Verticillium dahliae secreting type protein elicitor VdPEL1 and application thereof

A technology of secreted protein and Verticillium dahliae, which is applied in application, fermentation, fungi, etc., can solve many problems, and achieve the effect of low concentration, long action time and fast onset of action

Inactive Publication Date: 2018-07-24
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are not many active ingredients isolated and identified from Verticillium dahliae at home and abroad that can cause plant disease resistance

Method used

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  • Verticillium dahliae secreting type protein elicitor VdPEL1 and application thereof
  • Verticillium dahliae secreting type protein elicitor VdPEL1 and application thereof
  • Verticillium dahliae secreting type protein elicitor VdPEL1 and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1V

[0030] Eukaryotic expression and purification of embodiment 1VdPEL1 protein

[0031] (1) Isolation and identification of VdPEL1 protein elicitor

[0032] Pectin lyases are widely present in the secretions of plant pathogens and have been validated as protein elicitors to activate plant immune responses (Poinssot. Immune Response, 2003, 553–564.). Based on the completion of the genome sequencing of Verticillium dahliae and the proteomics data of Verticillium dahliae, combined with the previous screening system for protein elicitors in our laboratory, the CDD function in the NCBI database was used to analyze the The conserved sequences of the pectin lyases in the secretome were compared with the reported elicitors, and the phylogenetic tree analysis was carried out at the same time, and 10 candidate proteins were identified. The coding sequences of 10 candidate proteins were respectively constructed into the transient expression vector PYBA1132, transformed into Agrobacterium ...

Embodiment 2

[0041] Example 2 VdPEL1 induces plant defense response

[0042] (1) VdPEL1 induces tobacco hypersensitivity response (HR)

[0043] After the concentration of VdPEL1 protein was determined, its concentration was adjusted to 1 μM, 500nM, 300nM, 200nM and 100nM. Select the vigorously growing middle leaves of tobacco at the 5-6 leaf stage, and inject about 20uL of the recombinant protein VdPEL1 at a specified concentration into different tobacco leaves from the back of the leaves with a 1mL syringe without a needle. At the same time, 1 μM BSA (buffer solution: 20 mM Tris-HCl, pH 8.0) was used as a control, and allergic reactions were observed 48 hours after injection. At the same time, the treated leaves were photographed and stained with trypan blue to observe the staining condition.

[0044] Results: There were obvious allergic reactions on the leaves treated with recombinant protein VdPEL1, and the allergic reaction parts were stained blue after trypan blue staining. (Such a...

Embodiment 3

[0061] Example 3 VdPEL1 induces disease resistance in tobacco and cotton

[0062] (1) Inducing tobacco resistance to Botrytis cinerea

[0063] 50 uL of purified VdPEL1 (1 μM) was injected into different tobacco leaves from the back of the leaves. With the same concentration of BSA (20mM Tris-HCl buffer, pH 8.0) as the control, 20 strains were treated and repeated 3 times. After 72 hours of induction, the spore suspension of Botrytis cinerea (10 5 pc / ml), moisturized for 24h, and measured the lesion size on the 3rd day.

[0064] Control leaf lesion area = 31.15mm 2

[0065] Treated leaf lesion area = 20.94mm 2

[0066] Results: The elicitor protein VdPEL1 can induce tobacco resistance to Botrytis cinerea (such as Figure 5 ), and the lesion area was reduced by about 32.78%.

[0067] (2) To induce tobacco resistance to tobacco mosaic virus (TMV)

[0068] 50uL of purified VdPEL1 solution (500nM) was injected into different tobacco leaves from the back of the leaves, and ...

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Abstract

The invention discloses a verticillium dahlia secreting type protein elicitor VdPEL1 and application thereof. The protein can improve the plant disease resistance and can be used as an inductor for inducing the plant immunity and can be applied to the field of plant disease prevention and treatment. The amino acid sequence of the protein is shown as SEQ ID NO:2. The engineering strain expression is used for obtaining high small-to-big protein, so that the disease resistance of tobaccos and cotton can be improved; the related plant diseases include tobacco mosaic, gray mold and cotton verticillium wilt. The protein elicitor can obviously improve the plant disease resistance; the effect is fast; the action time is long. The VdPEL1 provides a new path for improving the plant disease resistance.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a secreted protein elicitor of Verticillium dahliae. Background technique [0002] At present, the control of plant diseases mainly adopts chemical control and selection of disease-resistant varieties. However, with the continuous evolution of the pathogenic races of pathogenic bacteria, resistance to pesticides has been produced and the control effect has been reduced, which in turn has led to a continuous increase in the amount of pesticides used. At the same time, the safety and ecological safety of agricultural products caused by the extensive use of chemical pesticides has attracted more and more attention. With the development of biotechnology, plant-induced resistance has received great attention. As the main factor inducing plant resistance, elicitors have attracted more and more attention, and have become an important development direction of biopesticides. Therefore, the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/37C12N15/31C12N15/81C12N1/19A01H5/00A01H6/82A01H6/60C12R1/84
CPCC07K14/37C12N15/8282C12N15/8283
Inventor 邱德文杨远坤杨秀芬曾洪梅郭立华袁京京
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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