CN114656541B

CN114656541B - Method for improving saline-alkali tolerance of plants by utilizing ZOG1 protein

Info

Publication number: CN114656541B
Application number: CN202011535560.9A
Authority: CN
Inventors: 杨永青; 郭岩; 刘晓
Original assignee: China Agricultural University
Current assignee: China Agricultural University
Priority date: 2020-12-23
Filing date: 2020-12-23
Publication date: 2023-05-12
Anticipated expiration: 2040-12-23
Also published as: CN114656541A

Abstract

The invention discloses a method for improving the salt and alkali resistance of plants by utilizing ZOG1 protein. The invention provides application of ZOG protein in regulating and controlling saline-alkali stress tolerance of plants. The invention also provides a method for cultivating transgenic plants, comprising the following steps: the ZOG gene is introduced into a receptor plant to obtain a transgenic plant with improved saline-alkali stress tolerance. ZOG1 protein is shown in a sequence 1 of a sequence table. ZOG1 gene is the gene encoding ZOG1 protein. The invention has great application value for cultivating saline-alkali tolerant plants.

Description

Method for improving saline-alkali tolerance of plants by utilizing ZOG1 protein

Technical Field

The invention belongs to the technical field of biology, and particularly relates to a method for improving saline-alkali tolerance of plants by utilizing ZOG protein.

Background

With the growth of population and the development of socioeconomic performance, the cultivated land area is gradually reduced, the land quality is also reduced, and the problem of land saline-alkali soil is more and more serious worldwide. Saline-alkali soil is an available resource, and improvement and utilization of the saline-alkali soil become important measures for ensuring that the red line of cultivated lands in China is not broken through and realizing sustainable development of agriculture. The promotion of ecological utilization and industrialization development of saline-alkali soil resources has important significance for building ecological civilization and promoting green development.

Saline-alkali soil refers to the phenomenon that the underground water level is increased, the mineralization degree is enhanced, and the salt content of deep soil is transferred to surface soil due to drought and strong evaporation of climate, so that the surface soil is salted or the alkalization degree is increased. The essence of the formation of saline-alkali soil is that soluble salts are redistributed in soil, and the salt accumulation in the soil surface exceeds a normal value. Slightly salinized soil with salt content of 0.1-0.2%; moderately salinized soil with a salt content of 0.2-0.4%; the salt content of the heavy salinized soil is 0.4-0.6%.

The harm of saline-alkali stress to plants is that Na with high concentration in saline-alkali soil ⁺ And too high a pH. Along with the accumulation of salt ions in plant cells, ion toxic effects are generated on the cells, so that the normal functions of the cells are influenced in various aspects. In alkaline earth environment, most of the metal elements except alkali metal elements form insoluble salts, na ⁺ As the alkali metal element with the highest content in nature, the alkali metal element is enriched in alkaline earth to cause salinization of soil.

Disclosure of Invention

The invention aims at providing a method for improving the saline-alkali tolerance of plants by utilizing ZOG1 protein.

The invention provides application of ZOG protein in regulating and controlling saline-alkali stress tolerance of plants. Specifically, the regulation is positive regulation, namely ZOG protein is increased, and the saline-alkali stress tolerance of plants is increased.

The invention provides application of ZOG protein in improving saline-alkali stress tolerance of plants.

The invention also protects the application of the ZOG gene or the recombinant vector containing the ZOG1 gene or the expression cassette containing the ZOG1 gene in cultivating transgenic plants with enhanced saline-alkali stress tolerance. The improvement of the stress tolerance of the saline and alkaline is reflected by greener leaves and/or higher plant height in the saline and alkaline environment.

The invention also provides a method for cultivating transgenic plants, comprising the following steps: the ZOG gene is introduced into a receptor plant to obtain a transgenic plant with improved saline-alkali stress tolerance. The ZOG gene is specifically introduced into a recipient plant by a recombinant vector containing the ZOG1 gene. The improvement of the stress tolerance of the saline and alkaline is reflected by greener leaves and/or higher plant height in the saline and alkaline environment.

