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WO2009018989A1 - Séquences nucléotidiques modulant l'expression de gènes dans des plantes - Google Patents

Séquences nucléotidiques modulant l'expression de gènes dans des plantes Download PDF

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Publication number
WO2009018989A1
WO2009018989A1 PCT/EP2008/006410 EP2008006410W WO2009018989A1 WO 2009018989 A1 WO2009018989 A1 WO 2009018989A1 EP 2008006410 W EP2008006410 W EP 2008006410W WO 2009018989 A1 WO2009018989 A1 WO 2009018989A1
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gene
seq
gene construct
expression
plant
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PCT/EP2008/006410
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Chiara Tonelli
Eleonora Cominelli
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Universita' Degli Studi Di Milano
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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)
    • C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
    • C12N15/8237—Externally regulated expression systems

Definitions

  • NUCLEOTIDE SEQUENCES MODULATING THE EXPRESSION OF GENES IN PLANTS
  • the present invention relates to the expression of recombinant nucleic acids in plants. More specifically the invention provides gene cassettes for the expression of nucleic acids in response to abiotic stresses such as drought and soil high salinity, regulatory sequences therefor, gene constructs and expression vectors carrying these sequences as well as plants transformed with the same.
  • ABA Abscisic acid
  • ABA Abscisic acid
  • ABA Abscisic acid
  • the ability of plants to survive cellular water deficit depends on the species and genotype, the length and severity of water loss, the age and stage of development and the organ and cell type. Responses to water deficit may occur within some seconds, such as modifications in membrane potential and in the phosphorylation status of proteins, or within minutes and hours, such as changes in protein composition and gene expression.
  • a first group of genes induced during water stress conditions, codes for proteins that directly function in protecting cell from dehydration like osmotins, chaperons and antifreeze, while a second group encodes for regulatory proteins like phosphatases and transcription factors that function in signal transduction pathways.
  • Proteins coded by these genes may be classified in two groups: the ones directly involved in protection from environmental stress, the others involved in gene expression regulation and signal transduction (Shinozaki and Yamaguchi-Shinozaki, Plant Physiol., 1997, 115: 327-334).
  • proteins belong to the first group such as channels involved in water movements through cell membranes, ubiquitins and proteases involved in protein turnover, detossifying enzymes like catalases, glutathione S-transferases, superoxide dismutases, desaturases, protective proteins like LEA class, osmotins, chaperons, enzymes necessary for osmoprotective compounds like sugars, proline, glycinebetaine.
  • Genes coding for kinases, PLC, 14-3-3 proteins, transcription factors belong to the second group. At least four independent pathways regulate gene expression in response to drought: two of them are ABA-dependent and two ABA-independent.
  • a c/s-element involved in one of the ABA-dependent ways is called "ABA-responsive element” (ABRE).
  • ABRE ABA-responsive element
  • AREB/ABF transcription factors belonging to the family of basic leucine-zipper, are induced in response to different stresses and bind to ABRE boxes present in promoter sequences of their target genes.
  • Another c/s-element called “dehydration responsive element/C-repeat” (DRE/CRT) is involved in one of the ABA-independent pathways.
  • Different "DRE/CRT-binding proteins” coding for ERF/AP2 transcription factors in some plant species are induced by desiccation, salt treatment, cold (Shinozaki and Yamaguchi-Shinozaki, Journal of Experimental Botany, 2007, 58: 221-227).
  • Some transcription factors belonging to MYB, MYC and NAC families are induced by osmotic stresses and are involved in other ABA-independent pathways. Genetic engineering approaches to obtain plants more tolerant to o
  • a "loss of function” strategy instead of a “gain of function” one in some case may be successful and it can be reached for example through antisense RNA or RNA interference approaches.
  • Inducible promoters allow to express a coding sequence or a transgene for antisense RNA or for RNA interference only in conditions in which the activation of the nucleic acid is required, while it results silenced when its function is not necessary.
  • Many examples of use of inducible promoters are known in plants.
  • Another example of application of this strategy concerns the rice OsNAC ⁇ gene coding for a transcription factor, involved in the response to abiotic stresses. Its over-expression under the control of CaMV35S, provokes a decrease in plant growth and productivity, while the expression of the same gene under the control of one of the two stress-inducible promoters LIP9 or OsNAC ⁇ improves stress tolerance, without affecting plant growth (Nakashima et al., Plant Journal, 2007, 51 :617-630).
