CN106893738B - The application of OsSGT1 albumen and its encoding gene in regulation plant salt tolerance resistance - Google Patents

Abstract

The invention discloses a kind of application of OsSGT1 albumen and its encoding gene in regulation plant salt tolerance resistance.The present invention provides a kind of method for cultivating genetically modified plants, includes the following steps: plant that the channel genes for encoding OsSGT1 albumen set out, obtains the genetically modified plants of salt tolerance reduction.The OsSGT1 albumen is following (a1) or (a2): the protein that (a1) amino acid sequence shown in sequence 2 in sequence table forms；(a2) by the amino acid sequence of sequence 2 by the substitution and/or deletion and/or addition of one or several amino acid residues and the protein as derived from sequence 2 relevant to plant salt endurance.The present invention understands the genesis mechanism of salt stress, improves plant salt endurance, cultivate the strong plant variety of salt tolerance, there is important theory directive significance and production application to be worth for excavating plant salt tolerance gene.

Description

The application of OsSGT1 albumen and its encoding gene in regulation plant salt tolerance resistance

Technical field

The present invention relates to a kind of application of OsSGT1 albumen and its encoding gene in regulation plant salt tolerance resistance.

Background technique

Abiotic stress is for example saline and alkaline, arid and low temperature severely impacts the high yield and stable yields of crop, is to threaten world food Safety the principal element soil salinization be limit crop growth, cause grain drop in production the main abiotic stress factor it One.In recent years, soil secondary salinization area is increasing year by year, and salt stress, which has become, influences biological husbantry production in world wide One of most important environmental stress factor.Salt stress is mainly shown as osmotic stress, influences plant pair Na⁺And K⁺Absorption and row The distribution and dynamic equilibrium of reprimand and the normal ion of cell.Rice is grass family model plant and a kind of pair of salt medium sensitivity Crop, salt stress has become the main restricting factor of the saline and alkaline rice region rice steady production of China.Salt stress patience related gene Relevant gene and transporter gene etc. are metabolized including participating in signal transduction and transcriptional modulatory gene, osmotic adjustment.Water The Study on Molecular Mechanism of rice salt stress response achieves some substantial progresses, is mainly manifested in multiple related to salt stress tolerance Gene and regulatory factor by successful identification, the biological function of portion gene also illustrates clearer.Though having identified at present The relevant molecular labeling of some salt stress and the site gene (or QTL), since salt stress patience is a quantitative character, by more A gene regulation.Therefore it also needs to increase the excavation and utilization of Rice Salt germ plasm resource, reinforces Rice Salt molecular labeling Position Research clones resistant gene of salt, and studies its mechanism of action.Plant salt tolerance gene is excavated, the genesis mechanism of salt stress is understood, Looking for plant responds the signal pathway node of salt stress, improves the disease resistance and salt tolerance of rice, cultivates the strong rice of salt tolerance There is kind important theory significance and production application to be worth.

Summary of the invention

The answering in regulation plant salt tolerance resistance the object of the present invention is to provide a kind of OsSGT1 albumen and its encoding gene With.

The present invention provides a kind of methods for cultivating genetically modified plants, include the following steps: that OsSGT1 albumen will be encoded Channel genes set out plant, obtain the genetically modified plants of salt tolerance reduction.

The OsSGT1 albumen is obtained from rice, is following (a1) or (a2):

(a1) protein that the amino acid sequence shown in sequence 2 in sequence table forms；

(a2) amino acid sequence of sequence 2 by the substitution of one or several amino acid residues and/or missing and/or is added Add and the protein as derived from sequence 2 relevant to plant salt endurance.

In order to make OsSGT1 albumen in (a1) convenient for purifying and detection, can in as sequence table amino shown in sequence 2 The amino terminal or carboxyl terminal of the protein of acid sequence composition connect upper label as shown in Table 1.