The invention also provides a plant breeding method, which comprises the following steps: increasing the content and/or activity of ZOG1 protein in the target plant, thereby increasing the saline-alkali stress tolerance of the plant. The improvement of the stress tolerance of the saline and alkaline is reflected by greener leaves and/or higher plant height in the saline and alkaline environment.

The invention also protects ZOG protein.

The invention also protects ZOG gene. ZOG1 gene is the gene encoding ZOG1 protein.

ZOG1 protein, obtained from corn (Zea mays l.), is (a 1) or (a 2) or (a 3) as follows:

(a1) A protein shown in a sequence 1 of a sequence table;

(a2) A fusion protein obtained by connecting a tag to the N-terminal or/and the C-terminal of the protein of (a 1);

(a3) The protein (a 1) is a protein derived from the protein which has the same function and is obtained by substituting and/or deleting and/or adding one or more amino acid residues.

The labels are specifically shown in table 1.

TABLE 1 sequence of tags

Label (Label)	Residues	Sequence(s)
			Poly-Arg	5-6 (usually 5)	RRRRR
Poly-His	2-10 (usually 6)	HHHHHH
			FLAG	8	DYKDDDDK
Strep-tag II	8	WSHPQFEK
			c-myc	10	EQKLISEEDL
HA	9	YPYDVPDYA

The ZOG gene is a DNA molecule of the following (b 1) or (b 2) or (b 3) or (b 4):

(b1) A DNA molecule with a coding region shown as a sequence 2 of a sequence table;

(b2) A DNA molecule shown in a sequence 3 of a sequence table;

(b3) A DNA molecule which hybridizes to (b 1) or (b 2) under stringent conditions and which encodes a protein having the same function;

(b4) A DNA molecule having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% homology to (b 1) or (b 2) and encoding said protein.

Recombinant vectors containing the ZOG gene can be constructed using existing plant expression vectors.

In constructing the recombinant vector, any one of the enhanced, constitutive, tissue-specific or inducible promoters may be added before the transcription initiation nucleotide thereof, and they may be used alone or in combination with other plant promoters. In addition, when constructing recombinant vectors, enhancers, including translational or transcriptional enhancers, may be used, which may be ATG initiation codons or adjacent region initiation codons, and the like, but must be in the same reading frame as the coding sequence to ensure proper translation of the entire sequence. The sources of the translational control signals and initiation codons are broad, and can be either natural or synthetic. The translation initiation region may be derived from a transcription initiation region or a structural gene. To facilitate the identification and selection of transgenic plants, the recombinant vectors used may be processed, for example by adding genes which express enzymes or luminescent compounds which produce a color change in the plants, antibiotic markers which are resistant or marker genes which are resistant to chemical agents, etc. From the viewpoint of transgenic safety, transformed plants can be screened directly phenotypically without adding any selectable marker gene.

The plant expression vector may specifically be a pBCXUN vector.

The recombinant vector can be specifically a recombinant plasmid pBCXUN-ZOG1 obtained by inserting a DNA molecule shown in a sequence 2 of a sequence table into a pBCXUN vector.

The plant is a monocotyledonous plant or a dicotyledonous plant.

The plant is Gramineae plant.

The plant is a maize plant.

The plant is corn.

The plant is maize B73-329.

The inventor finds that ZOG1 participates in the saline-alkali stress response process and regulates the saline-alkali stress tolerance of plants. The ZOG1 over-expressed line exhibited significantly increased saline-alkali stress tolerance compared to maize B73-329. Thus, the ZOG protein has the function of improving the stress tolerance of plants to high saline and alkaline. The invention has great application value for cultivating saline-alkali tolerant plants.

Drawings

FIG. 1 is a photograph of a plant in example 2.

