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Botrytis cinerea secreted protein elicitor BcXyl1 and application thereof

A secreted protein and elicitor technology, which is applied in application, fermentation, fungi, etc., can solve the problem of not many active ingredients, achieve broad application prospects, reduce disease occurrence, and improve plant disease resistance.

Inactive Publication Date: 2019-01-04
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 Botrytis cinerea at home and abroad that can cause plant disease resistance

Method used

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  • Botrytis cinerea secreted protein elicitor BcXyl1 and application thereof
  • Botrytis cinerea secreted protein elicitor BcXyl1 and application thereof
  • Botrytis cinerea secreted protein elicitor BcXyl1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1 Eukaryotic expression and purification of BcXyl1 protein

[0031] (1) Isolation and identification of BcXyl1 protein elicitor

[0032] Xylanases are ubiquitously present in the secretions of plant pathogens and have been validated as protein elicitors to activate plant immune responses (Barak Rotblat et al., The Plant Journal, EIX protein elicitor elicits an important component of the active site Identification, (2002) 32, 1049–1055). Based on the completed sequencing of the Botrytis cinerea genome and the proteomics data of Botrytis cinerea, combined with the screening system for protein elicitors, the CDD function in the NCBI database was used to compare the conserved sequences of the xylanases in the Botrytis cinerea secretome. Phylogenetic tree analysis identified 8 candidate proteins. The coding sequences of the eight candidate proteins were respectively constructed into the transient expression vector PYBA1132, transformed into Agrobacterium and inject...

Embodiment 2

[0041] Example 2 BcXyl1 induces plant defense response

[0042] (1) BcXyl1 induces tobacco hypersensitivity response (HR)

[0043] After the concentration of BcXyl1 protein was determined, the concentration was adjusted to 4 μM, 3 μM, 2 μM, 1 μM, 800 nM and 500 nM. Select the vigorously growing middle leaves of tobacco at the 5-6 leaf stage, and inject about 20uL of the recombinant protein 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, the treated leaves were photographed and stained with trypan blue to observe the staining condition.

[0044] Results: Obvious allergic reactions appeared on the leaves treated with recombinant protein BcXyl1, and after trypan blue staining, the allergic reaction parts were stained blue (such as figure 2 ).

[0045] (2) BcXyl1 induces ROS responses in tobacco and tomato leaves

[0046] The concentration of the BcXyl1 protein was determined to be a...

Embodiment 3

[0061] Example 3 BcXyl1 induces disease resistance in tobacco and tomato

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

[0063] 50 uL of purified BcXyl1 (1 μM) was injected into different tobacco leaves from the back of the leaves. With PEVC at the same concentration 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 = 40.09mm 2

[0065] Treated leaf lesion area = 23.94mm 2

[0066] Results: The elicitor protein BcXyl1 can induce tobacco resistance to Botrytis cinerea (eg Figure 5 ), and the lesion area was reduced by about 40.28%.

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

[0068] 50 uL of the purified BcXyl1 solution (500 nM) was injected into different tobacco leaves from the back of the leaves, and PEVC at the same concentratio...

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Abstract

The invention discloses botrytis cinerea secreted protein elicitor BcXyl1 and application thereof. The protein can increase plant disease resistance and can be used as the elicitor for inducing plantimmunity to apply to the prevention and treatment field of plant diseases. The amino acid sequence of the protein is as shown in SEQ ID No. 2. High-concentration purified protein can be obtained by using engineering strain expression. The BcXyl1 can be used for increasing the disease resistance of tobacco and tomatoes, and the related diseases are tobacco mosaic virus, tobacco gray mold and tomatogray mold. The protein elicitor is capable of evidently increasing the disease resistance of plants, quick in action and lasting in effect. The BcXyl1 provides a new approach for increasing the disease resistance of plants.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a secreted protein elicitor of gray mold (Botrytis cinere). 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. Therefor...

Claims

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Application Information

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IPC IPC(8): C07K14/37C12N15/31C12N15/81C12N1/19A01G7/06C12R1/84
CPCA01G7/06C07K14/37C12N15/815
Inventor 邱德文杨远坤杨秀芬曾洪梅郭立华李广悦袁京京王双超
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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