CN106957355A - It is a kind of to the resistance to low light of plant and low temperature resistant related PPR albumen and its encoding gene and application - Google Patents
It is a kind of to the resistance to low light of plant and low temperature resistant related PPR albumen and its encoding gene and application Download PDFInfo
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Abstract
The invention discloses a kind of resistance to low light of plant and low temperature resistant related PPR albumen and its encoding gene and application.The protein that the present invention is provided is following protein a) or b) or c)：A) amino acid sequence is the protein shown in sequence 2 in sequence table；B) fused protein that the N-terminal of the protein in sequence table shown in sequence 2 and/or C-terminal connection label are obtained；C) protein with identical function that substitution and/or missing and/or addition by the amino acid sequence shown in sequence in sequence table 2 by one or several amino acid residues is obtained.Proved by testing：The plant of PPR gene delections shows the phenotype to low temperature, low smooth tolerance reduction, illustrate that PPR genes have the function of the regulation and control resistance to low smooth low temperature of plant, for improve the low temperature of other regional improved seeds, low light intensity adaptability, it is rapid to expand the Northeast and In Middle And Lower Reaches of Changjiang River, cultivate more excellent, high yield rice varieties and provide the foundation.
Description
Technical field
The invention belongs to biological technical field, and in particular to a kind of resistance to low light of plant and low temperature resistant related PPR albumen and
Its encoding gene and application.
Background technology
The Northeast and In Middle And Lower Reaches of Changjiang River are China paddy rice main producing regions.In the Northeast's paddy growth initial stage and the Changjiang river
In The Middle And Lower Reaches early rice early growth period is frequently encountered the abiotic stress such as low temperature, low light intensity, especially early spring rain
Afterwards, temperature may drop to less than 10 DEG C, and light intensity is not enough 1/10th of normal light intensity, but adapt to the water of this area's plantation
Rice main breed resource is extremely limited.Low temperature, low light intensity not only slow down growing for plant, directly be led when serious
Cause the plant significantly underproduction, or even Large Scale Death.Many improved seeds are directly affects to promote in these areas.Therefore,
Need to further investigate the relation between plant and abiotic stress, disclose plant to low temperature, the environment stress signal of low light
Response and response mechanism.So as to improve theoretical base to cultivate the more extensive improved seeds of temperature, light intensity adaptability
Plinth, is that these paddy rice main producing regions introduce more excellent offer possibilities.
The content of the invention
The technical problems to be solved by the invention are how to regulate and control stress resistance of plant.
In order to solve the above technical problems, present invention firstly provides regulating and controlling plant stress-resistance with plant adversity resistance related protein
Application in property.
Entitled PPR provided by the present invention with plant adversity resistance related protein, be it is following a) or b) or c)
Protein：
The protein that the present invention is provided is following protein a) or b) or c)：
A) amino acid sequence is the protein shown in sequence 2 in sequence table；
B) fused protein that the N-terminal of the protein in sequence table shown in sequence 2 and/or C-terminal connection label are obtained；
C) by the amino acid sequence shown in sequence in sequence table 2 by one or several amino acid residues substitution and/or
The protein with identical function that missing and/or addition are obtained.
Wherein, sequence 2 is made up of 585 amino acid residues.
In order that the protein in a) is easy to purifying, the amino of protein that can be in sequence table shown in sequence 2
End or the upper label as shown in table 1 of carboxyl terminal connection.
The sequence of table 1, label
It is above-mentioned c) in protein PPR, the substitution of one or several amino acid residues and/or missing and/or add
Add as the substitution and/or missing and/or addition no more than 10 amino acid residues.
It is above-mentioned c) in protein PPR can be artificial synthesized, also can first synthesize its encoding gene, then carry out biological expression
Obtain.
It is above-mentioned c) in protein PPR encoding gene can by will in the DNA sequence dna shown in sequence 1 lack one
Or the codon of 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 the label shown in table 1 is obtained.
It is a further object to provide the biomaterial with above-mentioned albumen qualitative correlation.
