CN115161330A - Application of corn ZmGAPB gene in improving stress resistance of plants - Google Patents

Application of corn ZmGAPB gene in improving stress resistance of plants Download PDF

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CN115161330A
CN115161330A CN202210634801.8A CN202210634801A CN115161330A CN 115161330 A CN115161330 A CN 115161330A CN 202210634801 A CN202210634801 A CN 202210634801A CN 115161330 A CN115161330 A CN 115161330A
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zmgapb
gene
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李军
刘立功
李玉斌
郭新梅
赵美爱
裴玉贺
宋希云
王春晓
孙吉同
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Qingdao Agricultural University
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C12N15/8273—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for drought, cold, salt resistance

Abstract

The invention discloses application of a corn ZmGAPB gene in regulation and control of plant stress resistance, defines the relationship between the corn ZmGAPB gene and plant tolerance to high temperature, drought and salinity stress, and verifies the effect of the ZmGAPB gene on improvement of the plant stress resistance. The ZmGAPB gene is separated from the leaves of the maize seedlings, a pCambia1300-ZmGAPB vector is constructed, the vector is transformed into a plant for over-expression through an agrobacterium-mediated method, the high-temperature, drought and salinity stress tolerance of the transgenic plant is obviously improved, and the over-expression of the ZmGAPB gene is proved to be capable of obviously improving the stress resistance of the transgenic plant, so that the application prospect in the field of cultivating plant resistant varieties is wide.

Description

Application of corn ZmGAPB gene in improving stress resistance of plants
Technical Field
The invention relates to the technical field of agricultural biology, in particular to application of a corn ZmGAPB gene in improving the stress resistance of plants.
Background
In recent years, the problems of high-temperature heat damage frequently occurring in the growing season of crops, drought caused by the heat damage, soil salinization and the like bring a serious challenge to the corn production in China and even the world. In order to reduce the influence of the adverse factors, measures such as sowing in a staggered period, spraying chemical prevention and control agents, selecting varieties with strong resistance and the like are commonly adopted in agricultural production. Among them, the cultivation of new varieties of crops with strong resistance is the most economical and effective method for coping with various adverse environmental conditions.
Through a great deal of research for many years, people accumulate abundant experience and data on stress resistance mechanisms of crops in the aspects of physiology, biochemistry, metabolism, genetic evolution and the like, and along with the deepening of people on gene understanding, the combination of genes and traditional breeding for cultivating heat-resistant and drought-resistant varieties is an urgent problem to be solved.
Disclosure of Invention
The invention aims to provide application of a corn ZmGAPB gene in improving the stress resistance of plants, and experiments verify that the corn ZmGAPB gene is positively regulated by high temperature, drought and salt stress, so that the discovery has important significance in regulating the stress resistance of plants or cultivating plant resistant varieties.
In order to achieve the purpose, the invention provides the following scheme:
the invention provides application of a corn ZmGAPB gene, which comprises application in regulating and controlling plant stress resistance or cultivating plant resistance varieties.
Further, the nucleotide sequence of the ZmGAPB gene is shown as SEQ ID NO.1 or SEQ ID NO. 3.
Further, the stress tolerance includes the ability to withstand high temperature, drought and salt stress; the resistant varieties comprise heat-resistant, drought-resistant and salt stress-resistant varieties.
Further, the plant comprises maize or arabidopsis thaliana.
The invention also provides a method for improving the stress resistance of plants, which is used for obtaining plants with improved stress resistance by up-regulating the expression of the ZmGAPB gene in the plants.
Further, the up-regulating expression of the ZmGAPB gene in the plant specifically comprises: designing an amplification primer of the ZmGAPB gene to carry out PCR amplification of the gene, constructing a recombinant plasmid containing the ZmGAPB gene by using an amplification product and a vector, transforming the recombinant plasmid into a plant by using agrobacterium mediation, and carrying out ZmGAPB gene overexpression, thus obtaining the plant with strong stress resistance.
