modificar a capacidade regenerativa de uma planta, para produzir uma planta apomíctica e para selecionar uma planta modificada
US5824872A (en) A constitutive promoter from tobacco
JP2001524315A (ja) サイレント導入遺伝子の活性化のために有用な方法及び組成物
US7183109B2 (en) Embryo preferred promoter and method of using same
US7112723B2 (en) Globulin 2 regulatory region and method of using same
WO2000023578A2 (en) Methods of suppressing flowering in transgenic plants
WO2001031017A2 (en) Plants with a modified flower and seed development
CA2168617C (en) Constitutive promoter from tobacco
JP2003092936A (ja) タペート層特異的ジンクフィンガー転写因子の遺伝子を用いて花粉稔性を低下させる方法
Zhao Structure and regulation of 4-coumarate: CoA ligase genes in rice (Oryza sativa)

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20010802

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU MC NL PT SE

AX Request for extension of the european patent

Free format text: AL;LT PAYMENT 20010801;LV PAYMENT 20010801;MK;RO PAYMENT 20010801;SI PAYMENT 20010801

17Q First examination report despatched

Effective date: 20030613

RAP1 Party data changed (applicant data changed or rights of an application transferred)

Owner name: BIOGEMMA

17Q First examination report despatched

Effective date: 20030613

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20060901