KR101728889B1 - Nf-ya7 gene and transformants with enhanced tolerance to drought stress - Google Patents

Abstract

The present invention relates to a plant having enhanced resistance to arthritis by transformation with a recombinant expression vector comprising the NF-YA7 gene having the nucleotide sequence shown in SEQ ID NO: 1. The present invention also relates to a method for enhancing the tolerance of a plant by overexpressing a gene comprising the nucleotide sequence of SEQ ID NO: 1 encoding the NF-YA7 protein comprising the amino acid sequence of SEQ ID NO: 2. In this way, it is possible to develop drought tolerant plants that do not cause productivity problems due to climate change.

Description

[0001] The present invention relates to a novel NF-YA7 gene and a transgenic plant,

The present invention relates to a NF-YA7 gene and a potato transformant using the same to improve the resistance to dryness of potatoes.

Agriculture has unique risks and instabilities that are not found in other industries. Agriculture is not only uncertain because of the large loss of catastrophes as well as large fluctuations in agricultural prices and the relatively low price of agricultural produce compared to the purchase price of production materials. Agriculture is a natural hazard, and it is more vulnerable to damage than any other industry by a pair of floods, frosts, and hail. This is because agricultural production fields and production are constrained because they are closely connected to land that can not be moved, and because the objects of production are plants and plants, they are dominated by natural conditions depending on the constraints of life phenomena .

A pair of crops (drought, dry) causes crop loss due to lack of precipitation or there is no constant precipitation period, so crops are grown depending on irrigation water. A pair of crops can occur at any time during the cultivation period. In the early stage of growth, the flowering time is delayed, and when the long-term flowering is continued, the flowering time is delayed and the yield is seriously affected. Korea has abundant annual precipitation of 1,250 mm, but it is biased in July-August, so it has a semi-dry climate in spring and autumn. Therefore, spring and autumn habitually take a pair, sometimes even in July-August, due to shortage of rainfall (害 害).

With the development of plant transgenic technology, it became possible to express foreign genes far away from each other. Therefore, introduction of genes resistant to poor environment into susceptible plants has made it possible to develop inherently degradable crops. Therefore, the separation and development of cold tolerant and one strain resistant genes are indispensable requirements for the development of these crops.

In particular, a pair of plants causes moisture deficiency due to the osmotic pressure of crops, causing morphological and metabolic changes in the plants. This lack of water causes not only morphological changes of plants but also clogging of pores, reduction of photosynthesis rate and photo respiration rate, increase of small molecules and changes of plant hormones, and changes of gene expression.

As the population grows, food production continues to be demanded, but agricultural and agricultural land is declining. In addition, since the crops are in a stable state, when the surrounding environment changes, there is a great deal of stress during the growing period, resulting in a large difference in yield. Although Korea is in a temperate climate, unusual droughts and high temperature phenomena occur frequently due to global climate change. In the past 10 years (1995-2004), the weather disaster area due to flood, Thousand ha, corresponding to 8.87% of the total land area of 1,836 thousand ha in Korea. Such a decrease in agricultural productivity due to natural disasters is serious, and the development of crops that can withstand these poor environments is absolutely required to increase food production.

 The inventors of the present invention found the NF-YA7 gene of the present invention by testing the possibility of drought-tolerant plants using a gene resistant to drought. By using the gene of the present invention, It is possible to produce a crop which does not cause a problem in the productivity according to the present invention.

Korean Patent Publication No. 2013-0126068

An object of the present invention is to provide a plant having enhanced tolerance by transforming with a recombinant expression vector containing the NF-YA7 gene having the nucleotide sequence of SEQ ID NO: 1.

Another object of the present invention is to provide a method for promoting tolerance of a plant by overexpressing a gene comprising the nucleotide sequence of SEQ ID NO: 1 encoding the NF-YA7 protein comprising the amino acid sequence of SEQ ID NO: 2.

It is still another object of the present invention to provide a composition for promoting tolerance of a plant comprising a gene consisting of the nucleotide sequence of SEQ ID NO: 1 encoding the NF-YA7 protein comprising the amino acid sequence of SEQ ID NO: 2.