Further, the nucleotide sequence of the amplification primer is shown as SEQ ID NO. 5-6.
Further, the stress tolerance includes the ability to withstand high temperature, drought and salt stress.
Further, the plant comprises maize or arabidopsis thaliana.
The invention discloses the following technical effects:
the ZmGAPB gene is separated from leaves of maize seedlings, the forward regulation and control characteristics of the gene under high temperature, drought and salt stress are found through experiments, the gene is transferred into arabidopsis thaliana to culture a transgenic plant, and the germination rate, the survival rate, the root length and the dry fresh weight of seedlings and adult seedlings of transgenic lines (L1, L2 and L3) are remarkably improved under the stress condition compared with a control (WT) through experimental verification; chlorophyll content, soluble sugar content, soluble protein content and oxidoreductase activities such as catalase, peroxidase and superoxide dismutase in the transgenic lines are all obviously improved compared with the control, and the fact that the over-expression of ZmGAPB improves the high temperature, drought and salt stress tolerance of arabidopsis thaliana.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.
FIG. 1 is an electrophoretogram of ZmGAPB gene amplification product, in which M is Takara DL 2000marker, and 1 is amplification product;
FIG. 2 shows the PCR identification results of pMD19T-ZmGAPB colonies, where M is Takara DL 2000marker, and 1-2 is the PCR amplification product of white single colonies after transformation plating;
FIG. 3 shows the comparison of the cloned sequence with the nucleotide sequence of ZmGAPB gene published by NCBI;
FIG. 4 shows the comparison of the cloned sequence with the amino acid sequence of ZmGAPB gene published by NCBI;
FIG. 5 shows the expression of ZmGAPB gene in different organs of maize;
FIG. 6 shows the expression of ZmGAPB gene in maize leaves under different stress treatments;
FIG. 7 is a flow chart of plant expression vector construction;
FIG. 8 is the expression analysis of the ZmGAPB gene in different transgenic Arabidopsis lines, where WT is the control line and L1-L3 are Arabidopsis transgenic lines;
FIG. 9 shows the identification of the resistance phenotype of transgenic Arabidopsis at seedling stage;
FIG. 10 shows the germination rate and survival rate of transgenic Arabidopsis at seedling stage;
FIG. 11 shows the phenotypic identification of resistant roots in the seedling stage of transgenic Arabidopsis;
FIG. 12 shows the root length, fresh weight and dry weight of transgenic Arabidopsis at seedling stage;
FIG. 13 shows the phenotypic identification of the mature-stage resistant plant height of transgenic Arabidopsis;
FIG. 14 shows the plant height, fresh weight and dry weight of transgenic Arabidopsis at the mature period;
FIG. 15 shows the chlorophyll content, soluble sugar content and soluble protein content of leaves of transgenic Arabidopsis at the maturation stage;
FIG. 16 shows the hydrogen peroxide content, MDA content and relative conductivity of leaves of transgenic Arabidopsis at the maturation stage;
FIG. 17 shows leaf peroxidase activity assay of transgenic Arabidopsis at maturity.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in the present disclosure, it is understood that each intervening value, to the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless defined otherwise, 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. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the present disclosure without departing from the scope or spirit of the disclosure. Other embodiments will be apparent to those skilled in the art from consideration of the specification. The specification and examples are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.
Example 1
Application of corn ZmGAPB gene in improving heat resistance, drought resistance and salt stress of plants
The sequence information of the ZmGAPB full-length cDNA was obtained by searching GenBank accession number (NM-001305859) through NCBI website (https:// www.ncbi.nlm.nih.gov /), which Open Reading Frame (ORF) contains 1350bp, encoding 449 amino acids. The nucleotide sequence and the amino acid sequence published by the NCBI website are shown as SEQ ID NO. 1-2.
SEQ ID NO.1：
Figure BDA0003681666840000041
Figure BDA0003681666840000051
Note: the thick marked part is designed ZmGAPB gene full-length fragment amplification primer, and the underlined part is designed qPT-PCR specific primer.