Detailed Description

The following detailed description of the invention is provided in connection with the accompanying drawings that are presented to illustrate the invention 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 invention 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. Maize (Zea mays L.) B73-329 is described in: wang Fang, cui Pengjuan, huang Yun, wang Zhiwen, wang Haifeng, chen Yifang. Molecular mechanisms of corn phosphorus absorption and redistribution [ A ],2018 university of plant biology treatise [ C ], 2018. The pBCXUN vector is an expression vector obtained by replacing the HYG gene (hptII, hygromycin resistance gene) of the pCXUN vector (GenBank: FJ905215.1, 06-JUL-2009) with the Bar gene (encoding phosphinothricin acetyltransferase) shown in sequence 4 of the sequence table, and keeping the other nucleotides of pCXUN unchanged.

Example 1, ZOG Gene and discovery of its encoded protein

A novel protein is found from corn, and is named as ZOG1 protein as shown in a sequence 1 in a sequence table. In the corn genome DNA, ZOG gene is shown as sequence 3 in the sequence table. In the maize cDNA, the open reading frame of the ZOG gene is shown as a sequence 2 in a sequence table.

Example 2 acquisition of transgenic plants and saline-alkali stress tolerance identification

1. Construction of recombinant plasmids

The DNA molecule shown in the sequence 2 of the sequence table is inserted into a pBCXUN vector to obtain a recombinant plasmid pBCXUN-ZOG1, and sequencing verification is carried out. In the recombinant plasmid pBCXUN-ZOG1, transcription of the DNA molecule shown in sequence 2 of the sequence table is started by the Ubi promoter and terminated by the Nos terminator, thereby expressing ZOG protein.

2. Obtaining of ZOG1 Gene overexpression plants

1. Recombinant plasmid pBCXUN-ZOG1 was introduced into Agrobacterium EHA105 to give recombinant Agrobacterium.

2. Infecting embryogenic callus of corn B73-329 by adopting recombinant agrobacterium prepared in step 1, then sequentially performing co-culture and resistance screening (herbicide glufosinate is adopted for resistance screening), and then sequentially performing pre-differentiation, differentiation and rooting to obtain T ₀ Regenerating plants.

3、T ₀ Carrying out PCR identification on the generation regeneration plants, and screening to obtain transgenic plants; will T ₀ The transgenic plant is selfed to obtain seeds which are T ₁ Seed generation, T ₁ The plant grown from the seed generation is T ₁ Generating plants; will T ₁ The seed obtained by selfing the plant is T ₂ Seed generation, T ₂ The plant grown from the seed generation is T ₂ Generating plants; will T ₂ The seed obtained by selfing the plant is T ₃ Seed generation, T ₃ The plant grown from the seed generation is T ₃ And (5) replacing plants.

4. Will T ₁ Substitution plants and sampled T ₂ And carrying out PCR identification on the generation plants. For a certain T ₁ Plants of the generation, if the plants and T obtained by selfing the plants ₂ The generation plants are all transgenic plants, and the plants are self-grownThe cross offspring are homozygous transgenic lines. Three homozygous transgenic lines were obtained, ZOG-OX 1, ZOG-OX 2 and ZOG-OX 3, respectively.

The PCR identification method in the step 3 and the step 4 is as follows: extracting genomic DNA of plant leaves, carrying out PCR amplification by using primer pairs consisting of UbiP-seq (corresponding to the Ubi promoter) and NosR-seq (corresponding to the Nos terminator), and obtaining a specific amplification product if the specific amplification product is obtained, wherein the plant is a transgenic plant.

3. Saline-alkali stress tolerance identification of ZOG1 gene over-expressed plant

Test seed: t of ZOG-OX 1 Strain ₃ T of the seed of the generation, ZOG-OX 2 strain ₃ T of the seed of the generation, ZOG-OX 3 strain ₃ Seed generation, seed of corn B73-329.

Test seeds were planted in vermiculite, watered with 1/2Hoagland's nutrient solution, and normally cultured to 3 leaf stage. Then start with 120mM NaHCO ₃ 1/2Hoagland's nutrient solution (irrigated every 7-10 days) was used for normal culture for 28 days. Then observing, photographing and counting the plant height.

Five replicates were performed, with at least 5 biological replicates per sample set.

The plant photograph is shown in FIG. 1.

