EP0528826A1 - Adn, constructions d'adn, et cellules et plantes ainsi derivees - Google Patents

Adn, constructions d'adn, et cellules et plantes ainsi derivees

Info

Publication number
EP0528826A1
EP0528826A1 EP91908064A EP91908064A EP0528826A1 EP 0528826 A1 EP0528826 A1 EP 0528826A1 EP 91908064 A EP91908064 A EP 91908064A EP 91908064 A EP91908064 A EP 91908064A EP 0528826 A1 EP0528826 A1 EP 0528826A1
Authority
EP
European Patent Office
Prior art keywords
dna
cellulase
plants
plant
tomato
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Withdrawn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
EP91908064A
Other languages
German (de)
English (en)
Inventor
Colin Roger Bird
John Anthony 30 Sylvanus Wooden Hill Ray
Wolfgang Walter 14 Greenfinch Close Schuch
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.)
Syngenta Ltd
Original Assignee
Zeneca Ltd
Imperial Chemical Industries Ltd
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 Zeneca Ltd, Imperial Chemical Industries Ltd filed Critical Zeneca Ltd
Publication of EP0528826A1 publication Critical patent/EP0528826A1/fr
Withdrawn legal-status Critical Current

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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
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C12N15/1137—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against enzymes
    • 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
    • C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
    • C12N15/8243—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
    • C12N15/8249—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine involving ethylene biosynthesis, senescence or fruit development, e.g. modified tomato ripening, cut flower shelf-life
    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2405—Glucanases
    • C12N9/2434—Glucanases acting on beta-1,4-glucosidic bonds
    • C12N9/2437—Cellulases (3.2.1.4; 3.2.1.74; 3.2.1.91; 3.2.1.150)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
    • C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
    • C12Y302/01004—Cellulase (3.2.1.4), i.e. endo-1,4-beta-glucanase

