KR101335440B1 - OsRINGC2 gene from rice for enhancing salt stress resistance of plant and uses thereof - Google Patents

Abstract

The present invention is a method for controlling the resistance to salt tolerance of a plant comprising the step of transforming a recombinant vector containing the rice-derived OsRINGC2 OsRINGC2 -1 or -2 gene in the plant cell, the OsRINGC2 -1 or -2 gene OsRINGC2 a produced by the method of producing a transgenic plant salt tolerance is controlled, comprising transforming a plant cell, the method comprising the recombinant vector containing the salt tolerance control transgenic plants and their seeds or the OsRINGC2-1 or OsRINGC2 - Provided is a composition for controlling salt resistance of a plant comprising two genes.

Description

OsringC2 gene from rice for enhancing salt stress resistance of plant and uses approximately}

The present invention includes the step of transforming the present invention relates to OsRINGC2 gene derived from rice to increase the salt tolerance of a plant and use thereof, and more particularly a recombinant vector containing the rice-derived OsRINGC2 OsRINGC2 -1 or -2 gene in the plant cell method for controlling the resistance to salt tolerance of plants, the OsRINGC2 OsRINGC2 -1 or -2 transfected plant cell with a recombinant vector comprising the gene switch salt tolerance is controlled method of producing a transformed plant comprising the step of switching, the method of a salt tolerance of the control transgenic plant and seed thereof produced by and relates to a flame retardant for adjusting the composition of the plant including the OsRINGC2 OsRINGC2 -1 or -2 gene.

All living organisms are constantly exposed to biological and abiotic stresses throughout their lifetime, and plants are more likely to be affected than other organisms because they are sticky organisms and are unable to avoid various stresses. Therefore, plants can quickly detect abiotic stresses (eg, moisture, temperature, light, salts, metals and soils) and biological stresses (eg, viruses, bacteria, fungi and predators) to protect themselves. And respond. Plants have developed sophisticated networks for defense and response to environmental stress (Yang Y. et. al . , Genes Dev . 11 , 1621-1639, 1997), which has a mechanism for quickly recognizing and responding to environmental stimuli.

Ubiquitination by ubiquitin (ubiquitination) is a mechanism that all higher organisms, as well as plants, is known to have a variety of functions, but little is known about genes involved in abiotic stress. RING (Really Interest New Gene) finger proteins play an important role in moving ubiquitin proteins to other target proteins and contribute to post-transcriptional regulatory responses such as proteolytic degradation, structural and functional degeneration, and position changes (Deshaies and Joazeiro, Annu Rev Biochem 78 , 399-434, 2009). The attachment of ubiquitin protein to the target protein is achieved by the sequential reaction of three proteins (E1, E2, E3). Most RING finger proteins in eukaryotes have E3 ligase activity. Representative monocotyledonous rice contains protein with 425 RING domains, and dicotyledonous Arabidopsis contains protein with 469 RING domains (Lim et al. al . , Plant Mol Biol 72 , 369-380, 2010). Recently, 663 proteins in the apple genome have been identified as having a RING domain (Li et al. al . , Mol Genet Genomics 286 , 81-94, 2011). The RING motif consists of cysteine and histidine residues and has a site to which two zinc ions bind (Freemont et. al . , Cell 64 , 483-484, 1991). The RING-H2 and RING-HC types of RING finger proteins are 50.5% and 39% in Arabidopsis, 55.5% and 25.9% in rice, 53.3% and 29.3% in apple, respectively (Li et al. al . , Mol Genet Genomics 286 , 81-94, 2011). It is also known that the RING gene family has a similar proportion in many plants. Types modified from the existing RING finger protein types include RING-v, RING-D, RING-S / T, RING-G and RING-C2. Specifically, there are two RING-C2 types in rice, ten in the Arabidopsis, and ten in apples. RING finger protein inhibits light inactivation (McNellis et al . , Plant Cell 6 , 1391-1400, 1994), cell cycle progression (Liu et al . , Plant Mol Biol 68 , 17-30, 2008), positive regulation of ABA signaling, chromatin reconstitution (Bu et al . , Plant Physiol 150 , 463-481, 2009) and cell survival (Koiwai et al, Plant J 51, 92-104, 2007). However, the function of the modified RING type mentioned above is still unknown. The present invention was performed to isolate two RING - C2 type genes from Korean rice species and to identify expression patterns, intracellular expression sites, and overexpressed plant characteristics.

