CN110278959A - A kind of application of rape small peptide BnPEP4 - Google Patents
A kind of application of rape small peptide BnPEP4 Download PDFInfo
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- CN110278959A CN110278959A CN201910518849.0A CN201910518849A CN110278959A CN 110278959 A CN110278959 A CN 110278959A CN 201910518849 A CN201910518849 A CN 201910518849A CN 110278959 A CN110278959 A CN 110278959A
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- bnpep4
- 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
- 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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- Life Sciences & Earth Sciences (AREA)
- Agronomy & Crop Science (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Dentistry (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
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- Peptides Or Proteins (AREA)
Abstract
One rape of present invention offer (Brassica napus) small peptide BnPEP4 excitation crucifer generate to sclerotinite (Sclerotinia sclerotiorum) resistance application.The rape small peptide BnPEP4 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 BnPEP4 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.BnPEP4 small peptide provided by the invention is the product of plant itself synthesis, using BnPEP4 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, BnPEP4 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, be related to application of the rape small peptide BnPEP4 in immune stimulating, it can be achieved that
It carries out prevention and control to sclerotiniose.
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 rape (Brassica napus) small peptide BnPEP4 excitation crucifer
Generate to sclerotinite (Sclerotinia sclerotiorum) resistance application.The rape small peptide BnPEP4 sequence that the present invention applies
As shown in SEQ ID:1, which is made of 27 amino acid column altogether, is the C of full-length proteins (sequence is as shown in SEQ ID:2)
End part.Before making the present invention, the function of small peptide BnPEP4 does not have any open report.First passage of the present invention is artificial synthesized short
Peptide BnPEP4 illustrates the small peptide to the excitation of sclerotinite resistance, and provides the small peptide and pass through processing crucifer
It is excited 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 BnPEP4 small peptide obtains
Professional Peptide systhesis company can be entrusted to synthesize.Small peptide BnPEP4 can also be obtained by biological expression: BnPEP4 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 BnPEP4 small peptide
With the small peptide BnPEP4 solution immersion or spraying treatment plant leaf blade of suitable concentration, plant immune is excited.Infiltration and spray
Small peptide BnPEP4 concentration of aqueous solution needed for mist the two and treatment scale are different.BnPEP4 concentration needed for infiltration is handled is lower, and one
As in 100nM ~ 200nM range, be suitble to small-scale plant treatment, and BnPEP4 concentration needed for spraying treatment is higher, generally exists
200nM ~ 1000nM range is suitble to extensive plant treatment.
(3) BnPEP4 small peptide excitation plant generates the resistance to sclerotinite
Plant is excited to generate the immune response to sclerotinite rapidly after small peptide BnPEP4 processing.It can be examined by pathogen inoculation analysis
Survey the intensity of excitation.Plant, which generates, to be shown as small peptide BnPEP4 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, BnPEP4 induces the excitation of the immune response with it
Rapid a large amount of accumulation of active oxygen are related.
Beneficial effects of the present invention: (1) BnPEP4 small peptide provided by the invention is the product of plant itself synthesis, is utilized
The excitation of BnPEP4 small peptide it is immune to prevent and treat sclerotiniose with natural green, it is free from environmental pollution, to person poultry harmless, disease will not be induced
The advantages that original develops drug resistance.(2) it is latest theories research achievement that DAMP, which excites plant immune,.BnPEP4 is as a kind of rape
DAMP, which is applied to crop in cruciferae immune stimulating and disease prevention and control, sufficient theoretical foundation and theoretical direction practical application
Example.(3) BnPEP4 small peptide only has 27 amino acid, is easy for workers to synthesize, and obtains simple and easy to do.(4) BnPEP4 small peptide
Disease resistance excitation is strong, and required concentration is low, therefore cost is relatively low for disease control.
Detailed description of the invention
Fig. 1 show BnPEP4 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 BnPEP4 small peptide aqueous solution that concentration is 100 nM
Blade (b), or with BnPEP4 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 BnPEP4 processing blade is formed may be significantly smaller.
Show that BnPEP4 processing excitation rape produces the resistance to sclerotinite.
Fig. 2 shows that BnPEP4 small peptide excitation rape leaf accumulates rapidly active oxygen.Concentration is the BnPEP4 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 BnPEP4 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 BnPEP4 small peptide aqueous solution that concentration is 100 nM to plant
Strain blade (b), or with BnPEP4 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), BnPEP4 handles scab that blade is formed obviously more
It is small.Show that BnPEP4 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 BnPEP4 small peptide of embodiment rape (Brassica napus) application in anti-sclerotiniose
The embodiment of the present invention utilizes artificial synthesized BnPEP4 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) BnPEP4 small peptide is artificial synthesized
BnPEP4 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 BnPEP4 does not have any open report.
The present embodiment entrusts professional Peptide systhesis company (Qiang Yao Biotechnology Co., Ltd) to synthesize BnPEP4 small peptide, is used for
The analysis and application of rape disease-resistant excitation and sclerotiniose prevention and control function.
2) processing of the BnPEP4 small peptide to rape
Infiltration processing: rapeseed plants leaf is infiltrated by sterile one-shot injector with the BnPEP4 small peptide aqueous solution that concentration is 100 nM
Piece；
Spraying treatment: gun spraying to rapeseed plants blade is passed through with the BnPEP4 small peptide aqueous solution that concentration is 200 nM.
3) BnPEP4 small peptide excitation rape generates the resistance to sclerotinite
BnPEP4 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 BnPEP4 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 BnPEP4 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 BnPEP4 small peptide (100 nM) induction rape leaf disk is strongly long-pending rapidly
Tired active oxygen (Fig. 2).Show that BnPEP4 small peptide excites rape to generate the resistance to sclerotinite by inducing oxidative burst.