Definitions

  • This application relates to novel DNA constructs, plant cells containing them and plants derived therefrom.
  • it involves the use of recombinant DNA technology to control gene expression in plants.
  • a cell manufactures protein by transcribing the DNA of the gene for that protein to produce messenger RNA (mRNA), which is then processed (eg by the removal of introns) and finally translated by ribosomes into protein.
  • mRNA messenger RNA
  • antisense RNA an RNA sequence which is complementary to a sequence of bases in the mRNA in question: complementary in the sense that each base (or the majority of bases) in the antisense sequence (read in the 3' to 5' sense) is capable of pairing with the corresponding base (G with C, A with U) in the mRNA sequence read in the 5' to 3' sense.
  • RNA Ribonucleic acid
  • RNA Ribonucleic acid
  • Such antisense RNA may be produced in the cell by transformation with an appropriate DNA construct arranged to transcribe backwards part of the coding strand (as opposed to the template strand) of the relevant gene (or of a DNA sequence showing substantial homology therewith) .
  • the use of this technology to downregulate the expression of specific plant genes has been described, in for example European Patent publication no 271988 to ICI (corresponding to US serial 119614).
  • the cell walls of tomato and melon fruit predominantly consist of polysaccharides which have been sub-divided into cellulose, hemicellulose and pectin fractions. During fruit ripening there are considerable changes in the composition of the cell walls. In tomato pericarp tissue the proportion of cellulose in cell wall fractions increases slightly (Huber, Horticultural Science ⁇ 2fJ, 442-443, 1985).
  • the absolute level of cellulose may decline during ripening.
  • the proportion of cellulose in the cell wall fractions decreases during ripening.
  • Increases in cellulase activity have been correlated with avocado fruit ripening (Awad and Young, Plant Physiology ⁇ 4, 306-308, 1979).
  • the role of cellulase in tomato and melon fruit softening is unclear. There have been several reports of cellulase activity in tomato fruit (eg Hall, Nature 200 1010-1011,1963 and Hobson, Journal of food science 33, 588-592, 1968). Cellulase activity has been detected in mature green tomato fruit at the onset of ripening.
  • cellulase activity increases approximately ten fold as ripening progresses (Poovaiah and Nukaya, Plant Physiology 64 , 534-537, 1979 and Huber, 1985 cited above). Cellulase activity is higher in the locular gel than in the pericarp and also increases during ripening. It has been suggested that cellulase activity may particularly be an important feature of gel formation.
  • DNA constructs comprising a transcriptional initiation region operative in plants positioned for transcription of a DNA sequence homologous to some or all of a gene encoding cellulase or a like enzyme, said gene showing substantial homology to DNA encoded by either of the constructs TCELB6 and MCELE2.
  • the present invention provides such DNA constructs comprising a transcriptional initiation region operative in plants positioned for transcription of a DNA sequence encoding RNA complementary to a substantial run of bases showing substantial homology to an mRNA encoding cellulase or a like enzyme.
  • the invention also includes plant cells containing constructs of the invention; transformed plants derived therefrom showing modified ripening characteristics; and fruit and seeds of such plants
  • constructs of the invention may be inserted into plants to regulate the production of cellulase or like enzymes.
  • the production of the enzyme may be increased, or reduced, either throughout or at particular stages in the life of the plant.
  • production of the enzyme is enhanced only by constructs which contain DNA homologous to the substantially complete gene. What is more surprising is that constructs containing an incomplete DNA sequence substantially shorter than that corresponding to the complete gene generally inhibit the expression of the enzyme, whether they are arranged to express sense or antisense RNA.
  • the genes used in the invention typically derive from DNA from tomato or melon cellulase genes; or from DNA which is fully or partly homologous thereto.
  • This invention may be put into effect using the clones TCELB6 or MCELE2 that have been deposited.
  • such clones may be used as probes to identify other homologous cellulase or like genes (or parts thereof) in plant DNA, thereby obtaining DNA in longer lengths or having variant sequences. It is possible to screen
  • EET in this way DNA derived from tomato or melon, or from other plant material known to obtain cellulase genes.
  • the plants to which the present invention can be applied include commercially important fruit-bearing plants, in particular tomato and melon.
  • plants can be generated which express RNA from recombinant DNA and which may have one or more of the following characteristics:
  • DNA constructs according to the invention preferably comprise a base sequence at least 20 bases in length for transcription into RNA. If enhancement of expression of the enzyme is the objective, then substantially the whole of the gene sequence should be included in the construct. For inhibition by sense RNA, a shorter sequence is used. Where antisense RNA is used for inhibition, there is no theoretical upper limit to the length of the base sequence - it may be as long as the relevant mRNA produced by the cell - but for convenience it will generally be found suitable to use sequences of at least 50 and preferably between 100 and 1000 bases in length. The preparation of such constructs is described in more detail below.
  • TCELB6 tomato cellulase
  • MCELE2 melon cellulase
