EP0789773A1 - Procede permettant de modifier la floraison de plantes - Google Patents

Procede permettant de modifier la floraison de plantes

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Publication number
EP0789773A1
EP0789773A1 EP95938378A EP95938378A EP0789773A1 EP 0789773 A1 EP0789773 A1 EP 0789773A1 EP 95938378 A EP95938378 A EP 95938378A EP 95938378 A EP95938378 A EP 95938378A EP 0789773 A1 EP0789773 A1 EP 0789773A1
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EP
European Patent Office
Prior art keywords
plants
sucrose transporter
flowering
plant
dna
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Withdrawn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
EP95938378A
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German (de)
English (en)
Inventor
Georg Leggewie
Jörg Riesmeier
Wolf-Bernd Frommer
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Bayer CropScience AG
Original Assignee
Hoechst Schering Agrevo GmbH
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Publication of EP0789773A1 publication Critical patent/EP0789773A1/fr
Withdrawn legal-status Critical Current

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    • 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)
    • 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 present invention relates to processes for the production of plants with a different flowering behavior compared to wild-type plants, in particular premature flowering and increased flowering, and to the plants obtainable from the process. Such plants are produced by increasing the sucrose transporter activity in the plants.
  • the invention further relates to the use of DNA molecules which encode sucrose transporters to change the flowering behavior in plants.
  • flowers in plants are a prerequisite for sexual reproduction and is therefore essential for the multiplication of plants which cannot be propagated vegetatively, and for the formation of seeds and fruits.
  • the point in time at which plants pass from purely vegetative growth to flower formation is of central importance, for example in agriculture, gardening and plant breeding.
  • the number of flowers is often of economic interest, e.g. in the case of various useful plants (tomato, cucumber, zucchini, cotton, etc.) in which a higher number of flowers may mean a higher yield, or in the production of ornamental plants and cut flowers.
  • early flowering of the plants is advantageous.
  • early flowering of various crop plants would allow a shortening of the time between sowing and harvesting and thus the spreading of two sowings per year or an extension of the period between flowering and harvesting, which may result in an increase in yield .
  • Can also be used in plant breeding premature flower formation contributes to a considerable shortening of the breeding process and thus brings about an improvement in economy.
  • the economic benefits of early flowering are also evident for horticulture and ornamental plant production.
  • the efforts to date to elucidate the mechanisms which determine the time of flower formation in plants do not allow a clear conclusion to be drawn about the factors involved and the decisive factors.
  • This process therefore does not appear to be suitable for producing useful plants which have changed their flowering behavior.
  • the present invention is therefore based on the object of providing simple methods which make it possible to modify plants to the extent that their flowering behavior is changed, in particular in that they show premature flower formation and / or an increased flower set .
  • the invention therefore relates to the use of DNA molecules which encode proteins with the biological activity of a sucrose transporter to change the flowering behavior in plants.
  • sucrose transporters A function of sucrose transporters in regulating the flowering behavior had not previously been considered.
  • Sucrose has been discussed several times as a potential signal for flower induction in the apical meristem (Bernier et al., Plant Cell 5 (1993), 1147-1155; Lejeune et al., Planta 190 (1993), 71-74; Lejeune et al ., Plant Physiol. Biochem. 29 (1991), 153-157), however, an influence of a sucrose transporter on the flowering behavior as a result of an increased activity of this transporter has so far not been known and was also not to be expected for various reasons.
  • an increased activity of the sucrose transporter is understood to mean that the total sucrose transporter activity in the transgenic plants is increased, in particular by at least 30%, preferably by at least 50%, particularly preferably by, in comparison with non-transformed plants at least 100%, and in particular by at least 200%.
  • Sucrose transporters are understood to be proteins that are able to transport sucrose across biological membranes. The activity of such transporters can be according to the Riesmeier et al. (EMBO J. 11 (1992), 4705-4713) determine the method described.
