KR20100090135A - Cahb1 gene involved in growth enhancement, salt tolerance and senescence regulation of capsicum annuum and uses thereof - Google Patents

Abstract

PURPOSE: A Capsicum annuum-derived homeobox 1(CaHB1) protein and gene are provided to ensure plant growth, aging delay and inflammation resistance. CONSTITUTION: A Capsicum annuum-derived homeobox 1(CaHB1) protein comprises an amino acid of sequence number 2. A gene encoding the CaHB1 comprises a sequence of sequence number 1. The gene expression is increased by drought, salinity, mannitol, ethephon or salicylic acid. A method for producing transgenic plant comprises: a step of transforming plant cells with a recombinant plant expression vector containing CaHB1 gene; and a step of redifferentiating the transgenic plant from the transformed plant cells.

Description

CaHB1 gene involved in growth enhancement, salt tolerance and senescence regulation of Capsicum annuum and uses}

The present invention relates to homeobox 1 gene ( CaHB1 ) derived from red pepper ( Capsicum annuum ) and its properties.

Plants have been cultivated as crops. Because environmental conditions are constantly changing and affecting organisms, breeders have focused on improving the ability of organisms to survive stress in order to increase yields. Among environmental factors, salt stress, drought, cold water, and pathogen attack adversely affect plant survival and crop productivity. To overcome these problems, the genetic traits affecting stress tolerance have been identified and used in breeding for the development of stress resistant plants. In the last decades, the molecular mechanism by which plants adapt to stress has been identified and used in the development of transgenic plants with improved resistance to stress.

Korean Patent Publication No. 2003-0010744 discloses a plant lipase encoding diene, a transgenic plant, and a method for controlling aging in a plant. Korean Patent Registration No. 0499272 discloses a plant having a aging activity of a plant and a gene encoding the same. Korean Patent Registration No. 0877730 discloses a PBZ1 protein that is involved in aging and apoptosis of monocotyledonous plants, and Korean Patent Registration No. 0871591 discloses a pepper P.C. Although CLPLA1 gene is disclosed, it is different from the gene of the present invention.

The present invention has been made in accordance with the above requirements, CaHB1 under the development process and various stress conditions of pepper plants The expression patterns of the genes were observed, the genes were cloned and the transformation vector was prepared, and the expression patterns of the genes in the overexpression were analyzed to determine the role of the genes.

In order to solve the above problems, the present invention provides CaHB1 protein derived from pepper which is differentially expressed during the development of the plant and under stress.

The present invention also provides a gene encoding a CaHB1 protein.

The invention also provides CaHB1 A recombinant vector comprising a gene is provided.

The present invention also provides a plant transformed with the recombinant vector.

The present invention also provides a method for producing a transgenic plant using the vector.

The present invention also provides a method of increasing the growth and flame resistance of a plant using the vector and a method of controlling aging.

According to the present invention, CaHB1 protein and gene derived from red pepper of the present invention was strongly increased in expression by drought and salt, it was confirmed that the expression is strongly induced by plant hormones such as etepon, salicylic acid related to dry stress. In addition, it was confirmed that overexpression of CaHB1 promotes growth, promotes aging, and has strong flame resistance.

Therefore, the CaHB1 gene is expected to be able to develop plants that are resistant to drought and salt stress, as well as plants with enhanced growth.

In addition, since the protein according to the present invention has the aging activity of the plant, the protein and the gene encoding the same may be usefully used to search for the aging regulating material of the plant, and also useful for searching the aging control gene or protein of the plant Can be used.

In order to achieve the object of the present invention, the present invention is Capsicum annuum) derived CaHB1 (Capsicum annuum homeobox 1 ) provides protein.

The range of CaHB1 protein according to the present invention includes proteins having the amino acid sequence represented by SEQ ID NO: 2 isolated from red pepper and functional equivalents of the proteins. "Functional equivalent" means at least 70%, preferably at least 80%, more preferably at least 90%, even more preferably at least 70% of the amino acid sequence represented by SEQ ID NO: 2 as a result of the addition, substitution, or deletion of the amino acid Is 95% or more of sequence homology, and refers to a protein that exhibits substantially homogeneous physiological activity with the protein represented by SEQ ID NO: 2.