  • the base sequences of TCELB6 and MCELE2 are set out in Figure 1. Searches in DNA and protein data bases indicate that these clones show homology to clones for avocado fruit cellulase (Tucker et al , Plant Molecular Biology 9_, 197-203, 1987) and bean abscission cellulase (Tucker et al Plant Physiology 8 , 1257-1262, 1988).
  • TCELB6 and MCELE2 have been deposited on 29 March 1990 with the National Collections of Industrial and Marine Bacteria, Aberdeen, Scotland under Accession Nos. NCIB 40268 and 40269, respectively. DNA fragments similar to those cloned in
  • TCELB6 and MCELE2 may be generated in polymerase chain reactions using appropriate synthetic oligonucleotide primers and either tomato or melon genomic DNA as appropriate.
  • cDNA clones showing homology to TCELB6 and MCELE2 may be obtained from the mRNA of ripening tomatoes or melons by methods similar to those described by Slater et al, Plant Molecular Biology 5_, 137-147, 1985. In this way may be obtained sequences coding for the whole, or substantially the whole, of the mRNA produced by tomato or melon cellulase genes. Suitable lengths of the cDNA so obtained may be cut out for use by means of restriction enzymes.
  • the source of DNA fragments for the base sequence for transcription may be derived from
  • HEET either mRNA or from a gene encoding a cellulase or like enzyme. If the DNA is derived from a gene it may differ from that derived from mRNA in that introns may be present. The introns are not transcribed into mRNA (or, if so transcribed, are subsequently cut out). When using such a gene as the source of the base sequence for transcription it is possible to use either intron or exon regions. A further way of obtaining a suitable DNA base sequence for transcription is to synthesise it ab initio from the appropriate bases, for example using Figure 1 as a guide.
  • oligonucleotide synthesiser such as the Applied Biosystems oligonucleotide synthesiser are available which will synthesise lengths of single stranded DNA in any desired base sequence up to a maximum of around 100 bases. Complementary strands of DNA are annealed and subsequently ligated to form a double stranded DNA fragment of the desired length and sequence.
  • the strand that was formerly the template strand becomes the coding strand, and vice versa.
  • the new vector will thus encode RNA in a base sequence which is complementary to the sequence of the cellulase gene.
  • the two RNA strands are complementary not only in their base sequence but also in their orientations (5' to 3').
  • Recombinant DNA and vectors according to the present invention may be made as follows.
  • a suitable DNA source containing the desired base sequence for transcription for example TCELB6 or MCELE2 is treated with restriction enzymes to cut the sequence out.
  • a suitable fragment may be generated by polymerase chain reaction (PCR) from a suitable DNA source (for example TCELB6 or MCELE2) using synthetic oligonucleotide primers.
  • the DNA strand so obtained is cloned (if desired in reverse orientation) into a second vector containing the desired promoter sequence (for example cauliflower mosaic virus 35S RNA promoter or the tomato polygalacturonase gene promoter sequence - Bird et al., Plant Molecular Biology, 1JL, 651-662, 1988) and the desired terminator sequence (for example the 3' of the Agrobacterium tumefaciens nopaline synthase gene, the nos 3' end).
  • the desired promoter sequence for example cauliflower mosaic virus 35S RNA promoter or the tomato polygalacturonase gene promoter sequence - Bird et al., Plant Molecular Biology, 1JL, 651-662, 1988
  • the desired terminator sequence for example the 3' of the Agrobacterium tumefaciens nopaline synthase gene, the nos 3' end.
  • constitutive promoters such as cauliflower mosaic virus 35S
  • inducible or developmentally regulated promoters such as the ripe-fruit-specific polygalacturonase gene
  • Use of a constitutive promoter will tend to affect functions in all parts of the plant: while by using a tissue-specific promoter, functions may be controlled more selectively.
  • the promoter of the PG gene e.g. to tomatoes and melons
  • Use of this promoter, at least in tomatoes has the advantage that the production of antisense RNA is under the control of a ripening-specific promoter.
  • the antisense RNA is only produced in the organ in which its action is required.
  • Other tomato ripening-specific promoters that could be used include the E8 promoter (Diekman & Fischer, EMBO Journal 7_, 3315-3320, 1988).
  • Vectors according to the invention may be used to transform plants as desired, to make plants according to the invention.
  • Dicotyledonous plants, such as tomato and melon, may be transformed by
  • EET Agrobacterium Ti plasmid technology for example as described by Bevan (1984) Nucleic Acid Research, 12, 8711-8721. Such transformed plants may be reproduced sexually, or by cell or tissue culture.
  • the degree of production of antisense RNA in the plant cells can be controlled by suitable choice of promoter sequences, or by selecting the number of copies, or the site of integration, of the DNA sequences according to the invention that are introduced into the plant genome. In this way it may be possible to modify ripening or senescence to a greater or lesser extent.
  • the constructs of our invention may be used to transform cells of both monocotyledonous and dicotyledonous plants in various ways known to the art. In many cases such plant cells (particularly when they are cells of dicotyledonous plants) may be cultured to regenerate whole plants which subsequently reproduce to give successive generations of genetically modified plants. Examples of genetically modified plants according to the present invention include, as well as tomatoes, fruits of such as mangoes, peaches, apples, pears, strawberries, bananas and melons.