Any of what the present invention was provided is following A 1 with the biomaterial of above-mentioned albumen qualitative correlation) to A12)：
A1 the nucleic acid molecules of above-mentioned protein) are encoded；
A2 A1) is contained) expression cassettes of the nucleic acid molecules；
A3 A1) is contained) recombinant vectors of the nucleic acid molecules；
A4 A2) is contained) recombinant vector of the expression cassette；
A5 A1) is contained) recombinant microorganisms of the nucleic acid molecules；
A6 A2) is contained) recombinant microorganism of the expression cassette；
A7 A3) is contained) recombinant microorganism of the recombinant vector；
A8 A4) is contained) recombinant microorganism of the recombinant vector；
A9 A1) is contained) the transgenic plant cells systems of the nucleic acid molecules；
A10 A2) is contained) the transgenic plant cells system of the expression cassette；
A11 A3) is contained) the transgenic plant cells system of the recombinant vector；
A12 A4) is contained) the transgenic plant cells system of the recombinant vector.
In above-mentioned relevant biological material, A1) nucleic acid molecules for it is following 1) or 2) or 3) shown in gene：
1) its coded sequence is the cDNA molecules or DNA molecular shown in sequence 1 in sequence table；
2) there is 75% or more than 75% homogeneity with the nucleotide sequence of 1) restriction, and encodes above-mentioned protein
CDNA molecules or genomic DNA molecule；
3) under strict conditions with 1) or 2) nucleotide sequence hybridization limited, and the cDNA of the above-mentioned protein of coding
Molecule or genomic DNA molecule.
Wherein, the nucleic acid molecules can be DNA, such as cDNA, genomic DNA or recombinant DNA；The nucleic acid point
Son can also be RNA, such as mRNA or hnRNA.
Wherein, sequence 1 is made up of 1758 nucleotides, the amino acid sequence shown in coded sequence 2.
Those of ordinary skill in the art can be easily using known method, such as side of orthogenesis and point mutation
Method, is mutated to the coding PPR of present invention nucleotide sequence.Those by manually modified, with this hair
Bright isolated PPR nucleotide sequence 75% or the nucleotides of higher homogeneity, as long as encoding PPR and having
Identical function is the nucleotide sequence derived from the present invention and is equal to the sequence of the present invention.
Term " homogeneity " used herein refers to the sequence similarity with native sequence nucleic acid." homogeneity " includes and this
Shown in the coded sequence 2 of invention amino acid sequence composition protein nucleotide sequence have 75% or higher, or
85% or higher, or 90% or higher, or 95% or higher homogeneity nucleotide sequence.Homogeneity can with the naked eye or
Computer software is evaluated.Using computer software, the homogeneity between two or more sequences can use percentage
(%) represents that it can be used to evaluate the homogeneity between correlated series.
Above-mentioned 75% or more than 75% homogeneity, can be 80%, 85%, 90% or more than 95% homogeneity.
In above-mentioned biomaterial, the stringent condition be in 2 × SSC, it is miscellaneous at 68 DEG C in 0.1%SDS solution
Hand over and wash film 2 times, each 5min, and in 0.5 × SSC, 0.1%SDS solution, hybridize and wash at 68 DEG C
Film 2 times, each 15min；Or, in 0.1 × SSPE (or 0.1 × SSC), 0.1%SDS solution, 65 DEG C of bars
Hybridize under part and wash film.