The sequence of 1 label of table

Label	Residue	Sequence
			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

OsSGT1 albumen in above-mentioned (a2) can be artificial synthesized, can also first synthesize its encoding gene, then carry out biological expression It obtains.The encoding gene of OsSGT1 albumen in above-mentioned (a2) can be by will lack in DNA sequence dna shown in sequence 1 in sequence table The codon of one or several amino acid residues, and/or carry out the missense mutation of one or several base-pairs, and/or its 5 ' The coded sequence that end and/or 3 ' ends connect label shown in table 1 obtains.

" gene (abbreviation OsSGT1 gene) of coding OsSGT1 albumen " is following (b1) or (b2) or (b3):

(b1) DNA molecular shown in the nucleotide of 5 ' end 1-1104 of sequence 1 of code area such as sequence table；

(b2) the DNA sequence dna hybridization limited under strict conditions to (b1) and coding protein relevant with plant salt endurance DNA molecular；

(b3) there is 90% or more homology to the DNA sequence dna that (b1) or (b2) is limited and coding is related with plant salt endurance Protein DNA molecular.

Above-mentioned stringent condition can be for 0.1 × SSPE (or 0.1 × SSC), the solution of 0.1%SDS be miscellaneous in DNA or RNA It hands over and hybridizes at 65 DEG C in experiment and wash film.

In the method, the OsSGT1 gene can import the plant that sets out by recombinant expression carrier.The recombination table Ti-plasmids, Ri plasmid, plant viral vector, directly delivered DNA, microinjection, conductance, mediated by agriculture bacillus etc. can be passed through up to carrier Conventional biology methods are transformed into plant cell or tissue.

The recombinant expression carrier of the gene can be contained with existing plant expression vector construction.The plant expression vector Including double base agrobacterium vector and the carrier etc. that can be used for plant micropellet bombardment.Use the gene constructed recombinant expression carrier When, any enhanced, composing type, organizing specific type or inducible promoter can be added before its transcription initiation nucleotide, They can be used alone or are used in combination with other plant promoters；In addition, using the gene constructed recombinant expression carrier When, also enhancer, including translational enhancer or transcriptional enhancer, these enhancer regions can be used to can be ATG initiation codon Son or neighboring region initiation codon etc., but must be identical as the reading frame of coded sequence, to guarantee correctly turning over for entire sequence It translates.The source of the translation control signal and initiation codon be it is extensive, can be natural, be also possible to synthesis.It turns over Translating initiation region can come from transcription initiation region or structural gene.For the ease of reflecting to transgenic plant cells or plant Fixed and screening, can process plant expression vector used, be such as added the expression in plant can produce color change enzyme or The gene of luminophor, resistant antibiotic marker or anti-chemical reagent marker gene etc..

The recombinant expression carrier will concretely insert double-strand shown in the sequence 1 of sequence table in carrier pGWB18 The recombinant plasmid that DNA molecular obtains.

Any description above sets out plant as monocotyledon or dicotyledon.The monocotyledon can be Poales Plant.The Poales plant can be gramineae plant.The gramineae plant can be oryza plant.The oryza plant is specific It can be rice, such as OryzasativaLcv.Nipponbare rice.

The salt tolerance reduction is embodied as at least one of following (d1)-(d3):

(d1) under high salt conditions, survival rate is lower than the plant that sets out；

(d2) under high salt conditions, peroxidase activity is lower than the plant that sets out；

(d3) under high salt conditions, mda content is higher than the plant that sets out.

Concretely 100mM NaCl is handled the high salt conditions.

The present invention also protects the application of OsSGT1 albumen, for as follows (c1) or (c2):

(c1) regulate and control plant salt endurance；

(c2) plant salt endurance is reduced.

The present invention also protects application of any description above method in plant breeding.

The purpose of the breeding is the low plant of breeding salt tolerance.

The present invention also protects a kind of plant breeding method, includes the following steps: the expression for increasing OsSGT1 albumen in plant Amount and/or activity, obtain the rice of salt tolerance reduction.

