CN111424042A - Application of immune negative regulatory factor NbMORF8 gene and protein thereof in phytophthora resistance of plants - Google Patents
Application of immune negative regulatory factor NbMORF8 gene and protein thereof in phytophthora resistance of plants Download PDFInfo
- Publication number
- CN111424042A CN111424042A CN202010396024.9A CN202010396024A CN111424042A CN 111424042 A CN111424042 A CN 111424042A CN 202010396024 A CN202010396024 A CN 202010396024A CN 111424042 A CN111424042 A CN 111424042A
- Authority
- CN
- China
- Prior art keywords
- pro
- nbmorf8
- asn
- gly
- gene
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
- C12N15/8218—Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
- C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
- C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
- C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
- C12N15/8279—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance
- C12N15/8282—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for biotic stress resistance, pathogen resistance, disease resistance for fungal resistance
Landscapes
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Cell Biology (AREA)
- Virology (AREA)
- Botany (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses an application of an immune negative regulatory factor NbMORF8 gene and a protein thereof in plant phytophthora resistance, belonging to the technical field of biology, wherein the NbMORF8 gene negatively regulates the resistance of a plant to phytophthora by interfering a plant salicylic acid immune signal pathway and active oxygen burst; the nucleotide sequence of the NbMORF8 gene is shown as SEQ ID No: 1 or a homologous sequence with more than 70% of sequence homology with the sequence; the expression level of the protein coded by the NbMORF8 gene is reduced through genetic engineering, so that the resistance of the plant to phytophthora can be enhanced; the amino acid sequence of the protein is shown as SEQ ID No: 2 or homologous sequence with more than 70% of sequence homology with the sequence; the negative regulation factor has the potential to be applied to genetic engineering for cultivating new disease-resistant varieties and has important application value for changing the situation of pathogenic bacteria drug resistance enhancement in the prior art.
Description
Technical Field
The invention relates to the technical field of biology, in particular to an application of an immune negative regulatory factor NbMORF8 gene and a protein thereof in phytophthora resistance of plants.
Background
Oomycetes (Oomycotes) are a group of eukaryotic microorganisms including Phytophthora that are similar in morphology to fungi. Although morphologically similar to fungi, but evolutionarily homologous to algae, are independent populations with uniquely classified positions, including Phytophthora (Phytophthora), Saprolegniales (Saprolegniales), Peronosporales (Peronosporales), and the like. Many members of the phytophthora genus cause devastating plant diseases. For example, late blight of potato caused by Phytophthora infestans has caused the grand famine of Ireland in the 19 th century, causing the Ireland population to sharply decline by about one quarter and about one million people to move overseas due to hunger, an event that has profound effects on politics, economics, and culture in Ireland, and even throughout Europe. Tobacco black shank caused by phytophthora parasitica (p.parasitica) is one of the major tobacco diseases worldwide, and occurs to different degrees in each major tobacco production area in china every year, thus being a serious threat to the global tobacco industry. At present, the prevention and control of oomycetes such as phytophthora are mainly dependent on the utilization of the existing disease-resistant varieties and chemical prevention and control methods. However, phytophthora has a high rate of evolution, so that resistance of disease-resistant varieties is easily lost through mutation and resistance to chemical agents is easily generated. Therefore, the development of a novel disease-resistant way to excavate new disease-resistant resources is imperative.
During the affinity interaction between plants and pathogenic bacteria, the pathogenic bacteria rely on a series of nutrient transmission, molecular exchange and hormone signal transduction processes which are involved by plant susceptible factors to realize successful infection and colonization. When the disease-sensitive factors are deleted, the plants show resistance to pathogenic bacteria, which opens a new idea for breeding the plants for disease resistance. Strategies for enhancing the resistance of plants by finding natural mutants or site-directed mutagenesis of plant susceptible factors to loss of function have been applied in the breeding of plants against fungi.
Many immune negative regulators of plant resistance to oomycetes have been identified at present, but most of them focus on anti-oomycetes of downy mildew, and the research on immune negative regulators of phytophthora is less, and the specific action mechanism of many immune negative regulators is not clear. Although the application of the negative oomycete immune regulation factor in potatoes is explored in recent years, the negative oomycete immune regulation factor is not applied to crop disease-resistant breeding.
Disclosure of Invention
An object of the present invention is to solve at least the above problems and to provide at least the advantages described later.
The invention also aims to provide application of the plant immune negative regulatory factor NbMORF8 gene in phytophthora resistance, and provides a new biological control way for controlling potato late blight.