In above-mentioned biomaterial, A2) described in the nucleic acid molecules containing coding PPR expression cassette (PPR gene expressions
Box), it is the DNA for referring to express PPR in host cell, the DNA not only may include the startup for starting PPR transcriptions
Son, may also include the terminator for terminating PPR transcriptions.Further, the expression cassette may also include enhancer sequence.Can
Promoter for the present invention includes but is not limited to：Constitutive promoter；Tissue, organ and the special promoter of development
And inducible promoter.The example of promoter includes but is not limited to：The constitutive promoter 35S of cauliflower mosaic virus：
Wound-inducible promoter from tomato, leucine aminopeptidase (" LAP ", Chao et al. (1999) Plant
Physiol 120：979-992)；Chemical inducible promoter from tobacco, pathogenesis correlation 1 (PR1) (by
Salicylic acid and BTH (diazosulfide -7- carbothioic acid S-methyl esters) inductions)；Tomato protease inhibitors II
Promoter (PIN2) or LAP promoters (can use methyl jasmonate induction)；(U.S. is special for heat-shock promoters
Profit 5,187,267)；Tetracycline inducible promoter (United States Patent (USP) 5,057,422)；Seed specific promoters,
Such as Millet Seed specificity promoter pF128 (CN101063139B (Chinese patent 200710099169.7)), seed
The special promoter of storage protein is (for example, phaseolin, napin, oleosin and soybean beta
(Beachy et al. (1985) EMBO is J.4 for conglycin promoter：3047-3053)).They can be independent
It is used in combination using or with other plant promoters.All references cited herein is quoted in full.Suitably
Transcription terminator includes but is not limited to：Agrobacterium nopaline syntase terminator (NOS terminator), cauliflower mosaic
Malicious CaMV 35S terminators, tml terminators, pea rbcS E9 terminators and nopaline and octopine synthase are whole
Only sub (see, e.g.：Odell et al. (I985)Nature 313:810；Rosenberg et al. (1987)
Gene,56:125；Guerineau et al. (1991) Mol.Gen.Genet, 262:141；Proudfoot
(1991)Cell,64:671；Sanfacon et al. Genes Dev., 5:141；Mogen et al. (1990) Plant
Cell,2:1261；Munroe et al. (1990) Gene, 91:151；Ballad et al. (1989) Nucleic
Acids Res.17:7891；Joshi et al. (1987) Nucleic Acid Res., 15:9627).
The recombinant vector of the PPR expression casettes can be contained with existing expression vector establishment.The plant expression is carried
Body includes double base agrobacterium vector and the carrier available for plant micropellet bombardment etc..As pAHC25, pBin438,
PCAMBIA1302, pCAMBIA2301, pCAMBIA1301, pCAMBIA1300, pBI121, pCAMBIA1391-Xa or
PCAMBIA1391-Xb (CAMBIA companies) etc..The plant expression vector can also include the 3 ' of foreign gene and hold non-turn over
Translate region, i.e., the DNA fragmentation comprising polyadenylation signals and any other participation mRNA processing or gene expression.It is described
The bootable polyadenylic acid of polyadenylation signals is added to 3 ' ends of mRNA precursor, such as Agrobacterium crown gall nodule induction (Ti)
Plasmid gene (such as rouge alkali synthetase gene Nos), the transcription of the end of plant gene (such as soybean storage protein genes) 3 '
Non-translational region is respectively provided with similar functions.During using gene constructed plant expression vector of the invention, enhancer is it is also possible to use,
Including translational enhancer or transcriptional enhancer, these enhancer regions can be that ATG initiation codon or neighboring region rise
Beginning codon etc., but must be identical with the reading frame of coded sequence, to ensure the correct translation of whole sequence.It is described to turn over
It is extensive to translate the source of control signal and initiation codon, can be natural or synthesis.Translate
Beginning, region can come from transcription initiation region or structural gene.For the ease of being reflected to transgenic plant cells or plant
Fixed and screening, can be processed to plant expression vector used, and face can be produced as added the coding that can be expressed in plant
The enzyme of color change or the gene (gus gene, luciferase genes etc.) of luminophor, the marker gene of antibiotic are (such as
Assign to kanamycins and the nptII genes of associated antibiotic resistance, assign to herbicide phosphinothricin resistance
Bar genes, assign the hph genes to antibiotic hygromycin resistance, and assign the dhfr to methotrexate resistance
Gene, is assigned to the EPSPS genes of glyphosate) or (such as anti-herbicide such as anti-chemical reagent marker gene
Gene), provide metabolism mannose ability mannose-6-phosphate isomerase gene.From the security of genetically modified plants
Consider, any selected marker can be not added with, transformed plant is directly screened with adverse circumstance.
In above-mentioned biomaterial, the carrier can be plasmid, sticking grain, bacteriophage or viral vectors.
In above-mentioned biomaterial, the microorganism can be yeast, bacterium, algae or fungi, such as Agrobacterium.
In above-mentioned biomaterial, the transgenic plant cells system, Transgenic plant tissue and genetically modified plants organ are equal
Do not include propagating materials.
