CN110301454A - A kind of application of rape small peptide BnPEP5 - Google Patents
A kind of application of rape small peptide BnPEP5 Download PDFInfo
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- CN110301454A CN110301454A CN201910518850.3A CN201910518850A CN110301454A CN 110301454 A CN110301454 A CN 110301454A CN 201910518850 A CN201910518850 A CN 201910518850A CN 110301454 A CN110301454 A CN 110301454A
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- bnpep5
- small peptide
- rape
- resistance
- sclerotinite
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/44—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a nitrogen atom attached to the same carbon skeleton by a single or double bond, this nitrogen atom not being a member of a derivative or of a thio analogue of a carboxylic group, e.g. amino-carboxylic acids
- A01N37/46—N-acyl derivatives
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N47/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom not being member of a ring and having no bond to a carbon or hydrogen atom, e.g. derivatives of carbonic acid
- A01N47/40—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom not being member of a ring and having no bond to a carbon or hydrogen atom, e.g. derivatives of carbonic acid the carbon atom having a double or triple bond to nitrogen, e.g. cyanates, cyanamides
- A01N47/42—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom not being member of a ring and having no bond to a carbon or hydrogen atom, e.g. derivatives of carbonic acid the carbon atom having a double or triple bond to nitrogen, e.g. cyanates, cyanamides containing —N=CX2 groups, e.g. isothiourea
- A01N47/44—Guanidine; Derivatives thereof
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- 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
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Environmental Sciences (AREA)
- Agronomy & Crop Science (AREA)
- Dentistry (AREA)
- Plant Pathology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Pest Control & Pesticides (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Gastroenterology & Hepatology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
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Abstract
One rape of present invention offer (Brassica napus) small peptide BnPEP5 excitation crucifer generate to sclerotinite (Sclerotinia sclerotiorum) resistance and application.The rape small peptide BnPEP5 that the present invention applies is made of 27 amino acid, is the C-terminal part of full-length proteins.The artificial synthesized small peptide BnPEP5 of first passage of the present invention, illustrates the small peptide to the excitation of sclerotinite resistance, and provide the small peptide by infiltration and spraying treatment crucifer rape and leaf mustard (Brassica juncea) it is excited to generate to sclerotinite resistance, to the application of prevention and control rape and leaf mustard sclerotiniose.BnPEP5 small peptide provided by the invention is the product of plant itself synthesis, using BnPEP5 small peptide excite it is immune to prevent and treat sclerotiniose have many advantages, such as natural green, it is free from environmental pollution, to person poultry harmless, pathogen will not be induced to develop drug resistance.In addition, BnPEP5 small peptide is easy for workers to synthesize, simplicity is obtained, disease-resistant excitation is strong, and required concentration is low, is a kind of novel ecological and environmentally friendly crop disease anti-control agent.
Description
Technical field
The invention belongs to field of biotechnology, it is related to the immune stimulating function of rape small peptide BnPEP5 a kind of and its in sclerotiniose
Application in prevention and control.
Background technique
1, plant immune excitation technique
Plant immune is defense response of the plant by Receptor recognition pathogen molecule to activation.Plant immune system has multiple
Level, wherein the first level is transmembrane molecule pattern recognition receptors (the pattern recognition of surface of Plant callus cell
Receptors, PRRs) identification is originated from pathogen and the conservative molecular pattern of plant itself generates immune, referred to as mode touching
Hair property is immune (pattern-triggered immunity, PTI).Generally, plant can perceive pathogen and infect etc. and cause
The destruction to plant cell integrality, and generate injury associated molecular pattern (damage-associated molecular
Patterns, DAMPs), these DAMPs excite plant to generate PTI immune response by PRR Receptor recognition, including active oxygen spouts
Hair, the activation of kinase cascade, expression of defense response genes etc., finally show the resistance to pathogen.PTI prevent it is non-adaptive
The non―technological factors (nonhost resistance) and limitation adaptibility pathogen of property microbial infection plant infect susceptible post
It plays an important role in the basal resistance (basal resistance) of main plant.The PTI of DAMP excitation is immunized with wide spectrum, by force
The advantages that strong, stable.Further, since DAMP is the natural products of plant itself, therefore is excited using DAMP and be immunized to reach
The purpose of controlling disease is a kind of environmental ecology friendly green disease prevention and control new method.