The present invention also provides a gene encoding the CaHB1 protein. Genes of the invention include both genomic DNA and cDNA encoding the CaHB1 protein. Preferably, the gene of the present invention may include a nucleotide sequence represented by SEQ ID NO: 1. In addition, the CaHB1 gene is a multigene family, and there are many homologous genes in pepper. The CaHB1 gene of the present invention may include not only the CaHB1 gene having the nucleotide sequence of SEQ ID NO: 1, but also the multigene family of the CaHB1 gene. .

Variants of the above base sequences are also included within the scope of the present invention. Specifically, the gene has a base sequence having a sequence homology of at least 70%, more preferably at least 80%, even more preferably at least 90%, most preferably at least 95% with the nucleotide sequence of SEQ ID NO: 1, respectively. It may include. The "% sequence homology" for a polynucleotide is identified by comparing two optimally arranged sequences with a comparison region, 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 total length of the CaHB1 cDNA gene of the present invention is 819 bp cDNA encodes 272 amino acids. The blasting of the genes in the database shows that the HD-Zip domain and CPSCE motif are similar to those of HD-Zip class II genes and are well conserved in other HD-Zip class II proteins.

CaHB1 gene of the present invention was strongly expressed in the stem and leaves of the plant (see Figure 2). The gene is characterized in that the expression is increased by drought, salinity, mannitol, etepon, or salicylic acid. Specifically, the CaHB1 gene of the present invention may be strongly induced by plant hormones such as etepon or salicylic acid. The CaHB1 gene of the present invention showed that expression was induced after 1 hour when treated with etepon and persisted after 3 hours, but weakened after 6 hours. In the case of salicylic acid treatment, the expression was induced after 6 hours, persisted after 9 hours, and weakened after 24 hours (see FIG. 4).

The present invention also provides a recombinant vector comprising the CaHB1 gene according to the present invention. The recombinant vector is preferably a recombinant plant expression vector.

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, a heterologous peptide, or a heterologous nucleic acid. Recombinant cells can express genes or gene fragments that are not found in their natural form in either the sense or antisense form. Recombinant cells can also express genes found in natural cells, but the genes are modified and reintroduced into cells by artificial means.

The term "vector" is used to refer to a DNA fragment (s), nucleic acid molecule, which is transferred into a cell. The vector replicates the DNA and can be independently regenerated in the host cell. The term "carrier" is often used interchangeably with "vector". The term “expression vector” refers to a recombinant DNA molecule comprising a coding sequence of interest and a suitable nucleic acid sequence necessary to express a coding sequence operably linked in a particular host organism.

Preferred examples of plant expression vectors are Ti-plasmid vectors which, when present in a suitable host such as Agrobacterium tumerfaciens, can transfer part of themselves, the so-called T-region, into plant cells. Another type of Ti-plasmid vector (see EP 0 116 718 B1) is used to transfer hybrid DNA sequences to protoplasts from which current plant cells or new plants can be produced that properly insert hybrid DNA into the plant's genome. have. A particularly preferred form of the Ti-plasmid vector is a so-called binary vector as claimed in EP 0 120 516 B1 and U.S. Patent No. 4,940,838. Other suitable vectors that can be used to introduce CaHB1 according to the invention into a plant host are viral vectors, such as those which can be derived from double stranded plant viruses (eg CaMV) and single stranded viruses, gemini viruses, etc. For example, it may be selected from an incomplete plant viral vector. The use of such vectors can be advantageous, especially when it is difficult to properly transform a plant host.

The expression vector preferably comprises one or more selectable markers. The marker is typically a nucleic acid sequence having properties that can be selected by chemical methods, and all genes that can distinguish transformed cells from non-transformed cells. Examples include, but are not limited to, herbicide resistance genes such as glyphosate or phosphinothricin, antibiotic resistance genes such as kanamycin, G418, bleomycin, hygromycin, and chloramphenicol It doesn't happen.

In the plant expression vector according to an embodiment of the present invention, the promoter may be CaMV 35S, actin, ubiquitin, pEMU, MAS or histone promoter, but is not limited thereto. The term "promoter" refers to a region of DNA upstream from a 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. Constructive promoters may be preferred in the present invention because the choice of transformants can be made by various tissues at various stages. Thus, the constitutive promoter does not limit the selection possibilities.