SEQ ID NO.2：
MATHAALAASRIPAGARLHSRAAASSRHGVQRLDFADFSGLRPGSCSVSAAAREASFSDVLGAQLVAKATGENAVRAPAEAKLKVAINGFGRIGRNFLRCWHGREDSPIDVVVVNDSGGVRNASHLLKYDSMLGTFKADVKIVDDTTISVDGKPITVVSSRDPLKLPWGELGIDIVIEGTGVFVDGPGAGKHIQAGAKKVIITAPAKGADIPTYVVGVNEGDYDHSVADIISNASCTTNCLAPFVKILDEEFGIVKGTMTTTHSYTGDQRLLDASHRDLRRARAAALNIVPTSTGAAKAVALVLPQLKGKLNGIALRVPTPNVSVVDLVINTVKTGITADDVNAAFRKAAAGPLQGILDVCDVPLVSVDFRCSDVSCTIDASLSMVMGDDMVKVVAWYDNEWGYSQRVVDLAHLVAAKWPGAAAAGSGDPLEDFCKDNPETDECKVYEA。
1. Materials and treatments
The experimental material was maize inbred line E28 (provided by the molecular breeding laboratory of maize, university of Qingdao agriculture).
The seeds were sown in pots (1 grain/pot) containing nutrient soil and vermiculite (3:1) and placed in an intelligent greenhouse (25 ℃) of Qingdao university of agriculture for growth. When the seedlings grow to the three-leaf stage, respectively carrying out drought (20% PEG600), salt (200 mM NaCl) and high-temperature (37 ℃) treatment, taking the corn roots, stems and leaves after 0h, 12h, 24h, 36h and 48h of treatment, and quickly storing in liquid nitrogen for later use. Each treatment contained 5 seedlings, which were replicated three times.
2. Cloning and identification of genes
Extracting total RNA by using an RNA extraction kit, removing DNA by using DNAse, and detecting the integrity of the RNA and the concentration of the NanoDrop One trace nucleic acid by using a 1.2% agarose gel electrophoresis detector respectively. Then, the cDNA is reversely transcribed by a reverse transcription kit according to the operation instruction, frozen and stored at-80 ℃ for standby, and used for ZmGAPB full-length gene sequence cloning and qRT-PCR analysis. The RNA extraction kit and the reverse transcription kit are purchased from TaKaRa company.
The primers Premier 5.0 software was used to design amplification primers (Table 1), and the reverse transcribed cDNA was used as template for PCR full-length fragment amplification with high fidelity TaqDNA polymerase. The total volume of the reaction system was 25. Mu.L: 2 μ LcDNA +1 μ L P +1 μ L P +12.5 μ L Taq DNA Mix, finally ddH 2 And (4) supplementing oxygen. The PCR reaction conditions are as follows: pre-denaturation at 95 ℃ for 5min; 30 cycles of 94 ℃ 30s,58 ℃ 30s,72 ℃ for 1 min; extension at 72 ℃ for 10min. The result of gel electrophoresis of the amplified product is shown in FIG. 1, and the size of the PCR fragment obtained by amplification is 1350bp, which is in line with the expectation, indicating that the target gene has been successfully cloned.