The ZOG gene over-expressed plant shows stronger saline-alkali stress tolerance than that of the corn B73-329, and is particularly characterized by greener leaves and higher plant height. The plant height of the ZOG1 gene over-expression plant is 1.4 to 1.5 times that of the maize B73-329 plant.

The results show that the ZOG1 protein positively regulates the saline-alkali stress tolerance of plants.

The present invention is described in detail above. It will be apparent to those skilled in the art that the present invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with respect to specific embodiments, it will be appreciated that the invention may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention 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> method for improving saline-alkali tolerance of plant by ZOG1 protein

<130> GNCYX203352

<160> 4

<170> SIPOSequenceListing 1.0

<210> 1

<211> 226

<212> PRT

<213> Zea mays L.

<400> 1

Met Glu Ala Thr Leu Cys Pro Lys Pro His Phe Val Val Ile Pro Trp

1 5 10 15

Pro Ala Thr Ser His Met Ile Pro Ile Val Asp Ile Gly Cys Leu Leu

20 25 30

Ala Ala His Gly Ala Ala Val Thr Ile Ile Thr Thr Pro Ser Ser Ser

35 40 45

Gln Leu Val Gln Ser Arg Val Asp Arg Ala Gly Gln Gly Ser Ala Gly

50 55 60

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

65 70 75 80

Asp Gly Cys Glu Arg Thr Asp His Ile Pro Ser Pro Asp Leu Val Pro

85 90 95

Asn Phe Phe Val Ala Thr Ala Arg Phe Gly Glu Ala Val Ala Arg His

100 105 110

Cys Arg Arg Leu Pro Thr Ala Thr Ala Ala His Pro Arg Pro Ser Cys

115 120 125

Val Val Ala Gly Met Cys His Thr Trp Ala His Gly Val Ala Arg Glu

130 135 140

Leu Gly Ala Pro Cys Phe Ile Phe His Gly Phe Cys Ala Phe Ala Leu

145 150 155 160

Leu Cys Cys Glu Tyr Leu Asn Thr His Arg Pro His Glu Ala Val Gly

165 170 175

Ser Pro Asp Glu Leu Phe Asp Leu Pro Ala Leu Pro Pro Phe Glu Phe

180 185 190

Arg Phe Gln Glu Arg Cys Arg Pro Gly Lys Leu Cys Pro Gly Leu Arg

195 200 205

Val Asp Arg Thr Gly Ala Glu Glu Lys Arg Gln Gln Lys Arg Gln Asn

210 215 220

Glu Glu

225

<210> 2

<211> 681

<212> DNA

<213> Zea mays L.

<400> 2

atggaggcca ccctgtgccc aaagccccat ttcgtggtca tcccatggcc agccaccagc 60

cacatgatcc ccatcgtgga catcggctgc ctcctcgccg cgcacggcgc cgcggtcacg 120

atcatcacca cgccctccag ctcgcaactc gtccagagcc gcgtggaccg cgccgggcaa 180

ggctcagcgg gggtcacggt caccgcgatc ccgttcccgg gcgcggaggc cggcctgccg 240

gacggctgcg agaggacgga ccacatcccc tcccctgacc tcgtgcccaa cttcttcgtc 300

gccaccgcgc ggttcggcga ggccgtggcg cggcactgcc gccgcctgcc cacggccacg 360

gcggcgcacc cccggccgag ctgcgtcgtc gccgggatgt gccacacgtg ggcgcacggc 420

gtggcgcgcg agctcggcgc cccctgcttc atcttccacg gcttctgcgc gttcgctctg 480

ctgtgctgcg agtacctcaa cacgcacagg ccgcacgagg cggtcgggtc gccggacgag 540

ctcttcgacc tccccgccct gccgccgttc gagttcaggt ttcaggaacg ctgcaggccg 600

ggcaaacttt gtccaggatt gcgagtggac aggacaggag cagaggaaaa acggcaacag 660

aagcgacaaa atgaagagta a 681

<210> 3

<211> 2915

<212> DNA

<213> Zea mays L.

