CN114656544A - Protein GH3.9 and its biological material and method for cultivating plants with high stress tolerance - Google Patents

Abstract

The invention discloses protein GH3.9 and its biological material and a method for cultivating plants with high stress tolerance. The present invention provides a method for cultivating plants with high stress tolerance, comprising increasing the content and/or activity of protein GH3.9 in a target plant to obtain plants with high stress tolerance, wherein the stress tolerance of the high stress tolerance plants is higher than that of target plants. The invention provides a theoretical basis for elucidating the molecular mechanism of protein GH3.9 in the response of plants to saline-alkali stress, and provides genetic resources for cultivating and improving new varieties of stress-resistant plants.

Description

Protein GH3.9 and its biological material and method for cultivating plants with high stress tolerance

technical field

The present invention relates to the field of biotechnology, in particular to protein GH3.9 and its biological material and a method for cultivating plants with high stress tolerance.

Background technique

Because soil salinization and alkalization often occur together, people often refer to the increase of soil soluble salinity as "soil salinization". However, with the continuous in-depth research on the effects of salt stress and alkali stress on plant physiology and ecology, researchers have found that salt stress and alkali stress have different harms to plants, and the physiological responses to salt stress and alkali stress are also different. There are significant differences. Salt stress is mainly caused by neutral salts such as NaCl, Na ₂ SO ₄ , while alkaline stress is mainly caused by alkaline salts such as bicarbonate (HCO ₃ ^- ) and carbonate (CO ₃ ^2- ), comparing the two In terms of alkali stress, the damage is more serious and complex. The damage of salt stress to plants mainly includes ionic stress, osmotic stress and oxidative stress, while alkali stress increases HCO ₃ ^- or CO ₃ ^2- ion stress and high pH caused by bicarbonate or carbonate on this basis. coercion.

With the rapid development of molecular biology, genomics, genetics, biochemistry and gene editing technology, the research on the molecular mechanism of plant resistance to saline-alkali stress has been deepened, and many new genes or proteins have been found to be involved in regulating the process of saline-alkali stress response. middle.

SUMMARY OF THE INVENTION

The technical problem to be solved by the present invention is how to regulate plant stress tolerance and/or how to cultivate plants with high stress tolerance. The highly stress-tolerant plant may be a plant that is more stress-tolerant than a wild-type plant.

In order to solve the above technical problems, the present invention provides a method for cultivating plants with high stress tolerance, comprising increasing the content and/or activity of protein GH3.9 in a target plant to obtain plants with high stress tolerance. The stress tolerance is higher than that of the target plants.

The protein GH3.9 is as follows (1) or (2):

(1) a protein consisting of the amino acid sequence shown in SEQ ID No.2 in the sequence listing;

(2) A protein derived from (1) with the amino acid sequence shown in SEQ ID No. 2 in the sequence listing subjected to substitution and/or deletion and/or addition of one or several amino acid residues and having the same function.

Said substitution and/or deletion and/or addition of one or several amino acid residues is a substitution and/or deletion and/or addition of not more than 10 amino acid residues.

Optionally, according to the above method, the improving the content and/or activity of protein GH3.9 in the target plant is to introduce a substance that regulates the expression of the protein GH3.9 encoding gene into the target plant.

Optionally, according to the above method, the substance that regulates the expression of the protein GH3.9 encoding gene is any one of biological materials in B1)-B4);

B1) a nucleic acid molecule encoding the protein GH3.9 of claim 1;

B2) an expression cassette containing the nucleic acid molecule of B1);

B3) a recombinant vector containing the nucleic acid molecule described in B1) or a recombinant vector containing the expression cassette described in B2);

B4) a recombinant microorganism containing the nucleic acid molecule of B1), or a recombinant microorganism containing the expression cassette of B2), or a recombinant microorganism containing the recombinant vector of B3).

In the above, the nucleic acid molecule can be DNA, such as cDNA, genomic DNA or recombinant DNA; the nucleic acid molecule can also be RNA, such as mRNA, siRNA, shRNA, sgRNA, miRNA or antisense RNA.