TABLE 1 cloning amplification primers
Figure BDA0003681666840000061
And (3) adding a tail after the PCR product is purified, wherein the reaction system is 20 mu L:14.5 u LPCR product +3 u L dNTP +2 u L Taq buffer solution +0.5 u L Taq DNA polymerase, 72 degrees C extension 10min after connectionOnto the pMD19T vector (FIG. 7). The total volume of the ligation reaction system was 10uL:1 μ L of PCR tailing product +1 μ L of pMD19T vector +5 μ L of Solution I +3 μ L of ddH 2 O, ligation overnight at 10 ℃. Screening positive clones by blue-white spot method. The method comprises the following specific steps: and (2) taking 2 mu L of the ligation product to transform an escherichia coli competent cell (DH 5 alpha), gently mixing the ligation product and the cell, placing the mixture on ice for 30min, transferring the tube to a constant-temperature water bath at 42 ℃, thermally shocking the tube for 90 s, quickly placing the tube in the ice bath to cool the tube for 2min, adding 800 mu L of LB liquid culture medium, and incubating the tube at 37 ℃ at a rotating speed of less than 200rpm for 1hr to resuscitate the cell. Finally, 200. Mu.l of the transformed bacteria solution, 40. Mu. l X-gal (50 mg/ml) and 4. Mu.l of IPTG (100 mM) solution are mixed uniformly, a sterile glass rod is used for uniformly coating the mixture on a screening culture medium, the flat plate is inverted after the liquid is absorbed, and the bacterial colony appears after the liquid is cultured at 37 ℃ for 12-16 hours. The mixture is placed at 4 ℃ for several hours to ensure complete color development. White single colonies were picked with a sterile pipette tip for PCR validation. FIG. 2 shows the positive clones screened, and the PCR fragment size is 1350bp, which is expected. Then sending the DNA sequence to Beijing Nonsula genome research center, inc. for sequencing to confirm whether the cloned nucleic acid sequence is consistent with the sequence published on NCBI website. The nucleotide sequence and the amino acid sequence of the clone product are shown in SEQ ID NO. 3-4.
Sequencing results show that the overall length of the ZmGAPB gene nucleotide sequence is 1350bp, and 449 amino acids are coded. By comparison, the nucleotide sequence of the cloned ZmGAPB gene is found to have 4 base differences with the sequence published on the NCBI network, the homology is as high as 99.7 percent (FIG. 3), the amino acid sequence has 2 differences with the sequence published on the NCBI network (respectively, 23 rd amino acid residue and 359 th amino acid residue), and the homology is as high as 99.55 percent (FIG. 4). These differences may be due to differences in maize varieties, and therefore, we have succeeded in cloning the ZmGAPB gene from maize.
3. Expression characteristics of corn ZmGAPB gene under different organs and different stress treatments
Based on the above full-length gene sequence of ZmGAPB, qRT-PCR specific primers (Table 2) were designed using Primer Premier 5.0 software for analyzing the expression characteristics of ZmGAPB under different organs and different stress treatments. qPT-the PCR reaction system is 25 μ L: mu.L of cDNA, 0.5. Mu. L P1, 0.5. Mu. L P2, 12.5. Mu.L of 2 XSSYBR Green PCR Master Mix (Vazyme Biotech Co., ltd., ch.)ina), finally with ddH 2 And (4) supplementing oxygen. The method is carried out by a real-time fluorescent quantitative PCR instrument (Rotor gene Q) and the reaction procedure is as follows: pre-denaturation at 95 ℃ for 10min; denaturation at 95 ℃ for 10s, annealing at 56 ℃ for 30s, and extension at 72 ℃ for 30s for 40 cycles. Using corn ZmActin1 as reference gene, detecting relative expression of ZmGAPB and using formula 2 -△△CT To analyze the calculations.
TABLE 2qRT-PCR specific primers
Figure BDA0003681666840000071
ZmGAPB was expressed in maize roots, stems and leaves, but the relative expression was highest in leaves (FIG. 5). Under high temperature stress, the expression of ZmGAPB in leaves increases with the increase of treatment time, but under drought and salt stress conditions, the expression of ZmGAPB tends to increase and decrease with the increase of treatment time. These results indicate that ZmGAPB is involved in plant responses to high temperature, drought and salt stress (fig. 6).
4. Arabidopsis thaliana transformation experiment
Constructing a transformation arabidopsis vector pCambia1300-ZmGAPB, after enzyme digestion confirmation, electrically transforming the recombinant plasmid into agrobacterium tumefaciens GV3101, and transforming arabidopsis by using a flower dipping method. Plants that survived (L1, L2, and L3) were selected as transgenic positive plants on kanamycin-containing medium and confirmed by PCR. Uses corn ZmActin1 as an internal reference gene and adopts 2- △△Ct The relative expression amount of ZmGAPB in transgenic Arabidopsis is calculated. qPT-PCR used the same primers, reaction system and reaction conditions as above (Table 2).