The plant expression vector of the present invention may use a conventional terminator, and examples thereof include nopalin synthase (NOS), rice α-amylase RAmy1 A terminator, phaseoline terminator, Agrobacterium tumefaciens ( Agrobacterium tumefaciens) Terminator of the octopine gene, but is not limited thereto. With regard to 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 terminators is highly desirable in the context of the present invention.

The present invention also provides a plant transformed with the recombinant vector according to the present invention. The plant according to the present invention is Arabidopsis, potato, eggplant, tobacco, pepper, tomato, burdock, garland chrysanthemum, lettuce, bellflower, spinach, chard, sweet potato, celery, carrot, buttercup, parsley, Chinese cabbage, cabbage, mustard, watermelon, melon , Dicotyledonous plants such as cucumbers, pumpkins, gourds, strawberries, soybeans, green beans, kidney beans, peas, or monocotyledonous plants such as rice, barley, wheat, rye, corn, sugar cane, oats, and onions. Plant, more preferably tomato.

The invention also comprises the steps of transforming a plant cell with a recombinant vector according to the invention; And

It provides a method for producing a transformed plant comprising the step of regenerating a transformed plant from the transformed plant cells.

Transformation of a plant means any method of transferring DNA to a plant. Such transformation methods do not necessarily have a period of regeneration and / or tissue culture. Transformation of plant species is now common for plant species, including both terminal plants as well as dicotyledonous plants. In principle, any transformation method can be used to introduce hybrid DNA according to the invention into suitable progenitor cells. Methods include calcium / polyethylene glycol methods for protoplasts (Krens et al. (1982) Nature 296: 72-74; Negrutiu et al. (1987) Pl Mol Biol 8: 363-373), electroporation of protoplasts (Shillito et. (1985) Bio / Technol 3: 1099-1102), microscopic injection into plant elements (Crossway et al. (1986) Mol Gen Genet 202: 179-185), (DNA or RNA-coated of various plant elements) Particle bombardment (Klein et al. (1987) Nature 327: 70), infection with (incomplete) virus in Agrobacterium tumerfaciens mediated gene transfer by plant infiltration or transformation of mature pollen or vesicles (EP 0 301 316) and the like. A preferred method according to the present invention comprises Agrobacterium mediated DNA delivery. Especially preferred is the use of the so-called binary vector technology as described in EP A 120 516 and US Pat. No. 4,940,838.

The method of the present invention comprises the step of transforming a plant cell with a recombinant vector into which the gene according to the present invention is introduced, wherein the transformation can be mediated by Agrobacterium tumefaciens . The method also includes the step of regenerating the transgenic plant from said transformed plant cell. The method of regenerating the transformed plant from the transformed plant cell may use any method known in the art.

The present invention also provides a method of increasing plant growth, comprising the step of transforming plant cells with the recombinant vector according to the present invention, overexpressing the CaHB1 gene.

The present invention also provides a method of increasing plant flame resistance, which comprises overexpressing a CaHB1 gene by transforming plant cells with the recombinant vector according to the present invention.

The present invention also provides a method for regulating plant aging comprising transforming plant cells with the recombinant vector according to the present invention to regulate expression of CaHB1 gene. In a method according to an embodiment of the present invention, plant senescence may be delayed by inhibiting the expression of CaHB1 gene.

Plants used in the above methods include Arabidopsis, potato, eggplant, tobacco, pepper, tomato, burdock, garland chrysanthemum, lettuce, bellflower, spinach, chard, sweet potato, celery, carrot, buttercup, parsley, cabbage, cabbage, mustard, watermelon, It may be a dicotyledonous plant such as melon, cucumber, pumpkin, gourd, strawberry, soybean, mung bean, kidney bean, pea, or monocotyledonous plant such as rice, barley, wheat, rye, corn, sugar cane, oats, onion, and the like. Dicotyledonous plant, more preferably tomato.

Solanum lycopersicon cv. 'MicroTom' transformants overexpressed CaHB1 increased resistance to salt stress and accelerated aging. Nicotiana benthamiana with transient overexpression of CaHB1 Plants promoted pathogen-induced apoptosis.

