CN114656543B

CN114656543B - Application of protein ATNDX and DNA molecule encoding protein ATNDX in regulation and control of salt and alkali tolerance of plants

Info

Publication number: CN114656543B
Application number: CN202011542254.8A
Authority: CN
Inventors: 杨永青; 郭岩; 李钦沛
Original assignee: China Agricultural University
Current assignee: China Agricultural University
Priority date: 2020-12-23
Filing date: 2020-12-23
Publication date: 2023-10-20
Anticipated expiration: 2040-12-23
Also published as: CN114656543A

Abstract

The application discloses a protein ATNDX and application of a DNA molecule encoding the protein ATNDX in regulating and controlling salt and alkali tolerance of plants. The protein ATNDX provided by the application is as follows (1) or (2): (1) A protein consisting of an amino acid sequence shown as SEQ ID No.2 in a sequence table; (2) Protein which has 80% or more identity with the amino acid sequence defined by SEQ ID No.2 in the sequence table, is derived from corn and has the same biological function. The application provides a theoretical basis for elucidating the molecular mechanism of the protein ATNDX in plant saline-alkali stress signal response, and provides gene resources for cultivating and improving new varieties of stress-resistant plants.

Description

Application of protein ATNDX and DNA molecule encoding protein ATNDX in regulation and control of salt and alkali tolerance of plants

Technical Field

The application relates to the field of biotechnology, in particular to protein ATNDX and application of DNA molecules encoding the protein ATNDX in regulating and controlling saline-alkali tolerance of plants.

Background

Salinization of cultivated lands is a major abiotic stress leading to reduced crop yields due to the high salt content in cultivated lands and the alkaline salts (mainly Na ₂ CO ₃ And NaHCO ₃ ) The specific gravity is high. The salinized soil has wide distribution in China, and is widely distributed in North China, northwest China, northeast China, west China, coastal China and the north of Yangtze river, and the total area is about 1 multiplied by 10 ⁸ hm ² . In addition to naturally occurring saline-alkali soil, misuse of fertilizers and pesticides by people and improper cultivation methods can also cause accumulation of salt in cultivated layers, thereby leading to formation of secondary salinization of the soil. The saline-alkali stress has more serious negative effect on crops than the salt stress, and one of the main reasons is H under the saline-alkali stress ⁺ Transmembrane gradient dependent Na ⁺ Transporter (e.g. SOS 1) function is inhibited, na ⁺ The poisoning is aggravated.

So far, the molecular mechanism of salt and alkali resistance of crops such as corn is not clear, and the molecular mechanism becomes a theoretical bottleneck of salt and alkali resistance breeding of crops.

Disclosure of Invention

The application provides an application of protein ATNDX in regulating and controlling salt and alkali tolerance of plants or cultivating high salt and alkali tolerance plants.

Alternatively, the protein ATNDX is (1) or (2) as follows:

(1) A protein consisting of an amino acid sequence shown as SEQ ID No.2 in a sequence table;

(2) Protein which has 80% or more identity with the amino acid sequence defined by SEQ ID No.2 in the sequence table, is derived from corn and has the same biological function.

The protein can be synthesized artificially or obtained by synthesizing the coding gene and then biologically expressing.

The DNA molecule for encoding the protein ATNDX is named as ATNDX gene, and the application of the DNA molecule belongs to the protection scope of the application. The application is particularly applied to regulating and controlling the salt and alkali tolerance of plants or cultivating high salt and alkali tolerance plants.

Alternatively, the DNA molecule encoding the above protein ATNDX is a DNA molecule of any one of the following 1) -4):

1) The nucleotide sequence is a DNA molecule shown as SEQ ID No.3 in a sequence table;

2) The coding region is a DNA molecule shown as SEQ ID No.1 in the sequence table;

3) A DNA molecule which hybridizes under stringent conditions with the DNA molecule defined in 1) or 2) and which encodes a protein having the same function;

4) A DNA molecule derived from maize and encoding the protein ATNDX of claim 1 or 2 having 90% or more identity to the DNA molecule defined in 1) or 2).