Meanwhile, Korean Patent No. 0767100 discloses a pepper ring-finger protein gene (CaRFP1) and a method for detecting disease resistance and transformed plants of plants using the same, and Korean Patent No. 0900928 discloses plant stress resistance. An increasing CaRma1H1 (Capsicum annuum Ring finger protein with Membrane Anchor1 Homolog 1) gene and a transgenic plant into which the gene is introduced are disclosed. However, as in the present invention, there is no disclosure about the OsRINGC2 gene derived from rice which increases the flame resistance of plants.

The present invention was derived by the above requirements, the present inventors confirmed that the expression of OsRINGC2 gene in rice plants is increased under salt stress conditions, by confirming that the transgenic Arabidopsis plants overexpressing the OsRINGC2 gene exhibit flame resistance. The present invention has been completed.

In order to solve the above problems, the present invention provides a method for adjusting the salt tolerance of a plant comprising the step of transforming a recombinant vector comprising a gene of rice OsRINGC2-2 OsRINGC2 -1 or derived from a plant cell.

The present invention also provides a method for producing the OsRINGC2 -1 or salt tolerance is regulated transgenic plant comprising transforming a plant cell with a recombinant vector comprising a gene OsRINGC2 -2.

The present invention also provides a flame retardant transgenic plant and seeds thereof prepared by the above method.

The present invention also provides a flame retardant for adjusting the composition of the plant including the OsRINGC2 OsRINGC2 -1 or -2 gene.

According to the invention, rice-derived OsRINGC2 -1 or OsRINGC2-2 gene expression is strongly induced under salt stress conditions, OsRINGC2 -1 or transgenic Arabidopsis thaliana plants which overexpress OsRINGC2 2 gene showed the salt tolerance. In particular, it was confirmed that the flame retardant is further increased in transgenic Arabidopsis plants that overexpress genes OsRINGC2 -1. Therefore it expected to have a salt tolerance OsRINGC2 -1 gene is useful for the development of resistant transgenic plants.

Figure 1 shows the result of separating the OsRINGC2 gene using a restriction enzyme. A. pBIN35S-RINGC2 schematic diagram of the construct, B. pBIN35S-RINGC2 handle structures with Hind III and Kpn I restriction enzymes and the results of electrophoresis.
2 shows the expression patterns of OsRINGC2-1 and OsRINGC2-2 genes in various tissues. A. semi-quantitative RT-PCR method and various OsRINGC2-1 OsRINGC2-2 expression pattern of the gene in the tissue, B. Expression of OsRINGC2-1 and OsRINGC2-2 gene according to the developmental stage of the running water (Stage I using: leading Length less than 1.5 cm, Stage II: 1.5-5.0 cm, Stage III: 5.0-10.0 cm, Stage IV: 10.0-20.0 cm).
3 shows the expression patterns of OsRINGC2-1 and OsRINGC2-2 genes in rice plants according to various abiotic stress and hormone treatments. C represents a control and T represents a stressed sample. A. OssalT , OsLIP19 and OsHsp90-1 genes are significantly increased in expression by treatment of abiotic stresses (200 mM NaCl), dry stress, cold stress (4 ° C.) and high temperature stress (45 ° C.). . B. OssalT , OsPRIb, and OsPBZI genes show genes with significantly increased expression levels by treatment with 0.1 mM absic acid (ABA), 1 mM salicylic acid (SA), and 0.1 mM jasmonic acid (JA).
Figure 4 shows the intracellular expression sites of OsRINGC2-1-EYFP and OsRINGC2-2-EYFP. OsRINGC2-1-EYFP and OsRINGC2-2-EYFP are controlled by the CaMV 35S promoter and observed by confocal microscopy of fluorescence signals transiently expressed in tobacco leaf epidermal cells. EYFP is kept bonded. A. 35S: EYFP, B. 35S: OsRINGC2-1-EYFP, C. 35S: OsRINGC2-2-EYFP.
Figure 5 is a result of comparing the root growth of wild-type plants (WT), 35S: OsRINGC2-1 and 35S: OsRINGC2-2 overexpressing plants by salt stress treatment. 5a. Expression of wild type plants, OsRINGC2-1 and OsRINGC2-2 overexpressed plants using RT-PCR assay was confirmed. 5b. 7 days a wild-type, three OsRINGC2-1 overexpressing plant and three root growth phenotype of plants over-expressing OsRINGC2-2 of lines of the line incubated under various NaCl concentrations. 5c. Root length statistically analyzed using the Image J program. (The experiment was repeated three times, with mean and standard errors indicated. * And ** indicate significant differences between wild-type and overexpressing transgenic plants, with p -values representing p <0.05 and p <0.01, respectively. (Student's t-test)).

