WO2004001040A1 - Promoteur specifique par rapport a un tissu / une contrainte d'environnement - Google Patents

Promoteur specifique par rapport a un tissu / une contrainte d'environnement Download PDF

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Publication number
WO2004001040A1
WO2004001040A1 PCT/JP2003/007784 JP0307784W WO2004001040A1 WO 2004001040 A1 WO2004001040 A1 WO 2004001040A1 JP 0307784 W JP0307784 W JP 0307784W WO 2004001040 A1 WO2004001040 A1 WO 2004001040A1
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Prior art keywords
promoter
gene
plant
expression
rice
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PCT/JP2003/007784
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English (en)
Japanese (ja)
Inventor
Naoko Nishizawa
Satoshi Mori
Haruhiko Inoue
Hiromi Nakanishi
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Japan Science And Technology Agency
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Priority to AU2003244306A priority Critical patent/AU2003244306A1/en
Publication of WO2004001040A1 publication Critical patent/WO2004001040A1/fr

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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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/1085—Transferases (2.) transferring alkyl or aryl groups other than methyl groups (2.5)
    • 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
    • C12N15/8222—Developmentally regulated expression systems, tissue, organ specific, temporal or spatial regulation
    • C12N15/8223—Vegetative tissue-specific promoters
    • C12N15/8227—Root-specific