In order to achieve the above object,

The present invention provides a plant having enhanced tolerance by transforming with a recombinant expression vector comprising NF-YA7 (nuclear factor-Y) gene having the nucleotide sequence shown in SEQ ID NO: 1.

The NF-YA7 gene may be isolated from the potato. In the embodiment of the present invention, the abiotic stress-related gene is isolated from the potato, and the expression level is confirmed by stress treatment such as salt, drying, Gene.

The plant cell into which the recombinant expression vector of the present invention is introduced is not particularly limited to a specific form as long as the cell can be regenerated as a plant. These cells include, for example, cultured cell suspension, protoplasts, leaf sections and callus. Such plants include, but are not limited to, potatoes, rice, cabbage, cabbage, mustard, rapeseed, radish, Brassica napobrassica , and turnip ( Brassicarapa / Brassica campestris ), and preferably one or more selected from the group consisting of potatoes, especially subspecies.

More specifically, the plant having improved resistance to dryness according to the present invention can be obtained by transforming a plant with a recombinant expression vector containing the NF-YA7 gene, followed by induction of callus, rooting and soil purification according to a conventional method have.

The present invention also provides an NF-YA7 protein having an amino acid sequence represented by SEQ ID NO: 2. The present invention includes homologous proteins having homology with the above proteins.

The "homologous protein" has a sequence homology of preferably 80% or more, more preferably 90% or more, still more preferably 95% or more, most preferably 99% or more with the amino acid sequence of SEQ ID NO: 2 Quot; refers to a protein that exhibits substantially the same function as the NF-YA7 protein of the present invention.

The present invention also encompasses the NF-YA7 gene having the nucleotide sequence shown in SEQ ID NO: 1 and its equivalent. The equivalents include homologous genes having homology.

The NF-YA7 gene having the nucleotide sequence shown in SEQ ID NO: 1 or its homologous gene is encoded by a protein.

The "homologous gene" is a gene having a sequence homology of preferably 80% or more, more preferably 90% or more, still more preferably 95% or more, most preferably 99% or more with the nucleotide sequence of SEQ ID NO: 1 Refers to a gene that exhibits substantially the same function as the NF-YA7 gene of the present invention. Sequence homology can be analyzed by methods known in the art.

The "substantially homogeneous function" means that it is involved in improving the weatherability. Such functional equivalents include, for example, amino acid sequence variants in which some of the amino acids of the amino acid sequence are substituted, deleted or added. Substitution of amino acids is preferably conservative substitution. Examples of conservative substitutions of amino acids present in nature are as follows: (Gly, Ala, Pro), hydrophobic amino acids (Ile, Leu, Val), aromatic amino acids (Phe, Tyr, Trp), acidic amino acids (Asp, Glu), basic amino acids (His, Lys, Arg, Gln, Asn ) And sulfur-containing amino acids (Cys, Met). Deletion of the amino acid is preferably located at a site that is not directly involved in the activity of NF-YA7 of the present invention. The functional equivalents also include protein derivatives in which the basic skeleton of NF-YA7 and its chemical structure has been modified while maintaining its physiological activity. These include, for example, fusion proteins made by fusion with other proteins such as GFP, while retaining the structural modification and physiological activity to change the stability, storage stability, volatility or solubility of the protein of the present invention.

The present invention also provides a method for enhancing the tolerance of a plant by overexpressing a gene consisting of the nucleotide sequence of SEQ ID NO: 1 encoding the NF-YA7 protein comprising the amino acid sequence of SEQ ID NO: 2.

A method of overexpressing the gene may include transforming the plant with a recombinant expression vector in which the gene is operably linked to a promoter. The target plants according to the present invention include, but are not limited to, potatoes, rice, cabbage, cabbage, mustard, rapeseed, radish, Brassica napobrassica and turnip ( Brassicarapa / Brassica campestris ), preferably potatoes, especially subspecies.

In an embodiment of the present invention, the present inventors prepared recombinant expression vectors containing the NF-YA7 gene having the nucleotide sequence shown in SEQ ID NO: 1 and transformed the potato cells, whereby the expression level of NF-YA7 protein in potato cells Respectively.

