WO1992005261A1 - Composes organiques ameliores - Google Patents

Composes organiques ameliores Download PDF

Info

Publication number
WO1992005261A1
WO1992005261A1 PCT/EP1991/001786 EP9101786W WO9205261A1 WO 1992005261 A1 WO1992005261 A1 WO 1992005261A1 EP 9101786 W EP9101786 W EP 9101786W WO 9205261 A1 WO9205261 A1 WO 9205261A1
Authority
WO
WIPO (PCT)
Prior art keywords
wunl
promoter
dna
gene
plants
Prior art date
Application number
PCT/EP1991/001786
Other languages
English (en)
Inventor
Jürgen Logemann
Jeff Schell
Barbara Siebertz
Original Assignee
MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V.
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 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. filed Critical MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V.
Publication of WO1992005261A1 publication Critical patent/WO1992005261A1/fr

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)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8287—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for fertility modification, e.g. apomixis
    • C12N15/8289—Male sterility
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants

Definitions

  • the invention relates to a DNA sequence, an anther-specific promoter and the use thereof.
  • wunl gene A gene with a DNA sequence called "wunl gene", which is isolated from Solanu tuberosum, is known from literature references such as The Plant Cell, Vol. 1, 1989, p. 151-158; Proc. Natl. Acad.Sci.USA, Vol. 85, p. 1136-1140, Fbr.1988, as well as DE-OS 38 37 752. It is known that the wunl gene leads to the expression of gene products in the case of wounding or a pathogenic infection.
  • wunl gene a promoter region of 1022 bp length (+178bp wunl 5'untranslated region) maintains its wound- inducible activity even when the actual structural gene region is replaced by another structural gene (e.g. CAT, GUS). In this way, vound-inducible wunl promoter activity could be detected in leaf, stem and root of transgenic tobacco plants.
  • transgenic plants carrying fragments of the 5' region of the wound-inducible wunl gene in reverse orientation have constitutive anther-specific promoter activity.
  • bp are designated (-1022 +x)wunl (e.g. -571wunl).
  • Vectors having such promoter (e.g. -571wunl) attached at the 5'end of the GUS gene are designated (-1022 + x) wunl-GUS (e.g. -571 wunl-GUS).
  • DNA sequences of -1022 wunl cloned in reverse orientation are identified by r (e.g. 571r wunl resp. when cloned in front of the GUS reporter gene -571r wunl-GUS).
  • the invention therefore provides constitutive anther specific promoters, comprising a DNA fragment from the 5' region of the wunl gene, but which is in reverse orientation of said 5' region, DNA sequences homologous thereto, parts thereof or combinations of such parts.
  • vector as used herein relates to any vehicle by means of which DNA fragments can be introduced into a host organism.
  • the promoters of the invention are DNA fragments from the -1022 wunl promoter.
  • the DNA sequences of the invention have a bp range of from -571 of the wunl promoter to +178 of the 5' untranslated region of the wunl gene, or are homologous thereto, parts thereof or combinations of such parts, whereby the sequences are transcriptionally fused in the reverse orientation with gene.
  • a suitable example of such vector is a vector comprising the -571r wunl-GUS construct.
  • the DNA sequences of the invention have a bp range of from -571 to +1, or are homologous thereto, parts thereof or combinations of such parts and are transcriptionally fused in the normal or reverse orientation with a structural gene.
  • DNA sequence homologous to a DNA sequence of a constitutive anther specific promoter of the invention refers to a DNA sequence of a constitutive anther specific promoter of the invention wherein a number of nucleotides have been deleted and/or added but is still capable of hybridization to a nucleotide sequence having at least 50 nucleotides complementary to a DNA sequence of a constitutive anther specific promoter of the invention under appropriate hybridization conditions.
  • hybridization conditions conveniently include an incubation for 16 hours at 42°C, in a buffer system comprising 5 x standard saline citrate (SSC), 0.5 X sodium dodecylsulphate (SDS), 5 x Denhardt's solution, 50 X formamide and 100 ⁇ g/ml carrier DNA (hereinafter the buffer system), followed by washing 3 times with a buffer comprising 1 x SSC and 0.1 X SDS at 65 P C for approximately one hour each time.
  • SSC standard saline citrate
  • SDS sodium dodecylsulphate
  • 5 x Denhardt's solution 50 X formamide
  • carrier DNA hereinafter the buffer system