In the above method, the increase of the expression of the gene can be achieved by methods well known in the art, such as multiple copies, changing promoters, regulatory factors, and transgenes. For example, the protein GH3.9 encoding gene overexpression vector is introduced into the target plant, and the protein GH3.9 encoding gene overexpression vector may specifically be the recombinant plasmid pBCXUN-GH3.9 mentioned in the examples.

Optionally, according to the above-mentioned method, the nucleic acid molecule of B1) encoding the protein GH3.9 of claim 1 is the nucleic acid molecule of any one of the following 1)-4):

1) the nucleotide sequence is the nucleic acid molecule shown in SEQ ID No.3 in the sequence listing;

2) the coding sequence is the nucleic acid molecule shown in SEQ ID No.1 in the sequence listing;

3) Under stringent conditions, it hybridizes with the DNA molecule defined in 1) or 2) and encodes a nucleic acid molecule with the same functional protein;

4) have at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% homology with the DNA molecule as defined in 1) or 2) and encode a protein with the same function nucleic acid molecules.

One of ordinary skill in the art can easily mutate the nucleotide sequence of the nucleic acid molecule encoding the protein GH3.9 of the present invention using known methods, such as directed evolution and point mutation. Those artificially modified nucleotides with 90% or higher identity to the nucleotide sequences provided by the present invention, as long as they encode protein GH3.9 and are derived from corn, are all derived from the nucleotides of the present invention sequence and is equivalent to the sequence of the present invention. The term "identity" as used herein refers to sequence similarity to a native nucleic acid sequence.

For example, the nucleotide sequence of the genome gene of the nucleic acid molecule encoding the protein GH3.9 is shown in SEQ ID No. 3, named GH3.9 gene, derived from maize B73-329. The gene encodes protein GH3.9, the amino acid sequence of which is shown in SEQ ID No. 2 of the sequence table, and the CDS of the cDNA gene of the protein is shown in SEQ ID No. 1.

Protein GH3.9 and the above-mentioned biological materials also fall within the protection scope of the present invention.

The present invention also provides the application of the protein GH3.9 and the above-mentioned biological materials in regulating plant stress tolerance or cultivating plants with high stress tolerance.

Optionally, in the above, the plant is a dicotyledonous plant or a monocotyledonous plant. The monocotyledonous plant may be a grass plant such as maize.

Optionally, in the above, the stress tolerance is salinity and alkali tolerance, and the regulation of plant stress tolerance is improving the salinity and alkali tolerance of plants.

In the present invention, the white GH3.9 gene is introduced into maize B73-329 to obtain a transgenic overexpression line of GH3.9; It is proved that the function of protein GH3.9 is to improve the stress tolerance of plants to high salinity. The invention provides a theoretical basis for elucidating the molecular mechanism of protein GH3.9 in the response of plants to saline-alkali stress, and provides genetic resources for cultivating and improving new varieties of stress-resistant plants.

Description of drawings

Figure 1 shows the growth of the plants under saline-alkali stress treatment in Example 2.

Detailed ways

The present invention will be further described in detail below with reference to the specific embodiments, and the given examples are only for illustrating the present invention, rather than for limiting the scope of the present invention. The examples provided below can serve as a guide for those of ordinary skill in the art to make further improvements, and are not intended to limit the present invention in any way.

The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are performed according to the techniques or conditions described in the literature in the field or according to the product specification. The materials, reagents, etc. used in the following examples can be obtained from commercial sources unless otherwise specified.

Example 1. Construction of GH3.9 gene overexpression plants

1. Construction of pBCXUN-GH3.9 gene overexpression vector

The CDS of the cDNA gene of the GH3.9 protein (ie, the DNA molecule shown in SEQ ID No. 1) was inserted into pBCXUN to obtain a recombinant plasmid pBCXUN-GH3.9. In the recombinant plasmid pBCXUN-GH3.9, the CDS of the cDNA gene of the GH3.9 protein driven by the Ubi promoter (maize ubiquitin-1 promoter) expresses the GH3.9 protein, and the transcription is terminated by the Nos terminator.

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

2. Construction of recombinant Agrobacterium pBCXUN-GH3.9/EHA105 and preparation of recombinant Agrobacterium suspension

The pBCXUN-GH3.9 of step 1 was transformed into competent Agrobacterium EHA105 strain by heat shock method, colony PCR identified positive clones, selected positive clones for sequencing, and obtained recombinant Agrobacterium pBCXUN-GH3.9/EHA105 (containing pBCXUN- positive clone of GH3.9).