  • a changed flowering behavior is understood in the context of this application that premature flowering and flowering take place in transformed plants compared to non-transformed plants a), prematurely meaning that the transformed plants compared to wild-type plants at least a few days , preferably form and bloom flowers one to several weeks, in particular 1-2 weeks earlier, and / or b) show an increased flower set, which means that, compared to wild-type plants, the transformed plants produce on average more flowers per plant, generally use at least 5% more flowers, in particular 10-100% and preferably 10-40% more flowers.
  • sucrose transporter activity compared to wild-type plants can be achieved by introducing DNA molecules into plants which encode a sucrose transporter. This leads to additional synthesis of proteins with sucrose in the transgenic cells sea transporter activity. As a result of this, transformed tissues in which the introduced DNA molecule is expressed have an increased sucrose transporter activity compared to non-transformed cells.
  • the coding region of a DNA sequence which codes for a sucrose transporter is preferably linked to DNA sequences which are required for transcription in plant cells, and in Plant cells introduced.
  • the regulatory sequences required for the transcription are, on the one hand, promoters and possibly enhancer elements which are responsible for the initiation of the transcription.
  • termination signals which lead to the termination of the transcription and to the addition of a poly-A tail to the resulting transcript, can optionally be added.
  • sequences are linked to one another in such a way that the coding region of a sucrose transporter gene is connected in sense orientation to the 3 'end of the promoter, so that an mRNA is synthesized which converts into a protein with the activity of a sucrose transporter can be translated, and the termination signal is connected to the 3 'end of the coding region.
  • the coding region can be linked to sequences which increase translation in plant cells, as described in the examples.
  • the DNA molecules which code for a sucrose transporter can originate from any organism which contains such sequences, in particular from any prokaryotic or eukaryotic organism.
  • the DNA molecules come from plants, fungi or bacteria. In plants, higher plants, in particular monocotyledonous or dicotyledonous plants, are preferred.
  • DNA molecules that are saccharo- Coding transporters are already known from various organisms and are given below. These are preferably used in the context of the invention.
  • the invention also relates to a method for changing the flowering behavior in plants, in which the changed flowering behavior is brought about by increasing the activity of the sucrose transporter in plants.
  • transgenic plants which have a different flowering behavior compared to non-transformed plants, in particular a premature flower formation and / or an increased flower set, is preferably carried out by a process which comprises the following steps: a) producing an expression cassette which comprises the following DNA sequences: i) a promoter which is functional in plant cells and which ensures the transcription of a subsequent DNA sequence, ii) at least one DNA sequence which codes a sucrose transporter and is sent to the 3 'in sense orientation End of the promoter is coupled, and iii) optionally a termination signal for the termination of the transcription and the addition of a poly-A tail to the resulting transcript which is coupled to the 3 'end of the coding region, b) transformation plant cells with the expression cassette produced in step a) and stable integration of the expressi on cassette in the plant genome, and c) regeneration of whole, intact plants from the transformed plant cells.
  • an expression cassette which comprises the following DNA sequences: i) a promoter which is functional in plant cells and which ensures the transcription
  • any promoter which is functional in plants is suitable for the promoter mentioned under i).
  • the 35S promoter of the Cauliflower mosaic virus (Odell et al., Nature 313 (1985), 810-812) is suitable, which ensures constitutive expression in all tissues of a plant and that in WO / 9401571 described promoter con structure.
  • promoters can also be used which lead to an expression of subsequent sequences only at a point in time determined by external influences (see, for example, WO / 9307279) or in a certain tissue of the plant (see, for example, Hadash et al., Plant Cell 4 (1992), 149-159, Stockhaus et al., EMBO J. 8 (1989), 2245-2251).
  • the DNA molecules which comprise a coding region for a sucrose transporter, can be of either native or homologous origin as well as foreign or heterologous origin with respect to the plant species to be transformed. Both DNA molecules derived from prokaryotic organisms and those derived from eukaryotic organisms, in particular plants, can be used. Prokaryotic sequences are known, for example, from E. coli (Bookman et al., Mol. Gen. Genet. 235 (1992), 22-32; EMBL library: accession number X63740). DNA molecules which encode vegetable sucrose transporters are preferably used.