The transformant according to the present invention is not only a gene having the nucleotide sequence of SEQ ID NO: 1 but also a modified sequence in which some bases of SEQ ID NO: 1 are substituted, deleted or added, A base sequence encoding a protein that exhibits an activity equivalent to the activity of the protein encoded by the plant, i.e., the plant resistance to the plant, can be used.

The NF-YA7 gene according to the present invention can be inserted into a suitable expression vector, that is, a recombinant expression vector to transform plant cells.

In another aspect, the present invention also provides a gene encoding a NF-YA7 protein having the amino acid sequence shown in SEQ ID NO: 2 or a gene encoding the NF-YA7 gene having the nucleotide sequence shown in SEQ ID NO: 1, Lt; RTI ID = 0.0 > expression vector. ≪ / RTI >

The expression vector means a plasmid, virus or other medium known in the art to which the gene according to the present invention can be inserted or introduced. A gene according to the present invention may be operably linked to an expression control sequence, wherein the operably linked gene sequence and expression control sequence are contained within an expression vector containing a selection marker and a replication origin .

The term "operably linked" means that one nucleic acid fragment is associated with another nucleic acid fragment so that its function or expression is affected by other nucleic acid fragments.

The expression control sequence is a DNA sequence that regulates the expression of a gene operably linked to a particular host cell. Such a regulatory sequence includes a promoter for transcription, an arbitrary operator sequence for regulating transcription , Sequences that encode suitable mRNA ribosome binding sites, and sequences that control the termination of transcription and translation.

The term "promoter" refers to a DNA sequence that controls the expression of a gene operably linked to a specific host cell. As the promoter, a constitutive promoter that induces the expression of the target gene at all times at any time, or an inducible promoter that induces the expression of the target gene at a specific position and time can be used. However, A promoter that induces expression of a gene is used, and examples thereof include, but are not limited to, a p35S promoter. In addition, a ubiquitin promoter can be used to overexpress genes in monocotyledonous plants or woody plants.

The introduction of the recombinant expression vector into a plant can be carried out by a method known in the art. For example, but not limited to, Agrobacterium sp. -Mediated methods, particle gun bombardment, silicon carbide whiskers, sonication, heat shock A heat shock, an electroporation method and a precipitation method using PEG (Polyethylenglycol) can be used. In one embodiment of the present invention, a plant cell is transformed into a recombinant vector of the present invention by a method of Agrobacterium-mediated method, that is, a method of transforming plant cells using Agrobacterium to which an expression vector according to the present invention is introduced, But the present invention is not limited thereto.

In an embodiment of the present invention, a recombinant expression vector containing the NF-YA7 gene was introduced into Agrobacterium and transformed into potato using the introduced Agrobacterium. To examine the expression characteristics of the resistance gene, NF-YA7 overexpressing transformants were subjected to treatment tolerance tests such as low temperature, salt, and drying stress. As a result, it was confirmed that the dry resistance was greatly increased in the transformant.

The present invention also provides a composition for enhancing the tolerance of a plant comprising a gene consisting of the nucleotide sequence of SEQ ID NO: 1 encoding the NF-YA7 protein consisting of the amino acid sequence of SEQ ID NO: 2. In the composition of the present invention, the NF-YA7 gene may comprise the nucleotide sequence of SEQ ID NO: 1. The composition of the present invention contains the NF-YA7 gene as an active ingredient, and the resistance to potatoes can be improved by transforming the gene into a plant.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In general, the nomenclature used herein is well known and commonly used in the art.

By developing transgenic plants with increased dryness resistance according to the present invention, it is possible to produce disaster tolerant crops that do not cause problems in productivity even in sudden climate change such as global warming. It can also contribute to the development of drought - resistant potatoes and farm income.