  • stress-inducible promoters present these characteristics that may render the system ineffective: - they are not completely silent in not-inducing conditions;
  • a promoter completely silenced in normal growth conditions, but active in response to drought, ABA and salt is that controlling the expression of the rice OsLEA3-1 gene (Xiao et al., Theor Appl Genet, 2007, 115:35-46).
  • this promoter presents a drawback: its activation in response to different stress is induced after six days of drought stress application and after 18 hours of salt treatment, times quite long for an efficient response.
  • AtMYB41 Arabidopsis thaliana
  • Object of the present invention is a gene construct or cassette for the selective expression in response to osmotic stress, carrying a gene different from AtMYB41 (later on “heterologous gene”) functionally linked to the promoter sequence of the AtMYB41 gene specified in SEQ ID NO:1 or to the intergenic sequence present in the Arabidopsis genome between the end of the AtMYB41 coding sequence and the beginning of the At4g28100 gene, here indicated as SEQ ID NO:2, or to fragments or variants thereof, said variants having at least 80%, preferably at least 90%, more preferably at least 95% sequence identity to SEQ ID N. 1 and SEQ ID N. 2, provided that said fragments or variants retain a regulatory activity on heterologous gene transcription in response to osmotic stress.
  • the fragment of the promoter sequence is identified in SEQ ID NO:3.
  • the gene construct can also contain elements involved in transcriptional regulation, as introns, polyadenilation signals at 3' side of the heterologous gene, transcriptional activators or enhancers sequences, termination sequences, selection markers and other regulatory sequences.
  • Each gene with the exception of AtMYB41 can be functionally linked to the sequences disclosed in the present invention.
  • the expressions "operatively linked” and “functionally linked” indicate that the promoter (SEQ ID N. 1) and the heterologous nucleic acid are in such a reciprocal orientation as to allow the promoter directing the expression of the nucleic acid, generally in 5'-3' orientation and upstream of the coding sequence of the heterologous gene.
  • the regulatory sequence that in the Arabidopsis genome is downstream of the AtMYB41 coding sequence (SEQ ID N.2), according to the present invention it is preferably fused downstream of the heterologous gene, generally in 5'-3' orientation to the coding sequence.
  • the genetic construct contains the promoter sequence and the heterologous nucleic acid functionally linked and oriented in the direction 5'-3'.
  • the gene construct contains the heterologous nucleic acid and the regulatory sequence SEQ ID N. 2 functionally linked and oriented in the direction 5'-3'.
  • the gene construct contains the promoter sequence SEQ ID N. 1 , the heterologous nucleic acid and the regulatory sequence SEQ ID N. 2, functionally linked and oriented in the 5'-3' direction.
  • the gene or the heterologous nucleic acid codes for a protein or an RNA transcript - for example an antisense RNA - that may favor a function, regulate an effect or improve a characteristic of the host plant. Based on the selectivity demonstrated by the expression cassette in response to abiotic stresses, the coded product is involved in this kind of response. Examples of such genes are those coding for:
  • P5CS ⁇ -pyrroline carboxylate synthase
  • proline is one of the most common osmolite accumulated in plants in response to osmotic stresses. It was shown that the over-expression of genes coding for P5CS in different species confers tolerance to osmotic stress (Kavi et al., Plant Physiology, 1995, 108:1387-1394; Yamada et al., Journal of Experimental Botany, 2005, 56: 1975-1981 ; Simon-Sarkadi et al., J. Agric. Food Chem., 2005, 53:7512-7517).
  • LEA early embryogenesis abundant proteins work as chaperons that stabilize vesicles, proteins and membranes in stress conditions, having a protective function.
  • ROS reactive oxygen species
  • ABA is an hormone extremely important in water stress response and it is possible to induce its synthesis and to obtain in this way plants more tolerant to stresses. This was achieved through the over-expression of the Arabidopsis AtNCED3 gene that codes for the 9-cis-epoxycarotenoid dioxygenase, whereby transgenic plants show reduced transpiration level and higher drought tolerance than wild type plants (luchi et al., Plant Journal, 2001 , 27:325-333).