A single colony of pBCXUN-GH3.9/EHA105 was inoculated into 2-3 mL of liquid medium containing 100 μg/mL kanamycin and 50 μg/mL rifampicin, and cultured overnight at 28°C with shaking. The next day, the cells were transferred to a large amount of liquid medium containing antibiotics for shaking culture. After transferring several times, the cells were collected and resuspended to an OD ₆₀₀ of 0.8-1.0 to obtain a recombinant Agrobacterium suspension.

3. Construction of GH3.9 overexpression transgenic plants

After the recombinant Agrobacterium suspension obtained in step 2 was used to infect B73-329 maize young embryos, callus was induced to form seedlings, and then co-cultivation, resistance screening (the herbicide glufosinate was used for resistance screening), and then successively Pre-differentiation, differentiation and rooting were carried out to obtain T ₀ generation regenerated plants. The transgenic plants were screened by PCR identification method from the regenerated plants of T ₀ generation. The transgenic plants in the T ₀ generation were selfed to obtain the T ₁ generation seeds, and the plants grown from the T ₁ generation seeds were the T ₁ generation plants; the plants were selfed until the T ₃ generation seeds were obtained, that is, the GH3.9 overexpression transgenic plants T ₃ generation seeds.

_The total RNA of GH3.9 overexpressing transgenic plant T3 was extracted, cDNA was reverse transcribed, and the overexpression of GH3.9 gene was detected by RT PCR. Maize B73-329 was used as a control. The results showed that the GH3.9 gene expression level in the GH3.9 overexpression transgenic plants was much higher than that in the non-transgenic control plant maize B73-329.

Example 2. Detection of GH3.9 gene overexpression maize saline-alkali stress treatment

Two treatment groups were set up in the experiment, the control group and the experimental group. The experiment was repeated 3 times. Each repetition consisted of 10 pots and 3-4 seeds per pot. The control group used B73-329 corn seeds, and the experimental group used the T ₃ generation seeds of the GH3.9 overexpression transgenic plants harvested in Example 1.

Planted with pure vermiculite without nutrient soil and watered with 1/2 Hoagland's nutrient solution. Plants were grown for 7 days (three-leaf stage) by watering with 1/2 Hoagland's nutrient solution containing 100 mM NaHCO ₃ (saline-alkali stress) (liquid obtained by adding NaHCO ₃ to 1/2 Hoagland's nutrient solution to a NaHCO ₃ content of 100 mM), and watering every 10 days Once, two liters per pot were grown for 40 days to observe the phenotype and measure the plant height.

Figure 1 shows part of the experimental results of saline-alkali stress treatment. After 40 days of saline-alkali stress treatment, the plants in the experimental group showed stronger stress tolerance than the plants in the control group, with greener leaves and higher plant height. It is 1.3-1.6 times of the plant height of the control group.

Experiments showed that GH3.9 positively regulates plant salinity resistance.

The present invention has been described in detail above. For those skilled in the art, without departing from the spirit and scope of the present invention, and without unnecessary experimentation, the present invention can be implemented in a wide range under equivalent parameters, concentrations and conditions. While the invention has been given particular embodiments, it should be understood that the invention can be further modified. In conclusion, in accordance with the principles of the present invention, this application is intended to cover any alterations, uses or improvements of the invention, including changes made using conventional techniques known in the art, departing from the scope disclosed in this application. The application of some of the essential features can be made within the scope of the following appended claims.

sequence listing

<110> China Agricultural University

<120> Protein GH3.9 and its biomaterial and method for cultivating plants with high stress tolerance