  • RNA or DNA sequences from Arabidopsis thaliana (suc 1 and suc 2 genes; EMBL library: accession numbers X75365 and X75382, as well as H36128, H36415, R64756, T76707 and T42333), Solanum tuberosum (Riesmeier et al ., Plant Cell 5 (1993), 1591-1598; EMBL genebank: accession numbers X69165 and WO 94/00547), Plantago major (EMBL genebank: accession numbers X75764 and X84379), L.
  • EMBL genebank access ⁇ number X82275
  • Nicotiana tabacum EMBL gene bank: access numbers X82276 and X82277
  • R. co munis EMBL gene bank: access number Z31561
  • B. vulgaris EMBL gene bank: accession number X83850
  • rice EMBL library: accession numbers D40522 and D40515), which encode sucrose transporters.
  • a particular embodiment of the present invention provides for the use of a DNA molecule from Spinacia oleracea which encodes a sucrose transporter (see also Riesmeier et al., EMBO J. 11 (1992), 4705-4713 and WO 94/00547).
  • a further preferred embodiment of the method according to the invention provides that DNA molecules are used which encode sucrose transporters with the lowest possible I ⁇ value.
  • a transporter activity is known, for example, from Candida albicans (Williamson et al., Biochem. J. 291 (1993), 765-771).
  • the molecules which encode sucrose transporters can be cDNA molecules as well as genomic sequences.
  • the DNA molecules can be isolated from the corresponding organisms using the common techniques known to the person skilled in the art, for example hybridization or polymerase chain reaction, or they can be prepared synthetically.
  • termination signals for transcription in plant cells mentioned under iii) are described and can be interchanged as desired.
  • the termination sequence of the nopaline synthase gene from AgroJbac ⁇ eriLim tumefaciens can be used (see e.g. Gielen et al., EMBO J. 8 (1989), 23-29).
  • the expression cassette described can also contain DNA sequences which enhance the translation of the coding region in plant cells.
  • the method according to the invention can in principle be applied to any flower-forming plant species. It is preferably applied to the plants specified below.
  • the invention further relates to transgenic plants which, owing to the increased activity of the sucrose transporter, have a different flowering behavior compared to wild-type plants, in particular premature flowering and flowering and / or an increased flower set.
  • transgenic plants are preferably obtainable by the process described above. This means that the increase in the sucrose transport Interactivity preferably on the fact that DNA molecules which encode a saccharose transporter are introduced and expressed in the plants. These are preferably DNA molecules that come from plants, fungi or bacteria.
  • the plants according to the invention are preferably monocotyledonous or dicotyledonous crops, for example cereals (such as barley, oats, rye, wheat etc.), maize, rice, vegetable plants (such as tomato, melon, zucchini etc.) , Cotton, rapeseed, soybean, types of fruit (such as plum, apple, pear etc.), ornamental plants or cut flowers.
  • cereals such as barley, oats, rye, wheat etc.
  • maize such as tomato, melon, zucchini etc.
  • vegetable plants such as tomato, melon, zucchini etc.
  • Cotton rapeseed
  • soybean types of fruit (such as plum, apple, pear etc.), ornamental plants or cut flowers.
  • cloning vectors which contain a replication signal for E. coli and a marker gene for the selection of transformed bacterial cells.
  • examples of such vectors are pBR322, pUC series, Ml3mp series, pACYC184 etc.
  • the desired sequence can be introduced into the vector at a suitable restriction site.
  • the plasmid obtained is used for the transformation of E. coli cells.
  • Transformed E. coli cells are grown in a suitable medium, then harvested and lysed.
  • the plasmid is recovered. Restriction analyzes, gel electrophoresis and other biochemical-molecular biological methods are generally used as the analysis method for characterizing the plasmid DNA obtained.
  • the plasmid DNA can be cleaved and linked to other DNA sequences.
  • Each plasmid DNA sequence can be cloned in the same or other plasmids.
  • Plasmids are preferably used to transform plant cells using the expression cassette described in the method.
