KR101683721B1 - Transgenic plant with enhanced tolerance and resistance to environmental stress by introducing grxB gene and preparation method thereof - Google Patents

Abstract

The present invention relates to a composition for improving the environmental stress resistance of a plant comprising the grxB gene, and a plant cell or plant transformed with the composition. In addition, the present invention relates to a method for producing a transgenic plant having enhanced resistance to environmental stress by transforming a plant of the grxB gene into a plant. The present invention also relates to a method of transforming a plant into a plant to enhance resistance to environmental stress.

Description

TECHNICAL FIELD The present invention relates to a transgenic plant having enhanced tolerance and resistance to environmental stress in which a grxB gene is transformed and a method for producing the same.

The present invention relates to a composition for enhancing environmental stress resistance of a plant containing a grxB (glutaredoxin B) gene, and a plant cell or plant transformed with said composition. In addition, the present invention relates to a method for producing a transgenic plant having enhanced resistance to environmental stress by transforming a plant of the grxB gene into a plant. The present invention also relates to a method of transforming a plant into a plant to enhance resistance to environmental stress.

Reactive oxygen species (ROS) generated through the activity of various oxidase and peroxidase enzymes act as an intracellular signaling substance in response to an internal developmental signal or an external environmental stimulus, The expression of genes necessary for defense is induced, but the increase of ROS concentration causes physiological disorder. ROS in plants may accumulate excessively due to various abiotic environmental stresses such as drought, high temperature, low temperature, salinity, dryness, pollution, wound, ozone, excessive ultraviolet ray exposure and osmotic shock, (macromolecules) and can lead to cell death.

Recently, interest in ROS-mediated signal transduction pathways has been increasing in plants. Regulation of these pathways may increase the expression of antioxidant enzymes and bio-defense proteins in cells, and ultimately resistance to various environmental stresses It is expected to be able to develop plants.

On the other hand, the physiological role of glutaredoxin (Grx) in plant antioxidant systems is not well known. Grx is known as a glutathione-dependent reductase present in almost all animals, plants and microorganisms and catalyzes glutathione-dependent reactions to protect specific cellular proteins from damage by reactive oxygen species. E. coli has five Grx ( EcGrxA , EcGrxB , EcGrxC , EcGrxD , and EcNrdH ), and Arabidopsis has 31 Grx.

The present inventors have studied the effect of the Grx genes of E. coli on the environmental stress tolerance of plants. In the case of the transformed plant, the grxB gene has been found to be resistant to high salt stress, high temperature stress, dry (drought) stress, Stress and ultraviolet stress, and completed the present invention.

Korea Patent Registration No. 10-1431124 (Aug. 18, 2014)

It is an object of the present invention to provide a composition for improving the environmental stress resistance of a plant, which comprises the grxB gene.

It is another object of the present invention to provide a plant cell transformed with the above composition.

It is another object of the present invention to provide a plant transformed with said composition.

It is another object of the present invention to provide a method for producing a transgenic plant having enhanced resistance to environmental stress, which comprises transforming a plant into a plant.

It is another object of the present invention to provide a method for enhancing the resistance of a plant to environmental stress, including a step of transforming a plant into a plant.

In order to achieve the above object, the present invention relates to a composition for improving the environmental stress resistance of a plant, which comprises the grxB gene.

In a preferred embodiment, the environmental stress includes environmental stress such as high salt stress, high temperature stress, dry (drought) stress, submergence stress, hypoxic stress or ultraviolet stress.

In a preferred embodiment, the grxB gene may be derived from E. coli.

Preferably, the grxB gene may comprise a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 1, and more preferably, the grxB gene may comprise the nucleic acid sequence of SEQ ID NO: 2.

Also preferably, the grxB gene may be in the form of a recombinant vector for plant expression.

In another aspect, the present invention relates to a plant cell transformed with the above composition and having enhanced environmental stress resistance.

In another aspect, the present invention relates to a plant transformed with the above composition and having enhanced environmental stress resistance.