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atgaatgggt ccgtagacat ttctgccttc aagagccagg ttccagtagt gacgtatgat 60

gtggtccaac cctacattgc aaggattgcc gctggtgaaa ggtcttcaat tttgtgtgga 120

gaacaaattg tagagctgct ccgaagctcg ggaacttctc agggtgaacc aagattgatg 180

ccatcaatct cagaggatat ttaccgccga atgtatctgt atagtctgat catgcctatc 240

atgagcaagt acatacgagg ccttggcgag gggaaggcaa tgtatcttct ctttgtgaaa 300

gctgaaacat tgacaaatgc tggcattcca gtccgatcag tcctcaccag ctactacaag 360

agccctcagt tcttgcagcg caagcacgac ctctacaaca gctacaccag tcccgacgag 420

gtcattcttt gccccgacag ccagcagagc atgtactgcc agctactctg cggcctggtg 480

gaacgccaga acgtcgtccg tctcggagca gtcttcgcct cagcattcct ccggtccatc 540

agcttcctcg agaagcactg gcatgaccta gtcaccgaca tccgaaccgg ccggatcaat 600

cccagcatca ccaacgccgc atgccgggtg gccacggaga gctttctcgc gcagcccaac 660

cccgagctag ctgacgaagt cgaggcgata tgcagctcgg agtcatggaa gggcgttctt 720

ggcaggctct ggcctaacgt caagtacatc gaggccgtcc tgacggggac gatggcgcag 780

tacgtcccca tgctcgagtt ctacagcgac ggcaggatcc ccctggtctg caccatgtac 840

gcttcctcgg agagctactt cggcgtcaac ctgcggccgc tgtgcagccc gaaggacgtg 900

tcctacacca tcctccccaa catggcctac ttcgagttcg tgccgttgga cgacggcctc 960

aggctggcgg aggatggtga ggctgtggag aaggagaagc ttgtcggcct cgtggacgtc 1020

aaggtcggat gctactacga gctcgtggtt accacatttg ccggtcttta caggtacagg 1080

gtaggcgatg tactccaggt gacaggcttc tacaaccgcg caccccagtt caaattcatc 1140

tgccggagga acgtgatcct gagcgtggac gcagacaaga cgaacgagga agacctccac 1200

ggcagcgtct cccgggccaa gaagatcctg gaggaccgag gccacatcct actggagtac 1260

accagcttca ccgacacgtc gacggtgccg ggccactacg tcctgttctg ggaggtgaag 1320

gccacgccca cctccggctc cgggggcgcg cggctggacg cgcaggtcct ggagagctgc 1380

tgcgtcgccg tggaggagtc gctggactgc gtgtacaggc ggtgcagggc gcacgacagg 1440

tccgtggggc cgctggagat acggctggtg gaagccggcg ccttcgacgc gctcatggac 1500

ctgttggtca gccagggcgg ctccatcaac cagtacaaga cgccacggtg catcgagccg 1560

ggcggcgccg cgctggagct gctcgactcc aaggccaccg cctcgttctt cagcccccgg 1620

gatcctgaag ctactggtgt ctga 1644

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Val Thr Tyr Asp Val Val Gln Pro Tyr Ile Ala Arg Ile Ala Ala Gly

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Glu Arg Ser Ser Ile Leu Cys Gly Glu Gln Ile Val Glu Leu Leu Arg

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Ser Ser Gly Thr Ser Gln Gly Glu Pro Arg Leu Met Pro Ser Ile Ser

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Glu Asp Ile Tyr Arg Arg Met Tyr Leu Tyr Ser Leu Ile Met Pro Ile

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Leu Phe Val Lys Ala Glu Thr Leu Thr Asn Ala Gly Ile Pro Val Arg

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Ser Val Leu Thr Ser Tyr Tyr Lys Ser Pro Gln Phe Leu Gln Arg Lys

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His Asp Leu Tyr Asn Ser Tyr Thr Ser Pro Asp Glu Val Ile Leu Cys

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Glu Arg Gln Asn Val Val Arg Leu Gly Ala Val Phe Ala Ser Ala Phe

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Leu Arg Ser Ile Ser Phe Leu Glu Lys His Trp His Asp Leu Val Thr

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Asp Ile Arg Thr Gly Arg Ile Asn Pro Ser Ile Thr Asn Ala Ala Cys

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Arg Val Ala Thr Glu Ser Phe Leu Ala Gln Pro Asn Pro Glu Leu Ala

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Asp Glu Val Glu Ala Ile Cys Ser Ser Glu Ser Trp Lys Gly Val Leu

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Gly Arg Leu Trp Pro Asn Val Lys Tyr Ile Glu Ala Val Leu Thr Gly