2, sclerotiniose Control Technology
Sclerotiniose by sclerotinite (Sclerotinia sclerotiorum) infect caused by.Sclerotinite is downright bad auxotype
(necrotroph) disease fungus, it is the Major Diseases of oil crops and vegetable crop that host's defence is extremely wide.It causes every year huge
Big economic loss.Due to lacking highly resistance kind, chemical prevention is still important means.Due to some pesticides there are ecological pollution,
Toxicity to man and livestock and easy the problems such as making pathogen develop drug resistance, novel green prevention and treatment preparation are urgently researched and developed.
Summary of the invention
The object of the present invention is to provide a kind of rape (Brassica napus) small peptide BnPEP5 excitation crucifer
Generate to sclerotinite (Sclerotinia sclerotiorum) resistance application.The rape small peptide BnPEP5 sequence that the present invention applies
Column are made of 27 amino acid altogether as shown in SEQ ID:1, are the C-terminal parts of full-length proteins (sequence is as shown in SEQ ID:2).
Before making the present invention, the function of small peptide BnPEP5 does not have any open report.The artificial synthesized small peptide of first passage of the present invention
BnPEP5 illustrates the small peptide to the excitation of sclerotinite resistance, and provides the small peptide and swashed by processing crucifer
It is sent out to generate to sclerotinite resistance, to the application of prevention and control sclerotiniose.It is realized by following steps:
(1) the artificial synthesized or biological expression of BnPEP5 small peptide obtains
Professional Peptide systhesis company can be entrusted to synthesize.Small peptide BnPEP5 can also be obtained by biological expression: BnPEP5 is corresponding
Nucleotide sequence is cloned into expression vector, conversion to eukaryon or prokaryotic micro-organisms, microorganism expands numerous, extraction and purifying purpose is short
Peptide.
(2) plant treatment of BnPEP5 small peptide
With the small peptide BnPEP5 solution immersion or spraying treatment plant leaf blade of suitable concentration, plant immune is excited.Infiltration and spray
Small peptide BnPEP5 concentration of aqueous solution needed for mist and treatment scale are different.BnPEP5 concentration needed for infiltration is handled is lower, generally exists
Within the scope of 100nM ~ 200nM, it is suitble to small-scale plant treatment, and BnPEP5 concentration needed for spraying treatment is higher, generally in 200nM
Within the scope of ~ 1000nM, it is suitble to extensive plant treatment.
(3) BnPEP5 small peptide excitation plant generates the resistance to sclerotinite
Plant is excited to generate the immune response to sclerotinite rapidly after small peptide BnPEP5 processing.It can be examined by pathogen inoculation analysis
Survey the intensity of excitation.Plant, which generates, to be shown as small peptide BnPEP5 processing to the immune response of sclerotinite and makes to generate after being inoculated with
Downright bad scab is than water control treatment Shi Geng little.In terms of mechanism of action angle, BnPEP5 induces the excitation of the immune response with it
Rapid a large amount of accumulation of active oxygen are related.
Advantages of the present invention: (1) BnPEP5 small peptide provided by the invention is the product of plant itself synthesis, utilizes BnPEP5
Small peptide excitation it is immune to prevent and treat sclerotiniose with natural green, it is free from environmental pollution, to person poultry harmless, cause of disease produce will not be induced
The advantages that raw drug resistance.(2) it is latest theories research achievement that DAMP, which excites plant immune,.BnPEP5 is answered as a kind of rape DAMP
There is the model of sufficient theoretical foundation and theoretical direction practical application for crop in cruciferae immune stimulating and disease prevention and control
Example.(3) BnPEP5 small peptide only has 27 amino acid, is easy for workers to synthesize, and obtains simple and easy to do.(4) BnPEP5 small peptide is disease-resistant
Property excitation is strong, and required concentration is low, therefore cost is relatively low for disease control.