In another aspect, the present invention relates to a method for producing a transgenic plant having enhanced resistance to environmental stress, comprising the step of transforming a plant into a plant.

In another aspect, the present invention relates to a method for enhancing the resistance of a plant to environmental stress, including the step of transforming a plant into a plant.

Hereinafter, the present invention will be described in more detail.

Glutaredoxin (Grx) is known as a glutathione-dependent reductase present in almost all animals, plants and microorganisms. It has CXXC and CXXS as active sites and catalyzes the glutathione-dependent reaction, Protects specific cellular proteins. E. coli has five Grx ( EcGrxA , EcGrxB , EcGrxC , EcGrxD , and EcNrdH ).

In the present invention, the grxB gene is preferably derived from Escherichia coli. Herein, the grxB gene derived from E. coli has the nucleic acid sequence of SEQ ID NO: 2, and the protein encoded thereby has the amino acid sequence of SEQ ID NO: 1.

Preferably, in the present invention, the grxB gene comprises a nucleic acid molecule comprising a functionally equivalent codon or a codon encoding the same amino acid (by codon degeneracy), or a codon encoding a biologically equivalent amino acid. In addition, considering a mutation having biological equivalent activity, the gene used in the present invention is interpreted to include a sequence showing substantial identity with the sequence described in the sequence listing. The above-mentioned substantial identity is determined by arranging the above-mentioned sequence of the present invention and any other sequence to correspond to each other as much as possible, and analyzing sequences arranged using algorithms commonly used in the art, at least 60% , More preferably 70% homology, even more preferably 80% homology, and most preferably 90% homology. Arrangement methods for sequence comparison are known in the art.

In a situation where it is not known how Grx affects environmental stress resistance in the plant antioxidant system, the inventors have for the first time demonstrated that the resistance to environmental stress of the plant transformed with the grxB gene is greatly improved. Therefore, a plant having enhanced environmental stress resistance can be produced using the grxB gene.

Introduction of the grxB gene into the plant may be accomplished by a variety of methods known in the art, and preferably an expression vector for plant transformation may be used. Suitable promoters to be included in the vector may be any of those conventionally used in the art for transgenic introduction of plants, such as SP6 promoter, T7 promoter, T3 promoter, PM promoter, ubiquitin promoter of corn, (CaMV) 35S promoter, nopaline synthase (nos) promoter, Piguet mosaic virus 35S promoter, water crane basil lipid viral promoter, comeline yellow mothball virus promoter, ribulose-1,5- (TPI) promoter of Arabidopsis, adenine phospholiposyl transferase (APRT) promoter of Arabidopsis, a tryptophan synthase promoter of Octopane synthase (ssRUBISCO), a rice cytosolic triosephosphate isomerase An azeotropic promoter and a blue copper binding protein (BCB) promoter. .

The vector may also comprise a poly A signal sequence which results in a 3'-terminal polyadenylation, for example from the nopaline synthase gene of Agrobacterium tumefaciens (NOS 3 ' end of the protease inhibitor I or II gene of the tomato or potato, the CaMV 35S terminator and the octopine synthase terminator sequence of Agrobacterium tumefaciens. can do.

In addition, the vector may optionally further comprise a gene encoding a reporter molecule (e.g., luciferase and beta -glucuronidase), and may further comprise an antibiotic (such as neomycin, carbenicillin, kanamycin, spectinomycin, hygromycin, etc.) or a herbicide (e.g., Basta) resistance gene (e.g., neomycin phospho-transferase dehydratase (nptⅡ), hygromycin phospho transferase dehydratase comprise the like) (hpt) .

According to a preferred embodiment of the present invention, the recombinant vector for plant expression of the present invention does not contain an Agrobacterium binary vector, a cointegration vector or a T-DNA site but is designed so as to be expressed in a plant Can be used. Of these, the binary vector refers to a vector having two plasmids containing a plasmid having a left border (RB) and a right border (RB) necessary for movement in a Ti (tumor inducible) plasmid and a gene necessary for transferring a target nucleotide , A vector containing a promoter region and a polyadenylation signal sequence for expression in a plant can be used.