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Thr Met Ala Gln Tyr Val Pro Met Leu Glu Phe Tyr Ser Asp Gly Arg

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Ile Pro Leu Val Cys Thr Met Tyr Ala Ser Ser Glu Ser Tyr Phe Gly

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

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Leu Pro Asn Met Ala Tyr Phe Glu Phe Val Pro Leu Asp Asp Gly Leu

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Arg Leu Ala Glu Asp Gly Glu Ala Val Glu Lys Glu Lys Leu Val Gly

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Leu Val Asp Val Lys Val Gly Cys Tyr Tyr Glu Leu Val Val Thr Thr

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Phe Ala Gly Leu Tyr Arg Tyr Arg Val Gly Asp Val Leu Gln Val Thr

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tactacgagt tttgatcatg gatggtgccg gtaacgcaag gtgttgatta ctgaaagtag 180

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gctttcagga gagaaaactg tcagtgagct gcgagcctgc gacaagtcgg acaaaaagaa 360

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ccgctttcct tgccgatatt ggctgctcgc tgatctttga gggccgcgat taccgcacct 600

cgaggaggcg aggtgaggcg taaaaaactc aaacaacgta aggtgttcga cacccggtca 660

gcggcctcga cctctttcgg tcgatccaac tacaccattg cgctgaatca atctgcccgc 720

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cttttacgca gcagggcgtc cccactcccc cacggcgtta tgccgcgccc accacctgtt 1200

cggcaaaatg gctagtatgt catatgtgca ataaactaat acacttgacg ggtcgcgagt 1260

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ttctgaattg accggaaaat atcacaaagg aagtacatca gctgcaatta tcacagacag 1800