FIG. 1 shows the results of RT-PCR analysis of the NF-YA7 gene of the present invention at various time points after 10% PEG, 250 mM NaCl, 50 μM ABA and low temperature (4 ° C.) stress treatment.
2 is a schematic diagram of an NF-YA7 over-transformed binary vector prepared using the NF-YA7 gene.
FIG. 3 shows the result of analysis of the expression pattern after 10% PEG treatment in NF-YA7 gene potato transformant by PCR method.
FIG. 4 shows the results of analysis of resistance to aging by PEG treatment of the transformant transformed with the NF-YA7 gene. (Left: photographs of transformants by period; right: comparison of growth state of transformants by period)
FIG. 5 shows the result of analysis of dry strength after NF-YA7 transgenic transgenic plants (A: one week after the dry stress treatment (above), three weeks after the dry stress treatment (below) A graph comparing the survival rate of the transgenic plants; a graph comparing the leaf moisture content of the NF-YA7 transgenic plants;
FIG. 6 is a graph comparing gene expression levels of the control and NF-YA7 gene transformants by the A: qPCR method; B: Intracellular expression of NF-YA7 gene transformant was confirmed by confocal microscopy (left: wild type; right: transformant).

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

Example 1. Isolation of NF-YA7 gene

The abiotic stress related genes were isolated from potatoes and confirmed to be unknown genes after sequencing. This gene was confirmed by stress treatment such as salt, drying, low temperature and ABA respectively and it was confirmed that it is a gene resistant to specific stress.

For the isolation of the NF-YA7 gene, we first extracted total RNA from potato leaves and synthesized first strand cDNA using sprint RT complete oligodT (Clontech).

Specifically, a primer (5'-GTCGACTTTGCGGCCGCATGTTTCAAAATCGGATGGTGAT-3 ') (SEQ ID NO: 3) and an antisense primer (5'-AGTATCGATGCGGCCGCAGAAATGTTCCGAGTGCATGCAA-3' 4) were synthesized and the gene was amplified using the Applied Biosystem 9700 machine in PCR reaction solution together with the Ex Taq polymerase of Takara Co. using the first strand cDNA of potato as a template. The PCR reaction conditions used were denaturation at 95 ° C for 10 min, followed by 25 cycles of 94 ° C for 1 min, 56 ° C for 1 min, and 72 ° C for 2 min, followed by treatment at 72 ° C for 10 min. After the PCR reaction, the amplified gene was electrophoresed on 1% agarose, and a band estimated as NF-YA7 gene was cut out and purified using a QIAquick gel extraction kit and then used for cloning. The obtained NF-YA7 anticipated gene was cloned into Topo 2.1 vector (Invitrogen, TOPO TA Cloning kit), plated on LB medium supplemented with X-gal, and the resulting white colonies were selected and subjected to colony PCR.

As a result, the plasmid of the band with the confirmed colony was extracted and confirmed to be the NF-YA7 gene using a sequencer ABI 3700 sequencer. As a result of the nucleotide sequence analysis, the NF-YA7 gene was obtained and confirmed to be a gene having the nucleotide sequence of 723 bp (SEQ ID NO: 1) and having 240 amino acid residues (SEQ ID NO: 2).

Example  2. Drying, salt, low temperature, after ABA treatment NF The RT- PCR  Analysis of Expression Patterns by

10% PEG, 250 mM NaCl, 50 μM ABA, Cold (4 ° C), and 10% PEG were added to the wild type potatoes in order to test resistance to salt resistance, low temperature, The first strand cDNA was synthesized by using Sprint RT Complete-OligodT (Clontech) after extracting total RNA from potato leaves after 30, 3, 6, 12 and 24 hours of treatment. Each gene forward primer (5'-GTCGACTTTGCGGCCGCATGTTTCAAAAATCGGATGGTGAT-3 ') (SEQ ID NO: 3) and an antisense primer (5'-AGTATCGATGCGGCCGCAGAAATGTTCCGAGTGCATGCAA-3' (SEQ ID NO: 4) was added to the PCR reaction solution and PCR was performed. Then, the expression level was confirmed by electrophoresis on 1% agarose.