  • Another application of the present invention regards the use of the AtMYB41 regulatory sequences to express an antisense RNA or an RNA interference to silence the expression of different genes in response to abiotic stress.
  • the constitutive silencing of different genes is effective in improving plant stress tolerance. It has to be taken into consideration the fact that in some cases the complete constitutive silencing of orthologous genes may be disadvantageous. Therefore, also in these cases the use of stress-inducible regulatory sequences as those described in the present invention is particularly convenient.
  • proline dehydrogenase enzyme which is involved in proline degradation and, as previously discussed, is a very important osmolite accumulated in response to osmotic stress, is inactivated in dehydration conditions and induced in rehydration conditions (Yoshiba et al., Plant Cell Physiology, 1997, 38:1095-1102). It was shown that Arabidopsis plants expressing an antisense RNA, that silences this enzyme constitutively, are more tolerant to salt and cold stresses compared with wild- type plants (Nanjo et al., FEBS Lett., 1999, 461 : 205-210).
  • Another application of the present invention regards silencing of PARP1 and PARP2 genes coding for poly-(ADP-ribose)-polymerases, active in response to oxidative damages induced by drought, high light intensity and heat stress. Their activation causes cell death. If they are inactivated, this reaction is blocked, and plants result more tolerant than wild type to different stresses (De Block et al., Plant Journal, 2005, 41 :95-106).
  • RNA interference constructs Concerning silencing of transcription factors involved in osmotic stress response, a further application of the present invention is the use of the described regulatory sequences to inactivate a gene orthologous to AtMYB ⁇ O through an antisense RNA or an RNA interference constructs.
  • Another application of the present invention consists in the fusion of the AtMYB41 regulatory sequences to reporter genes in order to produce biosensors able to perceive osmotic stresses.
  • Electronic sensors can supply data on plant growth conditions, but they cannot supply detailed data on the physiological perception in these conditions. Monitoring physiological plant state in response to environmental conditions may be particularly relevant and may be linked to systems able to change growth conditions, depending on specific plant needs.
  • a biosensor can be prepared fusing a reporter gene, whose expression can be simply monitored, downstream of the promoter described in the present invention.
  • a reporter gene frequently used is uidA (GUS; Jefferson et al., EMBO Journal, 1987, 6:3901-3907), coding for the ⁇ -glucuronidase enzyme.
  • This reporter was very useful, yet it is not possible to follow the expression of this reporter in a course time as it is necessary to sacrifice entire plant portions, so its use in some cases is limited.
  • Another reporter gene whose expression can be monitored in real time, without destructive methods, is the gene coding for GFP (green fluorescent protein; Haseloff and Amos, Trends Genet., 1995, 11 :328-329), present in jelly fish Aequorea aequorea.
  • GFP green fluorescent protein
  • the signal derived from the expression of this protein can be simply visualized using the correct combination between the excitation wavelength and filters for vision. Then the expression of GFP can be monitored through not destructive means.
  • the signal can be revealed by an equipment to monitor digital images, eventually connected to a machine able to change plant growth conditions based on measurements performed (for example through activation of an irrigation system, regulated in response to specific plant needs).
  • reporter genes that can be used are regulatory genes of anthocyanin biosynthesis.
  • Anthocyanins are red, blue and violet pigments, accumulated in different plant organs. They are involved in protection against UV-B radiation, in defense from pathogen attacks, and they attract pollinators insects. Their biosynthesis is induced by different stress conditions like wound, cold, nutrient starvation. Regulation of anthocyanin biosynthesis depends on transcription factors belonging to three different families: MYB, bHLH and WD40. Regulatory genes of this pathway were isolated in many species (for a review Koes et al., Trends in Plant Science, 2005, 10:236-242).
  • the invention relates to expression vectors carrying the gene constructs or cassettes herein provided.
  • the vectors can be bacterial plasmids, bacterial artificial chromosomes (BACs), yeast artificial chromosomes (YACs), viral vectors, vectors for direct DNA transfer, or, preferably, vectors for Agrobacterium-med ⁇ a ⁇ ed DNA transfer.