  • a variety of techniques are available for introducing DNA into a plant host cell. These techniques include the transformation of plant cells with T-DNA using Agrobacterium tumefaciens or Agrobacterium rhizogenes as transformation agent, the fusion of protoplasts, the injection, the electroporation of DNA, the introduction of DNA using the biolistic method and other possibilities.
  • plasmids When injecting and electroporation of DNA into plant cells, no special requirements are made of the plasmids used. Simple plasmids such as e.g. pUC derivatives can be used. However, if whole plants are to be regenerated from such transformed cells, the presence of a selectable marker gene is necessary.
  • the Ti or Ri plasmid is used for the transformation of the plant cell, at least the right boundary, but often the right and left boundary of the Ti and Ri plasmid T-DNA as the flank region, must be linked to the genes to be introduced .
  • the DNA to be introduced must be cloned into special plasmids, either in an intermediate vector or in a binary vector. Because of sequences which are homologous to sequences in the T-DNA, the intermediate vectors can be integrated into the Ti or Ri plasmid of the agrobacteria by homologous recombination. This also contains the vir region necessary for the transfer of the T-DNA. Intermediate vectors cannot replicate in agrobacteria. The intermediate vector can be transferred to Agrobacterium tumefaciens by means of a helper plasmid (conjugation). Binary vectors can replicate in E. coli as well as in Agrobacteria.
  • the agrobacterium serving as the host cell is said to be a plasmid which carries a vir region.
  • the vir region is necessary for the transfer of the T-DNA into the plant cell. Additional T-DNA can be present.
  • the agrobacterium transformed in this way is used to transform plant cells.
  • T-DNA for the transformation of plant cells has been intensively investigated and is sufficient in EP 120516; Hoekema, In: The Binary Plant Vector System Offset- drukkerij Kanters B.V., Alblasserdam, Chapter V; Fraley et al., Crit. Rev. Plant. Sci., 4 (1985), 1-46 and An et al., EMBO J. 4 (1985), 277-287.
  • plant explants can expediently be cultivated with Agrobacterium tumefaciens or Agrobacterium rhizogenes.
  • Whole plants can then be regenerated from the infected plant material (e.g. leaf pieces, stem segments, roots, but also protoplasts or suspension-cultivated plant cells) in a suitable medium, which can contain antibiotics or biocides for the selection of transformed cells.
  • the plants obtained in this way can then be examined for the presence of the introduced DNA.
  • the introduced DNA is integrated in the genome of the plant cell, it is generally stable there and is also retained in the progeny of the originally transformed cell. It normally contains a selection marker which gives the transformed plant cells resistance to a biocide or an antibiotic such as kanamycin, G 418, bleomycin, hygroycin or phosphinotricin and others. mediated.
  • the individually chosen marker should therefore allow the selection of transformed cells from cells that lack the inserted DNA.
  • the transformed cells grow within the plant in the usual way (see also McCormick et al., Plant '' Cell Reports 5 (1986), 8-1-84).
  • the resulting plants can are normally grown and crossed with plants that have the same transformed genetic makeup or other genetic makeup.
  • the resulting hybrid individuals have the corresponding phenotypic properties. Two or more generations should be grown to ensure that the phenotypic trait is stably maintained and inherited. Seeds should also be harvested to ensure that the appropriate phenotype or other characteristics have been preserved.
  • Fig. 1 shows the plasmid p ⁇ A7DE-S21-Myc8.
  • A Fragment A: CaMV 35S promoter, nt 6909-7437 (Franck et al., 1980, Cell 21, 285-294). In the 5 'region of the promoter, two 35S enhancer elements (330 bp HincII / EcoRV fragment) were inserted into the Nco I interface.
  • fragment C DNA fragment approx. 1600 bp long, which encodes nucleotides 70 to 1644 of the cDNA encoding the sucrose transporter from spinach (Riesmeier et al., EMBO J. 11 (1992), 4705-4713), includes
  • fragment D fragment D: 33 bp long DNA fragment which encodes the amino acid sequence EQKLISEEDLN-COOH
  • E fragment E: termination sequence of the octopine synthase gene; nt 11748-11939 the T-DNA of the Ti plasmid pTiACH5 (Gielen et al., EMBO J. 3 (1984), 835-846)
  • Fragments A, B, C, D and E are in the vector pUC18.