To achieve these objects and other advantages of the present invention, there is provided a use of an immune negative regulator gene NbMORF8 in plant resistance to phytophthora, said NbMORF8 gene negatively regulating resistance of a plant to phytophthora by interfering with a plant salicylic acid immune signal pathway and active oxygen bursts;
the nucleotide sequence of the NbMORF8 gene is shown as SEQ ID No: 1 or a sequence corresponding to SEQ ID No: 1, and homologous sequences with homology of more than 70%.
The invention also aims to provide the application of the protein coded by the NbMORF8 gene of the immune negative regulatory factor in resisting phytophthora in plants, the expression level of the protein coded by the NbMORF8 gene is reduced through genetic engineering, and the resistance of the plants to the phytophthora is enhanced;
the amino acid sequence of the protein is shown as SEQ ID No: 2 or a sequence similar to SEQ ID No: 2, and homologous sequences with more than 70% of sequence homology.
The invention at least discloses the following technical effects:
the invention discloses an immune negative regulatory factor NbMORF8 gene which is cloned as an immune negative regulatory factor for influencing the signal transmission of ROS and salicylic acid of plants for the first time. The NbMORF8 defect type bunny material is obtained by a virus-induced gene silencing (VIGS) technology, and an in vitro leaf inoculation experiment proves that a silencing plant of the NbMORF8 can obviously increase the resistance to phytophthora parasitica, phytophthora infestans and phytophthora capsici. The experimental results prove that the NbMORF8 gene is used as a negative regulatory factor to improve the resistance of phytophthora blight plants, and can be used for breeding phytophthora blight resistant varieties.
In addition, the NbMORF8 gene is analyzed and found to interfere with the plant endogenous salicylic acid signal path by a real-time fluorescent quantitative PCR technology, and the plant shows stronger ROS burst after NbMORF8 is deleted by active oxygen measurement. In conclusion, NbMORF8 is used as a novel plant immune negative regulator to negatively regulate the resistance of plants to phytophthora by interfering with downstream signaling of plant endogenous salicylic acid and ROS signaling.
In addition, the invention discovers that NbMORF8 participates in HR reaction of plants through analysis of a gene transient expression technology. Further, the NbMORF8 is found to participate in the normal accumulation of phytophthora effector proteins through western blotting and semi-quantitative PCR (polymerase chain reaction) technical analysis. The results show that NbMORF8, as a novel plant immune negative regulatory factor, negatively regulates the resistance of plants to phytophthora by influencing the accumulation of effector proteins.
Additional advantages, objects, and features of the invention will be set forth in part in the description which follows and in part will become apparent to those having ordinary skill in the art upon examination of the following or may be learned from practice of the invention.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.
FIG. 1 shows the result of the sequence alignment of the NbMORF8 protein of the immune negative regulatory factor of the invention with the homologous sequences StMORF8-1 and StMORF8-2 protein;
FIG. 2 shows the resistance of the immune negative regulator NbMORF8 gene-silenced plant to Phytophthora;
FIG. 3 shows that the immune negative regulator NbMORF8 gene silences the expression of NbPR1 and NbPR2 genes and enhances the burst level of active oxygen of plants.
Detailed Description
Reference will now be made in detail to various exemplary embodiments of the invention, the detailed description should not be construed as limiting the invention but as a more detailed description of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. Further, for numerical ranges in this disclosure, it is understood that each intervening value, between the upper and lower limit of that range, is also specifically disclosed. Every smaller range between any stated value or intervening value in a stated range and any other stated or intervening value in a stated range is encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference herein for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It is to be understood that, as used herein, terms such as "comprising," "including," "having," "containing," and the like are open-ended terms that mean including, but not limited to.
Example 1
Cloning of N.benthamiana immune negative regulatory factor NbMORF8 gene
1. Plant material: benzenbacco (which can be obtained through public channels), and RNA is extracted from leaves of Benzenbacco.
RNA was extracted using an RNA extraction kit (OMGA, &lTtTtransfer = L "&gTtL &lTt/T &gTtot #: R6827-01), the integrity of the RNA was identified by agarose gel electrophoresis, and the purity and concentration of the RNA were then determined on a spectrophotometer.