Detailed description of the invention
Fig. 1 show BnPEP5 small peptide solution immersion or spraying rape (Brassica napus) blade excitation rape generation
To the resistance of sclerotinite.Rapeseed plants are infiltrated by sterile one-shot injector with the BnPEP5 small peptide aqueous solution that concentration is 100 nM
Blade (b), or with BnPEP5 small peptide aqueous solution that concentration is 200 nM by gun spraying to rape leaf (d), with core after 1d
Cup fungi inoculated by hypha block blade is illustrated as the Lesion size of 2d after inoculation.The result shows that compared with aseptic double-distilled water processing control
(a, the infiltration of aseptic double-distilled water syringe；C, aseptic double-distilled water gun spraying), the scab that BnPEP5 processing blade is formed may be significantly smaller.
Show that BnPEP5 processing excitation rape produces the resistance to sclerotinite.
Fig. 2 shows that BnPEP5 small peptide excitation rape leaf accumulates rapidly active oxygen.Concentration is the BnPEP5 small peptide of 100 nM
Rape leaf sequin is induced rapidly largely accumulates active oxygen.Sterile water process is as control.Active oxygen, which uses, is based on luminol
Method detected by chemiluminescence detector.
Fig. 3 show BnPEP5 small peptide solution immersion or spraying leaf mustard (Brassica juncea) blade excitation leaf mustard production
The raw resistance to sclerotinite.Leaf mustard is infiltrated by sterile one-shot injector with the BnPEP5 small peptide aqueous solution that concentration is 100 nM to plant
Strain blade (b), or with BnPEP5 small peptide aqueous solution that concentration is 200 nM by gun spraying to mustard leaf (d), after 1d with
Nuclear fungal hyphae block is inoculated with blade, is illustrated as the Lesion size of 2d after inoculation.The result shows that with aseptic double-distilled water processing to photograph
Than (a, the infiltration of aseptic double-distilled water syringe；C, aseptic double-distilled water gun spraying), BnPEP5 handles scab that blade is formed obviously more
It is small.Show that BnPEP5 processing excitation leaf mustard produces the resistance to sclerotinite.
Specific embodiment
The present invention is further described in conjunction with the accompanying drawings and embodiments.
1 BnPEP5 small peptide of embodiment rape (Brassica napus) application in anti-sclerotiniose
The embodiment of the present invention utilizes artificial synthesized BnPEP5 small peptide, handles rape by infiltration or spray-on process, excites its generation
To the resistance of sclerotinite, for the control of sclerotinia sclerotiorum.Key step includes:
1) BnPEP5 small peptide is artificial synthesized
BnPEP5 short peptide sequence provided by the invention is made of 27 amino acid altogether as shown in SEQ ID:1, is full-length proteins (sequence
Arrange as shown in SEQ ID:2) C-terminal part.Before making the present invention, the function of small peptide BnPEP5 does not have any open report.
The present embodiment entrusts professional Peptide systhesis company (Qiang Yao Biotechnology Co., Ltd) to synthesize BnPEP5 small peptide, is used for
The analysis and application of rape disease-resistant excitation and sclerotiniose prevention and control function.
2) processing of the BnPEP5 small peptide to rape
Infiltration processing: rapeseed plants leaf is infiltrated by sterile one-shot injector with the BnPEP5 small peptide aqueous solution that concentration is 100 nM
Piece；
Spraying treatment: gun spraying to rapeseed plants blade is passed through with the BnPEP5 small peptide aqueous solution that concentration is 200 nM.
3) BnPEP5 small peptide excitation rape generates the resistance to sclerotinite
BnPEP5 small peptide solution immersion or spraying rape leaf can excite rape to generate the resistance to sclerotinite.Sclerotinite connects
The results show that after with BnPEP5 small peptide (100 nM) infiltration or spraying (200 nM) processing 1d, nuclear fungal hyphae block is connect for kind analysis
The scab of kind blade is significantly less than aseptic double-distilled water processing control (Fig. 1).Show that BnPEP5 infiltration or spraying treatment can be rapidly
Rape is excited to generate the resistance to sclerotinite.