It is a still further object of the present invention to provide above-mentioned protein or the new application of above-mentioned relevant biological material.
The invention provides the application of above-mentioned protein or above-mentioned relevant biological material in regulation and control plant stress tolerance.
The genetically modified plants of resistance of reverse reduction are being cultivated present invention also offers above-mentioned protein or above-mentioned relevant biological material
In application.
In above-mentioned application, the resistance of reverse is Heat pretreatment and/or low light intensity patience.
Final object of the present invention is to provide a kind of method for the genetically modified plants for cultivating resistance reduction.
The method of the genetically modified plants for the cultivation resistance reduction that the present invention is provided includes that above-mentioned protein expression will be suppressed
Material is imported in recipient plant, the step of obtaining genetically modified plants；The resistance of reverse of the genetically modified plants less than it is described by
Body plant.
The resistance of reverse of the genetically modified plants is embodied in less than the recipient plant：Low temperature (be specially 4 DEG C or
21 DEG C) or low light intensity (80 μ Em-2s-1) under the conditions of, the survival rate of genetically modified plants is less than recipient plant and/or turns base
Because two of plant the important regulating and controlling factor OsCAB2 and the OsCBF2 expression quantity for responding low temperature less than recipient plant and/
Or growth of transgenic plants speed, Development of Chloroplasts level are slower than recipient plant.
In the above method, the material for suppressing above-mentioned protein expression is the DNA molecular shown in sequence 3.
In an embodiment of the present invention, the material for suppressing above-mentioned protein expression is by p1300-Cas9-PPRsg
Import in the recipient plant, the p1300-Cas9-PPRsg conversion carriers are by the DNA shown in sequence 3 points
DNA fragmentation between son replacement carrier pCAMBIA1300 KpnI and EcoRI restriction enzyme sites, and keep carrier
The constant obtained carrier of pCAMBIA1300 other sequences.
In the above method, the resistance of reverse is Heat pretreatment and/or low light intensity patience.
In the above method, the recipient plant is monocotyledon or dicotyledon, and the monocotyledon is specially
Paddy rice, the paddy rice is specially Nipponbare.
The present invention is cloned into from paddy rice to low temperature, low smooth tolerance related gene PPR, and constructs PPR genes
The plant of missing.Proved by testing：The plant of PPR gene delections shows to reduce low temperature, low smooth tolerance
Phenotype, illustrates that PPR genes have the function of the regulation and control resistance to low smooth low temperature of plant, can improve excellent by being overexpressed the gene
Non-defective unit kind is to low temperature, low smooth tolerance, so that break the territory restriction of paddy rice improved seeds, it is excellent to improve other areas
The adaptability of the low temperature of non-defective unit kind, low light intensity, it is rapid to expand the Northeast and In Middle And Lower Reaches of Changjiang River, it is especially eastern
The variety source of the paddy rice main producing region such as backlands area, cultivates more excellent, high yield rice varieties and provides the foundation.
Brief description of the drawings
Fig. 1 is phenotype of the PPR gene delections paddy rice under different light intensity and condition of different temperatures.
Fig. 2 grows under normal operation for PPR gene delections paddy rice is transferred to survival condition after 4 DEG C, 48 hours after 10 days.
Fig. 3 is OsCAB2 and OsCBF2 expressions in PPR gene delection paddy rice.
Embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
Material, reagent used etc., unless otherwise specified, are commercially obtained in following embodiments.
Carrier OsU6-sk in following embodiments, carrier 35S-cas9-sk and carrier pCAMBIA1300 are in document
“Feng Z.,Zhang B.,Ding W.,Liu X.,Yang D,Wei P,Cao F.,Zhu S.,Zhang F.,Mao Y.,
Zhang J.(2013).Efficient genome editing in plants using a CRISPR/Cas system.Cell
Research 23:Mistake disclosed in 1229-1232. ", the public can obtain from Institute of Botany, Chinese Academy of Sciences.
The acquisition of embodiment 1, PPR genes
1st, the extraction of plant cDNA
The RNA of wild rice (Nipponbare) blade is extracted, reverse transcription obtains cDNA.