2. Cloning of genes
The cDNA of Nichoisy was obtained using a reverse transcription kit (TaKaRa, &lTtTtranslation = L "&gTtL &lTt/T &gTtot #: AHE3187A), and an upstream and downstream primer NbMORF8-F/R was designed based on the full-length coding sequence of NbMORF8(Niben101Scf20512g00013.1) provided by the Solanaceae genomic site Sol Genomics Network,
NbMORF8-F：5’-CCGCTCGAGATGGCGACTCGCTATCTGACT-3’
NbMORF8-R：5’-GCTCTAGATCAGTAATTAGGAGGGGGCATAT-3’，
amplification was performed using cDNA as a template. The PCR product is subjected to enzyme digestion, connection and bacterial liquid PCR verification to construct a vector pKANNibal-NbMORF8, and then the vector is sent to sequencing. The sequencing results were compared to published sequences and the correct plasmid was used in subsequent experiments.
Example 2
Sequence information and homology analysis of NbMORF8 gene of Nicotiana benthamiana
The full-length CDS sequence of the NbMORF8 gene of the invention is 1296bp, and the detailed sequence is NbMORF8(Niben101Scf20512g00013.1) provided by Solgenomics Network of solanaceae, namely the gene of the invention has the SEQ ID No: 1; the amino acid sequence has 431 amino acids, and the detailed sequence is NbMORF8(Niben101Scf20512g00013.1) provided by Sol Genomics Network, namely the SEQ ID No: 2.
the amino acid sequence of the protein coded by the NbMORF8 gene of Nicotiana benthamiana is subjected to homology search in a potato genome by using a B L AST program, and 2 homologous genes StMORF8-1 and StMORF8-2 of NbMORF8 are found in the result, the similarity of the amino acid sequences is more than 70 percent, the 2 homologous genes are presumed to have the same function as NbMORF8, the homology of the amino acid sequences coded by the 2 homologous genes and the amino acid sequence of NbMORF8 gene is more than 50 percent, the amino acid sequences coded by the NbMORF8 gene are presumed to have similar biological functions, the amino acid sequences coded by the homologous genes are shown as GenBank accession Number: XP-006358943.1 and XP-006360053.1, and SEQ ID Nos. 3 and SEQ ID No. 4 of the invention.
The alignment results of the amino acid sequences of the NbMORF8 gene of Nicotiana benthamiana of the invention and 2 homologous genes in potato are shown in FIG. 1.
Example 3
NbMORF8 silenced plants show resistance to Phytophthora parasitica, Phytophthora capsici and Phytophthora infestans.
Selecting NbMORF8 specificThe segment is about 300bp, and a pTRV2-NbMORF8 silencing vector is constructed. pTVR1, pTRV2-GFP and pTRV2 vector into which the fragment of interest has been ligated are electroporated into Agrobacterium. The agrobacterium transformed by pTRV1 and pTRV2-NbMORF8 are co-transiently expressed on Nicotiana benthamiana by using an agrobacterium infiltration method, and the final concentration is OD6000.25. And (3) selecting the leaf inoculation analysis of the experimental group when the positive control has a relatively obvious silencing effect (after 2-3 weeks) by using the PDS genes silenced together as a positive control and GFP as a negative control. Meanwhile, a specific primer is designed, and the expression condition of NbMORF8 in a silent plant is detected: collecting about 0.1g of the Nicotiana benthamiana leaf sample, extracting RNA, carrying out reverse transcription to obtain cDNA, designing a specific primer, and detecting the expression condition of NbMORF8 by using a real-time fluorescent quantitative PCR technology.
Collecting the tobacco leaves of the Bunshi of NbMORF8 silent, using TRV-GFP treatment as a control, inoculating about 2000 GFP-labeled zoospores of phytophthora parasitica, carrying out fluorescence observation after dark culture in an incubator at 23 ℃ for about 40 hours, and counting the diameters of scabs, adjusting the number of the zoospores to be about 800 when inoculating phytophthora capsici, adjusting the number of the zoospores to be about 800 when inoculating the phytophthora parasitica, observing and measuring the diameters of the scabs under the rest culture conditions similar to those of the phytophthora parasitica, stimulating the zoospores after about 10 days after the culture of the freshly cultured phytophthora infestans, adding about 5m L sterile water into the culture, stimulating for about 1-2 hours at 4 ℃, inoculating the bacteria after releasing a large number of the zoospores, and inoculating about 1500 zoospores at each inoculation point.