The nucleotide sequence of the ATNDX gene of the present application can be easily mutated by one of ordinary skill in the art using known methods, such as directed evolution and point mutation. Those artificially modified nucleotides having 90% or more identity to the nucleotide sequence of the ATNDX gene of the present application are derived from the nucleotide sequence of the present application and are equivalent to the sequence of the present application as long as they encode the protein ATNDX and are derived from corn.

The term "identity" as used herein refers to sequence similarity to a native sequence.

The application of the expression cassette, the recombinant vector, the transgenic plant cell line or the recombinant bacteria containing the ATNDX gene in regulating and controlling the salt and alkali tolerance of plants or cultivating high salt and alkali tolerance plants also belongs to the protection scope of the application.

The application also provides a method for cultivating transgenic plants, which comprises the following steps: introducing a substance for increasing the content and/or activity of the protein ATNDX into a starting plant to obtain a transgenic plant; the transgenic plant has higher saline-alkali tolerance than the starting plant A.

In the above method, the "increasing the content and/or activity of the protein ATNDX" can achieve the effect of expressing or overexpressing the protein, or increasing the activity of the protein, by multicopy, changing a promoter, a regulatory factor, a transgene, or the like, which are well known in the art.

In the above method, the "introduction of a substance that increases the content and/or activity of protein ATNDX into a starting plant" may specifically be introduction of a DNA molecule encoding the protein ATNDX into a starting plant.

In the above method, the "introducing a DNA molecule encoding the protein ATNDX into a starting plant a" is introduced into the starting plant by a recombinant expression vector; the recombinant expression vector can be specifically a gene overexpression vector mentioned in the example. Plant expression vectors carrying the ATNDX gene or other homologous sequences of the present application can be transformed into plant cells, tissues or organs by using any one or a combination of conventional biological methods such as protoplast-chemical-mediated method (Ca2+, PEG), ti plasmid, ri plasmid, plant viral vector, direct DNA transformation, pollen tube, microinjection, electric shock, gene gun, agrobacterium-mediated method, and the like, and the transformed plant cells, tissues or organs are cultivated into plants; the tissues and organs may include pods, calli, shoot tips, leaves, seeds, and the like of the host plant.

The application also provides a plant breeding method, which comprises the following steps: increasing the content and/or activity of said protein ATNDX in plants, whereby the saline-alkali tolerance of the plants is increased.

The protein ATNDX, the DNA molecule encoding the protein ATNDX, the expression cassette, the recombinant vector, the transgenic plant cell line or the recombinant bacterium containing the DNA molecule encoding the protein ATNDX also belong to the protection scope of the application.

In the above, the modulating plant stress tolerance may be increasing plant stress tolerance.

In the above, the high salt and alkali tolerant plant may be a plant having a higher salt and alkali tolerance than the wild type plant.

In the above, the plant may be a dicotyledonous plant or a monocotyledonous plant. The monocot plant may be a gramineous plant, for example maize.

The application introduces ATNDX gene into corn B73-329 to obtain ATNDX transgene over-expression strain; compared with the wild type B73-329, the over-expression strain shows remarkable saline-alkali stress tolerance, so that the protein ATNDX has the function of improving the stress tolerance of plants to high saline alkali. The application provides a theoretical basis for elucidating the molecular mechanism of the protein ATNDX in plant saline-alkali stress signal response, and provides gene resources for cultivating and improving new varieties of stress-resistant plants.

Drawings

FIG. 1 shows the growth of saline-alkali stress treated plants.

FIG. 2 shows the results of ATNDX gene overexpression detection, wherein M is MARKER, the left graph shows the results of ACTIN1 gene expression detection, and the right graph shows the results of ATNDX gene expression detection.

Detailed Description

The following detailed description of the application is provided in connection with the accompanying drawings that are presented to illustrate the application and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the application in any way.

The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are carried out according to techniques or conditions described in the literature in the field or according to the product specifications. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.

The inventor of the application discovers a plant salt and alkali tolerance related gene, named ATNDX gene, in maize B73-329, and the nucleotide sequence of the genome gene is shown as SEQ ID No. 3. The amino acid sequence of the protein coded protein named protein ATNDX is shown in a sequence table SEQ ID No.2, and CDS of cDNA genes of the protein is shown in a sequence table SEQ ID No. 1.