2nd, PCR is expanded
CDNA using step 1 acquisition enters performing PCR amplification as template using F and R primers, obtains PCR amplifications
Product.Primer sequence is as follows：F:5’-GAATTCGGATCCATGCCTCCTCCCACCGTCCCCTTCT-3’
And R:5’-GGGCCCCTCGAGGTCGACCCAGAAATCCCCGCAAGAGCATG-3’.
3rd, the acquisition of PPR genes
1% agarose gel electrophoresis detection is carried out to pcr amplification product, reclaims and is sequenced after purification.
Sequencing result shows：PCR amplifications obtain the amplified production that size is 1758bp, such as sequence table of its nucleotide sequence
It is PPR genes, the amino acid sequence such as sequence of PPR gene codes by the unnamed gene shown in sequence 1 shown in middle sequence 1
In list shown in sequence 2, the albumen shown in sequence 2 is named as PPR albumen.
Embodiment 2, the acquisition of PPR gene delection paddy rice and resistance of reverse analysis
First, the acquisition of PPR gene delections paddy rice
Reference literature " Feng Z., Zhang B., Ding W., Liu X., Yang D, Wei P, Cao F., Zhu S., Zhang
F.,Mao Y.,Zhang J.(2013).Efficient genome editing in plants using a CRISPR/Cas system.
Cell Research 23:Method in 1229-1232. ", design construction PPR gene C RISPR/Cas genome editors
System, obtains PPR gene delection paddy rice.Show through sequencing identification：Compared with wild rice, PPR gene delections
Paddy rice is to lack the 55-62 nucleic acid molecule of the PPR genes shown in the sequence 1 of wild rice, and is kept
The constant obtained rice plant of rice genome other sequences.Specific construction step is as follows：
1st, the structure of p1300-Cas9-PPRsg conversion carriers
(1) genomic DNA using wild rice (Nipponbare) enters performing PCR as template using primers F and R
Amplification, forms the dimer with cohesive end, primer sequence is as follows by way of being denatured annealing：
F:5’-GTGTGTGCGGCTTCGCGGCCGCAG-3’；
R:5’-AAACCTGCGGCCGCGAAGCCGCAC-3’；
(2) dimer with cohesive end that step 1 is obtained is connected to carrier OsU6-sk two BbsI enzymes
Between enzyme site, recombinant vector 1 is obtained；
(3) with restriction enzyme KpnI and HindIII digestion recombinant vector 1, it is 474bp's to reclaim and obtain size
OsU6-sk fragments；
(4) restriction enzyme HindIII and EcoRI digestion carrier 35S-cas9-sk is used, recovery obtains size and is
5476bp 35S-cas9 fragments；
(5) the 35S-cas9 fragments that the OsU6-sk fragments and step (4) obtained step (3) is obtained are connected to
Between carrier pCAMBIA1300 KpnI and EcoRI restriction enzyme sites, obtain p1300-Cas9-PPRsg conversions and carry
Body.And it is sequenced.
Sequencing result shows：P1300-Cas9-PPRsg conversion carriers are to replace the DNA molecular shown in sequence 3 to carry
DNA fragmentation between body pCAMBIA1300 KpnI and EcoRI restriction enzyme sites, and keep carrier
The constant obtained carrier of pCAMBIA1300 other sequences.
2nd, the acquisition of recombinant bacterium
In the p1300-Cas9-PPRsg conversion carriers importing Agrobacterium EHA105 that step 1 is obtained (EHA105,
Beijing CHMC ocean is biological, article No.：GX0133-100), recombinant bacterium EHA105-PPRsg is obtained.
3rd, the acquisition of PPR gene delections paddy rice
Acceptor material wild rice (Nipponbare) is infected with recombinant bacterium EHA105-PPRsg, PPR genes is obtained and lacks
Dehydration rice.And Molecular Identification is carried out to it.The primer of Molecular Identification is as follows：PF:5’-
ATGCCTCCTCCCACCGTCCC-3’；PR:5’-TGCATCCCCACGAAGAACTC-3’.