When a binary vector or a cointegration vector is used, it is preferable to use Agrobacterium - mediated transformation as a transformation strain for introducing the recombinant vector into a plant ( Agrobacterium - mediated transformation) Agrobacterium tumefaciens ) Or Agrobacterium ( Agrobacterium < RTI ID = 0.0 > rhizogenes ) Can be used. In the case of using a vector that does not contain a T-DNA region, electroporation, microparticle bombardment, polyethylene glycol-mediated uptake and the like are used to introduce a recombinant plasmid into a plant .

Plants to be transformed herein are understood to include both plant cells, plant tissues, and plant seeds that develop into adult plants as well as mature plants.

Plants to which the method of the present invention can be applied are not particularly limited. The plants to be transformed in the present invention include food crops selected from the group consisting of rice, wheat, barley, corn, soybean, potato, wheat, red bean, oats and sorghum; Vegetable crops selected from the group consisting of Arabidopsis, cabbage, radish, red pepper, strawberry, tomato, watermelon, cucumber, cabbage, melon, squash, onions, onions and carrots; Ginseng, tobacco, cotton, sesame, sugar cane, beet, perilla, peanut and rapeseed; Apple trees, pears, jujubes, peaches, sheep grapes, grapes, citrus fruits, persimmons, plums, apricots and banana; Roses, gladiolus, gerberas, carnations, chrysanthemums, lilies and tulips; And feed crops selected from the group consisting of Ryegrass, Red Clover, Orchardgrass, Alpha Alpha, Tall Fescue, and Fereniallaigrus. Preferably, the plant is selected from the group consisting of Arabidopsis, potato, eggplant, cigarette, pepper, tomato, burdock, ciliaceae, lettuce, bellflower, spinach, modern sweet potato, celery, carrot, parsley, parsley, cabbage, cabbage, Can be a dicotyledon such as melon, cucumber, amber, pak, strawberry, soybean, mung bean, kidney bean, or pea, and most preferably it can be a rice cob.

By introducing the grxB gene into plants, it is possible to increase tolerance and resistance to high salt, high temperature, drought, submergence, hypoxia and ultraviolet stress, so that it can tolerate abnormal environmental stress due to climate change, The productivity of crops or flower crops in the soil can be promoted.

FIG. 1 shows a process for producing a plasmid for transforming plants with a GrxB gene (hereinafter referred to as " EcGrxB "
Figure 2 shows the results of confirming the expression of EcGrxB in transgenic plants.
Figure 3 shows the shoot form of the EcGrxB transgenic plant.
Figure 4 shows that the wild type and EcGrxB And the root phenotype of transgenic plants.
FIG. 5 shows the results of a comparison between the wild type and EcGrxB The root length of the transgenic plants was measured. Root length measurements were performed independently three times (p-value <0.01, n = 6).
Figure 6 shows the sensitivity of EcGrxB transgenic plants to high salt stress. (A) was taken in the form of wild-type and EcGrxB transgenic plants after high salt stress treatment, and the pictures were taken after 7 days of recovery. (B) The survival rate of wild-type and EcGrxB transgenic plants, the experiment was repeated three times (n = 12: EcGrxB transgenic plants, n = 20: wild type).
Figure 7 shows the sensitivity of EcGrxB transgenic plants to thermal stress. (A) In the form of wild-type and EcGrxB transgenic plants after thermal stress treatment, the photographs were taken after 14 days of recovery. (B) The survival rate of wild-type and EcGrxB transgenic plants, the experiment was repeated three times (n = 12: EcGrxB transgenic plants, n = 20: wild type).
Figure 8 shows the sensitivity of EcGrxB transgenic plants to drought stress. (A) In the form of wild-type and EcGrxB transgenic plants after drought stress treatment, photographs were taken after 7 days of recovery. (B) The survival rate of wild-type and EcGrxB transgenic plants, the experiment was repeated three times (n = 12: EcGrxB transgenic plants, n = 20: wild type).
Figure 9 shows the sensitivity of EcGrxB transgenic plants to submergence stress. (A) In the form of wild-type and EcGrxB transgenic plants after immersion stress treatment, the photographs were taken after 7 days of recovery. (B) Survival of wild-type and EcGrxB transgenic plants, the experiment was repeated three times (n = 12, respectively).
Figure 10 shows the sensitivity of EcGrxB transgenic plants to hypoxic stress. (A) In the form of wild-type and EcGrxB transgenic plants after hypoxic stress treatment, the photographs were taken after 3 days of recovery. (B) Survival of wild-type and EcGrxB transgenic plants, the experiment was repeated three times (n = 19, respectively).
Figure 11 shows the sensitivity of EcGrxB transgenic plants to UV-B stress. (A) In the form of wild-type and EcGrxB transgenic plants after UV-B stress treatment, the photographs were taken after 3 days of recovery. (B) Survival of wild-type and EcGrxB transgenic plants, the experiment was repeated three times (n = 12, respectively).