Definitions

  • the present invention relates to a promoter capable of regulating and controlling the expression of a gene in an environment-specific and site-specific manner corresponding to the environment, a vector containing the promoter, and a plant. More specifically, the present invention shows weak expression in endothelial cells around the root canal of plants under iron-sufficient conditions, and extremely strong in roots and aerial parts under iron-deficient conditions, The present invention relates to a promoter that can be regulated to be very strongly expressed in companion cells adjacent to the phloem, a vector containing the promoter, and a plant. Background art
  • a chimeric gene vector is constructed by linking a structural gene involved in the expression of a target trait to a promoter that can be expressed in a plant, and is introduced into a plant to express the desired trait. Promote or suppress.
  • this method has been applied to, for example, pest-resistant plants that have introduced the insecticidal BT toxin gene from Bacillus thuringens is, or overripening of tomato fruits. Tomatoes with a good shelf life and transgenic polygalacturonidase gene antisense have been put to practical use.
  • Cauliflower mosaic virus (CaMV) 35S promoter is widely used as a promoter of the structural gene introduced into these plants.
  • This C a MV 35 S promoter has the function of strongly promoting the transcription of structural genes.
  • transcriptional promotion by the Ca MV35S promoter is non-specific, and is regulated under the control of the whole plant, regardless of the growth stage of the plant, the environment in which the plant is placed, and the expression site. It constantly promotes transcription of placed structural genes. Therefore, it does not significantly affect the purpose of efficiently producing industrially useful substances or the metabolic system of plants It is useful for performing gene expression. Disclosure of the invention
  • a promoter capable of controlling the expression amount and the expression site in accordance with desired conditions for gene expression is extremely useful in plant breeding, avoiding the above problems. Furthermore, by combining such a promoter that can control expression, the expression of a group of genes existing downstream thereof can be regulated and controlled.
  • an object of the present invention is to provide a promoter capable of controlling gene expression in a tissue-specific manner and in response to environmental stress in response to an environment in which a plant is placed.
  • FIG. 1 shows an example of a vector 1 construct 1 when a structural gene is introduced into a plant cell using the promoter of the present invention.
  • Fig. 2 shows the appearance of GUS gene expression in rice roots when rice transformed with the promoter of the present invention was grown under control and iron deficiency conditions. This is a color photograph that replaces the drawing, which is colored blue. The upper row in Fig. 2 is for control conditions, and the lower row is for iron deficiency conditions.
  • RH indicates root hair
  • EX indicates epidermal cells
  • C indicates cortex
  • EP indicates epidermal cells
  • S indicates thick film cells.
  • FIG. 3 is a color photograph replacing the drawing showing an enlarged view of the center pillar portion of FIG. A of FIG. 3 is an enlarged view of the central part of the control, and B of FIG. 3 is an enlarged view of the expression site (the portion enclosed by the square of A in FIG. 3).
  • C in Fig. 3 is an enlarged view of the center of the cross section of the root of the transformed rice plant grown under iron deficiency conditions. Is an enlarged view of the expression site (the portion surrounded by a square C in FIG. 3).
  • MX indicates metagenetic duct
  • PX indicates progenitor duct
  • P indicates endothelial cell
  • PP indicates progenitor phloem
  • CC indicates companion cell
  • EN indicates endothelial cell.
  • FIG. 4 shows the expression of the GUS gene in rice roots when rice transformed using the promoter of the present invention was grown under control and iron deficiency conditions. This is a color picture that is replaced by a drawing showing a vertical cross-section of the root.
  • FIG. 5 shows the appearance of the GUS gene in rice roots when rice transformed with the promoter of the present invention was grown under control and iron deficiency conditions (color photograph). Is a pale blue color.) This is a color photograph that replaces the drawing with a branch root.
  • Fig. 6 shows the expression of the GUS gene in rice roots when the rice transformed with the promoter of the present invention was grown under control and iron deficiency conditions (light blue color image). It is colored instead of a drawing showing root hairs.
  • RH indicates root hair
  • EX indicates epidermal cells
  • C indicates cortical layer
  • EP indicates epidermal cells
  • S indicates thick film cells.
  • FIG. 7 shows rice leaves grown using the promoter of the present invention and rice transformed under the control of the rice plant, and the latest leaf (-Fe new leaf) that exhibited black mouth cis due to iron deficiency. , And the appearance of the GUS gene in the most developed leaf (-Fe old leaf) at that time (colored in pale blue in the color photograph). is there.
  • the upper part of Fig. 7 shows the control, the middle part shows the latest leaf (-Fe new leaf) exhibiting black closis due to iron deficiency, and the lower part shows the most developed leaf (-Fe old leaf) at that time.
  • FIG. 8 is a color photograph replacing the drawing showing the large vascular bundle of the leaf blade in FIG. A in Fig. 8 shows a control leaf, B in Fig. 8 shows the latest leaf (-Fe new leaf) exhibiting clonal cis due to iron deficiency, and C in Fig. 8 shows the expression site (- FIG. 8D is an enlarged view of a portion surrounded by a square B in FIG. 8, D in FIG. 8 shows the maximum developed leaf (-Fe old leaf) at that time, and E in FIG. FIG. 8 is an enlarged view of a portion surrounded by a square D in FIG. 8).
  • ST indicates the phloem
  • MX indicates the metagenetic conduit
  • CC Indicates a companion cell.
  • FIG. 9 is a color photograph instead of a drawing showing the laterally running vascular bundle of the largest developing leaf ( ⁇ Fe old leaf) of a rice transformed with the promoter of the present invention, which exhibited chlorosis under iron deficiency conditions.