Qualification result shows：Compared with wild rice, PPR gene delections paddy rice is by the sequence 1 of wild rice
Shown PPR genes the 55-62 nucleic acid molecule missing, and keep rice genome other sequences it is constant
The rice plant arrived.The missing of 8 bases of PPR genes causes reading frame to occur frameshit, causes encoding histone to lift end
Only.Illustrate the PPR gene delections success in PPR gene delection paddy rice, and PPR gene delection paddy rice is named as
ppr。
2nd, the resistance of reverse analysis of the paddy rice of PPR gene delections
1st, phenotype
By the seedling of PPR gene delections rice plant (ppr) and wild rice plant (WT) at low temperature (21 DEG C)
With different light intensity (80 μ Em-2s-1, 250 μ Em-2s-1) under the conditions of growth 10 days after observe phenotype.
As a result it is as shown in Figure 1：(27 DEG C, the μ Em of light intensity 250 under normal operation-2s-1), PPR gene delection paddy rice
There was no significant difference for the phenotype of plant (ppr) and wild rice plant (WT).But in low temperature (21 DEG C) and low
Light intensity (80 μ Em-2s-1) under the conditions of, PPR gene delections rice plant growth is slow, Development of Chloroplasts obstacle, performance
For albefaction；And low temperature (21 DEG C) and high light intensity (250 μ Em-2s-1) under the conditions of, PPR gene delections rice plant (ppr)
Slow-growing and Development of Chloroplasts obstacle obtain part recovery.Thus, illustrate that PPR albumen is not only involved in paddy rice to low
The response of temperature, also participates in the response of light intensity.
2nd, survival rate
By the seedling of PPR gene delections rice plant (ppr) and wild rice plant (WT) under normal operation
Growth is handled under the conditions of low temperature (4 DEG C) is transferred to after 10 days, and survival rate is counted respectively after processing 48h.
As a result it is as shown in Figure 2：4 DEG C of low-temperature treatments are after 2 days, and the survival rate of wild rice plant (WT) is about 95%,
And the survival rate of PPR gene delections rice plant (ppr) is less than 10%.The survival of PPR gene delection rice plants
Rate significantly lowers, and shows as low-temperature sensitive.Illustrate that paddy rice PPR albumen responds the unfavorable rings such as low temperature, low light intensity in paddy rice
During border, key player is play, with regulation and control plant low temperature patience and low light intensity patience.
3rd, paddy rice low response correlation factor
OsCAB2 and OsCBF2 are the important regulating and controlling factors of two response low temperature in paddy rice, when paddy rice runs into low temperature,
The two gene transcription levels are raised rapidly, start the stress reaction to low temperature.By detecting PPR gene delection paddy rice
The change of OsCAB2 and OsCBF2 expressions in plant (ppr), and then probe into the function of PPR genes.Tool
Body step is as follows：
By the seedling of PPR gene delections rice plant (ppr) and wild rice plant (WT) under normal operation
Growth is handled under the conditions of low temperature (4 DEG C) is transferred to after 10 days, respectively after processing 1h and 3h, is extracted at low temperature
The RNA of PPR gene delections rice plant (ppr) and wild rice plant (WT) after reason, reverse transcription,
Obtain cDNA.With quantitative real time PCR Instrument, using the cDNA of acquisition as template, be respectively adopted OsCAB2 primers and
OsCBF2 primers enter performing PCR amplification, detect the expression quantity of OsCAB2 and OsCBF2 genes.With ACT2 primers
For reference gene, primer sequence is as follows：OsCAB2 primers：5 '-CTACGCCACCAACTTCGTC-3 ' and
5’-TGAGGTCCATTGCTTTCATG-3’；OsCBF2 primers：
5 '-GACTACGACTGCGAGGTCAA-3 ' and 5 '-AACGGCACACCATAAGTACG-3 '；ACT2
Primer：5 '-CCTTCAACACCCCTGCTATG-3 ' and 5 '-CAATGCCAGGGAACATAGTG-3 '.
The reaction system of above-mentioned PCR amplifications：7.5 microlitres of SYBR Premix Ex Taq mixtures, each 2 microlitres of primer,
1 microlitre of cDNA templates, plus double distilled water is to 15 microlitres.