Example 1 ATNDX Gene overexpression maize saline-alkali stress treatment assay

The experiment was set up with two treatment groups, a control group and a transgenic group, the experiment was repeated 3 times, each time repeated with 8 pots each, 3-4 seeds per pot. The control group adopts B73-329 corn seeds, 6 transgenes group adopts ATNDX over-expression transgene plant T ₃ Seed generation, 15030085, 15050256, 15070742, 15110036, 15110112, 15121724 respectively. Pure vermiculite without nutrient soil is used for planting, and 1/2Hoagland's nutrient solution is used for watering. Plants were irrigated 9 days after growth (trefoil phase) with 100mM NaHCO ₃ (saline-alkaline stress) 1/2Hoagland's nutrient solution (NaHCO was added to 1/2Hoagland's nutrient solution ₃ To NaHCO ₃ Liquid obtained at 100 mM), irrigated once every 10 days, grown two liters per pot, observed for 30 days for phenotype, and measured plant height.

FIG. 1 shows the results of a part of experiments of the saline-alkali stress treatment, wherein the left picture is the growth condition of the plant before the saline-alkali stress treatment, the right picture is the growth condition of the plant after the saline-alkali stress treatment for 30 days, the transgenic group plant (OE-ATNDX) shows stronger salt-tolerant alkalinity than the control group plant, the leaves are greener, the plant height is higher, and the plant height of the transgenic group plant is 1.3-1.6 times of the plant height of the control group plant.

Experiments prove that ATNDX positively regulates the salt and alkali resistance of plants.

Wherein, ATNDX over-expresses transgenic plant T ₃ The seed substitutes were prepared as follows.

1. Construction of ATNDX Gene overexpression vector

CDS of cDNA gene of ATNDX protein (i.e. DNA molecule shown in SEQ ID No. 1) was inserted into pBCXUN to obtain recombinant plasmid pBCXUN-ATNDX. In the recombinant plasmid pBCXUN-ATNDX, CDS of cDNA gene of ATNDX protein is driven by Ubi promoter (size ubquitin-1 promoter) to express ATNDX protein, and transcription is terminated by Nos terminator.

pBCXUN is an expression vector obtained by replacing the HYG gene (hptII, hygromycin resistance gene) of pCXUN (GenBank: FJ905215.1, 06-JUL-2009) with the Bar gene (encoding phosphinothricin acetyltransferase) (SEQ ID No. 4) and keeping the other nucleotides of pCXUN unchanged.

2. Construction of ATNDX Gene overexpressing plants

The pBCXUN-ATNDX (gene over-expression vector) constructed in the first step is transformed into competent agrobacterium tumefaciens EHA105 strain by a heat shock method, positive clones are identified by colony PCR, the positive clones are selected for sequencing, and the positive clone containing the pBCXUN-ATNDX is the recombinant agrobacterium tumefaciens and named as pBCXUN-ATNDX/EHA105.

A single colony of pBCXUN-ATNDX/EHA105 was inoculated into 2-3mL of liquid medium containing 100. Mu.g/mL kanamycin and 50. Mu.g/mL rifampicin, and cultured overnight at 28℃with shaking. Transferring a large amount of liquid culture medium containing antibiotics for shake culture the next day, collecting thallus after transferring several times, and re-suspending to OD _600nm Recombinant Agrobacterium suspensions were obtained between 0.8-1.0. Infecting young embryo of B73-329 corn with the obtained recombinant agrobacterium suspension, inducing callus to form seedling, co-culturing, resistance screening (herbicide glufosinate) and pre-differentiating, differentiating and rooting to obtain T ₀ Regenerating plants. From T ₀ And screening transgenic plants from the regenerated plants by adopting a PCR identification method. Will T ₀ The transgenic plants in the generation are selfed to obtain T ₁ Seed generation, T ₁ The plant grown from the seed generation is T ₁ Generating plants; plant selfing until T is obtained ₃ Seed generation, namely ATNDX over-expressed transgenic plant T ₃ Seed generation, in which different T is to be used ₀ T obtained after selfing of regeneration plant ₃ The generation lines were named 15030085, 15050256, 15070742, 15110036, 15110112, 15121724, respectively.