The present invention also protects a kind of method for reducing plant salt endurance, includes the following steps: to increase OsSGT1 egg in plant White expression quantity and/or activity, to reduce plant salt endurance.

Any description above plant is monocotyledon or dicotyledon.The monocotyledon can be planted for Poales Object.The Poales plant can be gramineae plant.The gramineae plant can be oryza plant.The oryza plant specifically may be used For rice, such as OryzasativaLcv.Nipponbare rice.

The present invention has the function of plant salt endurance by research discovery OsSGT1 albumen and its encoding gene.Up-regulation The salt tolerance of plant can be significantly reduced in the expression of OsSGT1 gene in plant.The present invention for excavate plant salt tolerance gene, The genesis mechanism of salt stress is solved, plant salt endurance is improved, the strong plant variety of salt tolerance is cultivated, anticipates with important theoretical direction Justice and production application value.

Detailed description of the invention

Fig. 1 is the PCR qualification result of partial transgenic plant.

Fig. 2 is the Western blot result of transgenic plant.

Fig. 3 is that WT lines and transgenic plant salt-tolerant phenotype observe result.

Fig. 4 is WT lines and transgenic plant survival rate statistical result.

Fig. 5 is WT lines and transgenic plant MDA (malonaldehyde) content statistical result.

Fig. 6 is WT lines and the active statistical result of transgenic plant POD (peroxidase).

Specific embodiment

Embodiment below facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments Method is unless otherwise specified conventional method.Test material as used in the following examples is unless otherwise specified certainly What routine biochemistry reagent shop was commercially available.Quantitative test in following embodiment is respectively provided with three repeated experiments, as a result makes even Mean value.

The coding region sequence of OsSGT1 gene is as shown in the sequence 1 of sequence table, albumen shown in the sequence 2 of polynucleotide Matter (OsSGT1 albumen).

Rice strain OryzasativaLcv.Nipponbare: bibliography: Li Lei, Xue's fragrance, a left side show quick, wait rice varieties OryzasativaLcv.Nipponbare most suitable blade volume Curvature research [J] Yangzhou University's journal agricultural and life science version, 2013 (2): 47-51.；The public can be from Chinese agriculture section Crop science research institute, institute obtains.

Carrier pDONR201:Invitrogen company, article No.: 11798-014.

Carrier pGWB18:BioVectorNTCC Type Tissue Collection.

Induced medium: CaCl₂·2H₂O440mg, KH₂PO₄170mg, MgSO₄·7H₂O370mg, NH₄NO₃1650mg, KNO₃1900mg, KI 0.83mg, CoCl₂·6H₂O0.025mg, H₃BO₃6.2mg, Na₂MoO₄·2H₂O0.25mg, MnSO₄· 4H₂O22.3mg, CuSO₄·5H₂O0.025mg, ZnSO₄·7H₂O8.6mg, Na₂-EDTA·2H₂O37.3mg, FeSO₄· 7H₂O27.8mg, VB10.1mg, VB60.5mg, niacin 0.5mg, inositol 100mg, glycine 2mg, 2,4-D 2mg hydrolyze junket egg White 2g, maltose 30g, agar 3g, deionized water add to 1L.

Infect culture medium: preparation method is referring to bibliography: Hiei Y, Ohta S, Komari T, et Al.Efficient transformation of rice (Oryza sativa, L.) mediated by Agrobacterium, and sequence analysis of the boundaries of the T-DNA [J] .Plant Journal, 1994,6 (2): 271-282..The concentration of acetosyringone in bibliography is replaced with 200 μM.

Co-culture medium: acetosyringone and glucose are added in induced medium, is cultivating acetosyringone Final concentration of 200 μM in base, the final concentration of 10g/L of glucose in the medium.