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

Tomato ( Solanum lycopersicon cv.MicroTom) seeds were surface sterilized with 1% (v / v) NaOCl at 25 ° C. for 10 minutes and washed three times for 5 minutes with sterile distilled water. Seeds were germinated on MS agar medium and grown for two weeks at 25 ° C. with a 16 hour photoperiod in a plant culture room before use for transformation.

Pepper plants (cultivated cultivars) and Nicotiana benthamiana plants were grown for 5-7 weeks at 25 ° C. with a 16-hour photoperiod in a flowerpot and incubation, respectively.

2. Temporary Overexpression (TOE)

For the TOE, Nicotiana benthamiana plants, 5-7 weeks old, were used. Agrobacterium tumefaciens strain GV2260 was transformed by the Freeze thaw method. pMBP-1 or pMBP-1: Agrobacterium tumerfaciens containing CaHB1 overnight at 200 ° C. at 30 ° C. at 30 ° C. in a test tube containing 50 μg / ml of kanamycin and 5 ml of YEP medium containing 50 μg / ml of rifampicin Incubated. The cells harvested after incubation were resuspended in 10 mM MgCl ₂ and exposed to 200 μM acetosyringone while shaking at 20 ° C. for 3-5 hours. Induced Agrobacterium The suspension was inoculated onto the leaves of Nicotiana Ventamiana plants with a needleless syringe.

3. Microscopic Analysis

One month old control or leaves of CaHB1 -transformed plants were observed by light microscopy and transmission electron microscopy (TEM).

4. Chemical treatment and pathogen inoculation

Corresponding to the leaves of pepper plants to see the expression patterns of CaHB1 transcripts after treatment with 5 mM salicylic acid (SA), 5 mM ethephon, 100 μM methyl jasmonate (MJ), or 100 μM absic acid (ABA), respectively. The material was sprayed.

Peppers and Nicotiana in Ventiana To inoculate pathogens, Xanthomonas axonopodis pv. Glycines ( Xanthomonas axonopodis pv. Glycines ( Xag ) 8ra or 8-13 cultures were inoculated onto the leaves of plants with a needleless syringe.

5. CaHB1  Intracellular location of proteins

To view the expression of CaHB1 in Arabidopsis protoplasts, a CaHB1-mGFP construct was prepared under the control of the CaMV 35S promoter. Fusion constructs were introduced into the protoplasts of Arabidopsis young leaves by a polyethylene glycol-mediated transformation method. Expression of the fusion construct was observed with a Zeiss Axioplan fluorescence microscope (Jena, Germany) 40 hours after transformation, and the image was captured with a chilled camera.

6. Gel Blot  analysis

시약(Gibco-BRL, http://www. invitrogen.com/)을 사용하여 추출되었다. 고추 식물체의 총 RNA 20　㎍이 1% 포름알데하이드 아가로즈 겔 상에서 분리되었다. 겔은 나일론 멤브레인으로 블럿되어 ³²P로 표지된 cDNA 프로브와 혼성화되었다. RNA 로딩의 양은 겔 상의 rRNA를 에티듐 브로마이드로 염색하여 증명하였다. Total RNA was determined according to the manufacturer's instructions

Extracted using reagent (Gibco-BRL, http: // www. Invitrogen.com/). Twenty micrograms of total RNA of the pepper plants was isolated on a 1% formaldehyde agarose gel. Gels were blotted with nylon membranes and hybridized with cDNA probes labeled with ³² P. The amount of RNA loading was demonstrated by staining rRNA on gels with ethidium bromide.

7. Reverse transcription - Polymerase Chain Reaction ( RT - PCR ) analysis

1 μg total RNA was mixed with RT master premix (ELPIS, Korea) containing MMLV reverse transcriptase, dNTPs, reaction buffer, and oligo (dT) primers. First strand cDNA was synthesized according to the manufacturer's protocol. RT-PCR was performed with gene specific primers.

Example  One.

Genes that regulate expression during pathogenic response in red pepper were identified by microarray experiments (Lee et al. (2004) Functional & Integrative Genomics 4: 196-205). Among the discovered genes, only transcriptional regulators were selected to select genes whose function was not found. The purpose of this study was to elucidate the function of homeodomain-leucinezipper (CaHB1) gene, and full-length gene was obtained from EST.