NF-YA7 As a result of analysis of gene expression characteristics, the expression level of 10% PEG treatment increased with time, but the expression level tended to decrease slightly after 12 hours. In the analysis of expression after 250mM NaCl treatment, the expression level was slightly increased compared to the untreated group and then decreased after 12 hours. In the case of 50 袖 M ABA and low temperature treatment (4 째 C), there was no significant difference in the expression level as compared with the untreated group, but it was confirmed that the gene expression amount was increased during the treatment for 12 hours (Fig.

Example 3: Construction of vectors for potato transformation

The present inventors constructed a vector for transforming the NF-YA7 gene into potato.

Specifically, the NF-YA7 gene isolated in Example 1 was specifically inserted into a pORE-R3 vector, which is a binary vector for transformation into potatoes, by connecting the NF-YA7 gene to a 35S constant expression promoter, To prepare a vector for transformation (Fig. 2).

Example 4. Production of potato transformed with NF-YA7 gene

The vector prepared in Example 3 was transformed into Agrobacterium , and transgenic plants were prepared using Agrobacterium .

Specifically, in order to transform a binary vector into which an NF-YA7 gene has been inserted into Agrobacterium (C58), freezing and thawing were repeated 2 to 3 times, followed by heat shock (Yeast 10 g, NaCl 5 g, peptone 10 g, Agar 15 g / 1 L) overnight, and colonies were confirmed. Colony transformed by colony PCR was transformed into AB medium (AB buffer (K ₂ HPO ₄ 60 g, NaH ₂ PO ₄ 20 g / 1 L), AB Salts (NH ₄ Cl 60 g, MgSO ₄ 7H ₂ O 6g, KCl 3g, CaCl ₂ 2H ₂ O 0.265g, FeSO ₄ .7H ₂ O 50mg / 1L), Glucose 5g / 1L).

In order to obtain potato transformants, Agrobacterium-transformed genes cultivated in potato culture medium and YEP medium cultured in MS medium for 2 days were incubated with MS liquid medium for 30 minutes, and seeded with cefotaxime and kanamycin ) Was added to the MS solid medium and cultured for 7 days. The cells were transferred to callus induction medium for 7 days and cultured for 7 days in fresh MS medium supplemented with cefotaxime and kanamycin. The shoots were transferred to a new medium every 2 weeks for induction of shooting. After repeated shots, shoots were transplanted to obtain an overexpressed transformant with the NF-YA7 gene inserted therein.

As a result, 20 lines of potato transformants using the NF-YA7 gene were obtained. Among them, lines lacking the phenotype or genes were removed, and 20 lines selected for stress tolerance were subjected to stress tolerance Respectively.

Example 5. Analysis of expression pattern of NF-YA7 gene in NF-YA7 gene transformant

NF-YA7 gene-inserted transformants were analyzed for expression pattern after osmotic stress treatment.

Specifically, in order to analyze the expression of the NF-YA7 gene in the transgenic potato comprising the NF-YA7 gene selected in Example 4, the NF-YA7 gene was treated with PEG and RT-PCR Were analyzed.

Specifically, 10% PEG was treated for 0, 3, 6, 12 and 24 hours in NF-YA7 transgenic potato lines 2, 5, 8, 14, 15 and 16 lines and control group (Sumi). Then, total RNA was extracted from the leaves of NF-YA7 transgenic potatoes and first-strand cDNA was synthesized using Sprint RT Complete-OligodT (Clontech). Then, a primer for amplifying the fragment was used as a sense primer 5'-GTCGACTTTGCGGCCGCATGTTTCAAAAATCGGATGGTGAT -3 ') (SEQ ID NO: 3) and an antisense primer (5'-AGTATCGATGCGGCCGCAGAAATGTTCCGAGTGCATGCAA-3' agarose) to confirm the expression of the NF-YA7 gene.

As a result, the expression level of NF-YA7 gene in NF-YA7 transgenic potatoes was increased in all lines used compared to wild type. (Fig. 3).

Example  6. NF After phenotypic analysis of OsMotic stress in -YA7 gene transformants

After 0.3% PEG treatment by the method of Example 5, the growth state of the potatoes was examined at intervals of one week. As a result, it was confirmed that the growth condition of the NF-YA7 gene transformant was better than that of supermath.