  • Such genetically modified plants may further contain other exogenous DNA expressible under the control of plant promoters, for example DNA expressing RNA antisense to other fruit ripening enzymes, for example polygalacturonase or pectin methylesterase, as described in our European Patent application 271,988 (US Serial 119,614).
  • plant promoters for example DNA expressing RNA antisense to other fruit ripening enzymes, for example polygalacturonase or pectin methylesterase, as described in our European Patent application 271,988 (US Serial 119,614).
  • Figure 1 shows the base sequences of tomato and melon cellulase genes in clones TCELB6 (Seq ID No: 1) and MCELE2 (Seq ID No: 2) respectively.
  • Figure 2 shows the oligonucleotides used for generation of fragments of tomato and melon cellulase gene by polymerase chain reaction.
  • Figure 3 shows a strategy for generation of fragments of a tomato cellulase gene by polymerase chain reaction.
  • Figure 4 shows the base sequence of tomato cellulase cDNA clone lambda cel-1 (Seq ID No: 3)
  • Figure 5 shows a strategy for construction of a cellulase antisense RNA vector pJRlTCl according to the invention.
  • a fragment of a tomato cellulase gene was generated in a polymerase chain reaction.
  • Synthetic oligonucleotide primers for the reaction (TCEL10 and TCELll) were designed from regions of homology between avocado (Tucker et al, 1987, cited above) and bean abscision (Tucker et al, 1988, cited above) cellulase ( Figure 2).
  • these primers would be expected to generate a 266 base pair fragment based on the sequence in Tucker et al, 1987.
  • the nucleotide sequence of one of the hybridising clones was determined ( Figure 1). This had significant similarity to the nucleotide sequence of the avocado cellulase gene (Tucker et al, 1987, cited above).
  • a fragment of a melon cellulase gene was generated, cloned and sequenced in a manner similar to that described in example 1 for the tomato cellulase gene fragment.
  • the same PCR primers (TCEL10 and TCELll) were used to generate a fragment of approximately 300 base pairs from DNA extracted from melon (Cucumis melo L cv. Western Shipper). This fragment was cloned into Ml3mpl8 cut with Hindi and hybridised with [32P] labelled TCEL10.
  • the nucleotide sequence of one hybridising clone was determined (Figure 1) and had significant homology to the tomato and avocado cellulase genes. - 12 -
  • the insert of the clone TCELB6 was used as a hybridisation probe to screen a commercially available ripe tomato (Lycopersicon esculentum Mill cv Ailsa Craig) cDNA library.
  • ripe tomato Licopersicon esculentum Mill cv Ailsa Craig
  • the nucleotide sequence of the clone was completely determined (figure 4).
  • the clone had an insert of 1415 base pairs and showed significant similarity to the avocado sequence (Tucker et al cited above).
  • Vectors may be constructed using cloned sequences from tomato or melon cellulase gene or cDNA fragments as shown in Figure 5.
  • pJRlTCl may be synthesised in vitro by cutting TCELB6 RF 30 DNA with Pstl the cut ends are then made flush with T4 DNA polymerase. The DNA is then cut with BamHI. The 283bp fragment from this reaction is then isolated and cloned into pJRl cut with Smal and BamHI.
  • pJRl (Smith et al Nature 334, 724-726, 1988) is a Binl9 (Bevan, Nucleic 35 Acids Research, 12, 8711-8721, 1984) based vector, which permits the expression of the antisense RNA under the control of the CaMV 35S promoter. This vector includes a nopaline synthase (nos) 3' end termination sequence.
  • vectors with the correct structure of pJRlTCl are identified by DNA sequence analysis.
  • the vector pJRlMCl is made similarly, following the construction schemes shown in Figure 5.
  • tomato and melon cellulase antisense RNA vectors with the gene fragments isolated by PCR and the tomato polygalacturonase gene promoter.
  • the tomato and melon gene fragments described in example 3 are also cloned by the same procedure into pJR2 to give the following clones:
  • TCELB6 (223bp tomato cellulase gene) - pJR2TCl
  • pJR2 is a Binl9 based vector, which permits the expression of the antisense RNA under the control of the tomato polygalacturonase promoter.
  • This vector includes a nopaline synthase (nos) 3' end termination sequence (see Figure 4) .
  • tomato and melon gene fragments described in example 3 are also cloned into pJRl in the sense orientation to give the following clones: 1. TCELB6 (223bp tomato cellulase gene) - pJRlTClS
  • pJRlTClS may be synthesised in vitro by cutting TCELB6 RF DNA with Pstl and Xbal, the cut ends are then made flush with T4 DNA polymerase. The 283bp fragment from this reaction is then isolated and cloned into the Hindi site of pJRl. After synthesis, vectors with the sense orientation of TCELB6 sequence are identified by DNA sequence analysis. The vector pJRlMClS is made similarly.
  • the tomato cellulase cDNA clone (lambda cel-1) was cut with EcoRI to excise the cDNA insert. The 1415 base pair fragment was isolated and the cut ends were made flush with T4 DNA polymerase. This fragment was then cloned into the Smal site of pJRl. After synthesis, vectors with the antisense orientation of the lambda cel-1 sequence were identified by both PCR and DNA sequence analysis. One clone that contained the cellulase cDNA sequence in the antisense orientation was designated pJRTCelA.
  • a vector pJRTCelS may be obtained by using a similar strategy and identifying a clone with the cellulase cDNA sequence in the sense orientation.
  • SHEET micro-organism widely available to plant biotechnologists and are used to transform tomato and melon plants. Transformation of tomato stem segments follows standard protocols (e.g. Bird et al Plant Molecular Biology 11, 651-662, 1988). Melon plants are transformed by a similar process. Transformed plants are identified by their ability to grow on media containing the antibiotic kanamycin. Plants are regenerated and grown to maturity. Ripening fruit are analysed for modifications to their ripening characteristics.