Micro-organisms base: the cephalosporin in induced medium makes cephalosporin in the medium final concentration of 500mg/L。

Screening and culturing medium: hygromycin and cephalosporin are added in induced medium, makes the end of hygromycin in the medium Concentration is 65mg/L, the final concentration of 500mg/L of cephalosporin in the medium.

Pre- regeneration culture medium: CaCl₂·2H₂O440mg, KH₂PO₄170mg, MgSO₄·7H₂O370mg, NH₄NO₃1650mg, KNO₃1900mg, KI 0.83mg, CoCl₂·6H₂O0.025mg, H₃BO₃6.2mg, Na₂MoO₄·2H₂O0.25mg, MnSO₄· 4H₂O22.3mg, CuSO₄·5H₂O0.025mg, ZnSO₄·7H₂O8.6mg, Na₂-EDTA·2H₂O37.3mg, FeSO₄· 7H₂O27.8mg, VB1 0.1mg, VB60.5mg, niacin 0.5mg, inositol 100mg, glycine 2mg, caseinhydrolysate 2g, malt Sugared 30g, agar 3g, kinetin 2mg, methyl α-naphthyl acetate 1mg, deionized water add to 1L；Hygromycin is added before inverted plate and keeps its dense Degree is 50mg/L.

Regeneration culture medium: CaCl₂·2H₂O440mg, KH₂PO₄170mg, MgSO₄·7H₂O370mg, NH₄NO₃1650mg, KNO₃1900mg, KI0.83mg, CoCl₂·6H₂O0.025mg, H₃BO₃6.2mg, Na₂MoO₄·2H₂O0.25mg, MnSO₄· 4H₂O22.3mg, CuSO₄·5H₂O0.025mg, ZnSO₄·7H₂O8.6mg, Na₂-EDTA·2H₂O37.3mg, FeSO₄· 7H₂O27.8mg, VB1 0.1mg, VB60.5mg, niacin 0.5mg, inositol 100mg, glycine 2mg, caseinhydrolysate 2g, malt Sugared 30g, agar 6g, kinetin 2mg, methyl α-naphthyl acetate 1mg, deionized water add to 1L；Hygromycin is added before inverted plate and keeps its dense Degree is 50mg/L.

POD reaction solution: 28 μ L of guaiacol, magnetic stirring apparatus are added in 50ml 0.1M phosphate buffer (pH 6.0) Heating stirring is allowed to be completely dissolved, and 30% (percent by volume) H is added after cooling₂O₂19 μ L of aqueous solution mixing.

MDA reaction solution: 0.6gTBA (thiobarbituricacidα-), first with a small amount of 1M NaOH aqueous dissolution, then with 10% (percent by volume) TCA (trichloroacetic acid) aqueous solution is settled to 100 milliliters.

The functional analysis of embodiment 1, OsSGT1 albumen and its encoding gene

One, the building of OsSGT1 gene overexpression carrier

1, the total serum IgE of OryzasativaLcv.Nipponbare rice leaf, reverse transcription cDNA are extracted.

2, cDNA is obtained as template using step 1, PCR amplification is carried out using primer attB-F1 and primer attB-R1, is obtained Amplified production.

AttB-F1:5 '-GGGGACAAGTTTGTACAAAAAAGCAGGCTGCATGGCAACCGCCGCC-3′；

AttB-R1:5 '-GGGGACCACTTTGTACAAGAAAGCTGGGTGAGTACTCCCATTTCTTAAGCTCC-3′。

3, it is reacted by BP, the amplified production that step 2 is obtained imports carrier pDONR201, obtains containing sequence The positive Entry clone plasmids pDONR201-OsSGT1 of double chain DNA molecule shown in 1.

BP reaction system: amplified production 2.7 μ L (50-100ng), carrier pDONR201 1.0 μ L (30-50ng), 5 × BP 1.0 μ L, BP Enzymemix of ReactionBuffer, 0.3 μ L.

BP reaction condition: 25 DEG C of warm bath 1h.