gcttgttcca ttgacaatta gcatatcctt ttgaaatatt atacaattct atttcgactg 1860

ggaaaatgtg tcttccaatt gcagccacca ctgcccttaa caatagcgtt tagctaaata 1920

tatagcttaa ctgagaaaaa tatgcatgca tatcctatac tttatacctg tggtgaagct 1980

agataaactt aggacggggt ggggtgggat gtgctcttgc cttgtggggc atataaattt 2040

tatggaggga gggtgaatat atggtaaaaa aatagacaaa atcactgctt tagcattttt 2100

tacagggtgc atttgcatcc cttactctca acgcagcctc acccctgctt tataggcagc 2160

atgcccatgt ttattgtcga gactaatcct ttaatgcttt atagtcagtg tgcccatttg 2220

gtgtttgatt acggtcatat gtcttattgg ttcagtggag acggagccta gctatgcaat 2280

cttaggcttt tttctgtttt gattccgcac aatcacttgc acggacggca taacgtgcgg 2340

aacacaattt gtgtttaggt tctggtgtag ctcttcatct tttctcataa tctcttatga 2400

aggatttagg tgctgatgca acatttcaag aataactttt gaagtggata aatgcgttct 2460

tatctttcag tcctgcacac tttgttggtt gcatgcaggg gcagacccaa gtagggccga 2520

gtgagggcac atgcccccgc tcatttcttt gttctaagta ctagagtcta aattttcatc 2580

atgaggtccc ctgctcagtc taatttagat gcctatattc tggtatttta gttctcgtcc 2640

ttacaagtat gctctcagtt atattttgtc ctgggtccgc ccctggttgc atgtgcccgt 2700

gttggcagat agtgtgcatt ccttaggggt gtattatcga taactttgtg ggtggttaga 2760

caattattat tgtctctaaa tctgcacctt ctgtctcatg agtgttggta gtttcttctg 2820

gacgttttaa tttgttgaag aaaatattct tcaaatatct tttagtagtc ataagaacaa 2880

aatgtccatt ctcattggtt tgtattttga tctttcattt tcagtagctc tgtggtaggg 2940

tccatgggat tgtcttgcag aggaaacatc aaagttgttt taggtgttct caatctctgg 3000

aaagtctggt aagtattact actcactaga atcagtatag ttatgataga aaaattcctg 3060

gatgaaagaa aataagggta tgccaaaagt accacttttt tcaacccttt tagggctagt 3120

ttggaaacta aaatcccctc cgggattccc gaggattgaa tgggaaaata gctaattttc 3180

ccctcaatcc ccgggaatct cagaggggat ttaagtttcc aaactagacc ttagtagttg 3240

ctttatcagg ttatcctatc ctttgaacta ctttagccat gcccttaaaa attatggttt 3300

tgacactgaa cagttcacct ttagaagata aaattgtcct acgttgcaca ttgtttctcc 3360

aaacctgcag aagtgtagtt tcacataacg agaacacaag ttaagtgtga caagacaaaa 3420

caatgtaaca gatgaagtac tgagtcgaga cagataattt aaagtagaac agtaatgaca 3480

gtggaacttc tagatatact gtaacatatt atcgcgtcca taacaattgt atgtggattg 3540

ttttacttgg gtgcaccatc caattagtcc agaatctctc ggataaggta gagagaaatt 3600

aagaagatat atgcttctcg ggtcatgcaa gatctataaa aggggtgcac ttgctccaga 3660

aaccgttact ttgagccatt gccttcctct tctaaataag acccatctct aattcttcta 3720

acatggatgc acataagtta ggttaacacag gaaatgatgt tcttcaagag cttgaaatac 3780

tgactataaa tgccaaagca gcacaagagg ttatattgag taaaattctt gagaagaatc 3840

aggctactga atacctgaac aaattcatga atgggtccgt agacatttct gccttcaaga 3900

gccaggttcc agtagtgacg tatgatgtgg tccaacccta cattgcaagg attgccgctg 3960

gtgaaaggtc ttcaattttg tgtggagaac aaattgtaga gctgctccga aggtgtgagt 4020

ctcttcctga ttatttttta gttaacattt ttgcatgcat atagtagagg tattgtatta 4080

ggtataagaa tgattataca tttatcagaa gaccctttgt gctggctgct agtagaagct 4140

ggttctattg tataatgcga atatgcgact aatatgaaga aaatgagcat taatgttaga 4200

tcatttcatc gtcaaaatgc atgctggtat ataatttttt tttctcaaac aaagattgag 4260

aagtgcacat cattgtatta aagaaggtga acacaagtcc gtgaaaagtg aaaacacagg 4320

ctaaaaagaa aaaaagaaga acaggaaaga acaacaaaca caaatagata ccaggcgtag 4380

ccagtaaact aaacagaact gctcaagcta ttacggtaac tcagaaggca gcataaggag 4440

tgctggacga caatgatagg catagactcc tagcacccgc tgtcttccaa atatttcgct 4500

cactagtgat gatagcccgt agaaccctgg tgagctgagg ggcgagctga ggttgtgcgc 4560

ctttgaacag ccgttcactc taattccaga tggaccagca gacaaccata atgatgagct 4620

tgttcagacc cttacgaagc tgtttgtcca cacttttatc ctcctggaac caccatctta 4680

tatataaatt ttgttttgcg agtgttttcc acttggttac agattctgcc aaccctgaga 4740

ggttaatggt agtttagggt tcatccgttc atgtggagta caatattgct cataatagct 4800

tgaagctttc ccgcttcaag ctattctacc tcccaggatc ttgttatcac ttccttaatg 4860

tgtttgcatc catggactat gaagcccatt aacctattta acttgacagg cattgactgc 4920

ttttatattg tgcttagcat gatttctcct gtatttttat ggaaagtaac tcttagggac 4980