  • Preferred hybridization conditions for the purpose of the invention involve incubation in the buffer system for 16 hours at 49°C and washing 3 times with a buffer comprising 0.1 x SSC and 0.1 X SDS at 55°C for approximately one hour each time. Most preferred hybridization conditions for the purpose of the invention involve incubation in the buffer system for 16 hours at 55°C and washing 3 times with a buffer comprising 0.1 x SSC and 0.1 X SDS at 65°C for approximately one hour each time.
  • DNA sequences having constitutive anther specific promoter activity may be derived from the -1022wunl promoter by deleting fragments thereof, cloning it in reverse orientation and screening such shortened promoter for constitutive anther specific promoter activity in a manner known per se, e.g. employing a reporter gene such as GUS, analogous to the procedure described herein.
  • the DNA sequences according to the invention comprise at least 50 base pairs of the wunl promoter sequence in reverse orientation or a DNA sequence homologous thereto.
  • the DNA sequences according to the invention comprise at least 70 base pairs of the wunl promoter sequence in reverse orientation or a DNA sequence homologous thereto.
  • the DNA sequences according to the invention comprise at least 80 base pairs of the vunl promoter sequence in reverse orientation or a DNA sequence homologous thereto.
  • the promoters of the invention are useful, in that they allow the selective expression of gene products in the anthers.
  • a promoter of the invention is operably linked to a gene of which the selective expression of the gene product in the anthers is desired, e.g. to inhibit or influence cell development in the anthers or for ornamental purposes.
  • Such vectors also form part of the invention.
  • genes of which anther specific expression may be useful include genes causing development aberrations including male sterility, e.g. the rolC gene, or genes encoding gene products which inactivate the plant cell, e.g. the thionin gene, genes encoding gene products capable of degrading DNA, e.g. the gene encoding the synthesis of the Eco RI endonuclease or by inactivation of genes essential for anther development, e.g. the wunl antisense gene. Anther specific expression of such genes will result in male sterile plants. Where the gene resulting in male sterility is recessive, male sterile plants are obtained by selfing the plants containing the recessive gene.
  • a restorer system For the production of Fl hybrid seed the availability of a restorer system is necessary if the Fl hybrid crop involved is grown for its fruits or its seeds.
  • such restorer system can be made by using the anti-sense gene of the gene that was used to induce male sterility in the transgenic plant. Methods to achieve anti-sense inhibition are known to a person skilled in the art ⁇ ind include e.g.
  • the anti-sense rol C gene if the wunl-rolC construct was used to create male sterility; a similar strategy can be used for the thionine gene; in cases of the endonuclease gene induced male sterility not only the anti-sense strategy may be used, but also the insertion of the corresponding methylase gene creating a restorer for the endonuclease induced male sterility may be employed.
  • Male sterility brought about by gene technology has the advantage that by incorporating a definite gene into the genetic code of a plant, the remaining properties of the plant do not change. This applies especially to cultivated plants which genetically can be manipulated relatively well, e.g. to dicotyledoneous plants which are suited for tissue culture and transformable, but also to other cultivated plants. Male sterile ornamental plants have also the advantage that they may flower for a longer period of time.
  • the invention also provides male sterile plants or plant material comprising a constitutive anther specific promoter according to the invention.
  • promoters of the invention can be operably linked to transcription enhancers, such as the 35 S enhance .
  • Such plants or plant material are obtained in a manner known per se employing conventional transformation techniques, and where desired, followed by conventional breeding of thus obtained transgenic plants.
  • Fig. 1 Arrangement of wunl in the genomic clone wunl-85.
  • Fig. 2 Deletion analysis of the wunl-genomic clone wunl-85.
  • Fig. 3 Determination of the wunl transcription start point.
  • Fig. 4 Nucleotide sequence of wunl and flanking regions from wunl-85. CAAT-Box, TATA-Box and PolyA-signal are shown on black backgrounds. The start and stop of transcription are marked by arrows.
  • Fig. 5 Arrangement and position of important regions in the wunl gene.
  • Fig. 6 Construction of vunl-GUS 5' deletions for expression analysis in stable tobacco transformants. Between the left and the right T-DNA border sequence of the vector pPR69, a