4, the positive Entry clone plasmids pDONR201-OsSGT1 that step 3 obtains is taken, it is anti-to carry out LR with carrier pGWB18 It answers, obtains the 35S::MYC-OsSGT1 expression vector containing double chain DNA molecule shown in sequence 1 and (be sequenced and tested Card).

LR reaction system: Entry clone plasmids pDONR201-OsSGT1 1 μ L (50-100ng), 1 μ L of carrier pGWB18 (50-100ng), 0.5 μ L of LR enzymemix.

LR reaction condition: 25 DEG C of warm bath 1h.

Two, it is overexpressed the acquisition of transgenic paddy rice

1, the mature seed of OryzasativaLcv.Nipponbare rice is taken, the seed of mechanical dejacketing, the full bright and clean no bacterial plaque of picking carries out disinfection.

2, the seed after step 1 disinfection is inoculated into 28 DEG C of induced medium, dark culture 14 days or so, it is good chooses appearance It is good, the good callus of growing power.

3, the expression vector 35S::MYC-OsSGT1 for taking step 1 to construct imports Agrobacterium tumefaciems EHA105, is recombinated Bacterium EHA105/35S::MYC-OsSGT1.

4, the recombinant bacterium EHA105/35S::MYC-OsSGT1 for taking step 3 to obtain is resuspended thallus with culture medium is infected, obtains Bacteria suspension (OD_600nm=1.0).

5, the callus for completing step 2 is soaked in bacteria suspension prepared by step 4, infects 20min.By bacterium after infecting Suspension is outwelled, and callus is taken, and with aseptic filter paper suck dry moisture, is subsequently placed in co-culture medium, and 28 DEG C of dark cultures are cultivated 50-55h。

6, after completing step 5, selecting surface does not have the callus of obvious Agrobacterium to move in micro-organisms base, and 28 DEG C dark Culture 3-4 days.

7, after completing step 6, callus is moved into 28 DEG C dark culture culture 30 days on screening and culturing medium, every 10 days subcultures Once.

8, after completing step 7, the callus of fresh hygromycin resistance is taken, is connected in pre- regeneration culture medium, 28 DEG C of dark trainings It supports 7 days, (12h illumination/12h is dark) continues to be transferred on regeneration culture medium after cultivating 7 days between being subsequently placed in illumination cultivation, continues light According to culture, until growing regeneration plant, T is obtained₀For plant.T₀For plant selfing, T is obtained₁For plant.T₁For plant selfing, Obtain T₂For plant.T₂For plant selfing, T is obtained₃For plant.

Three, it is overexpressed the identification of transgenic paddy rice

1, the T for each strain for taking step 2 to obtain₂For plant, extracts the total serum IgE of plant leaf and reverse transcription is cDNA. Using cDNA as template, PCR identification is carried out using primer Hyg-F and Hyg-R.Made using over-express vector 35S::MYC-OsSGT1 For positive control, using the cDNA of OryzasativaLcv.Nipponbare rice as negative control.

Hyg-F:5 '-CTATTTCTTTGCCCTCGGAC-3 '；

Hyg-R:5 '-CCTGACCTATTGCATCTCCC-3 '.

If for a certain T₀For plant, the T of sampling Detection₂PCR qualification result for plant is the positive, the T₀Generation Plant and its self progeny are a homozygous transgenic line excessively.

The PCR qualification result of plant part is as shown in Figure 1.In Fig. 1, M is DNA Maker ,+it is positive control ,-it is negative Control, swimming lane 1-17 are corresponding in turn to the T of 17 different strains₂For plant.The result shows that the plant of 17 strains can amplify With the consistent specific band of plasmid vector hygromycin gene, 17 strains are homozygous transgenic line excessively.