  • the latter can be either integrating or binary vectors and may contain selection markers, such as antibiotic- or herbicide-resistance genes, reporter genes facilitating the identification and selection of transformed cells, or sequences regulating gene expression in plants.
  • Direct transfer of DNA may include protoplast microinjection, electroporation and biolistic techniques based on plant bombardment with DNA-coated microparticles.
  • the invention provides transgenic plants, either monocotyledonous or dicotyledonous, as well as vegetative or reproductive parts thereof, or seeds, containing the gene constructs according to the invention.
  • the constructs or cassettes according to the invention are used to express proteins in response to osmotic stresses of closely related crop species, such as canola, in other dicotyledon plant, including soybean, tomato, tobacco, potato, cotton, or in monocotyledon species, such as corn, wheat, barley, rice.
  • AtMYB41 is a member of the large family of R2-R3 MYB transcription factors of Arabidopsis (Stracke et al., Curr. Opin. Plant Biol., 2001 , 4:447-456). To gain insight into the regulation of the AtMYB41 gene, quantitative RT-PCR analysis was performed on RNA obtained from several organs and at various stages of development of seedlings, rosette leaves, flowers and siliques ( Figure 1). AtMYB41 transcript was not detectable, under normal growth conditions in any organs or at any developmental stages analyzed.
  • AtMYB41 expression is rapidly induced (in the first hour of the treatment) in all the conditions, except for cold treatment.
  • salt treatment we observed a two-step induction kinetic, similar to that previously described for ERD10 and ERD14 in response to cold (Kiyosue et al., Plant Molecular Biology, 1994, 25:791-798).
  • the first one more rapid than the second one, does not require ABA biosynthesis, while the second one needs this hormone to be activated, so it is slower than the first one (Kiyosue et al., Plant Molecular Biology, 1994, 25:791-798).
  • the first peak after one hour of treatment, may depend on the activation of the transcription of the gene in an ABA-independent pathway, while the second peak, after 16 hours, may depend on ABA-dependent transcript accumulation.
  • This invention provides two complementary and inverted sequences, corresponding to the genomic region of Chromosome 4 - according to the nomenclature used in "The Arabidopsis Information Resource” accessible at http://www.arabidopsis.org - the first one comprised between nucleotide 13971558 (belonging to 5' UTR of At1g28130 gene, dep. n. NM179387) and nucleotide 13969391 (belonging to 5' UTR of AtMYB41 gene -At4g28110 dep. n.
  • genomic regions contain, respectively, the entire promoter of AtMYB41 and regulatory elements in the 3' region.
  • AtMYB41 gene was cloned upstream and, respectively, downstream of the GUS ( ⁇ -glucuronidase) and GFP reporter genes.
  • GUS ⁇ -glucuronidase
  • GFP reporter genes were fused upstream of the GUS/GFP reporter genes in the vector pBGWFS7.0.
  • This construct was called P41 v. GUS/GFP.
  • a second construct was prepared, starting from the one just described, cloning the entire SEQ ID N.2 of 1305 bp downstream of the GUS/GFP reporter genes. This construct was called P41 -3': GUS/GFP.
  • constructs thus obtained were introduced in Arabidopsis and the resulting transgenic lines were histologically analyzed to detect the reporter expression domains.
  • the reporter gene expression was detected in different plant organs such as leaves when desiccated for 7 hours but it was completely absent in untreated organs. Similar results were obtained in transgenic plants carrying the two different constructs. These results confirm the RT-PCR results previously described and reinforce the specificity of the AtMYB41 promoter only in response to stress and not in standard growth conditions. Bioinformatic analysis of the AtMYB41 promoter sequence Bioinformatic analysis on the genomic sequence upstream of the AtMYB41 coding sequence was performed starting from nucleotide -1476 to nucleotide -1 from the ATG codon.
  • FIG. 1 Analysis of AtMYB41 gene expression in different organs and at different developmental stages.
  • the TSB1 gene was used as a control.