  • NPT II gene lies under the control of the nopalin synthase promoter (nos). This is necessary for selection for kanamycin.
  • the GUS gene bears a nos-terminator sequence and is regulated by the various 5'deletion fragments of the wunl promoter. The end positions of the respective
  • the media used for the culture of bacteria are taken from data given by Maniatis et al. (1982).
  • the media employed are derived from the media (MS) given by Murashige and Skoog (1962).
  • Plasmids pUC8 (Vieira and Messing, 1982)
  • pPR69 a derivative of bin 19, Bevan 1984
  • Phages EMBL4 (Frischholz et al. , 1983)
  • RNA isolation of RNA from various organs of the potato and of tobacco are carried out as described by Logemann et al., (1987).
  • RNA is separated by electrophoresis on a 1.5% formaldehyde- agarose-gel (Lehrach et al. , (1977)). As described by Willmitzer et al., (1982), the RNA is subsequently transferred onto nitrocellulose, fixed and hybridized on - -? radioactively-labelled cDNA, washed and exposed.
  • Nuclear DNA is recovered by the method of Bedbrook (1981) from potato leaves and used for cloning in lambda phages EMBL4. DNA from transformed tissue is purified by combined lyses with Triton X 100, SDS, and proteinase K (Wassenegger, 1988). "Southern Blot" Analysis
  • DNA is separated by electrophoresis on 0.8 to 1.22 agarose gels, transferred onto nitrocellulose and fixed (Southern, 1975), also hybridized and washed as described by Willmitzer et al., (1981).
  • Genomic potato DNA is isolated by the method of Bedbrook, 1981 from leaf material of the haploid species AM 80/5793 and digested totally with EcoRI.
  • EMBL4 DNA is digested with EcoRI into three fragments, whereby the two vector arms are separated from the middle fragment by means of gel electrophoresis separation with subsequent fragment isolation.
  • commercially available, purified EMBL4 arms are also used (Amersham).
  • the ligated, high molecular weight DNA is packaged in vitro into phage heads (Hohn and Murray, 1977; Hohn, 1979) employing packaging material from a "Lambda in vitro packaging kit" from the company Amersham.
  • the genomic bank is plated out at a concentration of 25000 plaques per plate (25x25 cm).
  • a bacteria culture 20 ml of a bacteria culture, lysed with C600, are mixed with chloroform, in order to obtain a bacteria-free supernatant after subsequent centrifugation. Sedimentation of the phages from the supernatant is effected by centrifugation for 4 hours at 10000 rpm.
  • the phage sediment is taken up in 500 ul phage buffer (10 mM tris-HCl pH 8.0, 10 mM MgCl 2 ), and treated with DNase and RNase. After extraction of the DNA by means of phenolization several times, the phage-DNA is EtOH-precipitated, washed with 70 X EtOH and taken up in TE.
  • the fragment to be examined is cloned in M13mpl8 in both orientations. Since exonuclease III digests 5' projecting ends, but are spares 3' projecting ends, 20 ug of the DNA to be analysed are digested with two restriction enzymes which produce a 5' and a 3' end on one side of the cloned fragment. By successive stopping of this reaction, fragments which are 200 bp smaller are isolated, and the projecting ends thereof are transformed into ligatable smooth ends by means of subsequent Sl-treatment. Finally, transformation of this DNA into BMH 7118 cells takes place, with subsequent single-stranded DNA isolation.
  • Sequencing of single-stranded DNA is carried out by the chain- terminating method of Sanger et al., (1977). Separation of the reaction products is performed on 6X and QX sequencing gels (Ansorge and de Mayer, 1980; Ansorge and Barker, 1984). SI analysis
  • the starting point of transcription is effected according to Berk and Sharp (1987).
  • the 1.2 kb fragment is isolated from the plasmid pLSOOO with EcoRI and Xhol, and dephosphorylated by treatment with phosphatase. Then, radioactive labelling of the 5' OH ends takes place through the combination of polynucleotide kinase and gamma- 32 P-ATP. After denaturing of the DNA fragments, they are hybridized with 50 ug of whole RNA (hybridization buffer: 80% formamide, 0.4 M NaCl, 40 mM PIPES pH 6.4 and 1 mM EDTA.
  • RNA-DNA hybrids compared with DNA-DNA hybrids (Casey and Davidson, 1977).
  • the temperature at the start of hybridization starts at 80°C and is lowered over night to 40°C.
  • Sl-nuclease digestion 120 U/ml removes unpaired strands.
  • the radioactively labelled Xhol cutting site found in the 5' untranslated region of the wunl gene will be protected by its homology to the mRNA.
  • the SI protected DNA strand is separated by electrophoresis on a sequencing gel, whereby the sequence of a known DNA fragment is used for length comparison.