  • FIG. 10 is a photograph, instead of a drawing, showing a moving cell (MC) of a rice plant transformed using the promoter of the present invention under control conditions.
  • MC moving cell
  • Nicotianamine (hereinafter referred to as NA) is thought to play an important role in metal transport in plants because it is present in all higher plants and has various heavy metal and chelate binding activities. Especially in grasses, NA plays an important role as a precursor in the biosynthetic pathway of mugineic acids. Nicotianamine synthase (hereinafter referred to as NAS) is an enzyme that synthesizes NA by polymerizing S-adenosylmethionine into small molecules. First, a genomic fragment containing the NAS cDNA and the NAS gene was cloned from wheat (Horudeum vulgare L. cv. E imehadaka no. 1) (Higuchi et al., 1989).
  • the rice NAS gene (hereinafter referred to as OsNAS1) was cloned (Higuchi et al., 2001). It has been reported that the rice NAS gene is constantly weakly expressed in roots under iron-sufficient conditions, and strongly induced in roots and shoots by iron deficiency (Higuchi et al., 2000). 1). Also, when the iron deficiency stress is released, the expression of the gene is rapidly suppressed.
  • the present inventors used the promoter of the rice nicotianamine synthase gene (OsNASl) to strongly express the gene when the plant lacks iron, and suppress the gene expression when the iron is sufficient.
  • the promoter of the OsNASI gene also regulates the expression of genes other than the OsNAS1 gene. It is a promoter that can be regulated, that is, it shows weak expression in the inner sheath cells around the conduit of the root of the plant under iron-sufficient conditions, and shows non-expression in the root and above-ground parts under iron-deficient conditions.
  • the present inventors have found that the promoter can be regulated so as to be extremely strongly expressed, particularly in a companion cell adjacent to the phloem, and completed the present invention.
  • the present invention provides a DNA fragment containing the promoter of the ⁇ sNAS1 gene.
  • the present invention further provides a plant vector comprising the DNA fragment of the above-mentioned invention. More specifically, the present invention relates to a promoter which is bound upstream of a structural gene other than the nicotianamine synthase gene of rice, wherein the structural gene responds to an environment in which a plant is placed, The present invention relates to a promoter capable of regulating and controlling gene expression specifically in response to tissue-specific and environmental stress.
  • the present invention also relates to a promoter that is linked upstream of a structural gene other than the nicotianamine synthase gene of rice, wherein the expression of the structural gene is regulated by a plant root conduit under iron-sufficient conditions. It relates to a promoter that is weakly expressed in surrounding inner sheath cells, and can be regulated and regulated to be very strongly expressed in roots and shoots under iron deficiency conditions.
  • the present invention relates to a vector, preferably a plant vector, comprising the promoter of the present invention.
  • the present invention also relates to a plant comprising the promoter of the present invention, preferably a Gramineous plant, and a seed of the plant.
  • the present inventors isolated the genomic DNA containing the 0 s NAS1 gene and analyzed it, thereby isolating the promoter portion of the 0 s NAS1 gene.
  • the jS-Darc mouth nidase (GUS) gene was ligated downstream of this promoter, and this gene was introduced into a plant to examine the function of the promoter of the ssNAS1 gene in plants.
  • GUS jS-Darc mouth nidase
  • FIG. 2 shows the cross-section of the roots of the transformed rice plants grown under control and iron deficiency conditions using color photographs instead of drawings.
  • RH indicates root hair
  • EX indicates epidermal cells
  • C indicates cortical layer
  • EP indicates epidermal cells
  • S indicates thick film cells.
  • FIG. 3 is a color photograph replacing the drawing showing an enlarged view of the center pillar portion of FIG. A in FIG. 3 is an enlarged view of the central part of the control, and B in FIG. 3 is an enlarged view of the expression site (the portion enclosed by the square of A in FIG. 3). C in Fig.
  • FIG. 3 is an enlarged view of the center of the cross section of the root of the transformed rice plant grown under iron-deficiency conditions, and D in Fig. 3 is the site of its occurrence (circled in C in Fig. 3).
  • MX indicates an epigenetic duct
  • PX indicates a primordial duct
  • P indicates an inner sheath cell
  • PP indicates a primordial phloem
  • CC indicates a companion cell
  • EN indicates an endothelial cell.
  • weak GUS activity was observed in endothelial cells, parenchyma around the primordial xylem conduit, and companion cells around the primordial phloem.
  • strong GUS activity was seen in the entire central column and in companion cells around the protozoa.
  • strong GUS activity was observed around metazoic vessels in both the control and iron deficient conditions where branch roots appeared.
  • FIG. 4 is a photograph that replaces the drawing showing the longitudinal section of the root. Under the control condition, weak staining was observed in a part of the central column and in the outer skin cells. Under iron deficiency conditions, GUS activity was found throughout the root, including the fission zone and the elongation zone, as well as in the root cap and cells shed from the root cap.
  • FIG. 5 is a color photograph replacing a drawing showing a branch root. Under control conditions, no GUS activity was found in the branch roots. On the other hand, under iron deficiency conditions, strong activity was shown throughout the main and branched roots.
  • Figure 6 is a color photograph replacing the drawing showing root hair. In FIG.
  • RH indicates root hair
  • EX indicates epidermal cells
  • C indicates cortical layer
  • EP indicates epidermal cells
  • S indicates thick film cells.
  • GUS activity was not observed in root hairs, but strong GUS activity was often observed in some of the epidermal cells and epidermal cells.
  • strong GUS activity is observed throughout root hair cells, and other epidermal and epidermal cells also exhibit strong GUS activity. Indicated.