  • FIG. 3 Bioinformatic analysis of a portion of intergenic region upstream of the coding sequence of the AtMYB41 (from -1476 to -1 of the beginning of the coding sequence). All the motifs found using the program
  • Seeds of wild-type Arabidopsis thaliana ecotype Columbia were used. For in-plate growth, the seeds were sterilized as follows: 5 min in absolute ethanol, 5 min in 0.6% (v/v) sodium hypochlorite, 0.05% Tween 20, 2 washes in sterile water. The seeds were layered at 4°C in the dark for a period of 4 days to allow uniform germination and then placed at 22°C with 16 hr light (48 ⁇ E/m2) and 8 hr dark periods in Petri dishes or in liquid culture. The solid medium contained MS (Sigma M-5519), 0.7% agar and 1 % saccharose, pH 5.7.
  • the liquid medium contained MS (Sigma M-5519), 3% saccharose, MES (Sigma M-8652) 0.5g/L, pH5.7. Plants were grown in liquid culture at 22°C, in constant agitation at 120 rpm. For the growth in soil, the seeds were germinated in Procedureserde soil (VM-type, Manna-Italy) in Araflat plates (Arasystem, Betatech, Belgium) with a 16 hr light (48 ⁇ E/m2) - 8 hr dark cycle.
  • MS Sigma M-5519
  • MES Sigma M-8652
  • - cauline leaves from 9 weeks old flowering plants; - flower buds: closed buds corresponding approximately to the 12 stage of flower development (time 0);
  • MS culture After three week growth under continuous shaking (120 rpm) the following treatments were performed:
  • the reverse-transcription reaction was performed with Reverse Transcriptase SuperscriptTM Il (Invitrogen), according to the manufacturer's indications, using the oligo(dT) primer, formed by 17 dT residues and by the adapter ⁇ '-GGGAATTCGTCGACAAGC-S'.
  • the cDNA samples were amplified in a reaction mixture containing Red Taq PCR Reaction Buffer 1X (Sigma) and 0.1 mM dATP, dCTP, dGTP and dTTP, 0.5 ⁇ M specific primers, 1 unit Red TaqTM polymerase (Sigma) and sterile distilled water to a final volume of 25 ⁇ l.
  • the amplification was carried out under the following conditions: 1 min at 94°C; 20 cycles at 94°C for 15 sec, 60 0 C for 15 sec, 72°C for 1 min; 72°C for 10 min.
  • the primers used were MYB41 F2 ( ⁇ '-GTATCGACCCTGTTACTCATTCT-S 1 ) and MYB41 R3 ( ⁇ '-AAGCAAGACATGTGTATGCAAAT-S 1 ) for AtMYB41 gene and TSBF1 ( ⁇ '-CTCATGGCCGCCGGATCTTGA-S') and TSBR1
  • PCR products were separated on 1 % agarose gel and transferred to Hybond N+ filters (Amersham) in 0.4N NaOH. Filters were hybridized with TSB1-, AtMYB41 -specific probes tagged with digoxigenin using the DIG-High Prime kit (Roche), following the manufacturer's instructions. Genomic DNA extraction
  • the DNA was precipitated by centrifugation of the samples at 13000 rpm for 10 min. Isopropanol was removed and the pellet washed with 300 ⁇ l of 80% ethanol. After removal of ethanol, the DNA was resuspended in 40 ⁇ l of 50 mM Tris-HCI pH 8.0, 20 ⁇ g/ml of 5 mM EDTA. 2 ⁇ l of 20 mg/ml RNase A (Roche) were added and the samples were incubated at 37°C for 30 min. The extracted DNA was kept at -20 0 C.
  • the genomic region upstream and downstream of the AtMYB41 coding sequence was amplified using Arabidopsis genomic DNA as template.
  • the genomic region upstream of the AtMYB41 coding sequence was amplified by a 3-step PCR reaction using primers P412kbllF1 (GATTTAAATAATGGAGATCCTGAA) and P412kbllR1
  • primers P41GWF aaaaagcaggctGATTT AAAT AATGGAGATC
  • P41GWR aaaaagcaggctGATTT AAAT AATGGAGATC
  • amplification product was cloned in pDONR207 vector by a BP clonase reaction (Invitrogen) and an Entry Clone was obtained.
  • the primer design, the PCR amplifications and the cloning procedure was performed, following the manufacturer's indications (Invitrogen). Then a LR clonase reaction was performed between the Entry Clone and the pBGWFS7.0 binary vector (Karimi et al., Trends Plant Sci, 2002, 193-195), carrying Bar as plant selectable marker.
  • the obtained construct corresponds to P41 v.GUS/GFP.
  • the genomic region downstream of the AtMYB41 coding sequence was amplified by a PCR reaction using primers 41-3UTRF1 (GACGTCTTTTCAACATTTGCATACAC) and 41-3UTRR1
  • the amplification product was cloned into the pCR4-TOPO vector
  • Wild-type Arabidopsis thaliana plants belonging to the Columbia ecotype were grown at 22°C with a photoperiod of 16 hr ligth/ 8 hr dark. In order to increase seed production, the primary inflorescences were removed and the plants were grown for additional 5-6 days, until the secondary inflorescences appeared. All the siliques were eliminated prior to transformation. The plants were then transformed with the Agrobacterium strain GV3101 by "floral-dip", following the Clough-Bent protocol (Clough and
  • Sterilized T1 seeds from transgenic plants were layered at 4°C in the dark for 4 days, and subsequently germinated in MS soil (Sigma M-5519), added with 0.8% bactoagar (Difco 0141-01) pH 5.7 and 100 ⁇ g/ml kanamycin. The plants were grown at 22°C, under 16 hr light / 8 hr dark photoperiod.
  • the reporter expression profiles were examined with the OLYMPUS SZX12 stereoscope (7X - 9OX magnification).

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Abstract

L'invention porte sur une cassette génétique pour l'expression d'acides nucléiques hétérologues en réponse à des stress abiotiques tels que la sécheresse et la salinité élevée du sol, sur des séquences régulatrices utilisées dans la cassette d'expression, sur des vecteurs d'expression portant ces séquences et sur des plantes transformées par celles-ci.
PCT/EP2008/006410 2007-08-08 2008-08-04 Séquences nucléotidiques modulant l'expression de gènes dans des plantes WO2009018989A1 (fr)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106967161A (zh) * 2017-05-02 2017-07-21 北京林业大学 调控蓝莓果实中花青素含量的蛋白及其编码基因与应用

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002016655A2 (fr) * 2000-08-24 2002-02-28 The Scripps Research Institute Sequences nucleotidiques de plantes a stress regule, plantes transgeniques contenant ces sequences, et methodes d'utilisation stress-regulated nucleotide sequences of plants, transgenic plants containing same, and methods of use
US20030229915A1 (en) * 1999-02-18 2003-12-11 James Keddie Plant gene sequences II

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030229915A1 (en) * 1999-02-18 2003-12-11 James Keddie Plant gene sequences II
WO2002016655A2 (fr) * 2000-08-24 2002-02-28 The Scripps Research Institute Sequences nucleotidiques de plantes a stress regule, plantes transgeniques contenant ces sequences, et methodes d'utilisation stress-regulated nucleotide sequences of plants, transgenic plants containing same, and methods of use

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
DATABASE EMBL [online] 14 April 2006 (2006-04-14), "Bst027158 lambda03 Boechera stricta genomic clone SAD-EX-38-T7_N17, genomic survey sequence.", XP002506235, retrieved from EBI accession no. EMBL:DU694628 Database accession no. DU694628 *
DATABASE EMBL [online] 27 September 2002 (2002-09-27), "Sequence 4924 from Patent WO0216655.", XP002506234, retrieved from EBI accession no. EMBL:AX510229 Database accession no. AX510229 *
DATABASE Geneseq [online] 22 April 2004 (2004-04-22), "cDNA encoding A. thaliana novel transcription factor (TF) #74.", XP002506236, retrieved from EBI accession no. GSN:ADI61448 Database accession no. ADI61448 *
NUCCIO M L ET AL: "Metabolic engineering of plants for osmotic stress resistance", CURRENT OPINION IN PLANT BIOLOGY, QUADRANT SUBSCRIPTION SERVICES, GB, vol. 2, no. 2, 1 April 1999 (1999-04-01), pages 128 - 134, XP002216348, ISSN: 1369-5266 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106967161A (zh) * 2017-05-02 2017-07-21 北京林业大学 调控蓝莓果实中花青素含量的蛋白及其编码基因与应用
CN106967161B (zh) * 2017-05-02 2020-10-27 北京林业大学 调控蓝莓果实中花青素含量的蛋白及其编码基因与应用

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