Abstract

Des constructions d'ADN utiles pour modifier le comportement des fruits lors de leur maturation comprennent une région d'initiation transcriptionnelle, active dans les plantes et implantée pour la transcription d'une séquence d'ADN homologue à tout ou partie d'un gène codant pour la cellulase ou une enzyme semblable, ledit gène présentant une homologie considérable à l'ADN codé par l'une des constructions TCELB6 et MCELE2. On décrit également des cellules de plantes et des plantes transformées avec lesdites constructions.
EP91908064A 1990-04-25 1991-04-19 Adn, constructions d'adn, et cellules et plantes ainsi derivees Withdrawn EP0528826A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB9009307 1990-04-25
GB909009307A GB9009307D0 (en) 1990-04-25 1990-04-25 Dna,constructs,cells and plant derived therefrom

Publications (1)

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EP0528826A1 true EP0528826A1 (fr) 1993-03-03

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EP91908064A Withdrawn EP0528826A1 (fr) 1990-04-25 1991-04-19 Adn, constructions d'adn, et cellules et plantes ainsi derivees

Country Status (7)

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EP (1) EP0528826A1 (fr)
JP (1) JPH05506576A (fr)
AU (1) AU652362B2 (fr)
CA (1) CA2081454A1 (fr)
GB (1) GB9009307D0 (fr)
WO (1) WO1991016440A1 (fr)
ZA (1) ZA913048B (fr)