According to an aspect of the invention, the invention is derived from rice -1 OsRINGC2 (Oryza sativa Really Interest New Gene finger protein C2-type 1) gene or OsRINGC2-2 (Oryza sativa Really Interest New Gene finger protein type C2-2 It provides a method for controlling the flame resistance of a plant comprising the step of transforming a plant cell with a recombinant vector comprising a gene.

Method according to one embodiment of the present invention is one characterized by the over-expression by OsRINGC2 -1 or OsRINGC2-2 genes of the rice plant derived from a plant cell to increase the salt tolerance of the plants, but is not limited thereto.

In the process according to one embodiment of the invention, the OsRINGC2 OsRINGC2 -1 or -2 gene may be comprised of the base sequence of SEQ ID NO: 1 or SEQ ID NO 2, respectively. In addition, variants of each of the above nucleotide sequences are included within the scope of the present invention. Specifically, the gene has at least 70%, more preferably at least 80%, even more preferably at least 90%, and most preferably at least 95% sequence homology with the nucleotide sequences of SEQ ID NO: 1 or SEQ ID NO: 2, respectively. Branches can include base sequences. The "% sequence homology" for a polynucleotide is determined by comparing the comparison regions with two optimally arranged sequences, wherein part of the polynucleotide sequence in the comparison region is the reference sequence (addition or deletion) for the optimal alignment of the two sequences. It may include the addition or deletion (ie, gap) compared to).

The term "recombinant" refers to a cell in which a cell replicates a heterologous nucleic acid, expresses the nucleic acid, or expresses a protein encoded by a peptide, heterologous peptide or heterologous nucleic acid. The recombinant cell can express a gene or a gene fragment that is not found in the natural form of the cell in one of the sense or antisense form. In addition, the recombinant cell can express a gene found in a cell in its natural state, but the gene has been modified and reintroduced intracellularly by an artificial means.

In the present invention, the OsRINGC2 OsRINGC2 -1 or -2 gene sequence can be inserted into a recombinant expression vector. The term "recombinant expression vector" means a bacterial plasmid, a phage, a yeast plasmid, a plant cell virus, a mammalian cell virus, or other vector. In principle, any plasmid and vector can be used if it can replicate and stabilize within the host. An important characteristic of the expression vector is that it has a replication origin, a promoter, a marker gene and a translation control element.

OsRINGC2 OsRINGC2 -1 or -2 gene each sequence and the expression vector containing the appropriate transcription / translation control signals can be constructed by methods well known to those skilled in the art. Such methods include in vitro recombinant DNA technology, DNA synthesis techniques, and in vivo recombination techniques. The DNA sequence can be effectively linked to appropriate promoters in the expression vector to drive mRNA synthesis. The expression vector may also include a ribosome binding site and a transcription terminator as a translation initiation site.

A preferred example of the recombinant vector of the present invention is a Ti-plasmid vector capable of transferring a so-called T-region to a plant cell when present in a suitable host, such as Agrobacterium tumefaciens. Other types of Ti-plasmid vectors (see EP 0 116 718 B1) are currently used to transfer hybrid DNA sequences to plant cells or protoplasts in which new plants capable of properly inserting hybrid DNA into the plant's genome can be produced have. A particularly preferred form of the Ti-plasmid vector is the so-called binary vector as claimed in EP 0 120 516 B1 and US Pat. No. 4,940,838. Other suitable vectors that can be used to introduce the DNA according to the invention into the plant host include viral vectors such as those that can be derived from double-stranded plant viruses (e. G., CaMV) and single- For example, from non -complete plant virus vectors. The use of such vectors may be particularly advantageous when it is difficult to transform the plant host properly.