2 BnPEP4 small peptide of embodiment leaf mustard (Brassica juncea) application in anti-sclerotiniose
The embodiment of the present invention utilizes artificial synthesized BnPEP4 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) BnPEP4 small peptide is artificial synthesized
BnPEP4 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)
BnPEP4 small peptide is synthesized, the analysis and application for leaf mustard disease-resistant excitation and sclerotiniose prevention and control function.
2) processing of the BnPEP4 small peptide to leaf mustard
The plant treatment mode of BnPEP4 small peptide is including infiltration and by spraying.
Infiltration processing: leaf mustard is infiltrated by sterile one-shot injector with the BnPEP4 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 BnPEP4 small peptide aqueous solution that concentration is 200 nM.
3) BnPEP4 small peptide excitation leaf mustard generates the resistance to sclerotinite
BnPEP4 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 BnPEP4 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 BnPEP4 infiltration or spraying treatment can be rapidly
Leaf mustard is excited to generate the resistance to sclerotinite.
From both examples above, one skilled in the art may determine that this BnPEP4 small peptide can effectively excite ten
Zi Hua section 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 BnPEP4
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 27
<212> PRT
<213>cabbage type rape (Brassica napus)
<400> 1
Arg Gly Leu Pro Pro Lys Lys Leu Gln Gln Lys Pro Arg Gly Ser Ser
1 5 10 15
Gly Lys Pro Gly Ser Ile Asn Lys Lys Pro Arg
20 25
<210> 2
<211> 86
<212> PRT
<213>cabbage type rape (Brassica napus)
<400> 2
Met Gln Arg Asp Gly Arg Arg Glu Gly Val Ser Tyr Tyr Leu Trp Leu
1 5 10 15
Pro Phe Lys Phe Ile His Gln Thr Leu Arg Ser Leu Leu Leu Lys Leu
20 25 30
Phe Gly Leu Arg Ser Pro Ser Arg His Thr Ala Ala Ser Leu Glu Glu
35 40 45
Glu Val Glu Val Glu Val Val Gln Val Thr Ser Arg Gly Leu Pro Pro
50 55 60
Lys Lys Leu Gln Gln Lys Pro Arg Gly Ser Ser Gly Lys Pro Gly Ser
65 70 75 80
Ile Asn Lys Lys Pro Arg
85
Claims (3)
1. a kind of rape (Brassica napus) small peptide BnPEP4 excitation crop in cruciferae generate to sclerotinite
(Sclerotinia sclerotiorum) resistance application, rape small peptide BnPEP4 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: with small peptide BnPEP4 solution immersion or
Spraying treatment crop in cruciferae blade excites plant immune.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910518849.0A CN110278959B (en) | 2019-06-16 | 2019-06-16 | Application of rape short peptide BnPEP4 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910518849.0A CN110278959B (en) | 2019-06-16 | 2019-06-16 | Application of rape short peptide BnPEP4 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110278959A true CN110278959A (en) | 2019-09-27 |
| CN110278959B CN110278959B (en) | 2020-10-13 |
Family
ID=68005035
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910518849.0A Active CN110278959B (en) | 2019-06-16 | 2019-06-16 | Application of rape short peptide BnPEP4 |
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| Country | Link |
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| CN (1) | CN110278959B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114375963A (en) * | 2021-12-29 | 2022-04-22 | 浙江大学 | Immune excitation function application of rape short peptide BnPEP6 |
| CN114621331A (en) * | 2022-04-07 | 2022-06-14 | 湖南农业大学 | Sclerotinia sclerotiorum small peptide and application thereof |
| CN116849228A (en) * | 2023-08-29 | 2023-10-10 | 浙江大学海南研究院 | Application of pear oligopeptide PdrPEP6 in plant disease prevention and control |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
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2019
- 2019-06-16 CN CN201910518849.0A patent/CN110278959B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
Non-Patent Citations (5)
| Title |
|---|
| ALISA HUFFAKER ET AL.: "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》 * |
| HUAJIAN ZHANG ET,AL.: "A novel protein elicitor (SsCut) from Sclerotinia sclerotiorum induces multiple defense responses in plants", 《PLANT MOL BIOL》 * |
| SHIV S. VERMA ET AL.: "A cysteine-rich antimicrobial peptide from Pinus monticola(PmAMP1)confers resistance to multiple fungal pathogens", 《PLANT MOL BIOL》 * |
| 无: "NCBI Reference Sequence: XP_013627550.1", 《NCBI PROTEIN》 * |
| 无: "NCBI Reference Sequence:NP_568223.1", 《NCBI PROTEIN》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114375963A (en) * | 2021-12-29 | 2022-04-22 | 浙江大学 | Immune excitation function application of rape short peptide BnPEP6 |
| CN114621331A (en) * | 2022-04-07 | 2022-06-14 | 湖南农业大学 | Sclerotinia sclerotiorum small peptide and application thereof |
| CN114621331B (en) * | 2022-04-07 | 2024-03-12 | 湖南农业大学 | Sclerotinia small peptide and application thereof |
| CN116849228A (en) * | 2023-08-29 | 2023-10-10 | 浙江大学海南研究院 | Application of pear oligopeptide PdrPEP6 in plant disease prevention and control |
| CN116849228B (en) * | 2023-08-29 | 2024-01-30 | 浙江大学海南研究院 | Application of pear oligopeptide PdrPEP6 in plant disease prevention and control |
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| Publication number | Publication date |
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| CN110278959B (en) | 2020-10-13 |
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