The expression level of ZmGAPB was significantly increased in transgenic arabidopsis lines L1, L2 and L3 compared to wild-type (WT) arabidopsis (fig. 8). This can provide materials for further analysis of the stress resistance function of ZmGAPB.
5. Identification of heat resistance, salt resistance and drought resistance of ZmGAPB overexpression arabidopsis thaliana
Mainly measures the growth index and physiological index of the seedling stage and the mature stage. Growth indexes for wild-type (WT) Arabidopsis and transgenic Arabidopsis lines L1, L2 and L3 include root length, survival rate, plant height, dry weight, fresh weight and the like; the physiological indexes comprise soluble sugar content, soluble protein content, malondialdehyde, peroxidase activity, catalase activity and superoxide dismutase activity. Wherein the content of soluble sugar and soluble protein is measured by anthrone method and Coomassie brilliant blue method; the malondialdehyde content is measured by adopting a Malondialdehyde (MDA) content detection kit (BC 0025), the peroxidase activity is measured by adopting a Peroxidase (POD) activity detection kit (BC 0095), the catalase activity is measured by adopting a Catalase (CAT) activity detection kit (BC 0205), the superoxide dismutase activity is measured by adopting a superoxide dismutase (SOD) activity detection kit (BC 0175), the kits are all provided by Beijing Solibao science and technology Limited, and the measurement method is carried out according to the corresponding kit instruction.
The results are as follows: germination rates, survival rates, root lengths and dry fresh weights of over-expressed transgenic lines (L1, L2 and L3) at seedling and seedling development were significantly improved under stress conditions over the control (WT) (fig. 9-14); physiological analysis showed that chlorophyll content, soluble sugar content, soluble protein content and oxidoreductase activities such as catalase, peroxidase and superoxide dismutase were all significantly improved in the transgenic lines compared to the controls, but hydrogen peroxide content, malondialdehyde content and relative conductivity were significantly reduced (fig. 15-17). Experimental results show that the over-expression of ZmGAPB improves the high temperature, drought and salt stress tolerance of Arabidopsis.
The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solutions of the present invention can be made by those skilled in the art without departing from the spirit of the present invention, and the technical solutions of the present invention are within the scope of the present invention defined by the claims.
Sequence listing
<110> Qingdao agricultural university
Application of corn ZmGAPB gene in improvement of plant stress resistance
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Ala
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tggcacggcc gcgaggactc gccgatcgac gtcgtcgtcg tgaacgacag cggaggcgtc 360
aggaacgcgt cccacctcct caagtacgac tcgatgctcg gcaccttcaa ggccgacgtc 420
aagatcgtgg acgacacgac catcagcgtc gacggcaagc ccatcacggt cgtctccagc 480
agggacccgc tcaagctccc atggggtgag ctcggcattg acatcgtcat cgagggcacc 540
ggagtcttcg tcgacggccc cggcgccggc aagcacatcc aggccggcgc gaagaaggtg 600
atcatcacgg ctcccgccaa gggcgccgac atcccgacct atgtcgtcgg cgtcaacgag 660
ggcgactacg accacagcgt cgccgacatc atcagcaacg cgtcctgcac gaccaactgc 720
ctcgcgccat tcgtcaagat cttggacgag gagttcggaa tcgtgaaggg gaccatgacg 780