The recombinant expression vector may preferably comprise one or more selectable markers. The marker is typically a nucleic acid sequence having a property that can be selected by a chemical method, and includes all genes capable of distinguishing a transformed cell from a non-transformed cell. Examples include herbicide resistance genes such as glyphosate or phosphinothricin, antibiotics such as kanamycin, hygromycin, chloramphenicol, G418, Bleomycin, Resistant gene, aadA gene, and the like, but are not limited thereto.

In the recombinant vector of the present invention, the promoter may be, but is not limited to, CaMV 35S, actin, ubiquitin, pEMU, MAS, histone promoter, Clp promoter. The term "promoter " refers to the region of DNA upstream from the structural gene and refers to a DNA molecule to which an RNA polymerase binds to initiate transcription. A "plant promoter" is a promoter capable of initiating transcription in plant cells. A "constitutive promoter" is a promoter that is active under most environmental conditions and developmental conditions or cell differentiation. Since the selection of the transformant can be carried out by various tissues at various stages, a constant promoter may be preferable in the present invention. Therefore, the constant promoter does not limit the selectivity.

In the recombinant vectors of the present invention, conventional terminators can be used, such as nopalin synthase (NOS), rice α-amylase RAmy1 A terminator, Agrobacterium tumefaciens ( Agrobacterium) but are not limited to, the Octopine gene terminator of the tumefaciens , the phaseoline terminator of E. coli, and the rrnB1 / B2 terminator of E. coli. Regarding the need for terminators, it is generally known that such regions increase the certainty and efficiency of transcription in plant cells. Therefore, the use of a terminator is highly desirable in the context of the present invention.

Methods of transporting vectors of the present invention into host cells include microinjection, calcium phosphate precipitation, electroporation, liposome-mediated transfection, DEAE-dextran treatment, Agrobacterium mediated transformation and gene balm The vector may be injected into the host cell by way of example, but is not limited thereto.

In addition, the present invention comprises the steps of transforming a plant cell with a recombinant vector comprising a OsRINGC2 OsRINGC2 -1 or -2 gene of rice origin; And it provides a method for producing a flame-resistant modified transgenic plant comprising the step of regenerating the plant from the transformed plant cell.

Method according to one embodiment of the present invention Salt Tolerance by by transforming each of the recombinant vector comprising the OsRINGC2 OsRINGC2 -1 or -2 gene derived from each of the rice plant cells overexpressing OsRINGC2 OsRINGC2 -1 or -2 gene in plant cells Characterized by producing this increased transgenic plant, but is not limited thereto.

Preferably the OsRINGC2 OsRINGC2 -1 or -2 gene may be comprised of the base sequence of SEQ ID NO: 1 or SEQ ID NO 2, respectively.

The method of the invention comprises the step of transforming a plant cell with a recombinant vector according to the present invention, the transformant is, for example, Agrobacterium tyumeo Pacific Enschede may be mediated by (Agrobacterium tumefiaciens). In addition, the method of the present invention comprises regenerating a transgenic plant from the transformed plant cell. Any of the methods known in the art can be used for regeneration of transgenic plants from transgenic plant cells.

Transformed plant cells must be regenerated into whole plants. Techniques for the regeneration of mature plants from callus or protoplast cultures are well known in the art for a number of different species (Handbook of Plant Cell Culture, Vol. 1-5, 1983-1989, Momillan, N. Y.).

The present invention also provides a flame retardant transgenic plant and seeds thereof prepared by the above method.

Preferably, the transgenic plant and its seeds are transgenic plants and seeds thereof characterized by increased salt resistance.

Preferably, the plant may be a dicotyledonous plant, more preferably Arabidopsis, potato, eggplant, tobacco, pepper, tomato, burdock, garland chrysanthemum, lettuce, bellflower, spinach, chard, sweet potato, celery, carrot, buttercup, It may be a dicotyledonous plant, such as parsley, Chinese cabbage, cabbage, cat radish, watermelon, melon, cucumber, pumpkin, gourd, strawberry, soybean, mung bean, kidney bean, pea, and most preferably, it may be a Arabidopsis plant, but is not limited thereto. .