Families Citing this family (26)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5585545A (en) * 1990-04-20 1996-12-17 The Regents Of The University Of California Endo-1,4-β-glucanase genes and their use in plants
US5168064A (en) * 1990-04-20 1992-12-01 The Regents Of The University Of California Endo-1,4-β-glucanase gene and its use in plants
DK115890D0 (da) * 1990-05-09 1990-05-09 Novo Nordisk As Enzym
US5767373A (en) 1994-06-16 1998-06-16 Novartis Finance Corporation Manipulation of protoporphyrinogen oxidase enzyme activity in eukaryotic organisms
US6084155A (en) 1995-06-06 2000-07-04 Novartis Ag Herbicide-tolerant protoporphyrinogen oxidase ("protox") genes
AU1316297A (en) * 1996-01-23 1997-08-20 Horticulture Research International Fruit ripening-related genes
GB9606906D0 (en) * 1996-04-02 1996-06-05 Zeneca Ltd Ripening-related dna from melon
GB9617184D0 (en) * 1996-08-15 1996-09-25 Danisco Enzyme
AU773779B2 (en) * 1996-09-12 2004-06-03 Syngenta Participations Ag Transgenic plants expressing cellulolytic enzymes
WO1998011235A2 (fr) 1996-09-12 1998-03-19 Novartis Ag Plantes transgeniques exprimant des enzymes cellulolytiques
EP1574580A3 (fr) * 1996-09-12 2009-07-01 Syngenta Participations AG Plantes transgéniques exprimant des enzymes cellulolytiques
US5981835A (en) * 1996-10-17 1999-11-09 Wisconsin Alumni Research Foundation Transgenic plants as an alternative source of lignocellulosic-degrading enzymes
US6818803B1 (en) 1997-06-26 2004-11-16 Wisconsin Alumni Research Foundation Transgenic plants as an alternative source of lignocellulosic-degrading enzymes
AUPP249298A0 (en) 1998-03-20 1998-04-23 Ag-Gene Australia Limited Synthetic genes and genetic constructs comprising same I
HU230353B1 (hu) 1998-03-20 2016-02-29 Benitec Australia Ltd A génexpresszió szabályozása
US8598332B1 (en) 1998-04-08 2013-12-03 Bayer Cropscience N.V. Methods and means for obtaining modified phenotypes
EP3214177A3 (fr) 1998-04-08 2017-11-22 Commonwealth Scientific and Industrial Research Organisation Procédés et moyens pour obtenir des phénotypes modifiés
US20040214330A1 (en) 1999-04-07 2004-10-28 Waterhouse Peter Michael Methods and means for obtaining modified phenotypes
US6423885B1 (en) 1999-08-13 2002-07-23 Commonwealth Scientific And Industrial Research Organization (Csiro) Methods for obtaining modified phenotypes in plant cells
DE10212892A1 (de) 2002-03-20 2003-10-09 Basf Plant Science Gmbh Konstrukte und Verfahren zur Regulation der Genexpression
EP1931789B1 (fr) 2005-09-20 2016-05-04 BASF Plant Science GmbH Methodes de regulation de l'expression genique utilisant ta-siarn
US9315818B2 (en) 2006-06-07 2016-04-19 Yissum Research Development Company Of The Hebrew University Of Jerusalem Plant expression constructs and methods of utilizing same
US20090205075A1 (en) 2008-01-30 2009-08-13 Stacy Miles Use of plastid transit peptides derived from glaucocystophytes
US8975470B2 (en) 2009-06-30 2015-03-10 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Introducing DNA into plant cells
WO2013184768A1 (fr) 2012-06-05 2013-12-12 University Of Georgia Research Foundation, Inc. Compositions et méthodes d'inactivation génique dans les plantes
US20150040268A1 (en) 2013-04-25 2015-02-05 Morflora Israel Ltd Methods and compositions for the delivery of nucleic acids to seeds

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
NZ219472A (en) * 1986-03-28 1990-08-28 Calgene Inc Regulation of phenotype in plant cells using dsdna constructs
GB8626879D0 (en) * 1986-11-11 1986-12-10 Ici Plc Dna
GB8811115D0 (en) * 1988-05-11 1988-06-15 Ici Plc Tomatoes

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9116440A1 *

Also Published As

Publication number Publication date
GB9009307D0 (en) 1990-06-20
AU7677091A (en) 1991-11-11
CA2081454A1 (fr) 1991-10-26
JPH05506576A (ja) 1993-09-30
WO1991016440A1 (fr) 1991-10-31
ZA913048B (en) 1992-10-28
AU652362B2 (en) 1994-08-25

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