Extracting total RNA of 15030085, 15050256, 15070742, 15110036, 15110112 and 15121724, reversely transcribing cDNA, detecting ATNDX gene over-expression by RT-PCR, taking corn B73-329 as a control plant and taking the ACTIN1 gene as a reference. The results are shown in FIG. 2, where ATNDX is overexpressed in transgenic plants compared to control plants B73-329,6 independent ATNDX overexpressing transgenic lines.

The present application is described in detail above. It will be apparent to those skilled in the art that the present application can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the application and without undue experimentation. While the application has been described with respect to specific embodiments, it will be appreciated that the application may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the application following, in general, the principles of the application and including such departures from the present disclosure as come within known or customary practice within the art to which the application pertains. The application of some of the basic features may be done in accordance with the scope of the claims that follow.

Sequence listing

<110> Chinese university of agriculture

<120> application of protein ATNDX and DNA molecule encoding protein ATNDX in regulation of salt and alkali tolerance of plants

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gcgaatgttg attatgatct gtttaagtca gctggatgga ttctaagtgt attttcatct 1260

tctgggcaat cagtcacacc tcaattcaag ctcagtttac aaaataacct taccatgtct 1320

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gttcccaacg tgtgccaaga acaggatagg aaccgtttca ttcagaatgt tatgagtgga 1440

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Leu His Trp Arg Asn Ser Val Glu Phe His Lys Leu Leu Lys Asp Asn

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Gly Asp Phe Ser Ile Cys Phe Asn Ser Glu Gln Val Leu Pro Gln Lys

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Ile Ser Val Glu Lys Met Val Lys Met Leu Pro Arg His Leu Ile Ala

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Val Val Met Thr Pro Asn Lys Asp Gly Lys Ser Arg Tyr Ile Leu Cys

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Gly Ile Arg Leu Leu Gln Thr Leu Cys Asp Leu Thr Pro Arg Asn Ala

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Ile Asp Leu Val Ile Leu Val Ile Ile Ala Leu Gly Arg Asn Arg Lys

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Glu Ser Cys Asn Ser Asn Lys Glu Ser Leu Leu Glu Ala Thr Leu Val

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Ala Ser Cys Leu His Leu Phe His Gly Phe Ile Ser Pro Asn Ser Gln

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Asp Leu Val Leu Val Leu Leu Ala His Pro Arg Val Asp Val Phe Ile

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Asp Ser Ala Phe Gly Ala Val Leu Asn Val Val Ile Ser Leu Lys Ala

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Lys Leu Leu Tyr Arg Gln Thr Asp Ser Pro Lys Lys Leu Gly Ala Ser

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Ser Val Glu Glu Val Asn Phe His Cys Gln Gln Ala Glu Ala Ala Leu

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Gln Phe Leu His Ser Leu Cys Gln His Lys Pro Phe Arg Glu Arg Val

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Ala Lys Asn Lys Glu Leu Cys Gly Lys Gly Gly Val Leu Arg Leu Ala

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Gln Ser Ile Leu Ser Leu Thr Ile Thr Pro Glu Phe Val Gly Ala Thr

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Val Thr Ile Ala Ser Thr Ser Arg Met Lys Ala Lys Val Leu Ser Ile

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Leu Gln His Leu Phe Glu Ala Glu Ser Val Ser Phe Leu Asp Glu Val

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Ala Asn Ala Gly Asn Leu His Leu Ala Lys Thr Val Ala Ser Glu Val

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Leu Lys Leu Leu Arg Leu Gly Leu Ser Lys Ala Ser Met Ala Thr Ala

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Ser Pro Asp Tyr Pro Met Gly Phe Val Leu Leu Asn Ala Met Arg Leu

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Ala Asp Val Leu Thr Asp Asp Ser Asn Phe Arg Ser Phe Phe Thr Glu

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His Phe Ser Met Val Leu Ser Ala Val Phe Cys Leu Ser His Gly Asp