In particular, the transformants into which the NF-YA7 gene was inserted showed that the growth state after 2 weeks of 0.3% PEG treatment was much better than that of Sumi (Fig. 4).

Example  7. NF -YA7 gene transformants after dry stress treatment Dry  Enhancement analysis

In order to confirm whether the NF-YA7 transgenic plants were resistant to drying, the drying stress was treated as follows. Wild type subspecies and NF-YA7 transgenic plants were subcultured in a culture room at 22 ° C for 3 weeks, followed by purification for 3-5 days. After that, it was cultivated again in the greenhouse for 3 weeks, grown about 20 cm, and then dried. Drying is done by weighing about 300-350g with enough water to the horticultural pollen in a greenhouse at about 23-25 ℃. When the weight of the pot is about 150g, Respectively.

As a result, it was confirmed that the NF-YA7 gene-inserted transformant was maintained in a healthy condition for about 3-5 days after the drying treatment. In addition, after drying stress, it was dried until almost to the end of the test and then rehydrated with water. As a result, it was confirmed that the transformant containing the NF-YA7 gene survived while the sumi was almost dead (Fig. 5A).

In addition, the survival rate of the NF-YA7 gene transfected with the NF-YA7 gene was found to be 50% or more in the transgenic lines 2 and 4 of the NF-YA7 gene, 30% survival rate (Fig. 5B)

In order to confirm the degree of water loss in the leaves of the NF-YA7 transgenic plants and the control group, the leaves were weighed on a line-by-line basis and weighed to determine the degree of water loss, 5C). As a result, it was confirmed that the degree of water loss was much lower than that of water.

Example 8. Identification of NF-YA7 gene expression in plants

Analysis of the expression level of the NF-YA7 transgenic plant using qPCR revealed that the expression level of the NF-YA7 transgenic plant was significantly increased as compared to the superior (Comparative Example) (Fig. 6A). For the qPCR reaction method, cDNA of each sample is synthesized using cDNA synthesis kit (cDNA EcoDry ™, Clontech cat # 639543), and then diluted 10-fold. The gene reaction is performed according to the protocol of the kit and repeated three times each. The qPCR machine used a BioRad MYiQ5 instrument and performed the reaction. The PCR reaction cycle was carried out at 95.0 ° C for 5 seconds, followed by 30 seconds at 95.0 ° C and 30 seconds at 58.0 ° C for 35 seconds. The reaction was repeated 40 times and finally at 72.0 ° C for 35 seconds. (Fig. 6A).

In addition, Confocal microscope was used to observe the expression pattern of NF-YA7 gene in the cells, and it was confirmed that the gene exists in the nucleus of the coculture of tobacco (Fig. 6B).