acgacgcact cgtacaccgg cgaccagagg ctgctggacg cgtcccaccg cgacctccgg 840
cgggcccgcg cggcggcgct gaacatcgtg ccgacgagca cgggcgccgc caaggccgtg 900
gcgctggtgc tgccgcagct caaggggaag ctgaacggca tcgcgctgcg ggtgccgacc 960
ccgaacgtgt ccgtggtgga cctggtgatc aacaccgtga agactggcat cacggccgac 1020
gacgtgaacg cggcgttccg caaggccgcc gccggcccgc tgcagggcat cctggaggtc 1080
tgcgacgtgc cgctcgtgtc cgtcgacttc aggtgctccg acgtgtcctg caccatcgac 1140
gcgtcgctca gcatggtcat gggcgacgac atggtcaagg tcgtcgcctg gtacgacaac 1200
gagtgggggt acagccaacg cgtggttgat ctggcgcacc tggtggcggc caagtggccc 1260
ggcgcggcgg cggccggcag cggcgaccct ctggaggact tctgcaagga caaccccgag 1320
accgacgagt gcaaggtgta cgaagcatga 1350
<210> 4
<211> 449
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 4
Met Ala Thr His Ala Ala Leu Ala Ala Ser Arg Ile Pro Ala Gly Ala
1 5 10 15
Arg Leu His Ser Arg Ala Pro Ala Ser Ser Arg His Gly Val Gln Arg
20 25 30
Leu Asp Phe Ala Asp Phe Ser Gly Leu Arg Pro Gly Ser Cys Ser Val
35 40 45
Ser Ala Ala Ala Arg Glu Ala Ser Phe Ser Asp Val Leu Gly Ala Gln
50 55 60
Leu Val Ala Lys Ala Thr Gly Glu Asn Ala Val Arg Ala Pro Ala Glu
65 70 75 80
Ala Lys Leu Lys Val Ala Ile Asn Gly Phe Gly Arg Ile Gly Arg Asn
85 90 95
Phe Leu Arg Cys Trp His Gly Arg Glu Asp Ser Pro Ile Asp Val Val
100 105 110
Val Val Asn Asp Ser Gly Gly Val Arg Asn Ala Ser His Leu Leu Lys
115 120 125
Tyr Asp Ser Met Leu Gly Thr Phe Lys Ala Asp Val Lys Ile Val Asp
130 135 140
Asp Thr Thr Ile Ser Val Asp Gly Lys Pro Ile Thr Val Val Ser Ser
145 150 155 160
Arg Asp Pro Leu Lys Leu Pro Trp Gly Glu Leu Gly Ile Asp Ile Val
165 170 175
Ile Glu Gly Thr Gly Val Phe Val Asp Gly Pro Gly Ala Gly Lys His
180 185 190
Ile Gln Ala Gly Ala Lys Lys Val Ile Ile Thr Ala Pro Ala Lys Gly
195 200 205
Ala Asp Ile Pro Thr Tyr Val Val Gly Val Asn Glu Gly Asp Tyr Asp
210 215 220
His Ser Val Ala Asp Ile Ile Ser Asn Ala Ser Cys Thr Thr Asn Cys
225 230 235 240
Leu Ala Pro Phe Val Lys Ile Leu Asp Glu Glu Phe Gly Ile Val Lys
245 250 255
Gly Thr Met Thr Thr Thr His Ser Tyr Thr Gly Asp Gln Arg Leu Leu
260 265 270
Asp Ala Ser His Arg Asp Leu Arg Arg Ala Arg Ala Ala Ala Leu Asn
275 280 285
Ile Val Pro Thr Ser Thr Gly Ala Ala Lys Ala Val Ala Leu Val Leu
290 295 300
Pro Gln Leu Lys Gly Lys Leu Asn Gly Ile Ala Leu Arg Val Pro Thr
305 310 315 320
Pro Asn Val Ser Val Val Asp Leu Val Ile Asn Thr Val Lys Thr Gly
325 330 335
Ile Thr Ala Asp Asp Val Asn Ala Ala Phe Arg Lys Ala Ala Ala Gly
340 345 350
Pro Leu Gln Gly Ile Leu Glu Val Cys Asp Val Pro Leu Val Ser Val
355 360 365
Asp Phe Arg Cys Ser Asp Val Ser Cys Thr Ile Asp Ala Ser Leu Ser
370 375 380
Met Val Met Gly Asp Asp Met Val Lys Val Val Ala Trp Tyr Asp Asn
385 390 395 400
Glu Trp Gly Tyr Ser Gln Arg Val Val Asp Leu Ala His Leu Val Ala
405 410 415
Ala Lys Trp Pro Gly Ala Ala Ala Ala Gly Ser Gly Asp Pro Leu Glu
420 425 430
Asp Phe Cys Lys Asp Asn Pro Glu Thr Asp Glu Cys Lys Val Tyr Glu
435 440 445
Ala
<210> 5
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 5
atggccaccc acgcagctct c 21
<210> 6
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 6
tcatgcttcg tacaccttgc 20
<210> 7
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 7
gacaacgagt gggggtacag 20
<210> 8
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 8
tgcttcgtac accttgcact 20

Claims (9)

1. The application of the corn ZmGAPB gene is characterized by comprising the application in regulating and controlling the stress resistance of plants or cultivating plant resistant varieties.