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Phe Leu Ser Met Leu Cys Ser Ser Asp Leu Ser Ser Arg Glu Asp Asp

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Leu Ile His Ser Glu Phe Glu Glu Ser Gln Val Gln Val Lys Val Lys

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Ser Gly Lys Leu Lys Glu Leu Leu Asn Leu Asn Asn Glu Glu Ala Ser

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Glu Asp Cys Asp Val Arg Val Glu Gly Val Met Thr Lys Gln Gly Val

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Asn Glu Glu Ile Asp Thr Val Glu Arg Leu Lys Glu Ser Asp Ala Asp

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Ala Ser Asn Leu Glu Thr Ser Gly Ser Asp Thr Ser Ser Asn Arg Gly

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Lys Gly Leu Val Glu Glu Gly Glu Leu Val Gln Asn Met Ser Lys Arg

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Phe Lys Gly Ser Ala Ser Gly Glu Val Lys Glu Asp Glu Lys Ser Glu

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Met Asn Ala Asp Gln Met Gly Met Ile Glu Lys Ala Leu Ala Glu Glu

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Pro Asp Leu Gln Arg Asn Ser Ala Ser Arg Gln Leu Trp Ala Asp Lys

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Ile Ser Gln Lys Gly Ser Glu Val Ile Thr Ser Ser Gln Leu Lys Asn

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Trp Leu Asn Asn Arg Lys Ala Lys Leu Ala Arg Ala Asn Lys Gln Thr

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Gly Pro Ala His Asp Asn Asn Ser Ser Gly Asp Leu Pro Glu Ser Pro

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Gly Asp Glu Asn Thr Trp Gln Gln Lys Pro Ser Thr Pro Ile Lys Asp

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Gln Thr Val Thr Glu Thr Pro Lys Thr Gly Glu Asn Leu Met Arg Thr

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Met Asp Glu Arg Gly Asp Glu Ile Gly Lys Gly Thr Val Leu Arg Thr

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Asp Gly Glu Trp Asn Gly Leu Ser Leu Glu Thr Arg Gln Ile Cys Val

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Val Asp Val Met Glu Leu Ser Glu Ser Tyr Asp Gly Ser Lys Lys Met

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Ser Arg Phe Gly Val Met Arg Val Ala Trp Asp Val Asn Lys Leu Gln

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atgccaccat accctgaagt agatttcact gcttttgatt ttcgtatctt cttctctgtc 180

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agaggctaca ttggtggctt cttgtctcca cctgtttcac gggtttatct ctcctaactc 1020