2, selecting step 1 identify correct 6 transgenic lines (be named as SA11, SA22, SA24, SA28, SA35 and SA36), each strain T is analyzed using western blot₂For OsSGT1 expressing quantity in plant (MYC antibody), identification turns base Because of the overexpression efficiency of strain.

Testing result is as shown in Figure 2.The result shows that: purpose band is detected in 6 transgenic lines, on total protein Under the premise of sample amount is consistent, the protein accumulation amount of OsSGT1 is significantly higher than transgenosis in transgenic line SA22, SA24 and SA28 The protein content of OsSGT1 in strain SA11, SA35 and SA36.

Four, turn the acquisition of empty carrier strain

It is operated, is obtained according to the 1-8 of step 2 using carrier pGWB18 substitution expression vector 35S::MYC-OsSGT1 Turn empty carrier strain.

Five, it is overexpressed the Salt-Tolerance Identification of transgenic line

Plant to be measured are as follows: OryzasativaLcv.Nipponbare rice (wild type), the T for being overexpressed transgenic line SA24₃Turn for plant, overexpression The T of gene strain SA28₃For plant, turn the T of empty carrier strain₃For plant.

1, the seed of plant to be measured is put in 50 DEG C of baking ovens and is dried 3 days, it is water-soluble with 0.5% (percent by volume) sodium hypochlorite Liquid, which impregnates 30 minutes, carries out surface sterilization to seed, and then seed soaking, vernalization are to showing money or valuables one carries unintentionally in 37 DEG C of incubators, and during which about 12h is changed Water.

2, it selects the step 1 consistent seed that germinates to be sowed, is cultivated one week with distilled water (pH=5.5), Zhi Houyong (preparation method sees reference document Yoshida nutrient solution: Yoshida S, Forno D A, Cock J H.Laboratory manual for physiological studies of rice.Laboratory manual for physiological Studies of rice.International Rice Research Institute, 1976.) continue culture to three for 28 DEG C Then plant is transferred to 28 DEG C of the Yoshida nutrient solution containing 100mM NaCl and continues to cultivate (salt treatment), existed respectively by Ye Qi The 3rd day, the 4th day, the 5th day, the 6th day, the 7th day, the 8th day and the 10th day of salt treatment counts its survival rate.

As a result as shown in Figure 3 and Figure 4.The Phenotypic Observation result that Fig. 3 is the 10th day.In Fig. 3, upper left and upper right are wild type (CK) Phenotypic Observation is as a result, lower-left is the Phenotypic Observation of transgenic plant SA28 as a result, bottom right is transgenic plant SA24 Phenotypic Observation result.Fig. 4 is survival rate statistical result.Start the result shows that 100mM NaCl is handled the 4th day, WT lines (CK) survival rate is higher than transgenic plant SA24 and SA28, and the 10th day, the survival rate of WT lines was respectively 60% He 80%, and the survival rate that the survival rate of transgenic plant SA24 is 13.3%, SA28 is 38%, i.e., transgenic plant is to salt stress Sensibility be significantly higher than wild type.Turning the survival rate of empty carrier plant, there was no significant difference with wild type.

3, it takes step 2 to handle the similar plant leaf blade of the 7th day Leaf Age bit length gesture using 100mM NaCl, weighs 0.5g is shredded and is placed in the mortar of pre-cooling, and the PBS (NaH of 21.25ml 0.2M of 3ml pre-cooling is added₂PO₄Aqueous solution with 228.25ml the Na of 0.2M₂HPO₄Aqueous solution mixes, and distilled water is settled to 1000ml, pH=7.8) it is fully ground, by lapping liquid It pours into centrifuge tube, then washs mortar with the PBS that 2ml is pre-chilled, and pour into centrifuge tube, 4 DEG C, 6,000rpm centrifugation 30min inhale It takes 5ml supernatant (i.e. zyme extract), carries out the measurement of POD (peroxidase) activity and MDA (malonaldehyde) content.