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

Example  1. Plant material and growth conditions

Arabidopsis thaliana ecotype Columbia seedlings were sterilized for 10 minutes with washing solution (0.05% Triton-X 100: Clorox: sterilized distilled water = 3: 2: 2) and washed 4-6 times with sterile distilled water. Cancer treatment. MS planted in the culture medium (0.8% phytoagar, 0.5x MS ( Murashige and Skoog medium, Duchefa), 1% sucrose), 100 μmolm -2 s - were grown under a light intensity of ^one. After 7 days, the plants were transferred to soil and grown in a 23-day growth chamber maintained for 9 days at 16 hr / 8 hr day-night cycle and 65% humidity.

Example  2. EcGrxB  Transgenic plant production

The process for producing EcGrxB transgenic plants is shown in Fig. (SEQ ID NO: 3) and reverse priem: 5'-AGC GGA TCC TTA AAT CGC CAT TGA-3 '(SEQ ID NO: 3) in the genomic DNA of the extracted E. coli. SEQ ID NO: 4) was used to amplify the EcGrxB gene. The amplified EcGrxB gene was inserted into the pSAT6.rbcP.MCS.rbcT entry vector and inserted into the pRCS2.ocs.bar using the nucleic acid internal hydrolase PI - Psp I. The inserted gene sequence was confirmed by DNA sequencing, and two kinds of rbc promoter forward primer (5'-gtccgttagatagcaaacaaca-3 '(SEQ ID NO: 5) and EcGrxB gene specific reverse primer were used. (75% viability) and T3 (100% viability) were grown in 50 μg ml ^-1 PPT (phosphinothricin) medium by the addition of 20 μg ml ^-1 BASTA, Survival rate).

Example  3. Abiotic  Sensitivity to stress

After treatment of the stresses described below, the sensitivity to stress through the number of surviving plants in total plants was measured.

3-1. Salt stress

To test for susceptibility to high salt, 350 mg NaCl solution was treated for 24 hours on a 16-day plant (grown in 0.5X MS medium for 7 days and then in soil for 9 days). One week later, the plants were again given water for 1-2 weeks.

3-2. Heat stress

To test for sensitivity to high heat, 16 plants (grown in 0.5X MS medium for 7 days and grown in soil for 9 days) were then treated at 50 ° C for 3 hours. One week later, the plants were again given water for two weeks. I put black plastic over the soil to avoid drying stress.

3-3. Drought (dry) stress

To test for susceptibility to drying, 13 plants (grown in 0.5X MS medium for 7 days and grown in soil for 6 days) were not watered for almost 17 days. Then, the plants were given water for one week to recover.

3-4. Immersion stress

To test for sensitivity to immersion, 21 plants (grown in 0.5X MS medium for 7 days and grown in soil for 14 days) were placed in a water-filled box capable of underwater stress. After almost a week, the plants were restored for 1-2 weeks.