The invention also OsRINGC2 -1 OsRINGC2 or rice derived each consisting of the nucleotide sequence of SEQ ID NO: 1 or SEQ ID NO: 2, there is provided a flame retardant for adjusting the composition of a plant containing the second gene. The composition consisting of the active ingredient in SEQ ID NO: 1 or the nucleotide sequence of SEQ ID NO: 2 or OsRINGC2 -1 OsRINGC2 includes 2 gene, respectively, to adjust the salt tolerance of a plant by transforming the gene in the plant (preferably, increasing You can do it.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Materials and methods

1. Plant materials and stress treatment

Germinated Rice ( Oryza sativa spp. japonica cv. Donganbyeo) was grown in a half MS medium maintained at a photoperiod and humidity of 70% at 16 hours of light (25 ℃) / 8 hours dark (23 ℃) conditions. Salt stress treatment was used 14 days old plants, MS medium with a concentration of 200mM NaCl. Samples were collected at 1, 6, 12 and 24 hours after treatment, respectively. Dry stress treatment was used for 14 days old plants and exposed to dry conditions. Cold stress treatment exposed the plants under dark conditions in a refrigerator at 4 ° C. Cold stress control control was used to prevent the light from entering the plant. Samples were collected at 1, 6, 12 and 24 hours after treatment, respectively. The hot stress treatment exposed the plants to a 45 ° C. temperature and collected samples at 1, 6, 12 and 24 hours after treatment respectively. Hormonal treatment was exposed to 14-day-old plants in medium containing 0.1 mM absic acid (ABA), 1 mM salicylic acid (SA) and 0.1 mM jasmonic acid (JA), respectively, and at 1, 6, 12, 24 and 48 hours. Each sample was collected. The prominent samples were harvested from experimental arable land at Chuncheon campus of Kangwon National University.

2. Osringc2 -One  And Osringc2 -2  Isolation of genes and Cloning

MSU rice genome annotation database 6.1 (Rice genome annotation database 6.1) -1 OsRINGC2 (LOC_Os11g01190) gene and OsRINGC2 -2 (LOC_Os12g01190) to obtain a coding sequence information about the gene in the (http://rice.plantbiology.msu.edu/). Total RNA was extracted using TRIzol reagent (Invitrogen) from 4 weeks old rice leaves and 1st cDNA (Takara) was prepared. PBIN35S plasmid after PCR using Pfu Turbo DNA polymerase (Stratagene) by constructing the forward (5'-GAGAGGATCCATGATGACCATGAGCGACGATG-3 ': SEQ ID NO: 3) and reverse (5'-GAGAGGTACCTCACAGTCCAAAAGCTCTGTTAT-3': SEQ ID NO: 4) primers Inserted in Remove the inserted plasmid pBIN35S after using the restriction enzymes Hind III and Kpn I did remove the OsRINGC2 OsRINGC2 -1 and -2 genes, respectively. Each isolated plasmid was analyzed after sequence identification.

3. RT - PCR

Total RNA was extracted using TRIzol reagent (Invitrogen) and 1st cDNA (Takara) was prepared. OsActinII gene (LOC_Os07g18100) was used to standardize the amount of each cDNA. Reference genes trusted from each treatment group are stress OssalT (LOC_Os01g24710), OsLIP19 (LOC_Os05g03860 ), OsHsp90 -1 (LOC_Os04g01740), OsPR1b (LOC_Os01g28450) and OsPBZ1 (LOC_Os12g36830) was used respectively. Specific primer design for each gene was used with the NCBI Primer BLAST tool (http://www.ncbi.nlm.nih.gov/tools/primer-blast/). PCR temperature conditions were denatured at 95 ° C. for 5 minutes, then repeated 30 ~ 33 times in a process of 30 seconds at 95 ° C., 30 seconds at 58 ° C., 1 minute at 72 ° C., and finally reacted at 72 ° C. for 5 minutes. .