CaHB1 and Arabidopsis (Arabidopsis), rice, and Chrysler Tero Stigma Planta Centipede help (Craterostigma plantagineum) Comparison of the amino acid sequence between the erroneous log in separate CaHB1 analysis from, CaHB1 other HD on HD-Zip domain and CPSCE motifs site It can be seen that the similarity with the zip class II genes, and similar to the genes of HAT22, HAT9, Kraterostigma plantageneum of the Arabidopsis (CAA06728) (Fig. 1).

Example 2.

As a result of confirming the gene expression during the various tissues and aging process, as a result of confirming which tissue or developmental process CaHB1 is relatively expressed in the stem and flowers (Fig. 2 (a)), specifically aging progresses It was confirmed that the expression is increased a lot (Fig. 2 (b)).

Example 3.

When pepper plants were subjected to pathogens and various stresses, the expression of CaHB1 was confirmed, and the function of the gene was inferred. Expression was decreased when inoculated with Xag 8ra and Xag 8-13 (Fig. 3 (a)), but expression did not increase or decrease in wounds, cold, and heat, but increased by drought and salt stress. (B) of FIG. 3. When treated with mannitol and NaCl it was confirmed that the expression is significantly increased by time zone (Fig. 3 (c)).

Example 4.

Since the expression of CaHB1 is regulated by stress, and it is generally known that the expression of stress is directly related to hormones, the expression of CaHB1 was confirmed by treating various stress-related hormones in plants. Expression was increased by SA and etepon, and it was confirmed that expression was not regulated by MJ and ABA (FIG. 4).

Example 5.

Since CaHB1 is a transcription factor belonging to the HD-Zip family, it was confirmed by similarity analysis between the known sequences. As a result, the position of CaHB1 protein was confirmed by tagging GFP on the C-terminus of CaHB1 protein (Fig. 5 (a)). Could see. GFP of the control was evenly distributed in the cytoplasm (Fig. 5 (b)).

Example 6.

In order to know the function of CaHB1, CaHB1 was overexpressed in a transgenic tomato cultivar microtome plant. When grown to adult, CaHB1 overexpressing plants were found to have improved growth than the control. In order to confirm whether such growth is caused by the size or number of cells, it was confirmed that the desk tissues or spongy tissue cells were larger than the control when observing the cut surface of the leaves. In addition, the cell walls were also relatively thick (Fig. 6).

Example 7.

Since the transcription of CaHB1 was increased by drought or salt stress, it was predicted that CaHB1 would function when the plant was under such stress. An experiment was conducted to determine whether CaHB1 actually functions by using a tomato cultivar 'Microtom' plant overexpressed CaHB1. Chlorophyll almost disappeared after 3 days in the control discs in 0.5 M or 1 M NaCl solution, but relatively small amounts of chlorophyll were released from CaHB1 overexpression. This means that the cells of CaHB1 overexpressing plants are resistant to salts (FIG. 7).

Example 8.

Since CaHB1 increased rapidly during the aging process, it was assumed that CaHB1 may function in relation to aging. Indeed, the CaHB1 overexpressed tomato cultivar 'microtom' plant was confirmed that aging proceeds relatively fast compared to the control plants (FIG. 8). From this, it can be seen that CaHB1 acts in a direction for promoting aging.

Example 9.

Since CaHB1 expression was reduced by invasion of pathogens such as Xag 8ra (Example 3), CaHB1 was thought to respond to invasion of pathogens. CaHB1 was temporarily overexpressed on leaves of Nicotiana Ventamiana and then inoculated with Xag 8ra, whereby apoptosis proceeded rapidly in CaHB1 overexpressed portions, whereas the control did not exhibit this reaction (FIG. 9 (a)). This process of cell death is closely related to reactive oxygen species, so DAB staining confirmed the accumulation of hydrogen peroxide, which resulted in the overexpression of CaHB1 and the accumulation of significant amounts of hydrogen peroxide at the inoculation of Xag 8ra, while in control. (FIG. 9B). Thus, it can be seen that CaHB1 is involved in the regulation of cell death process when plants are invaded by pathogens.