The results are shown in FIG. 2, where (a-c) indicate that NbMORF 8-silenced plants exhibit greater resistance to Phytophthora parasitica, Phytophthora infestans and Phytophthora capsici; carrying out trypan blue staining on the leaves infected by the phytophthora infestans to display the hypha expansion condition (b); (d, f and h) are respectively the statistical results of the diameters of the scabs after inoculation of phytophthora parasitica (d), phytophthora infestans (f) and phytophthora capsici (h); (e) detecting the biomass of the phytophthora parasitica on the silent plant by using real-time fluorescent quantitative PCR; (g) statistics of sporangium production on the leaves of silenced and control plants were carried out 10 days after inoculation.
Example 4
NbMORF8 silenced plant activates the expression of NbPR1 and NbPR2 genes and increases the active oxygen burst level of the plant.
About 2 weeks after VIGS treatment, about 0.1g of Nicotiana benthamiana leaf sample is collected to extract RNA and is reversely transcribed into cDNA, and specific primers are designed to detect the expression of NbPR1, NbPR2, NbPR3, NbPR4, WRKY7 and WRKY8 by using a real-time fluorescent quantitative PCR technology. For inoculated leaves, the inoculation method of phytophthora parasitica is similar to that in example 3, samples are collected 3, 6, 12 and 24 hours after inoculation, RNA is extracted and is reversely transcribed into cDNA, and the expressions of NbPR1 and NbPR2 are detected by using a real-time fluorescent quantitative PCR technology through specific primers.
Collecting the tobacco leaves with good silencing effect about two weeks after VIGS treatment, collecting leaf cakes with the diameter of 5 mm by using a puncher, and soaking in ultrapure water overnight. The ultrapure water is replaced by test solution (luminol solution and horseradish peroxidase solution), and the chemiluminescence value is directly detected by using a microplate reader, wherein the final concentration of the ultrapure water is 1 mu M flg 22.
The results are shown in figure 3, where (a) silencing of NbMORF8 can activate expression of the salicylic acid signaling pathways NbPR1 and NbPR2 in the absence of bacteria; (b) after PAMP flg22 treatment from bacteria, TRV-NbMORF8 plant leaves have higher active oxygen burst level; (c-d) in the early stage of phytophthora parasitica infection, the expression level of NbMORF8 silent plants NbPR1 and NbPR2 is obviously higher than that of a control group.
The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solutions of the present invention can be made by those skilled in the art without departing from the spirit of the present invention, and the technical solutions of the present invention are within the scope of the present invention defined by the claims.
Sequence listing
<110> northwest agriculture and forestry science and technology university
<120> application of immune negative regulatory factor NbMORF8 gene and protein thereof in phytophthora root rot resistance of plants
<160>4
<170>SIPOSequenceListing 1.0
<210>1
<211>1296
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>1
atggcgactc gctatctgac tcgttctcta ctcactgcct tcactcgctc atacacctct 60
atttcaccgc cgagcccatt atcttctttt tctctcctcc ggctgcgacc tcttatcacc 120
gtcgccgcca atctaaaaaa cgcctctccg ccgccgaggg cttttgctgt tagagggttt 180
gccactcgac agacgtcatc gtcgctcaat gatccgaatc ctaactggtc gaatcgaccg 240
ccgaaggaga cgattttgct tgacggttgt gattttgaac actggcttgt tgtgatggag 300
