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Ustilaginoidea virens elicitor protein SGP1, oligopeptide and application thereof

An elicitor, rice koji fungus technology, applied in the field of bioengineering, can solve the problem of less PAMP molecules, and achieve the effects of improving disease resistance, enhancing plant resistance to pathogen infection, and having broad application prospects

Active Publication Date: 2021-07-13
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there are very few PAMP molecules identified from fungi that are conserved in different fungi and that a short peptide segment of a few amino acids can induce a series of immune responses and mediate resistance

Method used

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  • Ustilaginoidea virens elicitor protein SGP1, oligopeptide and application thereof
  • Ustilaginoidea virens elicitor protein SGP1, oligopeptide and application thereof
  • Ustilaginoidea virens elicitor protein SGP1, oligopeptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Prokaryotic expression and purification of SGP1 protein

[0056] The protein elicitor SGP1 disclosed in this embodiment, its nucleotide sequence is shown in SEQ ID NO:1 in the sequence listing, and its amino acid sequence is shown in SEQ ID NO:2.

[0057] (1) Identification of SGP1 protein elicitor

[0058] According to the principle that the protein secreted by pathogenic bacteria can be recognized by plants to induce plant immune response, the laboratory collects the liquid culture solution of Aspergillus oryzae, extracts the protein, and sends the extracted protein to the company for mass spectrometry sequencing, and then analyzes it through bioinformatics Determining the secretome of Aspergillus oryzae. Proteins containing signal peptides, containing conserved domains, and present in pathogenic bacteria but not in plants were identified as candidate proteins. The coding sequences of 79 candidate proteins were respectively constructed on the plant transie...

Embodiment 2

[0068] Example 2 SGP1 Induces Plant Defense Response

[0069] (1) To induce tobacco allergic reaction

[0070] Adjust the SGP1 protein concentration to 1.5 nM, 7.5 nM, 15 nM, 30 nM, 150 nM, 300 nM, 1.5 μM. Select Nicotiana benthamiana about 4 weeks old, inject SGP1 recombinant protein from the back of Nicotiana benthamiana leaves into Nicotiana benthamiana leaves with a 1ml syringe without a needle, and at the same time, 1.5μM MBP is used as a control, and the allergic reaction is observed 48 hours after injection necrotic reaction. At the same time, after the leaves were photographed, placental blue staining was performed for observation.

[0071] Results: The protein at a concentration above 30nM can induce allergic cell death visible to the naked eye on Nicotiana benthamiana; placental blue staining stains the allergic reaction site blue, as shown in image 3 shown.

[0072] (2) Inducing ROS production in tobacco leaves

[0073] 1μM of SGP1 protein was injected into th...

Embodiment 3

[0088] Example 3 SGP1 induces disease resistance in tobacco and rice

[0089] (1) To induce the resistance of tobacco to Phytophthora nicotianae

[0090] After spraying the SGP1 recombinant protein and mock on the leaves of Nicotiana benthamiana for 24 hours, inoculate 100 zoospores of Phytophthora nicotiana for 48 hours, and count the diameter of the lesion.

[0091] Results: After SGP1 recombinant protein treated Nicotiana benthamiana leaves, the lesion diameter could be reduced by about 0.9 cm, which indicated that SGP1 recombinant protein could induce the resistance of Nicotiana benthamiana to Phytophthora nicotianae, such as Figure 6 shown.

[0092] (2) Inducing rice resistance to blast fungus

[0093] After spraying SGP1 recombinant protein and mock on rice leaves for 24 hours, inoculate the spore liquid of Magnaporthe oryzae (5×10 4 spores / mL), after 7 days of moisturizing culture at 28°C, count the number of five types of lesions. Type 1, pinhead-sized brown spots...

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Abstract

The invention discloses ustilaginoidea virens elicitor protein SGP1, oligopeptide and application thereof. The elicitor protein and the elicitor oligopeptide can obviously activate plant defense response, improve plant disease resistance, enhance plant resistance to pathogenic bacteria infection and effectively prevent or relieve diseases. The elicitor protein and the elicitor oligopeptide provide a new way for improving the disease resistance of plants, provide resources for development and application of biopesticides in the future, can be used as a novel microbial protein pesticide for defending the invasion of diseases and pests, also provide a basis for transgenic plants with the gene in the future, and have a wide application prospect in agricultural production.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to the aspergillus oryzae protein elicitor SGP1 and its application. Background technique [0002] At present, the main measures for plant disease prevention and control are chemical control and selection of disease-resistant varieties. However, the use of chemical agents often brings about environmental pollution and food safety problems; pathogen race variation brings drug resistance and overcomes disease-resistant genes, which reduces the control effect; the effects of the two disease control and control strategies are seriously affected, and it remains to be seen. Research and develop environmental protection, broad-spectrum, long-lasting and efficient disease prevention and control strategies. With the understanding of the interaction between plants and pathogens and the development of biotechnology, the use of pathogen-induced plant immunity and resistance provides the basis for...

Claims

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Application Information

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IPC IPC(8): C07K14/37C12N15/31C12N15/82C12N1/21A01H5/00A01H6/00C12R1/19
CPCC07K14/37C12N15/8282
Inventor 宋天巧刘永锋
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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