  • the DNA cloned in E. coli is transferred to A. tumefaciens LBA4404 (Hoeke a et al., 1983) by the method described by Van Haute et al., (1983), by conjugation .
  • the Agrobacterium strains containing the appropriate LBA4404 vector required for infection are grown in selective antibiotic medium (Zambrisky et al. , 1983), sedimented by centrifugation and washed in YEB medium without antibiotics. After further sedimentation and absorption in 3MS medium, the bacteria are used for infection.
  • Sterile leaf pieces of about 1 cm 2 size of tobacco species SNN and W38 are immersed into the Agrobacterium suspension described above, with subsequent transfer onto 3MS medium. After incubation for 2 days at 16 hours light and 25°C to 27°C, the pieces are transferred onto MSC16 for callus and shoot induction. Shoots appearing after 4-6 weeks are cut off and incubated on MSC15 medium.
  • the DNA from the plants is isolated by a varied method of Murray and Thompson (1980), cut with restriction enzymes and separated on a gel. After transfer of this DNA onto nitrocellulose, it is hybridized with a radioactively labelled sample, which indicates the presence of this specific DNA in the genome of the plant under examination. Detection of GUS activity
  • Plants regenerated from transformation are examined using the method of Jefferson (1987) for the degree of activity of the enzyme ⁇ -glucuronidase (GUS).
  • GUS ⁇ -glucuronidase
  • An additional histochemical test indicates localisation in the tissue.
  • thin sections of the plants are incubated with the substrate X-Gluc, 5-bromo-4-chloro-3-indolyl glucuronide. A blue precipitate forms at the site of enzyme activity.
  • the wunl-cDNA equivalent fragment in wunl-85 has a size of 4 Kb. Thus, it corresponds exactly to the fragment size of EcoRI digested DNA from the haploid potato, which hybridizes with wunl-cDNA. Based on the asymmetrical Xhol cutting site in the 5'region of the cDNA clone, and the use of radioactive cDNA samples which only cover the 5' region of the cDNA clone, the orientation of the gene in the 4 Kb fragment could be determined. Accordingly, 5' of the wunl gene there is a promoter region of ca. 1 Kb size, while at 3' from the gene there is a non-homologous part of ca. 2 Kb (fig. 5).
  • the EcoRI fragment of 8 Kb size additionally contain in wunl-85 could not be referred to for closer analysis of the wunl gene. Based on the digestion of the potato DNA with EcoRI, it is probable that during the subsequent ligation, two fragments not belonging together were brought into the EMBL4 vector, that is, neither can they enter a functional relationship with one another.
  • the genomic analysis of the wunl coding gene is carried out. Since in the restriction behaviour, no differences are established between wunl-85 and wunl-22, the 4 Kb fragment of wunl-85 is ligated into EcoRI cleaved pUC8 for further analysis (fig. 1). In order to sequence the wunl promoter and also the wunl gene, further re-cloning of the 4 Kb fragment into the Eco RI cutting site of M13mpl8 took place. Determination of its orientation is carried out using control digestion at the asymmetrical Xhol cutting site. The clones 85 * mpl8 and 85mpl8 represent both orientations of the fragment (fig. 2).
  • the method of SI nuclease mapping is used.
  • the principle of this method is based on the fact that as a result of hybridization of single-stranded DNA from the 5' region of the wunl gene with wunl-mRNA, only the regions which because of their homology may form a double strand are protected from degradation of the single-strand- specific nuclease SI.
  • the size of the protected DNA fragment can be determined on a sequencing gel and thus the start of transcription can be followed back.
  • a 1.2 Kb fragment is isolated from pLSOOO, and it contains the promoter and parts of the 5' untranslated region of the wunl gene.
  • This fragment is radioactively labelled with 32 P with polynucleotide kinase at the 5' end of the Xho I site, and hybridized on 50 ug of total RNA from wounded potato tubers.
  • the size of the protected fragment is determined on a sequencing gel; it varies between 162-178 bp (fig. 3).
  • the start of transcription begins 178 bp upstream from the Xho I site, i.e. with the sequence ACC ⁇ TAC.
  • This sequence conforms in the central region (CAT) with the consensus sequence, determined by Joshi et al., 1987, for the start of transcription CTCATCA. Further information is provided from the position of the start of transcription (figs. 4, 5):
  • CAAT box described by Benoist et al., 1980, in the region between -60 and -80 can be found in the wunl promoter at position -58 (CAAACT).
  • the 5' untranslated region of the wunl gene extends over 217 bp and is therefore comparatively large.