Active oxygen quantitative detecting analysis is the results show that BnPEP5 small peptide (100 nM) induction rape leaf disk is strongly long-pending rapidly
Tired active oxygen (Fig. 2).Show that BnPEP5 small peptide excites rape to generate the resistance to sclerotinite by inducing oxidative burst.
2 BnPEP5 small peptide of embodiment leaf mustard (Brassica juncea) application in anti-sclerotiniose
The embodiment of the present invention utilizes artificial synthesized BnPEP5 small peptide, handles leaf mustard by infiltration or spray-on process, excites its generation
To the resistance of sclerotinite, for the control of leaf mustard sclerotiniose.Key step includes:
1) BnPEP5 small peptide is artificial synthesized
BnPEP5 short peptide sequence provided by the invention is made of 27 amino acid altogether as shown in SEQ ID:1, is full-length proteins (sequence
Arrange as shown in SEQ ID:2) C-terminal part.The present embodiment entrusts professional Peptide systhesis company (Qiang Yao Biotechnology Co., Ltd)
BnPEP5 small peptide is synthesized, the analysis and application for leaf mustard disease-resistant excitation and sclerotiniose prevention and control function.
2) processing of the BnPEP5 small peptide to leaf mustard
The plant treatment mode of BnPEP5 small peptide is including infiltration and by spraying.Small peptide BnPEP5 concentration of aqueous solution and place needed for the two
Reason scale is different.BnPEP5 concentration needed for infiltration is handled is lower, is suitble to small-scale plant treatment, and BnPEP5 needed for spraying treatment
Concentration is higher, is suitble to extensive plant treatment.
Infiltration processing: leaf mustard is infiltrated by sterile one-shot injector with the BnPEP5 small peptide aqueous solution that concentration is 100 nM and is planted
Strain blade；
Spraying treatment: gun spraying to leaf mustard plant leaf is passed through with the BnPEP5 small peptide aqueous solution that concentration is 200 nM.
3) BnPEP5 small peptide excitation leaf mustard generates the resistance to sclerotinite
BnPEP5 small peptide solution immersion or spraying mustard leaf can excite leaf mustard to generate the resistance to sclerotinite.Sclerotinite connects
The results show that after with BnPEP5 small peptide (100 nM) infiltration or spraying (200 nM) processing 1d, nuclear fungal hyphae block is connect for kind analysis
The scab of kind blade is significantly less than aseptic double-distilled water processing control (Fig. 3).Show that BnPEP5 infiltration or spraying treatment can be rapidly
Leaf mustard is excited to generate the resistance to sclerotinite.
By above embodiments, one skilled in the art may determine that small peptide BnPEP5 can effectively excite Cruciferae
Plant generates the prevention and control to sclerotinite (Sclerotinia sclerotiorum) resistance, applied to sclerotiniose.
Sequence table
<110>Zhejiang University
<120>a kind of application of rape small peptide BnPEP5
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 27
<212> PRT
<213>cabbage type rape (Brassica napus)
<400> 1
Ser Leu Asn Val Ser Ser Lys Leu Thr Arg Lys Leu Pro Val Ser Ser
1 5 10 15
Gly Lys Arg Gly Gly Val Asn Asn His Asp Met
20 25
<210> 2
<211> 90
<212> PRT
<213>cabbage type rape (Brassica napus)
<400> 2
Met Gln Lys Glu Ser Asp Lys Lys Arg His Arg Cys Lys Leu Ile Thr
1 5 10 15
Gln Ser Val Met Ala Phe Leu Asp Cys Leu Asn Val Leu Asn Gln Lys
20 25 30
Lys Lys Lys Lys Lys Lys Leu Ser Glu Tyr Met Val Lys Ala Lys Ala
35 40 45
Arg Asn Glu Ala Glu Val Ala Ser Ser Val Glu Thr Ser Asp Arg Ser
50 55 60
Leu Asn Val Ser Ser Lys Leu Thr Arg Lys Leu Pro Val Ser Ser Gly
65 70 75 80
Lys Arg Gly Gly Val Asn Asn His Asp Met
85 90
Claims (3)
1. a kind of rape (Brassica napus) small peptide BnPEP5 excitation crop in cruciferae generate to sclerotinite
(Sclerotinia sclerotiorum) resistance application, rape small peptide BnPEP5 amino acid sequence such as SEQ ID:1 institute
Show.