  • FIG. 7 is a photograph instead of a drawing showing a cross section of a leaf blade.
  • the upper part of Fig. 7 shows the control, the middle part shows the latest leaf (-Fe new leaf) which exhibited clonal cis due to iron deficiency, and the lower part shows the most developed leaf (-Fe old leaf) at that time.
  • no GUS activity was observed under the control conditions.
  • the latest leaf (-Fe new leaf) which exhibited clos cis under iron deficiency conditions strong activity was observed in the whole vascular and mesophyll cells.
  • the most developed leaf (-Fe old leaf) strong activity was observed only in the vascular bundle and mesophyll cells around the vascular bundle.
  • FIG. 8 is a color photograph replacing the drawing showing the large vascular bundle of the leaf blade.
  • a in Fig. 8 shows the control
  • B in Fig. 8 shows the latest leaf (-Fe new leaf) that exhibited cloostosis due to iron deficiency
  • C in Fig. 8 shows the expression site (Fig. 8).
  • FIG. 8D is an enlarged view of the area of the maximum developed leaf (-Fe old leaf) at that time
  • FIG. It is an enlarged view of the part enclosed by the square D in the figure).
  • ST indicates a phloem
  • MX indicates an epigenetic duct
  • CC indicates a companion cell. No activity was observed under control conditions.
  • Fig. 9 is a color photograph instead of a drawing showing the transverse vascular bundle of the largest developing leaf (-Fe old leaf) exhibiting black mouth cis under iron deficiency conditions. No activity was observed in the control condition, but activity was also observed in the transverse vascular bundle in the largest developing leaf (-Fe old leaf) that showed clonal cis in the iron-deficient condition.
  • FIG. 10 is a color photograph in place of a drawing showing control cells. As shown in FIG. 10, there were cases where the leaf cell motility cells exhibited GUS activity under the control conditions.
  • the structural gene to be linked downstream of the promoter of the present invention is not limited to the GUS gene exemplified above, but may be any as long as it is necessary to regulate and control the expression in an environmental stress or site-specific manner. .
  • a gene that codes for a fluorescent protein or a structural gene that colors plant tissue it becomes possible to respond to iron deficiency and to sense the iron nutrient status of plants and soil. Dike overnight-A plant can be created.
  • Examples of the promoter of the present invention include preferably a promoter of rice nicotianamine synthase gene (OsNASI gene), and more specifically, a promoter having a base sequence represented by SEQ ID NO: 1 in the sequence listing.
  • OsNASI gene rice nicotianamine synthase gene
  • the present invention is not limited to this sequence.
  • gene transfer into a plant cell can be performed by a conventional method using the vector, by preparing a vector by connecting a promoter and a gene to be controlled by the promoter downstream as described above. .
  • viruses and bacteria that infect plants such as cauliflower mosaic virus, gemini virus, tobacco mosaic virus, brom mosaic virus, agrobacterium lemme faciliens, agrobacterium rhizogenes, etc.
  • Gene transfer to plant cells can be performed indirectly by using a physical or chemical method such as microinjection, electoral portation, polyethylene glycol, or fusion. Can also be introduced.
  • the plant can be re-differentiated by culturing them under appropriate conditions according to the type of the plant and the like. Can also be.
  • the plant cell thus obtained is a plant cell into which a structural gene controlled by the promoter of the present invention has been introduced, a plant, and a plant seed.
  • the plants of the present invention are not limited to the rice exemplified above, and include grass plants including rice of other varieties, cereal plants such as wheat, cultivated plants such as vegetables, buffer plants such as flowers, and the like. Can be applied to many plants.
  • the promoter of the present invention can control the expression of not only the nicotianamine synthase gene and the GUS gene, but also genes encoding various other proteins.
  • the present invention can provide not only plants to which various traits have been imparted, but also genes required for elucidation and research of plant gene expression mechanisms according to the present invention. It can be used for any purpose.
  • the contents described in Japanese Patent Application No. 2002-1777793 are incorporated herein in their entirety.
  • AB046401 was used as a type II, and a 1.6 kb DNA fragment upstream of OsNASl was amplified by the PCR method.
  • the following primers were designed and synthesized from the sequence of the upstream region of OsNAS1. The upstream sequence immediately before the translation start point was counted as -1.
  • the PCR was performed using the above genomic clone as type I, and the amplified DNA fragment was separated by agarose gel electrophoresis. As a result, amplification of a 1.6 kb DNA fragment was confirmed. Therefore, the PCR product was converted to a QIA Quick PCR purification kit.
  • T4 ligase T0Y0B0: 50 / L
  • 2 tL of 10X Buffer were placed in a microtube, and allowed to react for 16 hours for 1 hour.
  • ligation solution was placed in a competent cell (E. coli XL 1 -Blue) and placed on ice for 30 minutes. Thereafter, a heat shock of 42 ° C was applied for 90 seconds, and the plate was placed on ice for another 3 minutes, and then spread on a plate.
  • ampicillin Amp
  • Tet tetracycline
  • D-galactoviranoside (5-bromo-4-chloro-3-indoly 1-bD-galactopyranoside (X-gal)) is 20 ⁇ g final concentration, g / mL, isopropyl monob—D —Cho-galactopyranoside (Isopropy 1-bD-thio-galactopyranoside (IPTG)) was added to a final concentration of 0.1 mM. Cultured for ⁇ 16 hours. Plasmids were obtained by the alkali extraction method or an automatic DNA separator.
  • the binary vector PIG221Hm was introduced into a predetermined restriction enzyme site, and this was named plasmid pHI1.
  • the promoter of the 0 s NAS1 gene is located adjacent to the upstream of the) 3-Darctic nidase (GUS) structural gene, and if it functions as a promoter, It controls the expression of this GUS structural gene.
  • GUS 3-Darctic nidase
  • FIG. 1 shows a schematic diagram of a part of pHHI1, which is incorporated into plant genomic DNA.
  • HPT is a hygromycin resistance gene
