CN114375963B - Immune excitation function application of rape short peptide BnPEP6 - Google Patents

Immune excitation function application of rape short peptide BnPEP6 Download PDF

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CN114375963B
CN114375963B CN202111637124.7A CN202111637124A CN114375963B CN 114375963 B CN114375963 B CN 114375963B CN 202111637124 A CN202111637124 A CN 202111637124A CN 114375963 B CN114375963 B CN 114375963B
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short peptide
bnpep6
rape
sclerotinia sclerotiorum
resistance
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CN114375963A (en
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蔡新忠
陈松余
叶云峰
徐幼平
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Zhejiang University ZJU
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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
    • 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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  • Pest Control & Pesticides (AREA)
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  • Gastroenterology & Hepatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Botany (AREA)
  • Agronomy & Crop Science (AREA)
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  • Plant Pathology (AREA)
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Abstract

The invention provides application of a rape short peptide BnPEP6 in immune excitation function, and particularly relates to application of a rape (Brassica napus) short peptide BnPEP6 in sclerotiniose prevention and control. The invention firstly synthesizes the short peptide BnPEP6 artificially, clarifies the excitation effect on the sclerotinia sclerotiorum resistance, and provides the application of the short peptide BnPEP6 in the prevention and control of the sclerotinia sclerotiorum disease by exciting the short peptide BnPEP to generate the sclerotinia sclerotiorum resistance through the spray treatment of the rape, the Chinese cabbage and the cabbage. The BnPEP6 short peptide provided by the invention is a product synthesized by plants, and the short peptide is used for stimulating immunity so as to prevent and treat the sclerotinia sclerotiorum, and has the advantages of being natural, green, environment-friendly, incapable of inducing pathogens to generate drug resistance and the like. In addition, the short peptide is easy to artificially synthesize, simple and convenient to obtain, strong in disease-resistant excitation effect and low in required concentration, and is a novel ecological and environment-friendly crop disease prevention and control preparation.