ccatcactat tgctcttctc tacattgtgg acaggttgaa cacctatctc ccctttatgt 5040

tctgtgccaa caatattcat ctgtcacttt tgtgaaccac acagtgaagt tgtaaatgtg 5100

tattggaaaa cctatcactc ttctctgcct tgaaagaatt tattgatgtc agtattggtt 5160

tttgttcttt ttatggtctg tgactgatat taaccaaaaa attttgttta ttagctcggg 5220

aacttctcag ggtgaaccaa gattgatgcc atcaatctca gaggatattt accgccgaat 5280

gtatctgtat agtctgatca tgcctatcat gagcaagtaa gagtttgttt tattatagat 5340

ccattacttt taatgaaggc acttggaaag tcaaataaat gaatatacat caggatttat 5400

ctattctata aacctcttat tgtttgaccc acgtctttgt tcagtcaaga ggcactagct 5460

taaatcatga tgccatttat ccaaagtttc ttctcgtgat tgctatttga aagctgaaag 5520

agaaataatt cttatctctc cttagaagaa ggctagaagc taccgatctt ttctggtgat 5580

ttttttgttt tcattttggt gttagttata aaagttttggt caagttagag ttggagtaca 5640

cgaaagatag gacaatactt atcacaagct gaagtaaccc ccttcaaacc atagttagaa 5700

aggcttacaa attgaaatgt tagaattctg ctttagtaaa gagtttcagt atttcttaaa 5760

ggcttgttcg gttatttcca tgccatgtgg attggagtgg attgagaggg gttgtgatgg 5820

attttgactt gatatggatt taaaccgact caatctcatc caatccatat ggattaattt 5880

caaaacgaac cagccctaaa tatgcatttt aaagtctcga aagatatttt gctatatcat 5940

gatacttgga actgggataa actgcccact ttccttcatc ccttgtaatt atcacatctt 6000

tctgctcttg cttgcattca acataattga agaaaggtct tatagaatag cttatcagta 6060

catttcaaag gtttttagat tttaccatgt ctgtagtatt tttttttttt tgacttagtc 6120

aagtctccag gtacatacga ggccttggcg aggggaaggc aatgtatctt ctctttgtga 6180

aagctgaaac attgacaaat gctggcattc cagtccgatc agtcctcacc agctactaca 6240

agagccctca gttcttgcag cgcaagcacg acctctacaa cagctacacc agtcccgacg 6300

aggtcattct ttgccccgac agccagcaga gcatgtactg ccagctactc tgcggcctgg 6360

tggaacgcca gaacgtcgtc cgtctcggag cagtcttcgc ctcagcattc ctccggtcca 6420

tcagcttcct cgagaagcac tggcatgacc tagtcaccga catccgaacc ggccggatca 6480

atcccagcat caccaacgcc gcatgccggg tggccacgga gagctttctc gcgcagccca 6540

accccgagct agctgacgaa gtcgaggcga tatgcagctc ggagtcatgg aagggcgttc 6600

ttggcaggct ctggcctaac gtcaagtaca tcgaggccgt cctgacgggg acgatggcgc 6660

agtacgtccc catgctcgag ttctacagcg acggcaggat ccccctggtc tgcaccatgt 6720

acgcttcctc ggagagctac ttcggcgtca acctgcggcc gctgtgcagc ccgaaggacg 6780

tgtcctacac catcctcccc aacatggcct acttcgagtt cgtgccgttg gacgacggcc 6840

tcaggctggc ggaggatggt gaggctgtgg agaaggagaa gcttgtcggc ctcgtggacg 6900

tcaaggtcgg atgctactac gagctcgtgg ttaccacatt tgccggtgag aacaccatac 6960

ctcttgcgaa tactatgatc tgcaacaaca gttttacttc aaccaataca ttgctctctt 7020

ttttttgggg tggggtgcag gtctttacag gtacagggta ggcgatgtac tccaggtgac 7080

aggcttctac aaccgcgcac cccagttcaa attcatctgc cggaggaacg tgatcctgag 7140

cgtggacgca gacaagacga acgaggaaga cctccacggc agcgtctccc gggccaagaa 7200

gatcctggag gaccgaggcc acatcctact ggagtacacc agcttcaccg acacgtcgac 7260

ggtgccgggc cactacgtcc tgttctggga ggtgaaggcc acgcccacct ccggctccgg 7320

gggcgcgcgg ctggacgcgc aggtcctgga gagctgctgc gtcgccgtgg aggagtcgct 7380

ggactgcgtg tacaggcggt gcagggcgca cgacaggtcc gtggggccgc tggagatacg 7440

gctggtggaa gccggcgcct tcgacgcgct catggacctg ttggtcagcc agggcggctc 7500

catcaaccag tacaagacgc cacggtgcat cgagccgggc ggcgccgcgc tggagctgct 7560

cgactccaag gccaccgcct cgttcttcag cccccgggat cctgaagcta ctggtgtctg 7620

aggttgttct gctactctga aactaatcct ctgctggatc taaataatct gagaattttc 7680

gtctgggtta tccccgttta cattgcactt ttgtcaggct ttgctactat gcaaagaagc 7740

cttttttctt tatttgagat tgtgatcatc aattgtctat cacttgacga cttttgtgtg 7800

tgtgattgac ggagcgcatg gattatataa caataaaata aagtaaagag agaggccacg 7860

catgcttttc cgaaagcatc agtggtgagt gagctttcaa ccatggtaat catgcatggc 7920

ctgtacgtgc cattgtgcca gtgctaattg cttccaagct aaggactcga tcccgcccgc 7980

gtataccgta cctggctggc cggtcaaagc tacgtagctt aagagactga gctagcttag 8040

ctaagctaga gcaccacatg acacaccgtc agcaggtcag acaagaagca aagcggcgcg 8100

tggccgacgg ctcctacagt cctcctactg cactctgaag actgaacgtg tgaatggacc 8160