ccaagatctt gttctcgtct tgcttgcaca cccaagggta tgaaactatg accttatcat 1080

aatgtgtttg aaatgccaat gtgaggaaaa tatattaaga aaattacgct tatgctttgg 1140

gatctgggtt tctgctgcct tgggattagt ttgatctgat tctttttttt ttttcatcta 1200

ggttgatgtg tttatagaca gtgcttttgg agctgttctc aatgttgtga tatctttgaa 1260

agcgaagttg ctgtatagac aaactgactc cccaaaaaag ttaggcgcaa gttctgtaga 1320

ggaggttaac ttccactgcc aacaagctga agctgctttg cagttccttc attctctatg 1380

ccaacacaaa ccctttagag aacgtgtcgc taaaaacaag gtttctctat tgcttggtgt 1440

ttgtacatta atcacgagtt gttaataatt tgttttgctt aattgatgat tttatggagg 1500

acgaatctca gaattatgtt ttctggtatt ttctctcagg agctatgtgg aaaaggtggc 1560

gttcttaggc tagctcaatc catactatca ctaactatta cacctgaatt tgttggagca 1620

accgtaacta tagcttccac atctagaatg aaagcaaaag ttctttcaat tgtaagtcta 1680

ttgaaattta gcaatttttt tctctattta ctatctgcaa ctcataccaa agttcatgtg 1740

cctgcagttg cagcatctgt ttgaagcgga aagtgtctca ttccttgacg aggttgcaaa 1800

tgcaggaaac ttgcatttag ccaaaactgt tgcctcagag gttagtccta attagggtct 1860

ctgctctaaa atcgatagga cgttttcctt cttgattcga aaggtttacc actgagtgac 1920

gcttctattt ataaagattc catacaaatt gacaatctta tttgttttga tacgttgtaa 1980

ctagtcatag aacaaacctc actgcaaaaa gctcgtgttg gctaagtgat aatcagctct 2040

gctctagagt ctaaacctac agtttcctaa atggcatatg ttgttaaatt gacattttgt 2100

acttttctgt tcctcatttc tctccaggat agtttatcca tgtgaagtta agtgttgacc 2160

tattcagtga tgatatatct gtaatatagg ttcttaaatt attgaggctt ggcctttcta 2220

aagcttccat ggctactgct tctcctgact acccgatggg ttttgtgcta cttaacgcta 2280

tgcgcttggc tgacgtgctc actgatgact caaattttcg atcttttttc actgaacatt 2340

ttgtaagtat caagttattc ctgcctctgc caaagtctct tgttttattt attgttttgg 2400

cacatatgat tcatacaatt tatgagctgt cgtcggtatg gttataaatt aattgcgtta 2460

ctgaaattct gtaatttgtt gtgtgacgga tgttaaatct ttctccagag catggttctc 2520

agcgcggtat tttgtctctc tcatggagat ttcttgtcaa tgttgtgctc ttctgatctt 2580

tcttcaaggg aggatgatgc gaatgttgat tatgatctgt ttaagtcagc tggatggatt 2640

ctaagtgtat tttcatcttc tgggcaatca gtcacacctc aattcaagct cagtttacaa 2700

aataacctta ccatgtcttc atatgcacat caacgaacat ccttatttat taaaatgatt 2760

gcgaatcttc actgtttcgt tcccaacgtg tgccaaggtg atttttttgc ccgccactat 2820

cttctgcatt tccctgtatc atattgttgg tttgccttcg tttgttagtt actaagtgta 2880

ttcgctgtta atatctgcag aacaggatag gaaccgtttc attcagaatg ttatgagtgg 2940

attgcgaaaa gatccttcaa gcatattgat taagatgtta ccaggctctt catatactcc 3000

tgtggcacag agaggcactg gtgtttgcag aaacctaggt atgtatctct atcccgagac 3060

acatatatta tttcttatat acacatgttt tgctgatttg gttttaatgg atatttgtat 3120

atgtaggttc tctgttgcgc catgcagaat ccttgatccc tagttccctc aacgaggaag 3180

attttctgct tttgaggtaa cataaagcct cttcagtcat tctacttttc acctatgact 3240

acattaaagt tttgtagttc tcgagctgta ctattttatg tagccgttga tattattttt 3300

ttccggtata ttgtcttctg cagggtgttt tgtgaccagt tacagccgtt aatccattcc 3360

gagtttgagg aaagtcaagt acaggtgaag gtgaaaaagc tttttgctct cttatacatt 3420

ggttttacaa ttctctggct tatctgttta gttacactga tacaagtagg tgtaacaagg 3480

ttttactcat gtccatcgta tgttaaattt ctacgtaaga cagatctttt gctaaccttc 3540

aaatgttatg ttgtactagg atattgaagg taggggcggg aatttatctg gtaagctaaa 3600

agagcttctg aatcttaaca atgaggaagc ttcagaggat tgtgatgtcc gagttgaagg 3660

tgtgatgaca aagcaaggcg tgaacgagga gatagacaca gttgaaaggt tgaaagagag 3720

cgatgcagat gctagcaatc ttgaaaccag tggttcagat acaagctcta acagagggaa 3780

gggtctggtt gaagagggag agttggttca gaatatgagc aagcgattta aaggcagtgc 3840

atcaggagag gtgaaggagg atgagaaatc tgaaaccttc cttgtctttg agaagcagaa 3900