aagcccgaag gtgacccaac tagggatgag attattgata gttacatcaa aactcttgct 360
actgttgttg gaagtgagga agaagcaagg atgaagatat attctgtctc aaccaggcat 420
tactatgctt ttggtgccct tgtatcggaa gaactttcgt acaaactaaa agaggtgccc 480
agggttcgtt gggtgcttcc tgattcctat ctggatgtta gaaataaaga ttatggaggc 540
gaacctttta tcaatgggca ggctgtacct tatgacccga agtaccatga ggaatgggtg 600
agaaacaatg ctcgtgctaa tgagagaaac aggcgaaatg accgacctcg taattttgat 660
agatccagaa actttgagag aagaagagac atgcagacta accaaaacag tcagaaccct 720
ggatccaaca tgccaccggg cgggcctccc aacatgagga atccaccccc tcccaacatg 780
gtcgggatgc agcaacctaa catgggaggt cctcctaaca tgggcgggat gcaccagcaa 840
cctaacatgg gaggtccgcc taacatgggc gggatgcacc agcagggcat gggaggtcca 900
cccaacatgg gcgggatgca ccagcagggc atgggagggc cacccaaccc aggcgggatg 960
caccagccgg gcatgggagg gccacccaac ccaggcggga tgcaccagcc gggcatggga 1020
aggccacctc atgtgggtgg gatgcagcag ccaaacatgc ctcccaacac aggaggtgta 1080
ccaccaccga acatgggtgg agggcctccc cataactatg gaggagtgcc aaacaatgca 1140
cccaatttcc aatacaacag cggatcaaac agtggaggca cgccttaccc aacaggtccc 1200
aggccaaacc agagctactc tcctaatcca tctgatggaa acccttatca gaatcaaaac 1260
ttacccggaa gagatatgcc ccctcctaat tactga 1296
<210>2
<211>430
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>2
Met Ala Thr Arg Tyr Leu Thr Arg Ser Leu Leu Thr Ala Phe Thr Arg
1 5 10 15
Ser Tyr Thr Ser Ile Ser Pro Pro Ser Pro Leu Ser Ser Phe Ser Leu
20 25 30
Leu Arg Leu Arg Pro Leu Ile Thr Val Ala Ala Asn Leu Lys Asn Ala
35 40 45
Ser Pro Pro Pro Arg Ala Phe Ala Val Arg Gly Phe Ala Thr Arg Gln
50 55 60
Thr Ser Ser Ser Leu Asn Asp Pro Asn Pro Asn Trp Ser Asn Arg Pro
65 70 75 80
Pro Lys Glu Thr Ile Leu Leu Asp Gly Cys Asp Phe Glu His Trp Leu
85 90 95
Val Val Met Glu Lys Pro Glu Gly Asp Pro Thr Arg Asp Glu Ile Ile
100 105 110
Asp Ser Tyr Ile Lys Thr Leu Ala Thr Val Val Gly Ser Glu Glu Glu
115 120 125
Ala Arg Met Lys Ile Tyr Ser Val Ser Thr Arg His Tyr Tyr Ala Phe
130 135 140
Gly Ala Leu Val Ser Glu Glu Leu Ser Tyr Lys Leu Lys Glu Val Pro
145 150 155 160
Arg Val Arg Trp Val Leu Pro Asp Ser Tyr Leu Asp Val Arg Asn Lys
165 170 175
Asp Tyr Gly Gly Glu Pro Phe Ile Asn Gly Gln Ala Val Pro Tyr Asp
180 185 190
Pro Lys Tyr His Glu Glu Trp Val Arg Asn Asn Ala Arg Ala Asn Glu
195 200 205
Arg Asn Arg Arg Asn Asp Arg Pro Arg Asn Phe Asp Arg Ser Arg Asn
210 215 220
Phe Glu Arg Arg Arg Asp Met Gln Thr Asn Gln Asn Ser Gln Asn Pro
225 230 235 240
Gly Ser Asn Met Pro Pro Gly Gly Pro Pro Asn Met Arg Asn Pro Pro
245 250 255
Pro Pro Asn Met Gly Gly Met Gln Gln Pro Asn Met Gly Gly Pro Pro
260 265 270
Asn Met Gly Gly Met His Gln Gln Gly Met Gly Gly Pro Pro Asn Met
275 280 285
Gly Gly Met His Gln Gln Gly Met Gly Gly Pro Pro Asn Met Gly Gly
290 295 300
Met His Gln Pro Gly Met Gly Gly Pro Pro Asn Pro Gly Gly Met His
305 310 315 320
Gln Pro Gly Met Gly Gly Pro Pro Asn Pro Gly Gly Met His Gln Pro
325 330 335
Gly Met Gly Arg Pro Pro His Val Gly Gly Met Gln Gln Pro Asn Met
340 345 350
Pro Pro Asn Thr Gly Gly Val Pro Pro Pro Asn Met Gly Gly Gly Pro
355 360 365
Pro His Asn Tyr Gly Gly Val Pro Asn Asn Ala Pro Asn Phe Gln Tyr
370 375 380
Asn Ser Gly Ser Asn Ser Gly Gly Thr Pro Tyr Pro Thr Gly Pro Arg
385 390 395 400
Pro Asn Gln Ser Tyr Ser Pro Asn Pro Ser Asp Gly Asn Pro Tyr Gln
405 410 415
Asn Gln Asn Leu Pro Gly Arg Asp Met Pro Pro Pro Asn Tyr
420 425 430
<210>3
<211>403
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>3
Met Ala Ser Arg Ser Leu Leu Asn Ala Phe Lys Arg Ser Cys Asn Ser
1 5 10 15
Leu Ser Leu Ser Thr Thr Pro Ser Thr Ser Ser Leu His Arg Leu Arg
20 25 30