  • NOS is a polyadenylation signal of nopaline synthase.
  • Example 3 introduction to plant and breeding
  • Plasmid pHHI1 obtained in Example 2 was introduced into rice (Oryza Sativa L cv. Tsukinohikari) by the agrobacterium method. That is, Agrobacterium tumefaciens C58 (Agrobacterium tumefaciens C58) was cultured at 26 ° C with shaking for 2 ⁇ in 1 mL of B liquid medium containing an appropriate antibiotic, and transformed into E. coli having pH II. After incubating E. coli harboring Ruperplasmid pRK201 (helper plasmid pRK2013) for 1 3 with shaking at 37 t :, take 100 L of each, mix, and place on an L ⁇ plate without antibiotics. Cultured.
  • the method from the introduction of the gene into the plant cell to the regeneration of the plant was performed as follows. Ripe rice seeds were cultured in a scutellum callus induction medium to induce scutellum calli. Soak the agrobacterium in the scutellum callus for 30 seconds and use a co-culture medium with filter paper . And cultured for 3 days in the dark. After infection, the agrobacterium is washed with Claforan 250 mg gZL solution, and cultured in primary selection medium (hygromycin B 30 mg gZL) at 28 ° C and in the dark for 2 weeks, followed by secondary selection The cells were cultured in a medium (hygromycin B 50 mg / L) for 2 weeks.
  • the selected calli were cultured in a regeneration induction medium at 28 for 3 weeks in the light, and transplanted to a plant formation medium.
  • the regenerated plants grew vigorously, they were acclimated.
  • soil cultivation was performed.
  • Bonsol and vermiculite were mixed in equal proportions, and the regenerated plants were cultured in a natural light P2 greenhouse at 30 ° C in the day and 25 at night at night.
  • Izumi Kasei No. 15 (Sumitomo Chemical Industries) was given as top fertilizer.
  • Example 4 GUS gene expression
  • the expression test of the GUS gene was carried out according to the method of Jefferson et al. (1989). Was done.
  • the transformed rice used was cultivated hydroponically until the fifth leaf developed, and two types of expression tests were performed, a control plot grown with the same medium composition and an iron deficiency plot in which iron was removed from the medium.
  • Figure 2 shows the cross section of the roots of the transformed rice plants grown under control and iron deficiency conditions. Under control conditions, weak GUS activity was detected in a part of the central column. On the other hand, under iron deficiency conditions, strong GUS activity was observed in epidermal cells, epidermal cells, cortical cells, and central pillars.
  • Fig. 3 shows an enlarged view of the center column. Under the control conditions, weak GUS activity was observed in the inner sheath cells, parenchyma and progenitor cells around the virgin xylem conduit. Under iron deficiency conditions, strong GUS activity was seen in the entire central column and in the companion cells around the protozoa. In addition, strong GUS activity was observed around metazoic vessels under both control and iron deficiency conditions at the sites where branch roots appeared.
  • Figure 4 is a longitudinal section of the root. Under the control condition, weak staining was observed in a part of the central column and in the outer cells. Under iron deficiency conditions, GUS activity was found throughout the root, including the mitotic zone and elongation zone, as well as in the root cap and cells shed from the root cap. Figure 5 shows the branch roots. Under the control condition, no GUS activity was found in the branch roots. on the other hand, Under iron deficiency conditions, strong activity was shown throughout the main and branched roots. Figure 6 shows the root hairs.
  • FIG. 7 shows a cross section of the blade blade. As shown in FIG. 7, no GUS activity was observed under the control conditions. On the other hand, in the latest leaf (-Fe new leaf) which exhibited clos cis under iron deficiency conditions, strong activity was observed in the whole vascular and mesophyll cells. On the other hand, in the most developed leaf (-Fe old leaf), strong activity was observed only in the vascular bundle and mesophyll cells around the vascular bundle.
  • FIG. 8 shows the large vascular bundle of the leaf blade. No activity was observed under control conditions.
  • the latest leaf (-Fe new leaf) which exhibited clos cis under iron deficiency conditions, activity was observed in the entire vascular bundle and mesophyll cells, and particularly strong expression was observed in the companion cells around the phloem of the vascular bundle.
  • the largest developing leaf (-Fe old leaf)
  • strong activity was observed in companion cells around the phloem of the vascular bundle
  • weak activity was observed in duct parenchymal cells.
  • FIG. 9 shows the transverse vascular bundle. No activity was observed under the control conditions, but in the most developed leaf (-Fe old leaf), activity was also observed in the transverse vascular bundle.
  • FIG. 10 there was a case where the mobile cells of the leaf blade showed GUS activity under the control condition. .
  • the promoter of the present invention that is, the DNA having the nucleotide sequence of SEQ ID NO: 1 has the ability to control the expression of a structural gene in a tissue-specific and iron-deficiency-specific manner. In other words, by using this, it becomes possible to express a gene under its control in a strong and tissue-specific manner by being exposed to an iron deficiency state. Furthermore, this promoter can be used in any kind of gene. Therefore, if a target gene is selected, this is used to produce a vector under the control of the promoter of the present invention, and this vector is used to transform a desired plant, the plant becomes iron deficient.
  • this gene can be strongly expressed only when the gene is present, it avoids the problems caused by the gene introduced to give the desired form being expressed in unexpected tissues and situations. You can do it. Furthermore, by using a gene that can confer plant tolerance to iron deficiency with this promoter in plants susceptible to iron deficiency, it is possible to protect useful crops from iron deficiency stress, and it is possible to protect crops even in adverse environments. There is a possibility that things can be grown. Therefore, the present invention is considered to greatly contribute to agricultural or plant genetic engineering.