(1) POD (peroxidase) determination of activity: 20 μ L zyme extracts and 3ml POD reaction solution are added in cuvette, The light absorption value at 470nm is quickly mixed and measured immediately, read every 1 minute 1 time, reads 3 times altogether, with absorbance per minute change The size of change value (Δ A470/mingFW) expression enzyme activity.

The blank control that zyme extract is replaced using 20 μ L PBS, zeroing are set.

POD activity (Δ A470/mingFW)=Δ A470 × V × Va × W=Δ A470 × 5 × 0.02 × 0.5 =Δ A470 × 500

Difference of the Δ A470:A470 in 0-3min；V: extracting solution total volume (mL)；Va: measurement zyme extract volume (mL)； W: leaf dry weight (g)；

(2) MDA is measured: 1ml zyme extract and 2ml MDA reaction solution being added in centrifuge tube, boiling water bath is sealed 15min, rapidly after cooling, 12,000rpm centrifugation 10min, Aspirate supernatant is simultaneously measured under tri- 600,532,450nm wavelength Its light absorption value.

MDA(μmol·g^-1·F^-1W^-1)=(6.45 × (D532-D600) -0.56 × D450) × V/Va/W

V: extracting solution total volume (mL)；Va: measurement zyme extract volume (mL)；W: leaf dry weight (g)；

As a result as shown in Figure 5 and Figure 6.Fig. 5 is MDA content statistical result, and Fig. 6 is POD activity statistical result.As a result table Bright, under condition of salt stress, the content of MDA is significantly higher than wild type (CK) in rotaring gene plant blade, and cell membrane is by situation than wild Raw type is serious, and POD enzymatic activity is substantially less than wild type, shows effectively remove intracorporal oxygen in rotaring gene plant blade Free radical reduces oxygen radical peroxidative damage caused by cell membrane that salt stress generates.Turn the statistics knot of empty carrier plant There was no significant difference with wild type for fruit.

<110>Institute of Crop Science, Chinese Academy of Agricultural Science

Shenzhen Biology Breeding innovation research institute, the Chinese Academy of Agricultural Sciences

<120>application of OsSGT1 albumen and its encoding gene in regulation plant salt tolerance resistance

<160> 2

<210> 1

<211> 1104

<212> DNA

<213>rice (Oryza sativa)

<400> 1

atggcaaccg ccgccgcgtc ggatctggag agcaaggcca aggcggcctt cgtcgacgac 60

gacttcgagc tcgccgccga gctctacacg caggcaatcg aggccagccc cgccaccgcc 120

gagctctacg ccgaccgcgc ccaggcccat atcaagctag gcaactacac tgaggctgta 180

gctgatgcta acaaggccat tgaacttgac ccatcaatgc acaaggctta tcttcgtaaa 240

ggcgctgcat gtatacgact ggaggagtat caaactgcaa aagcagctct tgaattgggt 300

tactcgttcg catctggtga ctcaaggttt actcgcctaa tgaaggagtg tgatgagcgc 360

attgctgagg agcttagtga agtccctgtt aagaaggctg aagatggagc agctgccccc 420

tctgttgctt cttttgttga ggaaaaggat gatgctgcaa acatggataa tacaccacca 480

atggtagaag tgaagccaaa atacaggcac gacttctaca acagtgctac agaagttgta 540

ttgacaattt ttgcaaaggg tgttcctgct gagaatgttg ttgttgattt tggtgaacaa 600

atgttaagtg tgtcgattga agtccctgga gaggagccgt accattttca gcctcgtctg 660

ttttctaaga tcatccctga gaaaagcaga taccaagtgc tatccacgaa ggttgaaata 720

agactggcta aagctgaaca gattacatgg acctcacttg attatgataa aaaaccaaag 780

gctgttccac aaaagataat ccctccagct gaatcggccc agaggccatc atatccttcc 840

tcaaaatcca agaaagactg ggataaactg gaagctgaag ttaaaaagga ggagaaggag 900

gagaagcttg aaggcgatgc tgcattgaac aaatttttcc gtgacatcta cagtgatgct 960

gatgaagaca tgcgacgagc aatgatgaaa tcttttgttg aatctaacgg tactgttctg 1020

tcgaccaatt ggaaagatgt tggctcgaag aaggtagagg gaagcccacc tgatgggatg 1080

gagcttaaga aatgggagta ctaa 1104

<210> 2

<211> 367

<212> PRT

<213>rice (Oryza sativa)