Ser Leu Ser Ala Val Ala Ala Asn Leu Arg Asn Val Ser Pro Ala Pro
35 40 45
Thr Thr Phe Ser Arg Gly Phe Ala Thr Arg Gln Thr Ser Ser Ser Leu
50 55 60
Asn Asp Pro Asn Pro Asn Trp Ser Asn Arg Pro Pro Lys Glu Thr Ile
65 70 75 80
Leu Leu Asp Gly Cys Asp Phe Glu His Trp Leu Val Val Met Glu Lys
85 90 95
Pro Glu Gly Asp Pro Thr Arg Asp Glu Ile Ile Asp Ser Tyr Ile Lys
100 105 110
Thr Leu Ala Thr Ile Val Gly Ser Glu Glu Asp Ala Arg Met Lys Ile
115 120 125
Tyr Ser Val Ser Thr Arg His Tyr Phe Ala Phe Gly Ala Leu Val Ser
130 135 140
Glu Glu Leu Ser Tyr Lys Leu Lys Glu Leu Pro Lys Val Arg Trp Val
145 150 155 160
Leu Pro Asp Ser Tyr Leu Asp Val Arg Asn Lys Asp Tyr Gly Gly Glu
165 170 175
Pro Phe Ile Asn Gly Gln Ala Val Pro Tyr Asp Pro Lys Tyr His Glu
180 185 190
Glu Trp Val Arg Asn Asn Ala Arg Ala Asn Glu Arg Asn Arg Arg Asn
195 200 205
Asp Arg Pro Arg Asn Phe Asp Arg Ser Arg Asn Phe Glu Arg Arg Arg
210 215 220
Glu Met Gln Asn Gln Pro Ser Gln Asn Pro Gly Ser Asn Met Ala Ala
225 230 235 240
Gly Gly Pro Pro Asn Met Gly Gly Met Gln Gln Gln Gln Gly Met Gly
245 250 255
Gly Pro Ser His Thr Gly Gly Met His Gln Gln Gln Gly Tyr Gly Gly
260 265 270
Pro Pro Pro His Ala Gly Gly Met Gln Gln Pro Asp Met Arg Gly Pro
275 280 285
Pro Met Gln Gln Gln Gly Tyr Gly Gly Pro Pro Pro His Ala Gly Gly
290 295 300
Met Gln Gln Pro Ser Met Gly Gly Pro Pro Pro Arg Gly Gly Gly Met
305 310 315 320
Gln Gln Pro Asn Met Pro Pro Asn Ile Gly Gly Val Pro Pro Asn Met
325 330 335
Gly Gly Val Pro Pro Asn Asn Tyr Gly Val Pro Asn Asn Ala Pro Ser
340 345 350
Phe Gln Tyr Asn Gly Gly Gly Pro Asn Asn Gly Gly Thr Pro Tyr Gln
355 360 365
Thr Gly Pro Gly Pro Asn Gln Ser Tyr Ala Pro Asn Thr Ser Asp Gly
370 375 380
Asn Ser Tyr Gln Asn Pro Asn Met Pro Gly Arg Asp Met Ser Asn Pro
385 390 395 400
Asn Tyr Arg
<210>4
<211>411
<212>PRT
<213> Artificial Sequence (Artificial Sequence)
<400>4
Met Ala Thr Arg Tyr Leu Thr Arg Ser Leu Leu Thr Thr Leu Ser Arg
1 5 10 15
Ser Tyr Ile Pro Leu Ser Leu Ser Thr Pro Leu Pro Ser Ser Phe Ser
20 25 30
Leu Leu Arg Phe Arg Pro Leu Ile Ala Val Ala Ala Asn Leu Arg Thr
35 40 45
Phe Thr Pro Ala Thr Leu Thr Ala Ala Thr Ser Leu Arg Gly Phe Ala
50 55 60
Thr Arg Gln Thr Ser Ser Ser Leu Asn Asp Pro Asn Pro Asn Trp Ser
65 70 75 80
Asn Arg Pro Pro Lys Glu Thr Ile Leu Leu Asp Gly Cys Asp Phe Glu
85 90 95
His Trp Leu Val Val Met Glu Lys Pro Glu Gly Asp Pro Thr Arg Asp
100 105 110
Glu Ile Ile Asp Ser Tyr Ile Lys Thr Leu Ala Thr Ile Val Gly Ser
115 120 125
Glu Glu Glu Ala Arg Met Lys Ile Tyr Ser Val Ser Thr Arg His Tyr
130 135 140
Tyr Ala Phe Gly Ala Leu Val Ser Glu Glu Leu Ser Tyr Lys Leu Lys
145 150 155 160
Glu Leu Pro Lys Val Arg Trp Val Leu Pro Asp Ser Tyr Leu Asp Val
165 170 175
Lys Asn Lys Asp Tyr Gly Gly Glu Pro Phe Ile Asn Gly Gln Ala Val
180 185 190
Pro Tyr Asp Pro Lys Tyr His Glu Glu Trp Val Arg Asn Asn Ala Arg
195 200 205
Ala Asn Glu Arg Asn Arg Arg Asn Asp Arg Pro Arg Asn Phe Asp Arg
210 215 220
Ser Arg Asn Phe Glu Arg Arg Arg Glu Met Gln Asn Pro Gly Ser Asn
225 230 235 240
Met Gly Gly Gly Pro Pro Asn Met Arg Asn Ala Pro Pro Pro Asn Met
245 250 255
Gly Gly Met Gln Gln Gln Gln Pro Asn Met Ser Gly Val Gln Gln Gln
260 265 270
Gln Pro Asn Met Gly Gly Met Gln Gln Gln Gln Pro Asn Met Gly Gly
275 280 285
Met Gln Gln Gln Gln Pro Asn Met Gly Gly Met His Gln Pro Gly Met
290 295 300
Gly Gly Pro Pro Asn Met Gln Pro Asn Met Gly Gly Ala Pro Pro Asn
305 310 315 320
Tyr Gly Gly Gly Pro Pro Arg Asn Tyr Gly Gly Ala Pro Pro Asn Asn
325 330 335
Tyr Gly Gly Ala Pro Asn Asn Gln Tyr Asn Gly Gly Pro Asn Asn Gly
340 345 350