Description

Immune excitation function application of rape short peptide BnPEP6
Technical Field
The invention belongs to the technical field of plant immune biology, relates to application of an immune excitation function of rape short peptide BnPEP6, and particularly relates to application of rape short peptide BnPEP6 in prevention and control of sclerotinia rot.
Background
1. Plant immunity excitation technology
Plant immunity is a defense response activated by plants recognizing pathogen molecules through receptors. The plant immune system has multiple levels, the first of which is the immunity generated by Pattern Recognition Receptors (PRRs) on the surface of plant cells recognizing conserved molecular patterns from pathogens and the plant itself, called pattern-triggered immunity (PTI). The plant can sense the damage to the integrity of plant cells caused by pathogen infection and the like, and generate injury-associated molecular patterns (DAMPs), the DAMPs stimulate the plant to generate PTI immune response through PRR receptor recognition, including active oxygen burst, kinase cascade activation, defense-associated gene expression and the like, and finally express resistance to the pathogen. PTI plays an important role in non-host resistance (nonhost resistance) to prevent non-adaptive microorganisms from infecting plants and basal resistance (basal resistance) to limit infection of adaptive pathogens to host plants that are susceptible. The PTI immunity stimulated by the DAMP has the advantages of broad spectrum, strong, stable and lasting property and the like. In addition, because the DAMP is a natural product of the plant, the DAMP is utilized to stimulate immunity so as to achieve the purpose of preventing and controlling diseases, and the method is a novel environment-friendly green disease prevention and control method.
2. Plant sclerotinia preventing and controlling technology
Plant sclerotinioses are caused by infection with Sclerotinia sclerotiorum (sclerotirotirus). Sclerotinia sclerotiorum is a dead body nutritional (necrotrop) pathogenic fungus, has a very wide host range, and is a main disease of oil crops and vegetable crops. Causing huge economic losses every year. Chemical control remains an important tool due to the lack of high resistance varieties. Because some pesticides have the problems of ecological pollution, human and animal toxicity, easy generation of drug resistance of pathogens and the like, novel green prevention and control preparations are urgently needed to be researched and developed.
Disclosure of Invention
The invention aims to provide application of rape short peptide BnPEP6 in immune excitation function, and relates to application of rape (Brassica napus) short peptide BnPEP6 in sclerotinia sclerotiorum disease prevention and control. The amino acid sequence of the rape short peptide BnPEP6 applied by the invention is shown as SEQ ID:1, and the full-length protein C-terminal part has a full-length protein sequence shown in SEQ ID:2, respectively.
The invention uses rape (Brassica napus) short peptide BnPEP6 to excite crucifer crops such as rape, chinese cabbage (Brassica rapa) and cabbage (Brassica oleracea) to generate resistance to Sclerotinia sclerotiorum, thereby preventing and controlling the Sclerotinia sclerotiorum of the rape, the Chinese cabbage and the cabbage and playing a role in prevention and control.
Before the present invention, the function of the short peptide BnPEP6 was not reported in any way. The invention firstly synthesizes the short peptide BnPEP6 through manual synthesis, clarifies the excitation effect of the short peptide on the resistance of sclerotinia sclerotiorum, and provides the application of the short peptide in preventing and controlling sclerotinia sclerotiorum by processing cruciferous plants to excite the short peptide to generate the resistance of sclerotinia sclerotiorum. The application of the invention is realized by the following steps:
(1) Artificial synthesis or biological expression acquisition of BnPEP6 short peptide
The peptide can be synthesized by a professional polypeptide synthesis company. The short peptide BnPEP6 can also be obtained by biological expression: the nucleotide sequence corresponding to the BnPEP6 is cloned into an expression vector and is transformed into eukaryotic or prokaryotic microorganisms, and the target short peptide is amplified, extracted and purified by the microorganisms.
(2) Plant treatment of BnPEP6 short peptides
The plant leaves are treated by spraying with a short peptide BnPEP6 aqueous solution with a proper concentration to stimulate the plant immunity. The spray treatment enables large-scale plant treatment.
(3) BnPEP6 short peptide stimulates plants to generate resistance to sclerotinia sclerotiorum
After the short peptide BnPEP6 is treated, the plant is quickly stimulated to generate immune response to sclerotinia sclerotiorum. The intensity of the excitation can be detected by pathogen inoculation analysis. The immune response of the plants to sclerotinia sclerotiorum is shown by the short peptide BnPEP6 treatment, so that necrotic lesions generated after inoculation are smaller than those generated by the water control treatment.
The invention has the advantages that: (1) The BnPEP6 short peptide provided by the invention is a product synthesized by plants, and has the advantages of natural green, no environmental pollution, no harm to people and livestock, no induction of drug resistance of pathogens and the like by utilizing the BnPEP6 short peptide to stimulate immunity for preventing and treating sclerotinia sclerotiorum. (2) the stimulation of plant immunity by DAMP is the result of recent theoretical studies. The BnPEP6 serving as a rape DAMP has a sufficient theoretical basis for being applied to immunity excitation and disease control of cruciferous crops, and is also an example for guiding practice application by theory. (3) The BnPEP6 short peptide only has 23 amino acids, is easy to artificially synthesize and is simple and easy to obtain. (4) The BnPEP6 short peptide has strong disease resistance excitation effect and low required concentration, so the cost for preventing and treating diseases is low.