1 is an alignment and phylogenetic tree of the amino acid sequence of CaHB1 protein. (a) compares the orthologous amino acid sequences of CaHB1 and CaHB1 isolated from Arabidopsis , rice, and Craterostigma plantagineum , wherein the bars above the sequence compare the HD and Zip domains. Indicates. Conserved CPSCE motifs are marked with a red box. (b) CaHB1 and Arabidopsis And the phylogenetic tree of the subfamily II HD-Zip family gene of Kraterostigma plantageneum. HAT1, HAT2, HAT3, HAT9, HAT14, HAT22, ATHB2, ATHB4, and ATHB17 are isolated from Arabidopsis thaliana , and CAA06728 is isolated from Krarotstigma plantageneum.

2 shows the expression of CaHB1 transcripts during various tissues and aging. (a) shows CaHB1 expression patterns in leaves, stems, roots, and flowers. (b) shows the expression of Ca HB1 during aging. The amount of CaHB1 transcript was determined by RT-PCR in the leaves of the red pepper plants (cultivated oyster ). YL is a young leaf, NS is a fully unexpanded leaf, ES is a leaf of early aging, and LS is a leaf of late aging.

Figure 3 shows the expression pattern of CaHB1 in response to pathogens and abiotic stress. Pepper leaves of (a) were inoculated with Xag 8ra or Xag 8-13. Pepper leaves in (b) were treated with injury, cold, drought, heat, or salt. Pepper leaves of (c) were treated with 400 mM mannitol or NaCl.

Figure 4 shows the expression pattern of CaHB1 mRNA in response to plant hormones. CaHB1 after Processing of Plant Hormones Including Pepper , SA, Ethephon, MJ, and ABA Expression was monitored. CaPR1a , CaACO , CaPinII , or CaDhn genes were used as positive gene markers, respectively. As a control solution, water or 0.1% ethanol was treated.

5 shows the location of CaHB1 in the nucleus. (a) is a vector constructed for the expression of mGFP-fused CaHB1 protein. mGFP is fused to the C-terminal section of CaHB1 and constitutively expressed by the CaMV 35S promoter. (b) shows the position of CaHB1-mGFP protein in the nucleus, green fluorescence indicates that mGFP accumulated. Nuclei were stained with DAPI.

Figure 6 shows the tomato cultivar 'microtome' plants overexpressed CaHB1. Enhanced growth (first and second lines) of transgenic lines, cross sections of lobules (third line), and TEM images of cell walls (fourth line) compared to control plants are shown.

7 shows increased resistance of CaHB1 overexpression to salinity stress. Leaf disks were floated in 0.5 M or 1 M NaCl solution and photographed after 3 days. # 13, # 14, and # 18 represent lines of transformed tomato cultivars 'microtome' plants.

8 shows that overexpression of CaHB1 promotes accelerated aging. Compared to control plants, the transformed tomato cultivar 'microtome' plants overexpressing CaHB1 shows accelerated aging.

9 shows the enhancement of pathogen-induced apoptosis by transient overexpression of CaHB1. (a) shows that CaHB1 promotes pathogen-induced cell death by the TOE. Apoptosis was induced by inoculation of Xag 8ra to the leaves of Nicotiana Ventamiana . The circle of white dotted line represents the leaf disk used for DAB staining. The picture was taken 3 days after the pathogen inoculation. (b) Accumulation of hydrogen peroxide is shown by DAB staining. Leaf disks of pathogen-inoculated or untreated areas were examined.