3-5. Hypoxia  stress

To test for sensitivity to hypoxia, plants were grown in a hypoxic gas mixture (0.1% O ₂ / 99.9% N ₂ ) for 14 days in a plant (grown in 0.5X MS for 7 days and then grown in 0.5X MS without sucrose for 7 days) Cows were treated for 5 days. The plants were then restored under normal conditions for 3-5 days.

3-6. UV-rays( UV -B) stress

In order to test the sensitivity to ultraviolet light, the plants were grown on a 14-day plant (grown in 0.5X MS medium for 7 days and grown in 0.5X MS medium without sucrose for 7 days) with 15 μmol m ^-2 s ^-1 UV-B light (ETX-20M , Vilbert Lourmat) for 30 minutes. Then, the plants were restored for 3 days.

Experiment result

One. EcGrxB  Transgenic plant production

To investigate ectopic expression of the E. coli GrxB gene in Arabidopsis, an EcGrxB transgenic plant was constructed. Three T3 lines plants (named EcGRxB # 4-5, EcGRxB # 10-8 and EcGRxB # 11-7, respectively) were selected and the expression of EcGrxB gene in transgenic plants was determined by RT-PCR (Fig. 2).

2. EcGrxB  Characteristics of growth and development in transgenic plants

Most of the EcGrxB transgenic plants were similar to the wild type in leaf growth and root height. However, the leaf size of EcGrxB transgenic plants was slightly larger than that of the wild type (Fig. 3). The root length of the EcGrxB transgenic plants was slightly shorter than that of wild type (FIGS. 4 and 5).

3. EcGrxB  Of transgenic plants Abiotic  Sensitivity to stress

To see how the effect of EcGrxB expression is, EcGrxB We treated six different stresses on transgenic plants. On day 16, the plants grown and growing in the medium for 7 days gave high salt, heat, and drought stress, while the 21st plants treated immersion stress. Plants grown for 14 days in the medium were treated with hypoxic and UV-B stress.

(One) High salt  stress

All EcGrxB in Arabidopsis To test whether gene expression could enhance high salt resistance, three T3 lines of 16-day old wild-type and EcGrxB transgenic plants were treated with 350 mM NaCl for 24 hours. The high salt treatment was carried out with the surviving wild type and EcGrxB Resulting in typical damage, necrosis, and whitening in leaves of transgenic plants. Plant survival rates were observed after 2 weeks of recovery under normal conditions. Overall, the survival rate of wild-type was 22% based on t- test ( P <0.01) and 27.78% for EcGrxB (Fig. 6).

Number of plants surviving high salt treatment n = 4 test1 test2 test3 test4 test5 WT  (n = 20) 4 4 3 5 6 EcGrxB T3  # 4-5 2 One One One One EcGrxB T3  # 10-8 2 2 One 2 2 EcGrxB T3  # 11-7 One 2 One One One EcGrxB  (n = 12) 5 5 3 4 4

Survival rate of plants at high salinity (%) n = 4 test1 test2 test3 test4 test5 average stdev p- value WT  (n = 20) 20 20 15 25 30 22 5.70 EcGrxB T3  # 4-5 50 25 25 25 25 30 11.18 0.2560 EcGrxB T3  # 10-8 50 50 25 50 50 45 11.18 0.0036 EcGrxB T3  # 11-7 25 50 25 25 25 30 11.18 0.2560

(2) Heat stress

All EcGrxB in Arabidopsis To test whether gene expression could enhance thermal resistance, expression lines of 16-day-old wild-type and EcGrxB transgenic plants were treated at 50 ° C for 3 hours. The high-temperature treatment was carried out with the surviving wild type and EcGrxB Resulting in typical damage, necrosis, and whitening in leaves of transgenic plants. Plant survival rates were observed after 2 weeks of recovery under normal conditions. Overall survival rate was 36.11% on the basis of t- test ( P <0.01) and 61.11 % for EcGrxB transgenic plants, showing a much higher survival rate than the wild type (Fig. 7).