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Abstract

L'invention concerne un promoteur pouvant réagir à l'environnement d'une plante et réguler ainsi l'expression d'un gène de façon spécifique par rapport à un tissu / une contrainte d'environnement. L'invention concerne notamment un promoteur ligaturé en amont d'un gène de structure autre qu'un gène de nicotianamine synthase de riz, permettant au gène de structure de réagir à l'environnement d'une plante, ce qui permet de contrôler et de réguler l'expression du gène de façon spécifique par rapport à un tissu / une contrainte d'environnement. L'invention concerne plus particulièrement un promoteur pouvant contrôler et réguler l'expression d'un gène de telle sorte que son expression soit faible dans des cellules péricycliques autour des vaisseaux des racines d'une plante en présence d'un taux de fer élevé et que son expression soit forte dans les racines et les parties en surface en présence d'un taux de fer faible. L'invention concerne également un vecteur, de préférence un vecteur pour une plante, contenant ce promoteur.
PCT/JP2003/007784 2002-06-19 2003-06-19 Promoteur specifique par rapport a un tissu / une contrainte d'environnement WO2004001040A1 (fr)

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JP2002177943A JP2004016153A (ja) 2002-06-19 2002-06-19 組織特異的・環境ストレス特異的プロモーター

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004092380A1 (fr) * 2003-04-15 2004-10-28 Plant Functional Genomics Co., Ltd. Promoteur de gene hybride d'ubiquitine et son utilisation
CN103229713A (zh) * 2013-05-06 2013-08-07 福建农林大学 一种高营养水稻品种的选育方法

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Publication number Priority date Publication date Assignee Title
WO1999057249A1 (fr) * 1998-04-30 1999-11-11 Japan Science And Technology Corporation Nicotianamine synthase et gene codant cette derniere
WO2001001762A1 (fr) * 1999-07-05 2001-01-11 Japan Science And Technology Corporation Production de riz resistant aux carences en fer
JP2001017181A (ja) * 1999-07-09 2001-01-23 Japan Science & Technology Corp 鉄欠乏オオムギ根に出現する３６ｋＤａタンパク質の遺伝子

Patent Citations (3)

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Publication number Priority date Publication date Assignee Title
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