<110> Republic of Korea <120> NF-YA7 GENE AND TRANSFORMANTS WITH ENHANCED TOLERANCE TO DROUGHT          STRESS <130> P14R12D1381 <160> 4 <170> Kopatentin 2.0 <210> 1 <211> 723 <212> DNA <213> Solanum tuberosum <400> 1 atgtttcaaa aatcggatgg tgattccaag agagaaggca aacatgtaga atttatgcct 60 ccaagcatgg gtgaaaattt aagagcagca aatcaatttg aactaatggc accctcctca 120 attgcattta aatcataccc ctattcagaa gtaccacaat attctggtgg caatgttact 180 attgcttgtg gtgaacctat ggtaaatcaa aatatggaga gatcatcagc agttcatcat 240 aatggaagaa tgattttgcc agttgaagtg aaagaggagc caatgtatgt aaatgcaaaa 300 caataccatg gaattcttag acgaaggcaa cttcgtgcta aggctgtgtt gcaacaaaaa 360 gtggtcaaat ctcgaaagcc ttatcttcac gaatcgcgtc accgacacgc gatgaggaga 420 gctagagatg gtggaggtag atttctcaac acaaagaaga agacccaatc tactactaca 480 actaataaca acactactcc aagtagtaaa ggcaaaagtt caatggattc tgattcttct 540 ccaaattacc tactcaatta tgaacatgaa attggatcat ccaatcatag taattctgtt 600 gaaggatttc agttccaatc tggaatacat gatactacag aaaatcttca attgggttgt 660 cattatcagt ggaacctcaa tgacaacaac cattgcaatt gcatgcactc ggaacatttc 720 tga 723 <210> 2 <211> 240 <212> PRT <213> Solanum tuberosum <400> 2 Met Phe Gln Lys Ser Asp Gly Asp Ser Lys Arg Glu Gly Lys His Val   1 5 10 15 Glu Phe Met Pro Pro Ser Met Gly Glu Asn Leu Arg Ala Ala Asn Gln              20 25 30 Phe Glu Leu Met Ala Pro Ser Ser Ile Ala Phe Lys Ser Tyr Pro Tyr          35 40 45 Ser Glu Val Pro Gln Tyr Ser Gly Gly Asn Val Thr Ile Ala Cys Gly      50 55 60 Glu Pro Met Val Asn Gln Asn Met Glu Arg Ser Ser Ala Val His His  65 70 75 80 Asn Gly Arg Met Ile Leu Pro Val Glu Val Lys Glu Glu Pro Met Tyr                  85 90 95 Val Asn Ala Lys Gln Tyr His Gly Ile Leu Arg Arg Arg Gln Leu Arg             100 105 110 Ala Lys Ala Val Leu Gln Gln Lys Val Val Lys Ser Arg Lys Pro Tyr         115 120 125 Leu His Glu Ser Arg His His His Ala Met Arg Arg Ala Arg Asp Gly     130 135 140 Gly Gly Arg Phe Leu Asn Thr Lys Lys Lys Thr Gln Ser Thr Thr Thr 145 150 155 160 Thr Asn Asn Thr Thr Pro Ser Ser Lys Gly Lys Ser Ser Met Asp                 165 170 175 Ser Asp Ser Ser Pro Asn Tyr Leu Leu Asn Tyr Glu His Glu Ile Gly             180 185 190 Ser Ser Asn His Ser Asn Ser Val Glu Gly Phe Gln Phe Gln Ser Gly         195 200 205 Ile His Asp Thr Thr Glu Asn Leu Gln Leu Gly Cys His Tyr Gln Trp     210 215 220 Asn Leu Asn Asp Asn Asn His Cys Asn Cys Met His Ser Glu His Phe 225 230 235 240 <210> 3 <211> 41 <212> DNA <213> Artificial Sequence <220> <223> sense primer <400> 3 gtcgactttg cggccgcatg tttcaaaaat cggatggtga t 41 <210> 4 <211> 40 <212> DNA <213> Artificial Sequence <220> <223> antisense primer <400> 4 agtatcgatg cggccgcaga aatgttccga gtgcatgcaa 40

Claims (7)

A plant having enhanced resistance to arthritis by transformation with a recombinant expression vector comprising NF-YA7 (nuclear factor-Y) gene consisting of the nucleotide sequence shown in SEQ ID NO: 1,
Wherein the NF-YA7 gene is isolated from potatoes. delete The method of claim 1, wherein the plant is a potato, rice, Chinese cabbage, cabbage, mustard, rapeseed (rapeseed), radish, homu (Brassica napobrassica ) and turnips ( Brassicarapa / Brassica campestris ). &lt; / RTI &gt; A gene encoding a NF-YA7 protein consisting of the amino acid sequence of SEQ ID NO: 2 is over-expressed in a plant comprising the nucleotide sequence of SEQ ID NO: 1,
Wherein said gene is isolated from potatoes. 5. The method of claim 4, wherein the gene overexpression comprises transforming the plant with a recombinant expression vector in which the gene is operably linked to a promoter. The method according to claim 4, wherein the plant is selected from the group consisting of potato, rice, cabbage, cabbage, mustard, rapeseed, radish, Brassica napobrassica and turnip ( Brassicarapa / Brassica campestris ). &lt; / RTI &gt; 1. A composition for increasing the tolerance of a plant comprising a gene consisting of the nucleotide sequence of SEQ ID NO: 1 encoding the NF-YA7 protein consisting of the amino acid sequence of SEQ ID NO: 2,
Wherein the gene is isolated from potatoes.

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