Number of plants surviving high temperature treatment n = 4 test1 test2 test3 WT  (n = 20) 5 3 5 EcGrxB T3  # 4-5 2 2 2 EcGrxB T3  # 10-8 4 2 3 EcGrxB T3  # 11-7 2 2 2 EcGrxB  (n = 12) 8 6 7

Survival rate of plants at high temperature treatment (%) n = 4 test1 test2 test3 average stdev p- value WT  (n = 20) 25 15 25 21.67 5.77 EcGrxB T3  # 4-5 50 50 50 50 0.00 0.0136 EcGrxB T3  # 10-8 100 50 75 75 25.00 0.0447 EcGrxB T3  # 11-7 50 50 50 50 0.00 0.0136

(3) Drought stress

All EcGrxB in Arabidopsis To test whether gene expression could enhance resistance to drought (dry), the three expression lines of 16-day old wild-type and EcGrxB transgenic plants were drought-treated for 17 days without water. Plant viability was observed after one week of recovery under normal conditions. Overall, the survival rate of wild-type was 31.67% based on t- test ( P <0.01) and EcGrxB 41.67%, showing a significant difference (Fig. 8).

Number of plants surviving drought treatment n = 4 test1 test2 test3 WT  (n = 20) 6 4 9 EcGrxB T3  # 4-5 3 One 3 EcGrxB T3  # 10-8 3 2 One EcGrxB T3  # 11-7 2 0 2 EcGrxB  (n = 12) 8 3 6

Survival rate of plants in drought treatment (%) n = 4 test1 test2 test3 average stdev p- value WT  (n = 20) 30 20 45 31.67 12.58 EcGrxB T3  # 4-5 75 25 75 58.33 28.87 0.1496 EcGrxB T3  # 10-8 75 50 25 50.00 25.00 0.4493 EcGrxB T3  # 11-7 50 0 50 33.33 28.87 0.8995

(4) Immersion stress

All EcGrxB in Arabidopsis To test whether gene expression could enhance immersion resistance, the three expression lines of 21-day-old wild-type and EcGrxB transgenic plants were submerged for 1 week. Inundation treatments were carried out using surviving wild type and EcGrxB Resulting in typical damage, necrosis, and whitening in leaves of transgenic plants. Plant viability was observed after one week of recovery under normal conditions. Overall, the survival rate of wild-type was 33.33% based on t- test ( P <0.01) and 52.08% for EcGrxB . Underwater stress handling EcGrxB Transgenic plants showed higher survival rates compared to wild-type conditions (Figure 9).

Number of plants surviving in flood treatment n = 4 test1 test2 test3 test4 WT  (n = 12) 3 3 6 4 EcGrxB T3  # 4-5 2 2 3 2 EcGrxB T3  # 10-8 2 2 2 3 EcGrxB T3  # 11-7 One 2 2 2 EcGrxB  (n = 12) 5 6 7 7

Survival rate of plants in inundation (%) n = 4 test2 test3 test4 average stdev p- value WT  (n = 20) 25 50 33.33 36.11 12.73 EcGrxB T3  # 4-5 50 75 50 58.33 14.43 0.0153 EcGrxB T3  # 10-8 50 50 75 58.33 14.43 0.2079 EcGrxB T3  # 11-7 50 50 50 50.00 0.00 0.1994

(5) Hypoxia  stress

All EcGrxB in Arabidopsis To test whether gene expression could enhance hypoxic stress resistance, three expression lines of wild-type and EcGrxB transgenic plants grown for 14 days were treated with 0.1% O ₂ / 99.9% N ₂ for 5 days. Hypoxic stress treatment is the surviving wild type and EcGrxB Resulting in typical damage, necrosis, and whitening in leaves of transgenic plants. Plant survival rates were observed after 3 days of recovery under normal conditions. Overall, the wild-type survival rate was 47.37% based on t- test ( P <0.01) and 61.40% for EcGrB . That is, the transgenic plants showed a very high survival rate compared to the wild type condition (Fig. 10).