  • the plasmid has a size of approximately 5700 bp.
  • Fig. 2 shows the plasmid p ⁇ -S21.
  • A Fragment A: CaMV 35S promoter, nt 6909-7437 (Franck et al., Cell 21 (1980) 285-294). In the 5 'region of the promoter, two 35S enhancer elements (330 bp HincII / EcoRV fragment) were inserted into the Nco I interface.
  • B fragment B: 73 bp Nco I / Asp 718 fragment (TMV-Ul) with the translation enhancer from the tobacco mosaic virus
  • fragment C DNA fragment approx. 1600 bp long, which encodes nucleotides 70 to 1644 of the cDNA encoding the sucrose transporter from spinach (Riesmeier et al., EMBO J. 11 (1992), 4705-4713), includes
  • fragment D fragment D: 33 bp long DNA fragment which encodes the amino acid sequence EQKLISEEDLN-COOH
  • E fragment E: termination sequence of the octopine synthase gene; nt 11748-11939 the T-DNA of the Ti plasmid pTiACH5 (Gielen et al., EMBO J. 3 (1984), 835-846)
  • the plasmid has a size of approximately 12.6 kb
  • Fig. 3 shows the plasmid p35 S- ⁇ -OCS
  • 5 shows as a bar graph the average number of days between the transfer of the plants from the tissue culture into soil until the opening of the first flower. In each case 12 plants per genotype were grown in a plant growth chamber under the following light conditions. 7- 9 a.m. 300 ⁇ mol quantum m 2 sec -1
  • FIG. 6 shows as a bar chart the average number of days between the transfer of the plants from the tissue culture into soil until the opening of the first flower. In each case 12 plants per genotype were grown in a plant growth chamber under the following light conditions. Lines 32, 12, 5 and 1 are 4 independent transgenic lines which had been transformed with the plasmid p ⁇ -S21. 7 am-11pm 400-500 ⁇ mol quantum m "2 sec _1
  • Fig. 7 shows a bar chart of the average number of leaf attachments until the flowers are formed. For experimental conditions, see FIG. 6.
  • NSEB buffer 0.25 M sodium phosphate buffer pH 7.2
  • the vector pUCl ⁇ was used for cloning in E. coli.
  • the gene constructions were cloned into the binary vector pBinAR (Höfgen and Willmitzer, Plant Sei. 66 (1990), 221-230).
  • E.coli strain DH5 ⁇ (Bethesda Research Laboratories, Gaithersburgh, USA) was used for the pUC vectors and for the pBinAR constructs.
  • the transformation of the plasmids into the tobacco plants was carried out using the Agrobacterium tumefaciens strain C58C1 pGV2260 (Deblaere et al., Nucl. Acids Res. 13 (1985), 4777-4788).
  • the DNA was transferred by direct transformation according to the method of Höfgen & Willmitzer (Nucleic Acids Res. 16 (1988), 9877).
  • the plasmid DNA of transformed agrobacteria was isolated according to the method of Birnboim & Doly (Nucleic Acids Res. 7 (1979), 1513-1523) and analyzed by gel electrophoresis after suitable restriction cleavage. 4. Transformation of tobacco
  • the leaf pieces for shoot induction on MS medium (0.7% agar) with 1.6% glucose, 1 mg / 1 6-benzylaminopurine, 0.2 mg / 1 naphthylacetic acid, 500 mg / 1 claforan and 50 mg / 1 kanamycin.
  • the medium was changed every 7 to 10 days.
  • the leaf pieces were transferred to glass jars containing the same medium. Resulting shoots were cut off and placed on MS medium + 2% sucrose + 250 mg / 1 claforan and whole plants were regenerated from them.