2. The use according to claim 1, wherein the ZmGAPB gene has the nucleotide sequence shown in SEQ ID No.1 or SEQ ID No. 3.
3. The use of claim 1, wherein the stress tolerance comprises the ability to withstand high temperature, drought and salinity stress; the resistant varieties comprise heat-resistant, drought-resistant and salt stress-resistant varieties.
4. Use according to claim 1, wherein the plant comprises maize or arabidopsis.
5. A method for improving the stress resistance of plants is characterized in that a plant with improved stress resistance is obtained by up-regulating the expression of ZmGAPB gene in the plant.
6. The method according to claim 5, wherein the up-regulating expression of the ZmGAPB gene in the plant specifically comprises: designing an amplification primer of the ZmGAPB gene to carry out PCR amplification of the gene, constructing a recombinant plasmid containing the ZmGAPB gene by using an amplification product and a vector, transforming the recombinant plasmid into a plant by using agrobacterium mediation, and carrying out ZmGAPB gene overexpression, thus obtaining the plant with strong stress resistance.
7. The method of claim 5, wherein the nucleotide sequence of the amplification primer is shown in SEQ ID No. 5-6.
8. The method of claim 5, wherein the stress tolerance comprises the ability to withstand high temperature, drought and salt stress.
9. The method of any one of claims 5-8, wherein the plant comprises maize or Arabidopsis thaliana.
CN202210634801.8A 2022-06-07 2022-06-07 Application of corn ZmGAPB gene in improving stress resistance of plants Pending CN115161330A (en)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20110179523A1 (en) * 2008-09-30 2011-07-21 Basf Plant Science Gmbh Method for Producing a Transgenic Plant Cell, a Plant or a Part Thereof with Increased Resistance Biotic Stress
CN108424920A (en) * 2018-04-24 2018-08-21 吉林省农业科学院 The resistance to inversely related transcription factor ZmNAC33 genes of corn and its application
CN113355333A (en) * 2021-04-19 2021-09-07 青岛农业大学 Cloning method, application and application method of corn gene ZmNAC7

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20110179523A1 (en) * 2008-09-30 2011-07-21 Basf Plant Science Gmbh Method for Producing a Transgenic Plant Cell, a Plant or a Part Thereof with Increased Resistance Biotic Stress
CN108424920A (en) * 2018-04-24 2018-08-21 吉林省农业科学院 The resistance to inversely related transcription factor ZmNAC33 genes of corn and its application
CN113355333A (en) * 2021-04-19 2021-09-07 青岛农业大学 Cloning method, application and application method of corn gene ZmNAC7

Non-Patent Citations (5)

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Title
KEITH 等: ""NM_001305859.1"" *
KEITH 等: ""NP_001292788.1"" *
YANGXUAN LIU 等: ""Proteomic Analysis of Rice Subjected to Low Light Stress and Overexpression of OsGAPB Increases the Stress Tolerance"" *
卢倩;弭晓菊;崔继哲;: "植物甘油醛-3-磷酸脱氢酶作用机制的研究进展" *
李严曼;欧阳孟真;孙守如;朱磊;: "辣椒3-磷酸甘油醛脱氢酶基因CaGAPB的克隆及表达分析" *

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Application publication date: 20221011