4. 35S: Osringc2 -One- EYFP  And 35S: Osringc2 -2- EYFP Intracellular location analysis

OsRINGC2 to determine the location within the cell - the complete cDNA of the first and OsRINGC2 -2 pBIN-based 35S-EYFP vector (Lim et al . , unpublished). Each vector was infiltrated onto tobacco leaves using agroterium with gene silencing inhibitor p19 (Voinnet et al . , Plant J 33,949-956, 2003). Agrobacterium was grown to a concentration of 2 to 2.5 in A600, centrifuged for 20 minutes, rinsed three times using infiltration buffer (10 mM MES, 10 mM MgCl2, pH 5.6), and then aceto having a concentration of 200 μM. Siringon (Acetosyringone, 3 ', 5'-Dimethoxy-4'-hydroxyacetophenone) was added. The final concentration of the cells was adjusted to 0.5 at A ₆₀₀ and then infiltrated on the back of the tobacco leaf using a 1 ml syringe. Confocal microscopy was observed using tobacco leaves 3-5 days old. Fluorescence images were taken using the LSM 510 META NLO system (Carl Zeiss).

5. Osringc2  Observation of the Arabidopsis transformed with the gene

Each one of the cloned construct (35S: OsRINGC2 -1 and 35S: OsRINGC2 -2) was transformed in the Agrobacterium. Wild-type Arabidopsis larvae were grown in soil and transformed using soaking firearms (Zhang et. al . , Nat Protoc 1 , 641-646, 2006). T1 generation transgenic plants were selected in 1/2 MS medium with 50 mg / L kanamycin antibiotic concentration and T3 generation transgenic individuals less than one month after harvest were used. RT-PCR was used to confirm that the gene is expressed in the transgenic plant. MRNA expression of OsRINGC2 genes was confirmed using gene specific primers in each transgenic plant. Salt Stress In order to observe how stress affects transgenic plants, experiments were carried out at concentrations of 0, 100 and 150 mM NaCl, respectively. It was incubated at different OsRINGC2-1 and OsRINGC2 -2 1/2 MS medium having a concentration of 0, 100, 150mM NaCl with a seed of the transgenic plant after immersion in water for two days, and wild-type Arabidopsis thaliana. Growth conditions were adjusted to light conditions for 16 hours light / 8 hours at 22 ℃ temperature.

Example  One. Osringc2  Isolation of genes

Among the genes encoding the RING finger protein in rice, two RING-C2 type genes, LOC_Os11g01190 and LOC_Os12g01190, which are estimated to be generated by gene duplication, were selected and subjected to molecular biological analysis. These genes, produced by gene duplication, showed significantly higher sequence similarities of 99%. Full-length CDS was cloned into pBIN35S-RINGC2 for gene isolation, and each gene was isolated using Hind III and Kpn I (FIG. 1). These genes are, respectively, -1 OsRINGC2 (Oryza sativa RING finger protein C2-type 1) and OsRINGC2 -2 (Oryza sativa RING finger protein C2 type-2).

Example  2. RT - PCR  Through analysis Osringc2  Identify gene expression

RT-PCR was performed to identify the expression patterns of plant tissues and young seed stages, abiotic stress, and the expression of OsRINGC2 by plant hormones. First, it was confirmed that the amount of expression of more genes than OsRINGC2 -1 -2 OsRINGC2 by organization (A in Fig. 2). However, the developmental stage of young seed (Stage Stage I-II), it was confirmed that the amount of expression of the gene is higher than OsRINGC2-2 -1 OsRINGC2 (B in Fig. 2). Also abiotic stress in OsRINGC2 -1 gene (A in Fig. 3), of the plant hormone as compared to OsRINGC2-2 gene was found that the expression is increased by salt stress (200mM NaCl) it is only with the jasmonic acid treatment OsRINGC2 It was confirmed that the expression level of -1 gene slightly increased (B of FIG. 3). However, under different abiotic stress and plant hormone treatments, the expression patterns of the two genes were constant (FIG. 3).

Example  3. Confocal  Using a microscope Osringc2  Positional testing of proteins in plant cells

In order to determine the location of expression in OsRINGC2 proteins was performed OsRINGC2 -1 and Agrobacterium infiltration analysis (agro-infiltration assay) by cloning a gene OsRINGC2-2 the 35S :: EYFP vector using Agrobacterium. As a result, the 35S :: EYFP used as a control (A in FIG. 4) showed fluorescence signals in the nucleus and cytoplasm, and 35S :: OsRINGC2-1-EYFP showed numerous dot forms in the cells, and 35S :: OsRINGC2- Unlike 2-EYFP, fluorescent signals were observed in the nucleus and cytoplasm (Fig. 4 B, C).