Drawings
FIG. 1 shows that spraying of rape (Brassica napus) leaves with BnPEP6 aqueous short peptide solution elicits resistance to Sclerotinia sclerotiorum. Spraying 200nM BnPEP6 short peptide water solution to rape leaf via spray gun, inoculating the leaf with sclerotinia sclerotiorum mycelium block after 1d, and showing the size of disease spot after 1 d. The results show that the lesion spots formed by the BnPEP6 treated leaves are obviously smaller and the lesion spot expansion is severely inhibited compared with the sterile double distilled water (Mock). It is shown that BnPEP6 treatment strongly stimulates the rape to generate resistance to sclerotinia sclerotiorum.
FIG. 2 shows that the leaf blade of Chinese cabbage (Brassica rapa) is sprayed with BnPEP6 short peptide aqueous solution to stimulate the Chinese cabbage to generate resistance to sclerotinia sclerotiorum. Spraying 200nM BnPEP6 short peptide water solution onto leaf of Chinese cabbage via spray gun, inoculating the leaf with sclerotinia sclerotiorum mycelium block after 1d, and showing the size of lesion spot after 1 d. The results show that compared with the sterile double distilled water treatment (Mock), the lesion spots formed by the BnPEP6 treated leaves are obviously smaller, and the lesion spot expansion is seriously inhibited. The BnPEP6 treatment strongly stimulates the Chinese cabbage to generate resistance to sclerotinia sclerotiorum.
FIG. 3 shows that leaf discs of Brassica oleracea (Brassica oleracea) are stimulated to develop resistance to Sclerotinia sclerotiorum by BnPEP6 short peptide aqueous solution spraying. Spraying 200nM BnPEP6 short peptide water solution to cabbage leaf blade via spray gun, inoculating leaf blade with sclerotinia sclerotiorum mycelium block after 1d, and showing the size of lesion spot after 1 d. The results show that the lesion spots formed by the BnPEP6 treated leaves are obviously smaller and the lesion spot expansion is severely inhibited compared with the sterile double distilled water (Mock). It is shown that BnPEP6 treatment strongly stimulates cabbage to develop resistance to sclerotinia sclerotiorum.
Detailed Description
The invention is further explained by the accompanying drawings and examples.
Example 1 application of BnPPEP 6 short peptide in rape (Brassica napus) to resisting sclerotinia rot
According to the embodiment of the invention, the artificially synthesized BnPEP6 short peptide is utilized to treat the rape by a spraying method, so that the rape is stimulated to generate resistance to sclerotinia sclerotiorum, and the rape sclerotiniose control method is used for controlling sclerotinia sclerotiorum. The method mainly comprises the following steps:
1) Artificial synthesis of BnPEP6 short peptide
The BnPEP6 short peptide provided by the invention has a sequence shown in SEQ ID:1, which consists of 23 amino acids in total, is the C-terminal part of a full-length protein (the sequence is shown as SEQ ID: 2). Before the present invention, the function of the short peptide BnPEP6 was not reported in any way.
In the embodiment, professional polypeptide synthesis companies (Qiangyao biotechnology limited) are entrusted to synthesize the BnPEP6 short peptide for the analysis and the application of the functions of rape disease resistance excitation and sclerotinia disease prevention and control.
2) Treatment of rape by BnPEP6 short peptide
The plant treatment mode of the BnPEP6 short peptide is spraying. The BnPEP6 short peptide aqueous solution with the concentration of 200nM is sprayed to the leaves of rape plants through a spray gun, and the plants can be treated in a large scale.
3) BnPEP6 short peptide stimulates rape to generate resistance to sclerotinia sclerotiorum
The BnPEP6 short peptide aqueous solution sprayed on the rape leaves can obviously stimulate the rape to generate resistance to sclerotinia sclerotiorum. The result of sclerotinia sclerotiorum inoculation analysis shows that after 200nM BnPEP short peptide is sprayed and treated for 1d, the scab of the sclerotinia sclerotiorum mycelium block inoculated leaf is obviously smaller than that of the aseptic double distilled water treatment control, and the scab expansion is seriously inhibited (figure 1). The BnPEP6 spray treatment can rapidly and strongly stimulate the rape to generate resistance to sclerotinia sclerotiorum.
Example 2 application of BnPPEP 6 short peptide in cabbage (Brassica rapa) antibacterial nuclear disease
According to the embodiment of the invention, the artificially synthesized BnPEP6 short peptide is utilized to treat the Chinese cabbage by a spraying method, so that the Chinese cabbage is stimulated to generate resistance to sclerotinia sclerotiorum, and the Chinese cabbage is used for controlling sclerotinia sclerotiorum. The method mainly comprises the following steps:
1) Artificial synthesis of BnPEP6 short peptide
The BnPEP6 short peptide provided by the invention has a sequence shown as SEQ ID:1, which consists of 23 amino acids and is the C-terminal part of the full-length protein (the sequence is shown as SEQ ID: 2). Before the present invention, the function of the short peptide BnPEP6 was not reported in any way.