  • the radiocative labeling of DNA fragments was carried out using a DNA random primer labeling kit from Boehringer (Germany) according to the manufacturer's instructions.
  • sucrose transporter from spinach in the extracts of transgenic tobacco plants was identified using a "blotting detection kit for rabbit antibodies" (Amersham UK) according to the manufacturer's instructions.
  • the monoclonal antibody from mouse 9E10 (Kolodziej and Young, In: Methods in Enzy ology 194 (1991), 508-519), which is directed against the myc epitope shown as fragment D in FIG. 1, was used as the primary antibody .
  • telomere sequence With the help of PCR technology an approximately 1600 bp long DNA fragment was amplified, which contained nucleotides 70 to 1644 of the Riesmeier et al. (EMBO J. 11 (1992), 4705-4713) comprises the sequence of the clone pS21.
  • the oligonucleotide (1) introduced a Pst I and a Noc I interface at the 5 'end of the coding region.
  • the coding region was extended by the oligonucleotide (2) by an 11 amino acid long sequence (EQKLISEEDLN-COOH (Seq ID No. 3)) at the C-terminus and a Pst I site was also introduced.
  • This sequence originates from the c-Myc gene and represents the recognition epitope for the monoclonal antibody from mouse 9E10 (Kolodziej and Young, In: Methods in Enzymology 194 (1991), 508-519; commercially available from Dianova, Hamburg).
  • the resulting PCR product was cut with the restriction enduclease Pst I and ligated into a pUCl ⁇ vector cut with Pst I.
  • the resulting plasmid was named p-S21-Myc8.
  • An approximately 1600 bp fragment containing the PCR product was isolated from this by Nco I / Pst I partial digestion and ligated into the vector p35SDE- ⁇ -OCS cut with Nco I and Pst I.
  • the vector p35SDE- ⁇ -OCS was produced as follows: A 530 bp EcoR I / Asp718 fragment (nucleotides 6909-7439 (Franck et al.
  • the resulting plasmid was named p35SDE.
  • the 35S promoter contained in this fragment with two additional Hinc II / EcoR V fragments was designated 35SDE.
  • Another pUC plasmid was constructed, which is constructed as shown in FIG. 3. The following DNA fragments were inserted between the EcoR I and the Hind HI sites of the polylinker of a pUC18 vector:
  • This plasmid was called p35S- ⁇ -OCS.
  • the plasmid p35S- ⁇ -OCS was cut with EcoR I / Asp718, whereby the 35S promoter was removed. This was replaced by the promoter 35SDE isolated with EcoR I and Asp718 from the plasmid p35SDE. The resulting plasmid was named p35SDE- ⁇ -OCS.
  • This plasmid was cut with Nco I and Pst I in the polylinker.
  • the approximately 1600 bp fragment which contains the PCR product described above and was isolated from the plasmid p-S21-Myc8 by partial digestion with Nco I and Pst I was ligated into the interfaces. This resulted in the plasmid p ⁇ A7DE-S21-Myc8.
  • This plasmid is shown in Figure 1.
  • Plasmid p ⁇ A7DE-S21-Myc8 was digested with EcoR I and Hind III and the entire expression cassette comprising the promoter 35SDE, the translation enhancer, the region coding for the saccharose transporter from spinach, with the sequence containing the c-Myc epitope coded, and the termination sequence isolated.
  • the plasmid p ⁇ -S21 was used for the transformation of tobacco plants with the aid of the agrobacterium-mediated gene transfer.
  • RNA was isolated from leaf tissue of transgenic and non-transformed plants. 50 ⁇ g of this RNA were separated on an agarose gel, transferred to a nylon membrane and hybridized with the radioactively labeled cDNA, which encodes the sucrose transporter from spinach.
  • Such a Northern blot analysis showed that out of three transformants (transformants No. 5, 12 and 32) two transformants (No. 12 and No. 32) have a high expression of the sucrose transporter from spinach, one In comparison, transformants (No. 5) showed only a relatively low expression of the sucrose transporter from spinach, and no transcripts could be found in non-transformed potato plants which encoded the sucrose transporter from spinach.