gaagaaacgg aagcgtagta ttatgaatgc tgatcaaatg gggatgattg agaaggcgct 3960

tgctgaagaa cctgatttgc agcggaattc agcttcgaga cagttatggg ctgataaaat 4020

aagtcaaaag gtgagttcaa gaattctact ctttttaatt tccaaccacg tattacgaac 4080

gatgcgctcg ttgattacat gattttgcag ggttcggagg ttattacatc ttcgcagctg 4140

aaaaactggt acttgctttt atttgttcct cttcgtcagc tgcaaagaga atctcgtaat 4200

cactattcat tatctgacat tgaaagttta ttatcaggct gaataaccga aaagcgaaac 4260

tagctcgagc aaacaagcaa acggggccag ctcatgataa taacagctca ggggatctac 4320

cggagagtcc tggagatgaa aacacttggc agcagaaacc atcaacacca attaaagatc 4380

aaactgtaac agaaaccccg aaaacaggag agaatctgat gagaacatcg tcatcatcag 4440

aagaagggat aaagcaaggg caacaagtga ggcttatgga tgagagagga gacgagatcg 4500

ggaagggaac agtgttgaga acagacggtg aatggaacgg tttaagcttg gagacaagac 4560

agatatgtgt agttgatgta atggagctta gtgagtcata tgatgggagc aaaaagatga 4620

tcccttatgg atcagatgat gttgggagga ctttcacaga ggcgaattca aggtttggag 4680

ttatgagagt ggcttgggat gtgaataagc ttcagtatta gactcagttg cattgaattt 4740

gaccagtagg tttttgattt ttgtttattt ttaactgttt agagattctc ctctgttttg 4800

tgagctttgt ctctctctct ctgtggttat ttacaggcaa aggttgagac tttgagagga 4860

aggtaataac agaaggaata gtttgacttt tttaatctca gttagttgac tttacagcta 4920

tttaaagaaa caaa 4934

<210> 4

<211> 552

<212> DNA

<213> Streptomyces hygroscopicus (Streptomyces hygroscopicus)

<400> 4

atgagcccag aacgacgccc ggccgacatc cgccgtgcca ccgaggcgga catgccggcg 60

gtctgcacca tcgtcaacca ctacatcgag acaagcacgg tcaacttccg taccgagccg 120

caggaaccgc aggagtggac ggacgacctc gtccgtctgc gggagcgcta tccctggctc 180

gtcgccgagg tggacggcga ggtcgccggc atcgcctacg cgggcccctg gaaggcacgc 240

aacgcctacg actggacggc cgagtcgacc gtgtacgtct ccccccgcca ccagcggacg 300

ggactgggct ccacgctcta cacccacctg ctgaagtccc tggaggcaca gggcttcaag 360

agcgtggtcg ctgtcatcgg gctgcccaac gacccgagcg tgcgcatgca cgaggcgctc 420

ggatatgccc cccgcggcat gctgcgggcg gccggcttca agcacgggaa ctggcatgac 480

gtgggtttct ggcagctgga cttcagcctg ccggtaccgc cccgtccggt cctgcccgtc 540

accgagattt ga 552

Claims

1. The application of the protein ATNDX in improving the salt and alkali tolerance of corns or cultivating corns with high salt and alkali tolerance;

the protein ATNDX is a protein consisting of an amino acid sequence shown as SEQ ID No.2 in a sequence table.

2. Use of a DNA molecule encoding the protein ATNDX of claim 1 to increase salt and alkaline tolerance of corn or to cultivate high salt and alkaline tolerance corn.

3. Use according to claim 2, wherein the DNA molecule encoding the protein ATNDX of claim 1 is 1) or 2) as follows:

2) The coding region is a DNA molecule shown as SEQ ID No.1 in the sequence table.

4. Use of an expression cassette, recombinant vector, transgenic plant cell line or recombinant bacterium comprising the DNA molecule of claim 2 or 3 for increasing the salt and alkaline tolerance of maize or for cultivating high salt and alkaline tolerance maize.

5. A method of growing a transgenic plant, characterized by: the method comprises the following steps: a step of introducing a gene encoding the protein ATNDX of claim 1 into a starting plant to obtain a transgenic plant; the transgenic plant has higher saline-alkali tolerance than the starting plant; the plant is corn.

6. A method for plant breeding, characterized in that: the method comprises the following steps: increasing the amount of the protein ATNDX of claim 1 in a plant, thereby increasing the saline-alkali tolerance of the plant; the plant is corn.