  • the wunl-mRNA coding gene thus encloses 794 bp, which corresponds very exactly with the size determination of wunl-mRNA based on "Northern-Blot" analysis.
  • the wunl gene has no introns at its disposal.
  • the promoter -1022 wunl is recloned into vector pPR69 (Fig. 6).
  • the deletion fragments beyond the Xhol cutting site are cut out of M13mpl8.
  • the promoter -1022 wunl and the promoter fragment -571 wunl were cloned in reverse orientation in front of the GUS gene (1022r; 571r). This was done by filling up the 5' overlapping ends of the fragments with DNA polymerase I and recloning it in the reversed orientation into the vector. Correct integration was tested by hybridisation with wunl radioactively labelled probes and by restriction analysis.
  • the plasmids pLS034-1022, -1022r and -571r are transformed into the agrobacterial strain LBA4404. These agrobacteria are then used for transformation of the tobacco cultivar Wisconsin 38 (W38) by means of the leaf section infection method. Under kanamycin selection, the callus produced on the plant are regenerated and are examined on Southern Blot for correct integration of the pLS034-DNA into the plant genome.
  • the functional analysis of the transgenic tobacco plants is extended to the detection of GUS activity in greenhouse plants.
  • Non-transformed tobacco plants serve as a control (K).
  • the activity of the enzyme in various tissues of the plant is analysed by means of histochemical in-situ analysis. For this, thin sections of the tissue are incubated over night at 37°C with the substrate of the enzyme, 5-bromo-4-chloro-3-indolyl glucuronide (X-Gluc). A blue precipitate forms at the site of enzyme activity.
  • This analysis indicates that in plants of constructions pLS034, -571r and -1022r, no activity can be detected in leaves, stem or roots.
  • Plants carrying the full wunl promoter (-1022), in its natural orientation, in their construction show activity in all the said tissues.
  • Plants of construction pLS034-1022 show GUS activity in the stomium of the anthers and in the pollen grain. 30% of the pollen grain show blue colouring after incubation with X-Gluc. Anthers of pLS034-571r show the same pattern of activity. In control plants, as with plants of construction pLS034-1022r, no blue colouring is detected in the stomium, and colouring of the pollen grain is reduced to 0.75%.
  • Comparison of the measured GUS activity of anthers from the various constructions shows 2-10 times stronger activity of pLS034-1022 than in pLS034-571r.
  • the average enzyme activity of the full promoter in anthers reaches 1276 pmol MU/mg/min.
  • Construction pLS034-571r shows an average enzyme activity of 217 pmol MU/mg/min in the anthers.
  • the 35S promoter of cauliflower mosaic virus is active in anthers. Its activity is 500 pmol MU/mg/min. Since the 35S promoter of the CaMV is a strong promoter which is active in plants, the deletion fragments of the wunl promoter extend into this activity range throughout. Transformation of tobacco employing the rol C-gene
  • the purpose of the experiments was to obtain male sterility by targeting of rol C expression to anthers employing a -571r-rolC construct and a EN6-571r-rol C construct resp.
  • the -571r-rol C construct is the same construct as the -571r-GUS construct, except that the GUS has been replaced by the rol C-gene. Transformation with this construct results in male sterile plants.
  • the EN6-571r-rol C construct is a -571r-rol C construct having 5' thereof a fragment containing 6 times the 35 S enhancer (-96 to -420). Transformation with this construct results in male sterile plants.
  • Thionin is a small (5 Kd) protein which has been isolated from barley seeds and its activity of thionin is supposed to be similar to a detergent. Because of its bipolar character it is able to cause holes into membranes and therefore to destroy cells.
  • Thionin (5Kd) is synthesised as an inactive preprotein (15 Kd). It is flanked by a 3 Kd leader peptide at its N-terminus which is responsible for transport of the precursor protein to the endoplasmatic reticulum. At its C-terminus thionin is flanked by a 7 Kd acidic protein of unknown function.
  • thionin As a plant cell destroying agent the published sequencing data for hordothionin were used to synthesise an oligonucleotide coding for an active thionin.
  • Said "thionin” is translated as an active detergent directly into the cytoplasm employing a -571r-thionin construct.
  • wunl-gene is derived from potato plants, there is some cross homology with tobacco plants on Southern- and Northern- level. Anther-specific repression of wunl-gene products is sought employing the wunl-antisense constructs.