2. application according to claim 1, which is characterized in that the crop in cruciferae includes rape, leaf mustard
(Brassica juncea).
3. application according to claim 1, which is characterized in that the application are as follows: by small peptide BnPEP5 solution immersion or spray
Mist handles crop in cruciferae blade, excites plant immune.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910518850.3A CN110301454B (en) | 2019-06-16 | 2019-06-16 | Application of rape short peptide BnPEP5 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910518850.3A CN110301454B (en) | 2019-06-16 | 2019-06-16 | Application of rape short peptide BnPEP5 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110301454A true CN110301454A (en) | 2019-10-08 |
| CN110301454B CN110301454B (en) | 2020-10-16 |
Family
ID=68077210
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910518850.3A Active CN110301454B (en) | 2019-06-16 | 2019-06-16 | Application of rape short peptide BnPEP5 |
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| Country | Link |
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| CN (1) | CN110301454B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114317554A (en) * | 2021-12-29 | 2022-04-12 | 浙江大学 | Application of rape gene BnPEP5 in sclerotinia sclerotiorum prevention and control |
| CN114375963A (en) * | 2021-12-29 | 2022-04-22 | 浙江大学 | Immune excitation function application of rape short peptide BnPEP6 |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1251371A (en) * | 1999-08-10 | 2000-04-26 | 南京农业大学 | Exciter for inducing allergic reaction of plant and preparation method and use thereof |
| US20090300802A1 (en) * | 2005-01-26 | 2009-12-03 | Washington State University | Plant defense signal peptides |
| CN104530204A (en) * | 2014-12-19 | 2015-04-22 | 中国农业科学院油料作物研究所 | Rape antibacterial peptide BnPRP1 and application thereof |
| CN108883148A (en) * | 2016-04-06 | 2018-11-23 | 植物保健公司 | The peptide and application thereof in allergic reaction exciton source |
-
2019
- 2019-06-16 CN CN201910518850.3A patent/CN110301454B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1251371A (en) * | 1999-08-10 | 2000-04-26 | 南京农业大学 | Exciter for inducing allergic reaction of plant and preparation method and use thereof |
| US20090300802A1 (en) * | 2005-01-26 | 2009-12-03 | Washington State University | Plant defense signal peptides |
| CN104530204A (en) * | 2014-12-19 | 2015-04-22 | 中国农业科学院油料作物研究所 | Rape antibacterial peptide BnPRP1 and application thereof |
| CN108883148A (en) * | 2016-04-06 | 2018-11-23 | 植物保健公司 | The peptide and application thereof in allergic reaction exciton source |
Non-Patent Citations (3)
| Title |
|---|
| ABD57458.1: "At5g09990 [Arabidopsis thaliana]", 《GENBANK数据库》 * |
| ALISA HUFFAKER 等: "An endogenous peptide signal in Arabidopsis activates components of the innate immune response", 《PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA》 * |
| CDY21620.1: "BnaC09g46720D[Brassica napus]", 《GENBANK数据库》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114317554A (en) * | 2021-12-29 | 2022-04-12 | 浙江大学 | Application of rape gene BnPEP5 in sclerotinia sclerotiorum prevention and control |
| CN114375963A (en) * | 2021-12-29 | 2022-04-22 | 浙江大学 | Immune excitation function application of rape short peptide BnPEP6 |
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| CN110301454B (en) | 2020-10-16 |
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