Number of plants surviving the hypoxic stress treatment test1 test2 test3 WT  (n = 19) 9 8 10 EcGrxB T3  # 4-5 3 3 3 EcGrxB T3  # 10-8 5 4 5 EcGrxB T3  # 11-7 5 3 4 EcGrxB  (n = 15) 13 10 12

Survival rate of plants in hypoxic stress treatment (%) test1 test2 test3 average stdev p- value WT  (n = 19) 47.37 42.11 52.63 47.37 5.26 EcGrxB T3  # 4-5 (n = 5) 60 60 60 60.00 0.00 0.0533 EcGrxB T3  # 10-8 100 80 100 93.33 11.55 0.0087 EcGrxB T3  # 11-7 100 60 80 80.00 20.00 0.0878

(6) UV -B stress

All EcGrxB in Arabidopsis To test whether expression of the gene could enhance UV-B stress resistance, three expression lines of wild-type and EcGrxB transgenic plants grown for 14 days were irradiated with 15 μmol m ^-2 s ^-1 UV-B light for 30 minutes . The UV-B treatment is a survival of wild type and EcGrxB Resulting in typical damage, necrosis, and sudden whitening of the leaves of transgenic plants. Plant survival rates were observed after 3 days of recovery under normal conditions. Overall, the wild-type survival rate was 28.89 % based on t- test ( P <0.01) and 51.11% for EcGrxB . During UV-B treatment Transgenic plants had higher survival rates of EcGrxB transgenic plants compared to wild type (Fig. 11).

Number of surviving plants in ultraviolet treatment n = 4 test1 test2 test3 WT  (n = 12) 2 4 6 EcGrxB T3  # 4-5 2 3 3 EcGrxB T3  # 10-8 3 3 4 EcGrxB T3  # 11-7 3 3 3 EcGrxB  (n = 12) 8 9 10

Survival rate of plants in UV treatment (%) n = 4 test1 test2 test3 average stdev p- value WT  (n = 19) 16.67 33.33 50.00 33.33 16.67 EcGrxB T3  # 4-5 (n = 5) 40 60 60 53.33 11.55 0.0591 EcGrxB T3  # 10-8 60 60 80 66.67 11.55 0.0225 EcGrxB T3  # 11-7 60 60 60 60.00 0.00 0.1093

4. Conclusion

According to the results of previous physiological experiments, EcGrxB transgenic plants showed a high resistance to high salt, high temperature, drought (dry), submergence, hypoxic and UV stress. Therefore, introduction of EcGrxB gene into crops of flower crops is expected to be able to withstand abnormal environmental stress due to climate change or to grow in high salt soil such as reclaimed land.