Landscapes

  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Engineering & Computer Science (AREA)
  • Medicinal Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Zoology (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Cell Biology (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Botany (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

L'invention décrit des promoteurs permettant à des gènes se trouvant en liaison opérationnelle avec lesdits promoteurs, d'être exprimés spécifiquement en anthères. En utilisant des promoteurs spécifiques d'anthères, il est entre autres possible d'obtenir une stérilité mâle.
PCT/EP1991/001786 1990-09-21 1991-09-19 Composes organiques ameliores WO1992005261A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US58662790A 1990-09-21 1990-09-21
US586,627 1990-09-21

Publications (1)

Publication Number Publication Date
WO1992005261A1 true WO1992005261A1 (fr) 1992-04-02

Family

ID=24346510

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP1991/001786 WO1992005261A1 (fr) 1990-09-21 1991-09-19 Composes organiques ameliores

Country Status (3)

Country Link
AU (1) AU8634791A (fr)
IL (1) IL99522A0 (fr)
WO (1) WO1992005261A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994023043A2 (fr) * 1993-03-31 1994-10-13 Nickerson Biocem Limited Regulation de l'abscission et de la dehiscence des gousses dans les vegetaux
US5470359A (en) * 1994-04-21 1995-11-28 Pioneer Hi-Bred Internation, Inc. Regulatory element conferring tapetum specificity
WO1996040925A2 (fr) * 1995-06-07 1996-12-19 Pioneer Hi-Bred International, Inc. Elements regulateurs conferant une specificite au tapetum

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2042447C (fr) * 1990-06-12 1999-09-28 Marc C. Albertsen Controle de la microsporogenese par sequences promotrices inductibles de l'exterieur

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990005187A1 (fr) * 1988-11-07 1990-05-17 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Sequence d'adn de solanum tuberosum provoquee par des lesions et son utilisation
EP0420819A1 (fr) * 1989-09-25 1991-04-03 Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. Améliorations concernant des compositions organiques

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990005187A1 (fr) * 1988-11-07 1990-05-17 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Sequence d'adn de solanum tuberosum provoquee par des lesions et son utilisation
EP0420819A1 (fr) * 1989-09-25 1991-04-03 Max-Planck-Gesellschaft zur Förderung der Wissenschaften e.V. Améliorations concernant des compositions organiques