<400> 3

tgcattaatc tgcagtcgcc atcagtataa cggttacagg ccaaattgca acatccagga 60

tcagtgtaag ccagcaatgc cgtggcgtca aatccaaatc gaattgctta acctctccca 120

agaacggcca aagcctattt ctgtgacact ggcaacgagt gtagcaacag tcacactggc 180

aagcaagcaa gcagcaccag cgtttactca aagtcgcaaa ccaacgaaat gccatggcga 240

agctattgtg gaagctggcg tacgtcgagc gccagcgcta cgcccagcgt cggctcatcg 300

tgtggcaagt ctggacattg tgtaatttgg aggattgcag aaaaataaat aacttgtgga 360

gtttggacaa atccagccgg tccttatcag ttagttcagg ctatacaaag acgacaatgc 420

acacgtcgcc accaatcttg ttttacttgg ctttgtccac tgcacttctg ctgcggtata 480

tatataagcc gctctgctag cagaagcaaa acatgcatcg ggaaccctta cacaggtaga 540

taccacaacc ctgtgcagag ttcttgcatt ctcgtcaagg tgacaggaac gacataggaa 600

atggaggcca ccctgtgccc aaagccccat ttcgtggtca tcccatggcc agccaccagc 660

cacatgatcc ccatcgtgga catcggctgc ctcctcgccg cgcacggcgc cgcggtcacg 720

atcatcacca cgccctccag ctcgcaactc gtccagagcc gcgtggaccg cgccgggcaa 780

ggctcagcgg gggtcacggt caccgcgatc ccgttcccgg gcgcggaggc cggcctgccg 840

gacggctgcg agaggacgga ccacatcccc tcccctgacc tcgtgcccaa cttcttcgtc 900

gccaccgcgc ggttcggcga ggccgtggcg cggcactgcc gccgcctgcc cacggccacg 960

gcggcgcacc cccggccgag ctgcgtcgtc gccgggatgt gccacacgtg ggcgcacggc 1020

gtggcgcgcg agctcggcgc cccctgcttc atcttccacg gcttctgcgc gttcgctctg 1080

ctgtgctgcg agtacctcaa cacgcacagg ccgcacgagg cggtcgggtc gccggacgag 1140

ctcttcgacc tccccgccct gccgccgttc gagttcaggt tcgccaggag gcagctgccg 1200

atccacttcc agccgtcttc ctccatcccg gaggaccgcc accgggagct ccgggagttc 1260

gagctggccg tggacggcat cgtcgtgaac agcttcgagg agctggagca cggctcggtt 1320

tcgcgcctcg cggcggccac gggcaaggcc gtgctctccg tcggccccgt ctcgctgtgc 1380

ggcgccgccg cacctcctag cctcctcgac tcgcgggcca actcggacga tgccaggctg 1440

tgcatggcgt ggctggacgc caagagagcc gactccgttc tctacgtcag cttcggcagc 1500

gccgggcgca tgcctccagc gcagctgata cagctcgggc tggcgctcgt gtcgtgcccc 1560

tggcctgtcc tttgggtcat caagggcgca gacacgttgc ccgatgacgt caacgagtgg 1620

ttgcagcgca acaccgatgg aagcggcctc ccggagagcc agtgtcttgc gcttcgcggg 1680

tgggcgccgc aggtcgccat cctggagcac ccggccgtcg gcggctttct gacgcactgc 1740

gggtggggct cgacgctgga gagcgtcgcc gcgggcgtgc ccatggccac ctggccgttc 1800

tccgccgagc agttcctgaa cgagaagttg atcgtccacg tgcttggcat cggattgtct 1860

gtcggcgtga cgaagcccac ggagagcgtg ctgaccggtg ccaaagacgg cggtggcaag 1920

gcggacgcag atgtgggaat ggaacaggtg aagcaagcct tggatatgct catggatgga 1980

ggggcggatg gggaggcaag gaagacgaaa gctaaggaac tgaaggccaa atcaaagact 2040

gctctggagc atggaggatc gtcatatatg aacctggaga agctgataca gtttggtgct 2100

tgaaaatgga gtggtcgtgg actcgtggtc tgtttcggca cccaaactta gtttatttct 2160

ccctaataaa ggttaataaa ctaggactgc aagcaaaacg tggtacatgt aggtttcagg 2220

aacgctgcag gccgggcaaa ctttgtccag gattgcgagt ggacaggaca ggagcagagg 2280

aaaaacggca acagaagcga caaaatgaag agtaacaaag agccgttgga ttgaagatca 2340

atgactcatg tgcgttgcct aagaattaga catatagtca tatgagcgtc ttctctatga 2400

tacctgaggt ttcttcgatt ccacgttttt ccgtctagtt tctccactta gtacgcgttc 2460