Gly Met Pro Tyr Gln Thr Gly Pro Gly Pro Asn Gln Asn Tyr Ala Ser
355 360 365
Asn Thr Ser Gly Gly Asn His Tyr Gln Asn Pro Asn Met Pro Ser Pro
370 375 380
Ser Gln Asn Tyr Ala Pro Asn Thr Ser Gly Gly Asn Pro Leu Gln Asn
385 390 395 400
Gln Asn Met Pro Gly Arg Asp Pro Asn Tyr Gln
405 410
Claims (2)
1. An application of an immune negative regulatory factor NbMORF8 gene in plant phytophthora resistance is characterized in that the NbMORF8 gene negatively regulates the resistance of plants to phytophthora by interfering a plant salicylic acid immune signal pathway and active oxygen burst;
the nucleotide sequence of the NbMORF8 gene is shown as SEQ ID No: 1 or a sequence corresponding to SEQ ID No: 1, and homologous sequences with homology of more than 70%.
2. The application of an immune negative regulatory factor NbMORF8 gene in plant phytophthora resistance is characterized in that the expression level of protein encoded by the NbMORF8 gene is reduced through genetic engineering, so that the resistance of plants to phytophthora can be enhanced;
the amino acid sequence of the protein is shown as SEQ ID No: 2 or a sequence similar to SEQ ID No: 2, and homologous sequences with more than 70% of sequence homology.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010396024.9A CN111424042B (en) | 2020-05-12 | 2020-05-12 | Application of immune negative regulatory factor NbMORF8 gene and protein thereof in phytophthora resistance of plants |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010396024.9A CN111424042B (en) | 2020-05-12 | 2020-05-12 | Application of immune negative regulatory factor NbMORF8 gene and protein thereof in phytophthora resistance of plants |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111424042A true CN111424042A (en) | 2020-07-17 |
| CN111424042B CN111424042B (en) | 2021-11-09 |
Family
ID=71552592
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010396024.9A Active CN111424042B (en) | 2020-05-12 | 2020-05-12 | Application of immune negative regulatory factor NbMORF8 gene and protein thereof in phytophthora resistance of plants |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111424042B (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6858707B1 (en) * | 1998-10-05 | 2005-02-22 | Eden Bioscience Corporation | Hypersensitive response elicitor fragments which are active but do not elicit a hypersensitive response |
| CN107699579A (en) * | 2017-11-03 | 2018-02-16 | 南京农业大学 | A kind of gene for improving disease resistance of plant and its application |
| CN110468142A (en) * | 2019-09-27 | 2019-11-19 | 西北农林科技大学 | Negative regulatory factor AtRTP5 gene and its application on anti-plant phytophthora |
| CN110938637A (en) * | 2019-11-26 | 2020-03-31 | 西北农林科技大学 | Homologous gene of phytophthora resistance negative regulatory factor StMKK1 and application thereof |
-
2020
- 2020-05-12 CN CN202010396024.9A patent/CN111424042B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6858707B1 (en) * | 1998-10-05 | 2005-02-22 | Eden Bioscience Corporation | Hypersensitive response elicitor fragments which are active but do not elicit a hypersensitive response |
| CN107699579A (en) * | 2017-11-03 | 2018-02-16 | 南京农业大学 | A kind of gene for improving disease resistance of plant and its application |
| CN110468142A (en) * | 2019-09-27 | 2019-11-19 | 西北农林科技大学 | Negative regulatory factor AtRTP5 gene and its application on anti-plant phytophthora |