In the embodiment, a professional polypeptide synthesis company (Qiangyao biotechnology limited) is entrusted to synthesize the BnPEP6 short peptide for the analysis and the application of the functions of resisting and exciting the diseases of the Chinese cabbage and preventing and controlling the sclerotinia rot.
2) Treatment of Chinese cabbage with BnPEP6 short peptide
The plant treatment mode of the BnPEP6 short peptide is spraying. The BnPEP6 short peptide aqueous solution with the concentration of 200nM is sprayed to the leaves of the Chinese cabbage plants through a spray gun, and the method can be suitable for large-scale plant treatment.
3) BnPEP6 short peptide stimulates the Chinese cabbage to generate resistance to sclerotinia sclerotiorum
The BnPEP6 short peptide water solution sprayed on the leaves of the Chinese cabbage can obviously stimulate the Chinese cabbage to generate the resistance to sclerotinia sclerotiorum. The result of sclerotinia sclerotiorum inoculation analysis shows that after 200nM BnPEP short peptide is sprayed and treated for 1d, the scab of the sclerotinia sclerotiorum mycelium block inoculated leaf is obviously smaller than that of the aseptic double distilled water treatment control, and the scab expansion is seriously inhibited (figure 2). The BnPEP6 spray treatment can rapidly and strongly stimulate the Chinese cabbage to generate the resistance to the sclerotinia sclerotiorum.
Example 3 application of BnPEP6 short peptide in cabbage (Brassica oleracea) to resistance to sclerotinia rot
According to the embodiment of the invention, the artificially synthesized BnPEP6 short peptide is utilized to treat the cabbage by a spraying method, so that the cabbage is stimulated to generate resistance to sclerotinia sclerotiorum, and the method is used for controlling the cabbage sclerotiniose. The method mainly comprises the following steps:
1) Artificial synthesis of BnPEP6 short peptide
The BnPEP6 short peptide provided by the invention has a sequence shown as SEQ ID:1, which consists of 23 amino acids in total, is the C-terminal part of a full-length protein (the sequence is shown as SEQ ID: 2). Before the present invention, the function of the short peptide BnPEP6 was not reported in any way.
In the embodiment, professional polypeptide synthesis companies (Qiang Yao biological science and technology Limited companies) are entrusted to synthesize the BnPEP6 short peptide for analyzing and applying the functions of cabbage disease resistance excitation and sclerotinia disease prevention and control.
2) Treatment of cabbage with BnPEP6 short peptide
The plant treatment mode of the BnPEP6 short peptide is spraying. The BnPEP6 short peptide aqueous solution with the concentration of 200nM is sprayed to leaves of cabbage plants through a spray gun, and the method is suitable for large-scale plant treatment.
3) BnPEP6 short peptide stimulates cabbage to generate resistance to sclerotinia sclerotiorum
The BnPEP6 short peptide aqueous solution sprayed on the leaves of the cabbage can obviously stimulate the cabbage to generate resistance to sclerotinia sclerotiorum. The result of sclerotinia sclerotiorum inoculation analysis shows that after the sclerotinia sclerotiorum mycelium block is sprayed and treated for 1d by 200nM BnPEP short peptide, the scab of the sclerotinia sclerotiorum mycelium block inoculated leaf is obviously smaller than that of the aseptic double distilled water treatment control, and the scab expansion is seriously inhibited (figure 3). The BnPEP6 spray treatment can rapidly and strongly stimulate the cabbage to generate resistance to sclerotinia sclerotiorum.
Sequence listing
<110> Zhejiang university
<120> immune excitation function application of rape short peptide BnPEP6
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 23
<212> PRT
<213> rape (Brassica napus)
<400> 1
Met Val Ala Arg Leu Thr Arg Arg Arg Pro Arg Pro Pro Tyr Ser Ser
1 5 10 15
Gly Gln Pro Gly Gln Ile Asn
20
<210> 2
<211> 95
<212> PRT
<213> rape (Brassica napus)
<400> 2
Met Glu Val Asn Gly Glu Glu Lys Arg Ser Tyr Arg Arg Glu Asp Glu
1 5 10 15
Glu Lys Glu Val Tyr Tyr Pro Leu Leu Asn Ser Pro Cys Ser Ala Phe
20 25 30
His Lys Thr Val Gln Ala Ile Leu Lys Cys Leu Gly Leu Glu Ser Ser
35 40 45
Ser Ile Ser Pro Ser Ser Ser Ser Ser Gln Asp Pro Gly Thr Glu Thr
50 55 60
Val Gln Glu Thr Gly Phe Met Ala Met Val Ala Arg Leu Thr Arg Arg
65 70 75 80
Arg Pro Arg Pro Pro Tyr Ser Ser Gly Gln Pro Gly Gln Ile Asn
85 90 95

Claims (1)

1. The application of rape short peptide BnPEP6 in the prevention and control of sclerotinia sclerotiorum is characterized in that the rape short peptide BnPEP6The amino acid sequence of the peptide BnPEP6 is shown as SEQ ID:1, the application is to utilize rapeBrassica napusShort peptide BnPEP6 excites brassicaceous crops rape and Chinese cabbageBrassica rapaAnd cabbageBrassica oleraceaProduction of Sclerotinia sclerotiorumSclerotinia sclerotiorumResistance, and thus sclerotinia rot of rape, chinese cabbage and cabbage.
CN202111637124.7A 2021-12-29 2021-12-29 Immune excitation function application of rape short peptide BnPEP6 Active CN114375963B (en)

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Publication number Priority date Publication date Assignee Title
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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* Cited by examiner, † Cited by third party
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WO2006081301A2 (en) * 2005-01-26 2006-08-03 Washington State University Research Foundation Plant defense signal peptides
CN110278959B (en) * 2019-06-16 2020-10-13 浙江大学 Application of rape short peptide BnPEP4
CN110301454B (en) * 2019-06-16 2020-10-16 浙江大学 Application of rape short peptide BnPEP5

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