The reaction condition of above-mentioned PCR amplifications：The first step, 95 degree 30 seconds, second step, 95 degree 5 seconds, 60 degree 20
Second, 40 circulations；3rd step, 95 degree 1 minute, 55 degree 1 minute, 95 degree 1 minute.
As a result it is as shown in Figure 3：After low-temperature treatment, two low temperature response in PPR gene delections rice plant (ppr)
Regulatory factor OsCAB2 and OsCBF2 expression quantity are not raised not only, and have downward trend, so as to can not start
Stress reaction of the plant to low temperature.It is the expression that correlation factor is responded by adjusting and controlling rice low temperature to illustrate PPR genes, is come
Improve adaptability of the paddy rice to environment such as low temperature, low light.
Claims (10)
1. protein, is following protein a) or b) or c)：
A) amino acid sequence is the protein shown in sequence 2 in sequence table；
B) fused protein that the N-terminal of the protein in sequence table shown in sequence 2 and/or C-terminal connection label are obtained；
C) by the amino acid sequence shown in sequence in sequence table 2 by one or several amino acid residues substitution and/or
The protein with identical function that missing and/or addition are obtained.
2. the biomaterial with the albumen qualitative correlation described in claim 1, is following A 1) to A12) in any
Kind：
A1) the nucleic acid molecules of the protein described in coding claim 1；
A2 A1) is contained) expression cassettes of the nucleic acid molecules；
A3 A1) is contained) recombinant vectors of the nucleic acid molecules；
A4 A2) is contained) recombinant vector of the expression cassette；
A5 A1) is contained) recombinant microorganisms of the nucleic acid molecules；
A6 A2) is contained) recombinant microorganism of the expression cassette；
A7 A3) is contained) recombinant microorganism of the recombinant vector；
A8 A4) is contained) recombinant microorganism of the recombinant vector；
A9 A1) is contained) the transgenic plant cells systems of the nucleic acid molecules；
A10 A2) is contained) the transgenic plant cells system of the expression cassette；
A11 A3) is contained) the transgenic plant cells system of the recombinant vector；
A12 A4) is contained) the transgenic plant cells system of the recombinant vector.
3. relevant biological material according to claim 2, it is characterised in that：A1) nucleic acid molecules is such as
It is lower 1) or 2) or 3) shown in gene：
1) its coded sequence is the cDNA molecules or DNA molecular shown in sequence 1 in sequence table；
2) there is 75% or more than 75% homogeneity with the nucleotide sequence of 1) restriction, and described in coding claim 1
Protein cDNA molecules or genomic DNA molecule；
3) under strict conditions with 1) or 2) nucleotide sequence hybridization limited, and described in coding claim 1
The cDNA molecules or genomic DNA molecule of protein.
4. the relevant biological material described in protein or Claims 2 or 3 described in claim 1 is resistance in regulation and control plant
Application in inverse property.
5. the relevant biological material described in protein or Claims 2 or 3 described in claim 1 is cultivating resistance of reverse
Application in the genetically modified plants of reduction.
6. the application according to claim 4 or 5, it is characterised in that：The resistance of reverse be Heat pretreatment and/or
Low light intensity patience.
7. it is a kind of cultivate resistance reduction genetically modified plants method, including by suppress claim 1 described in albumen
The material of matter expression is imported in recipient plant, the step of obtaining genetically modified plants；The resistance of reverse of the genetically modified plants is low
In the recipient plant.
8. method according to claim 7, it is characterised in that：The protein suppressed described in claim 1
The material of expression is the DNA molecular shown in sequence 3.
9. the method according to claim 7 or 8, it is characterised in that：The resistance of reverse be Heat pretreatment and/or
Low light intensity patience.
10. according to any described method in claim 7-9, it is characterised in that：The recipient plant is unifacial leaf
Plant or dicotyledon.
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| CN110923244A (en) * | 2019-12-19 | 2020-03-27 | 中国烟草总公司郑州烟草研究院 | Tobacco mitochondrial RNA editing factor NtMEF1 and application thereof |
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| CN114605512B (en) * | 2020-12-09 | 2023-10-17 | 中国农业大学 | Application of corn PPR protein or coding gene thereof in regulation and control of plant cold resistance |
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