ggttagctct caatccatgt gctcgattcc acgtttttcc gtctagtttc tccacttagg 2520

acgcgttcgg ttagctctca atccatgtgg attaaatgta attgggtgga tttaaatctc 2580

aaataagtca aacttattct caatttttcc attcctatct aatccgtgtg taatgcgaat 2640

aaccaaataa tagttaaaaa ttattaatta aagattataa agttaatttg aatatagata 2700

tatgtatatg ctttgtaaat catgccttat ttggtaggct tcttcttctt cttctcaaaa 2760

aatgacttta gctcttgttt cttttgtcaa aggtgctttc ctagagaagc caaaggcata 2820

caaaacaatt tctatgctat tttaaaaaag aaactatatc tagttacatt agtaatatag 2880

ttaatctcaa ttgtaatttg ttgtgtgcct gtgta 2915

<210> 4

<211> 552

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<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

Application of ZOG1 protein in regulating alkali stress tolerance of plants;

ZOG1 protein is (a 1) or (a 2) as follows:

(a1) A protein shown in a sequence 1 of a sequence table;

(a2) A fusion protein obtained by connecting a tag to the N-terminal or/and the C-terminal of the protein of (a 1);

the regulation is that ZOG1 protein is increased, and the alkali stress tolerance of plants is increased;

the plant is Gramineae plant.
2. Use of a gene encoding a ZOG1 protein, a recombinant vector comprising a gene encoding a ZOG1 protein, or an expression cassette comprising a gene encoding a ZOG1 protein for breeding transgenic plants with increased alkali tolerance;

ZOG1 protein is (a 1) or (a 2) as follows:

(a1) A protein shown in a sequence 1 of a sequence table;

(a2) A fusion protein obtained by connecting a tag to the N-terminal or/and the C-terminal of the protein of (a 1);

the plant is Gramineae plant.
3. The use according to claim 2, wherein:

the gene encoding ZOG1 protein is the following (b 1) or (b 2):

(b1) A DNA molecule with a coding region shown as a sequence 2 of a sequence table;

(b2) A DNA molecule shown in a sequence 3 of a sequence table.
4. A method of growing a transgenic plant comprising the steps of: introducing a gene encoding ZOG protein into a recipient plant to obtain a transgenic plant with increased alkali stress tolerance;

ZOG1 protein is (a 1) or (a 2) as follows:

(a1) A protein shown in a sequence 1 of a sequence table;

(a2) A fusion protein obtained by connecting a tag to the N-terminal or/and the C-terminal of the protein of (a 1);

the plant is Gramineae plant.
5. The method of claim 4, wherein:

the gene encoding ZOG1 protein is the following (b 1) or (b 2):

(b1) A DNA molecule with a coding region shown as a sequence 2 of a sequence table;

(b2) A DNA molecule shown in a sequence 3 of a sequence table.
6. A plant breeding method comprising the steps of: increasing the content of ZOG protein in the target plant, thereby increasing the alkali stress tolerance of the plant;

ZOG1 protein is (a 1) or (a 2) as follows:

(a1) A protein shown in a sequence 1 of a sequence table;

(a2) A fusion protein obtained by connecting a tag to the N-terminal or/and the C-terminal of the protein of (a 1);

the plant is Gramineae plant.