catcaggatc tccatttcag aattcacctg tgccgctgtg cctgtgtgct ggccgctgcc 8220

tgctttttca cctgtccgtc tcgctgggtg ctctagctag atccgcggcc cccaggctag 8280

agattgttta gcacaatgac agccacagat ctctgccgcc gggattccat cgacgggact 8340

gatgataatt aaaatgtcag ttaaccccgt acgtacgtag agggtggcgt ggctagtcaa 8400

ac 8402

<210> 4

<211> 552

<212> DNA

<213> 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 (10)

1. A method for cultivating plants with high stress tolerance is characterized in that: comprises increasing the content and/or activity of a protein GH3.9 in a target plant to obtain a plant with high stress tolerance, wherein the stress tolerance of the plant with high stress tolerance is higher than that of the target plant;

the protein GH3.9 is (1) or (2) as follows:

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

(2) and (2) the protein which is derived from the protein (1) and has the same function and is obtained by substituting and/or deleting and/or adding one or more amino acid residues in the amino acid sequence shown by SEQ ID No.2 in the sequence table.

2. The method of claim 1, wherein: the content and/or activity of the protein GH3.9 in the target plant is improved by introducing a substance for regulating and controlling the expression of a gene coding the protein GH3.9 into the target plant.

3. The method of claim 2, wherein: the substance for regulating and controlling the expression of the protein GH3.9 coding gene is any one of biological materials B1) -B4);

B1) a nucleic acid molecule encoding a GH3.9 protein according to claim 1;

B2) an expression cassette comprising the nucleic acid molecule of B1);

B3) a recombinant vector containing the nucleic acid molecule according to B1) or a recombinant vector containing the expression cassette according to B2);

B4) a recombinant microorganism containing B1) the nucleic acid molecule, or a recombinant microorganism containing B2) the expression cassette, or a recombinant microorganism containing B3) the recombinant vector.

4. The method of claim 3, wherein: the B1) nucleic acid molecule for encoding the protein GH3.9 of claim 1 is any one of the following nucleic acid molecules 1) to 4):

1) the nucleotide sequence is a nucleic acid molecule shown as SEQ ID No.3 in the sequence table;

2) the coding sequence is a nucleic acid molecule shown as SEQ ID No.1 in the sequence table;

3) nucleic acid molecules which hybridize under stringent conditions with the DNA molecules defined in 1) or 2) and which code for proteins having the same function;

4) a nucleic acid molecule having at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% homology to the DNA molecule defined in 1) or 2) and encoding a protein having the same function.

5. A protein characterized by: the protein is the protein GH3.9 of claim 1.

6. A biomaterial characterized by: the biomaterial as claimed in claim 3 or 4.

7. The application is characterized in that: the application is the protein of claim 5 or the biological material of claim 6 in regulating and controlling the stress tolerance of plants.

8. The application is characterized in that: the application is the protein of claim 5 or the biological material of claim 6 in the cultivation of plants with high stress tolerance.

9. The method according to any one of claims 1-4, the use according to claim 7 or 8, characterized in that:

the plant is a dicotyledonous plant or a monocotyledonous plant;

the monocotyledon is a gramineous plant.

10. The method according to any one of claims 1-4, the use according to claim 7 or 8, characterized in that:

the stress tolerance is saline-alkali tolerance;

the regulation and control of the plant stress tolerance is to improve the saline-alkali tolerance of the plant.

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