<400> 2

Met Ala Thr Ala Ala Ala Ser Asp Leu Glu Ser Lys Ala Lys Ala Ala

1 5 10 15

Phe Val Asp Asp Asp Phe Glu Leu Ala Ala Glu Leu Tyr Thr Gln Ala

20 25 30

Ile Glu Ala Ser Pro Ala Thr Ala Glu Leu Tyr Ala Asp Arg Ala Gln

35 40 45

Ala His Ile Lys Leu Gly Asn Tyr Thr Glu Ala Val Ala Asp Ala Asn

50 55 60

Lys Ala Ile Glu Leu Asp Pro Ser Met His Lys Ala Tyr Leu Arg Lys

65 70 75 80

Gly Ala Ala Cys Ile Arg Leu Glu Glu Tyr Gln Thr Ala Lys Ala Ala

85 90 95

Leu Glu Leu Gly Tyr Ser Phe Ala Ser Gly Asp Ser Arg Phe Thr Arg

100 105 110

Leu Met Lys Glu Cys Asp Glu Arg Ile Ala Glu Glu Leu Ser Glu Val

115 120 125

Pro Val Lys Lys Ala Glu Asp Gly Ala Ala Ala Pro Ser Val Ala Ser

130 135 140

Phe Val Glu Glu Lys Asp Asp Ala Ala Asn Met Asp Asn Thr Pro Pro

145 150 155 160

Met Val Glu Val Lys Pro Lys Tyr Arg His Asp Phe Tyr Asn Ser Ala

165 170 175

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

180 185 190

Val Val Val Asp Phe Gly Glu Gln Met Leu Ser Val Ser Ile Glu Val

195 200 205

Pro Gly Glu Glu Pro Tyr His Phe Gln Pro Arg Leu Phe Ser Lys Ile

210 215 220

Ile Pro Glu Lys Ser Arg Tyr Gln Val Leu Ser Thr Lys Val Glu Ile

225 230 235 240

Arg Leu Ala Lys Ala Glu Gln Ile Thr Trp Thr Ser Leu Asp Tyr Asp

245 250 255

Lys Lys Pro Lys Ala Val Pro Gln Lys Ile Ile Pro Pro Ala Glu Ser

260 265 270

Ala Gln Arg Pro Ser Tyr Pro Ser Ser Lys Ser Lys Lys Asp Trp Asp

275 280 285

Lys Leu Glu Ala Glu Val Lys Lys Glu Glu Lys Glu Glu Lys Leu Glu

290 295 300

Gly Asp Ala Ala Leu Asn Lys Phe Phe Arg Asp Ile Tyr Ser Asp Ala

305 310 315 320

Asp Glu Asp Met Arg Arg Ala Met Met Lys Ser Phe Val Glu Ser Asn

325 330 335

Gly Thr Val Leu Ser Thr Asn Trp Lys Asp Val Gly Ser Lys Lys Val

340 345 350

Glu Gly Ser Pro Pro Asp Gly Met Glu Leu Lys Lys Trp Glu Tyr

355 360 365

Claims (2)

1. Application of the 1.OsSGT1 albumen in regulation plant salt endurance；

   The amino acid sequence of the OsSGT1 albumen is as shown in sequence 2 in sequence table；

   The plant is monocotyledon.
2. 2. application according to claim 1, it is characterised in that: the salt tolerance of the regulation plant is to reduce plant salt tolerance Property.