<110> SNU R & DB FOUNDATION <120> CaHB1 gene involved in growth enhancement, salt tolerance and          senescence regulation of Capsicum annuum and uses <130> PN08266 <160> 2 <170> KopatentIn 1.71 <210> 1 <211> 819 <212> DNA <213> Capsicum annuum <400> 1 atgggttttg atgatatttg caacactggc ctggttctag gattaggctt ttcctcaaca 60 accgatcaaa aatcaacaaa aataacacca ctagcaagca aaggaccagc atctcttacg 120 tttgaacctt cactaactct aagcctaatt tctggtgatc gtacctatga gcaacaagct 180 acaaagaagg ttaacgttac gaagccatcg aacgatcatc aatcggctga tttgtacaga 240 caagatagtg ctgcttcttc ctattcaaat gctagtgtga aaagggagag agatgttggt 300 agtgaagaga caactacaga ggtagaaaga gtttcctcca gagttattag cgatgaagat 360 gatgatggct ctaacgctag gaagaaactt aggctcacta aagcacaatc cgccctttta 420 gaggaaagct tcaagctaca cagcactctc aatcctaagc aaaaacagga tttagccatg 480 gaacttagtc taaggcctcg ccaagttgaa gtttggttcc agaacagaag agccagaaca 540 aagctgaagc aaacagaggt agactgtgaa ttcctgaaaa aatgctgtga gacactgaca 600 gaagaaaaca ggaggcttca caaagagcta caagaactaa aggcactaaa aatagcacaa 660 cctttgtaca tgcaattgcc ggcggcaacc ttaactatgt gtccttcatg cgaaaggatc 720 ggcggcggtg tcggagaaaa tccatcgaaa aatcctttta ctattgctca gaagcctcac 780 ttctacagtc catttaataa tccatcagca gcttgttaa 819 <210> 2 <211> 272 <212> PRT <213> Capsicum annuum <400> 2 Met Gly Phe Asp Asp Ile Cys Asn Thr Gly Leu Val Leu Gly Leu Gly   1 5 10 15 Phe Ser Ser Thr Thr Asp Gln Lys Ser Thr Lys Ile Thr Pro Leu Ala              20 25 30 Ser Lys Gly Pro Ala Ser Leu Thr Phe Glu Pro Ser Leu Thr Leu Ser          35 40 45 Leu Ile Ser Gly Asp Arg Thr Tyr Glu Gln Gln Ala Thr Lys Lys Val      50 55 60 Asn Val Thr Lys Pro Ser Asn Asp His Gln Ser Ala Asp Leu Tyr Arg  65 70 75 80 Gln Asp Ser Ala Ala Ser Ser Tyr Ser Asn Ala Ser Val Lys Arg Glu                  85 90 95 Arg Asp Val Gly Ser Glu Glu Thr Thr Thr Glu Val Glu Arg Val Ser             100 105 110 Ser Arg Val Ile Ser Asp Glu Asp Asp Asp Gly Ser Asn Ala Arg Lys         115 120 125 Lys Leu Arg Leu Thr Lys Ala Gln Ser Ala Leu Leu Glu Glu Ser Phe     130 135 140 Lys Leu His Ser Thr Leu Asn Pro Lys Gln Lys Gln Asp Leu Ala Met 145 150 155 160 Glu Leu Ser Leu Arg Pro Arg Gln Val Glu Val Trp Phe Gln Asn Arg                 165 170 175 Arg Ala Arg Thr Lys Leu Lys Gln Thr Glu Val Asp Cys Glu Phe Leu             180 185 190 Lys Lys Cys Cys Glu Thr Leu Thr Glu Glu Asn Arg Arg Leu His Lys         195 200 205 Glu Leu Gln Glu Leu Lys Ala Leu Lys Ile Ala Gln Pro Leu Tyr Met     210 215 220 Gln Leu Pro Ala Ala Thr Leu Thr Met Cys Pro Ser Cys Glu Arg Ile 225 230 235 240 Gly Gly Gly Val Gly Glu Asn Pro Ser Lys Asn Pro Phe Thr Ile Ala                 245 250 255 Gln Lys Pro His Phe Tyr Ser Pro Phe Asn Asn Pro Ser Ala Ala Cys             260 265 270  

Claims (11)

CaHB1 protein from Capsicum annuum consisting of the amino acid sequence of SEQ ID NO: 2. The gene encoding the CaHB1 protein of claim 1. The gene according to claim 2, which consists of the nucleotide sequence of SEQ ID NO: 1. The gene of claim 2, wherein the gene is increased in expression by drought, salinity, mannitol, etepon, or salicylic acid. A recombinant plant expression vector comprising the gene of claim 2. Plant transformed with the vector according to claim 5. Transforming the plant cell with the vector according to claim 5; And Regenerating a transformed plant from the transformed plant cells. A method of increasing plant growth, comprising the step of transforming plant cells with the vector according to claim 5, overexpressing the CaHB1 gene. A method for increasing plant flame resistance, comprising the step of transforming plant cells with the vector according to claim 5, overexpressing the CaHB1 gene. A method of controlling plant aging comprising transforming plant cells with the vector according to claim 5 to control expression of the CaHB1 gene. The method of claim 10, wherein the method of delaying plant aging by inhibiting the expression of the CaHB1 gene.

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