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
J. CELL. BIOCHEM. SUPPL., Vol. 13D, 1989, Meeting 1-9 April 1989, Abstract No. M331, page 306, LOGEMANN J. et al., "5' Upstream Sequences from the Wun 1 Gene are Responsible for Gene-Activation by Wounding and Tissue Specific Expression in Transgenic Plants". *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1994023043A2 (fr) * 1993-03-31 1994-10-13 Nickerson Biocem Limited Regulation de l'abscission et de la dehiscence des gousses dans les vegetaux
WO1994023043A3 (fr) * 1993-03-31 1994-11-24 Nickerson Biocem Ltd Regulation de l'abscission et de la dehiscence des gousses dans les vegetaux
US5907081A (en) * 1993-03-31 1999-05-25 Biogemma Uk Limited Control of plant abscission and pod dehiscence
US5470359A (en) * 1994-04-21 1995-11-28 Pioneer Hi-Bred Internation, Inc. Regulatory element conferring tapetum specificity
US5837850A (en) * 1994-04-21 1998-11-17 Pioneer Hi-Bred International, Inc. Regulatory element conferring tapetum specificity
WO1996040925A2 (fr) * 1995-06-07 1996-12-19 Pioneer Hi-Bred International, Inc. Elements regulateurs conferant une specificite au tapetum
WO1996040925A3 (fr) * 1995-06-07 1997-05-01 Pioneer Hi Bred Int Elements regulateurs conferant une specificite au tapetum

Also Published As

Publication number Publication date
IL99522A0 (en) 1992-08-18
AU8634791A (en) 1992-04-15

Similar Documents

Publication Publication Date Title
US6329570B1 (en) Cotton modification using ovary-tissue transcriptional factors
US7667097B2 (en) Rice pollen-preferential promoters and uses thereof
Yokoi et al. Tapetum-specific expression of the Osg6B promoter-β-glucuronidase gene in transgenic rice
US20050144667A1 (en) Plant polypeptides and polynucleotides encoding same
AU1681992A (en) Chimeric plant genes based on upstream regulatory elements of helianthinin
Twell et al. The 5′ flanking DNA of a patatin gene directs tuber specific expression of a chimaeric gene in potato
US5847102A (en) Cold induced promoter from winter Brassica napus
US7276370B2 (en) Rf2a and rf2b transcription factors
Sharada et al. Generation of genetically stable transformants by Agrobacterium using tomato floral buds
US5428146A (en) Wound-stimulated DNA-sequence from solanum tuberosum and its use
US20130091603A1 (en) Reproductive ablation constructs
AP181A (en) A DNA sequence, anther-specific promoter and the use thereof.
WO2002078438A2 (fr) Promoteur du mais prefere des tissus
WO1999043818A1 (fr) Nouveaux fragments d'adn regissant l'expression genique predominante dans les organes des fleurs
US20030233671A1 (en) Promoters for molecular farming and/or early detection of transgenic plants
WO1992005261A1 (fr) Composes organiques ameliores
AU6330496A (en) Use of ovary-tissue transcriptional factors
AU641373B2 (en) Wound-induced DNA sequence from solanum tuberosum and its use
US7557264B2 (en) Gossypium hirsutum tissue-specific promoters and their use
KR20060013382A (ko) 세포질 웅성 불임 고추에 특이 ｄｎａ 단편 및 이의 용도
WO2006080598A9 (fr) Genes specifiques des antheres derives de malus x domestica, promoteurs de ces genes, et procede permettant de preparer une plante transformee au moyen de ceux-ci
JP2005168470A (ja) 花粉特異的発現活性を有するプロモーター
JP2004534535A (ja) 新規の植物プロモーター
AU5187000A (en) Use of ovary-tissue transcriptional factors
AU5350900A (en) Ovary-tissue transcriptional factors

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU BR CA HU JP KR

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IT LU NL SE

NENP Non-entry into the national phase

Ref country code: CA

122 Ep: pct application non-entry in european phase
122 Ep: pct application non-entry in european phase