  • the tobacco plants transformed with the plasmid p ⁇ -S21 had a different flowering behavior compared to non-transformed tobacco plants.
  • transformants 12 and 32 which showed a strong expression of the spinach-sucrose transporter, premature flower formation and a slightly increased flower set were observed.
  • Table I shows how many leaf batches, approximately 128-day-old transformed or non-transformed tobacco plants which were kept in the phytotron, had on average before the apical meristem was differentiated into inflorescences.
  • FIGS. 4a and b show transformed tobacco plants of line 12 (FIG. 4a) and line 32 (FIG. 4b) kept in the phytron in comparison to non-transformed tobacco plants. Under the same cultivation conditions, the flowering and flowering of untransformed tobacco plants started significantly later, on average approx. 14 days for plants kept in the phytotron.
  • the transformed plants In addition to the premature flower formation, the transformed plants have more flowers in comparison to wild-type plants. This is shown in Table II.
  • GAGACTGCAG TCAGTTGAGG TCTTCTTCGG AGATTAGTTT TTGTTCATGA CCACCCATGG 60

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Abstract

L'invention concerne des procédés qui permettent de produire des plantes ayant une floraison modifiée par rapport au type sauvage, notamment une floraison précoce et une floraison plus abondante, ainsi que les plantes ainsi obtenues. L'invention concerne en outre l'utilisation de molécules d'ADN codant des transporteurs de saccharose pour modifier la floraison de plantes.
EP95938378A 1994-10-31 1995-10-30 Procede permettant de modifier la floraison de plantes Withdrawn EP0789773A1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
DE4439748A DE4439748A1 (de) 1994-10-31 1994-10-31 Verfahren zur Veränderung des Blühverhaltens bei Pflanzen
DE4439748 1994-10-31
PCT/EP1995/004257 WO1996013595A1 (fr) 1994-10-31 1995-10-30 Procede permettant de modifier la floraison de plantes

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EP0789773A1 true EP0789773A1 (fr) 1997-08-20

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US (1) US6025544A (fr)
EP (1) EP0789773A1 (fr)
JP (1) JPH10507920A (fr)
AU (1) AU701718B2 (fr)
CA (1) CA2204023A1 (fr)
DE (1) DE4439748A1 (fr)
HU (1) HU221005B1 (fr)
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WO1997025433A1 (fr) * 1996-01-09 1997-07-17 Eidg. Technische Hochschule Zürich Ethz Controle de la floraison de plantes
WO1999053068A2 (fr) 1998-04-09 1999-10-21 E.I. Du Pont De Nemours And Company Transporteurs de sucrose d'origine vegetale
DE19857654A1 (de) 1998-12-14 2000-06-15 Max Planck Gesellschaft Beeinflussung des Blühverhaltens von Pflanzen durch Expression Saccharose-spaltender Proteine
EP1268831A2 (fr) * 2000-03-31 2003-01-02 Institut Für Pflanzengenetik Und Kulturpflanzenforschung Procede de production de legumineuses a teneur en proteines et duree de remplissage du grain superieures
JP2002153283A (ja) * 2000-11-24 2002-05-28 National Institute Of Agrobiological Sciences 植物の開花を誘導する遺伝子Ｈｄ３ａおよびその利用
CN103880935B (zh) * 2012-12-19 2017-02-08 中国科学院植物研究所 蔗糖转运蛋白AtSUT2在培育高产转基因植物中的应用
CN105624171B (zh) * 2015-06-08 2020-12-15 南京农业大学 梨蔗糖转运蛋白基因PbSUT2及其应用

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JPH10507920A (ja) 1998-08-04
US6025544A (en) 2000-02-15
DE4439748A1 (de) 1996-05-02
HU221005B1 (hu) 2002-07-29
CA2204023A1 (fr) 1996-05-09
WO1996013595A1 (fr) 1996-05-09
AU3979495A (en) 1996-05-23
AU701718B2 (en) 1999-02-04
HUT77471A (hu) 1998-05-28

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