<110> Ewha University - Industry Collaboration Foundation <120> Transgenic plant with enhanced tolerance and resistance to          environmental stress by indroducing grxB gene and preparation          method thereof <130> DPP20146008KR <160> 5 <170> Kopatentin 1.71 <210> 1 <211> 215 <212> PRT <213> Escherichia coli <220> <221> PEPTIDE &Lt; 222 > (1) <223> amino acid sequence of grxB <400> 1 Met Lys Leu Tyr Ile Tyr Asp His Cys Pro Tyr Cys Leu Lys Ala Arg   1 5 10 15 Met Ile Phe Gly Leu Lys Asn Ile Pro Val Glu Leu His Val Leu Leu              20 25 30 Asn Asp Asp Ala Glu Thr Pro Thr Arg Met Val Gly Gln Lys Gln Val          35 40 45 Pro Ile Leu Gln Lys Asp Asp Ser Arg Tyr Met Pro Glu Ser Met Asp      50 55 60 Ile Val His Tyr Val Asp Lys Leu Asp Gly Lys Pro Leu Leu Thr Gly  65 70 75 80 Lys Arg Ser Ser Ala Ile Glu Glu Trp Leu Arg Lys Val Asn Gly Tyr                  85 90 95 Ala Asn Lys Leu Leu Leu Pro Arg Phe Ala Lys Ser Ala Phe Asp Glu             100 105 110 Phe Ser Thr Pro Ala Ala Arg Lys Tyr Phe Val Asp Lys Lys Glu Ala         115 120 125 Ser Ala Gly Asn Phe Ala Asp Leu Leu Ala His Ser Asp Gly Leu Ile     130 135 140 Lys Asn Ile Ser Asp Asp Leu Arg Ala Leu Asp Lys Leu Ile Val Lys 145 150 155 160 Pro Asn Ala Val Asn Gly Glu Leu Ser Glu Asp Asp Ile Gln Leu Phe                 165 170 175 Pro Leu Leu Arg Asn Leu Thr Leu Val Ala Gly Ile Asn Trp Pro Ser             180 185 190 Arg Val Ala Asp Tyr Arg Asp Asn Met Ala Lys Gln Thr Gln Ile Asn         195 200 205 Leu Leu Ser Ser Ale Ile     210 215 <210> 2 <211> 648 <212> DNA <213> Escherichia coli <220> <221> gene &Lt; 222 > (1) .. (648) <223> nucleic acid sequence of grxB <400> 2 ttaaatcgcc attgatgata acaaattgat ttgtgtctgt ttcgccatat tatcgcggta 60 atcagcaacg cggcttggcc agttgattcc ggctaccagc gtcagattac gcagtagcgg 120 aaatagctga atatcatctt ccgaaagttc gccattcacg gcgttcggtt tgacgatcag 180 tttgtccagc gcacgcaaat catcgctgat attcttaatc agaccgtcag agtgggccag 240 caggtcggca aaattacccg cgctggcctc tttcttgtcg acgaaatatt tgcgcgcggc 300 gggagtagaa aactcatcaa atgccgattt ggcaaaacgc ggcaacagca gtttgttggc 360 gtagccattg actttgcgca gccactcttc aatggcaggg gaacgtttgc cggtcagtaa 420 cggtttgccg tcgagtttat cgacatagtg aacgatatcc atgctttctg gcatatagcg 480 gctgtcatct ttttgcagaa tgggaacctg tttttgaccg accatccggg tgggtgtttc 540 tgcgtcgtcg ttgagcagaa catgtaattc gacggggata tttttcaggc cgaaaatcat 600 gcgggctttg aggcagtaag ggcagtgatc gtaaatgtat agcttcac 648 <210> 3 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> forward primer <400> 3 acgctgcagg tgaagctata cat 23 <210> 4 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> reverse primer <400> 4 agcggatcct taaatcgcca ttga 24 <210> 5 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> rbc promoter forward primer <400> 5 gtccgttaga tagcaaacaa ca 22

Claims (12)

1. A composition for enhancing environmental stress resistance of a plant, which comprises a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 1 and a glxB (glutaredoxin B) gene derived from E. coli.
The composition of claim 1, wherein the environmental stress is high salt stress, high temperature stress, dry stress, submergence stress, hypoxic stress or ultraviolet stress.
delete delete 2. The composition of claim 1, wherein the grxB gene comprises the nucleic acid sequence of SEQ ID NO: 2.
The composition of claim 1, wherein the grxB gene is included in the form of a recombinant vector for plant expression.
An environmental stress resistance enhanced plant cell transformed with the composition of any one of claims 1 to 2 and 5 to 6.
An environmental stress resistance enhanced plant transformed with the composition of any one of claims 1 to 2 and 5 to 6.
(Glutaredoxin B) gene derived from E. coli consisting of a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 1,
A method for producing a transgenic plant having enhanced resistance to environmental stress.
10. The method of claim 9, wherein the environmental stress is high salt stress, high temperature stress, dry stress, submergence stress, hypoxic stress or ultraviolet stress.
(Glutaredoxin B) gene derived from E. coli consisting of a nucleic acid sequence encoding the amino acid sequence of SEQ ID NO: 1,
A method of enhancing the resistance of a plant to environmental stress.
12. The method of claim 11, wherein the environmental stress is high salt stress, high temperature stress, dry stress, submergence stress, hypoxic stress or ultraviolet stress.

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