Example  4. Salt Stress Resistance Assay Using Arabidopsis Transgenic Plants

The molecular function of the protein OsRINGC2 see their effect on the salt stress tolerance of the actual plants, the gene overexpression OsRINGC2 to make about how we react under salt stress, such as genes and OsRINGC2-1 OsRINGC2-2 created by the overlapping phenomena Transformed Arabidopsis was produced. After confirming each gene expression amount in each independent lines by RT-PCR, three lines with strong expression were selected and the salt stress resistance experiment was conducted (FIG. 5A).

Salt stress treatment proceeded with plant growth in medium of 0, 100 and 150 mM NaCl from the initial germination, and root length was checked after one week. First OsRINGC2 -1 transgenic plants grown in MS medium in the control medium Despite that the rapid root growth, OsRINGC2-2 transgenic plants did not show any differences and root length of the wild-type plants. 2 OsRINGC2 -1 transgenic plant of independent lines grown in 100mM NaCl added medium showed a difference (p <0.01) which is very significant and the root length of the wild-type plant. Also OsRINGC2 -2 transgenic plants showed a significant difference in the three independent line (p <0.05). OsRINGC2 -1 transgenic plants in 150 mM NaCl medium had similarly significant differences between two independent lines, which showed a significant difference in 100 mM NaCl medium ( p <0.01). On the other hand the OsRINGC2 -2 transgenic plant grown in 150mM NaCl added medium There was no difference between the wild type (Fig. 5). In conclusion, as shown E3 ligase which OsRINGC2 OsRINGC2 -1 and -2 genes in rice represents the resistance from the root is first subjected to salt stress, in particular to more role OsRINGC2 -1 gene is a salt stress tolerance. OsRINGC2 -1 gene conferring resistance to salt stress this will be able to be applied to a crop as a material for the molecular genetic breeding environment changed corresponding crops.

Attach an electronic file to a sequence list

Claims (13)

Recombination comprising OsRINGC2-1 (Oryza sativa RING finger protein C2 type-2) or OsRINGC2-2 (Oryza sativa RING finger protein C2 type-2) genes derived from rice, each consisting of the nucleotide sequences of SEQ ID NO: 1 or SEQ ID NO: 2 A method of controlling the flame resistance of a plant comprising the step of transforming the vector into plant cells. According to claim 1, characterized in that that by overexpressing OsRINGC2 OsRINGC2 -1 or -2 gene in the rice plant derived from a plant cell to increase the salt tolerance of plants. delete Recombination comprising OsRINGC2-1 (Oryza sativa RING finger protein C2 type-2) or OsRINGC2-2 (Oryza sativa RING finger protein C2 type-2) genes derived from rice, each consisting of the nucleotide sequences of SEQ ID NO: 1 or SEQ ID NO: 2 Transforming the plant cell with the vector; And
A method for producing a flame resistant modified transgenic plant comprising the step of regenerating a plant from the transformed plant cell. The method of claim 4, wherein the OsRINGC2 OsRINGC2 -1 or -2 of the gene derived from rice characterized in that for producing a transgenic plant with increased salt tolerance by over-expression in plant cells. delete A flame resistant modified transgenic plant prepared by the method of claim 4. 8. The plant of claim 7, wherein the plant has increased salt resistance. 8. The transgenic plant of claim 7, wherein the plant is a dicotyledonous plant. 10. The transgenic plant of claim 9, wherein the dicotyledonous plant is Arabidopsis thaliana. Seeds of plants according to claim 7. A composition for controlling salt resistance of a plant, comprising a rice-derived OsRINGC2-1 (Oryza sativa RING finger protein C2 type-1) gene consisting of the nucleotide sequence of SEQ ID NO: 1. A composition for regulating flame resistance of a plant comprising OsRINGC2-2 (Oryza sativa RING finger protein C2 type-2) gene derived from rice, consisting of the nucleotide sequence of SEQ ID NO: 2.

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