| CN110938637A (en) * | 2019-11-26 | 2020-03-31 | 西北农林科技大学 | Homologous gene of phytophthora resistance negative regulatory factor StMKK1 and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| GENBANK: "NCBI Reference Sequence: XM_016608536.1", 《GENBANK》 * |
| 张美祥等: "效应因子PsCRN77基因在本氏烟中的表达降低其对寄生疫霉的抗性", 《植物病理学报》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111424042B (en) | 2021-11-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111235164B (en) | Negative regulatory factor AtPPR1 gene for phytophthora resistance and application of homologous gene thereof | |
| CN109705201B (en) | Cotton verticillium wilt resistance related gene GhABC and coding protein and application thereof | |
| CN110938637B (en) | Homologous gene of phytophthora resistance negative regulatory factor StMKK1 and application thereof | |
| CN114164217B (en) | Application of rice OsSTE24 gene in improving rice blast fungus resistance | |
| US20230272411A1 (en) | Heat-shock related gene zmhsf11 and application of zmhsf11 in regulating heat-resistence of plant | |
| CN112662692A (en) | Peanut cysteine protease coding gene AhRD21A, and expression vector and application thereof | |
| Czajkowski et al. | Genome-wide identification of Dickeya solani transcriptional units up-regulated in response to plant tissues from a crop-host Solanum tuberosum and a weed-host Solanum dulcamara | |
| CN109706132B (en) | Cotton verticillium wilt resistance-related protein GhMAPK13 as well as coding gene and application thereof | |
| CN113337536B (en) | Application of RS2Z32 gene as plant immune negative regulatory factor in improving crop resistance | |
| CN108588041B (en) | Gossypium barbadense cytochrome P450 gene, and coding protein and application thereof | |
| CN116286724B (en) | Lectin receptor protein TaLecRLK2 and encoding gene and application thereof | |
| CN111424042B (en) | Application of immune negative regulatory factor NbMORF8 gene and protein thereof in phytophthora resistance of plants | |
| CN112813083A (en) | Application of OsCIPK31 gene and coding protein in regulation and control of rice sheath blight disease resistance | |
| CN109207485B (en) | Application of OsAPS1 gene in improving disease resistance of rice | |
| CN111704659A (en) | Root-knot nematode RALF protein, coding gene and application thereof | |
| CN111187779A (en) | Disease-resistant gene OsRLR1, transcription factor OsWRKY19 and application in breeding of rice resistant to bacterial blight | |
| CN115976052A (en) | Wheat stem basal rot resistance gene TaHSP18.6, expression product, recombinant vector and application thereof | |
| CN114807187A (en) | Ural chart wheat receptor protein kinase gene TuRLK1 and application thereof | |
| CN107287169A (en) | A kind of eggplant cytochrome P450 gene StCYP84A and its preparation method and application | |
| CN111088239B (en) | Corn high-temperature response protein kinase ZmCDPK7, and coding gene and application thereof | |
| CN113046366A (en) | StCAD7 gene and application of protein encoded by gene as negative regulator in improving potato late blight resistance | |
| CN111424043B (en) | Positive regulatory factor NbPRO19C55-1 gene and application of protein thereof in phytophthora parasitica resistance | |
| CN111850012A (en) | Soybean bacillus subtilis protease gene GmSub and application thereof | |
| Hucklenbroich et al. | Rhizobiales commensal bacteria promote Arabidopsis thaliana root growth via host sulfated peptide pathway | |
| CN116751769B (en) | Pc-CL protein of Caesalpinia aphelenchoides, coding gene and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |