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Penicillium protein elicitor EPTP and application thereof in improving plant disease resistance

An elicitor and disease resistance technology, applied in the fields of application, plant genetic improvement, plant growth regulators, etc., can solve the problem of less protein elicitors found, and achieve the effects of inhibiting proliferation, alleviating symptoms, and reducing morbidity.

Inactive Publication Date: 2016-11-23
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, protein elicitors have shown good application prospects in inducing plant resistance. However, most of the reported protein elicitors are derived from bacterial hapin protein, and few protein elicitors derived from fungi have been found.

Method used

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  • Penicillium protein elicitor EPTP and application thereof in improving plant disease resistance
  • Penicillium protein elicitor EPTP and application thereof in improving plant disease resistance
  • Penicillium protein elicitor EPTP and application thereof in improving plant disease resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Detect the physicochemical properties of the protein elicitor EPTP:

[0046] (1) thermal stability identification

[0047] Take EPTP (artificially synthesized according to the sequence shown in SEQ ID NO.2, 10 μg / ml) and incubate at 40°C, 60°C, 70°C, 80°C, 90°C, and 100°C for 5 min, put them on ice quickly, and wait until the solution After cooling down to room temperature, the allergic reaction test was carried out. The thermal stability of EPTP was examined.

[0048] The subject of the experiment is Yunyan 87, which grows healthily. The healthy and complete leaves are taken, and the half-leaf injection method is used. The left side of the leaves is the water control, and the right side is EPTP. Inject 20 μL of EPTP or water at each point using a microsyringe without a needle. After 6 days, the leaves were cut and photographed to observe whether allergic spots appeared on the leaves and the shape of allergic spots. The presence of hypersensitivity plaques indicates...

Embodiment 2

[0054] Tobacco systemic resistance response induced by EPTP:

[0055] Take 2-month-old tobacco plants, generally 5-7 true leaves are advisable, inject 400 μl (10 μg / ml) protein EPTP solution from the back of the leaves with a 1ml syringe without a needle, and use PB buffer as a control, every One plant per treatment was repeated 3 times. 24, 48, 72, 96, 120, 144, 168, and 192 hours after treatment, the upper leaves of the treated leaves were taken respectively, and the changes of defense-related enzyme activities were measured.

[0056] (1) Determination of phenylalanine ammonia lyase (PAL) activity

[0057] The PAL activity was determined according to the Hano literature method with slight changes (Hano 2008). Take 0.25g of the treated leaf tissue, add 3ml of buffer solution (5mmol / Lβ-mercaptoethanol, 1mmol / L EDTA, 1%PVP, pH 8.4), add quartz sand, grind in ice bath in a mortar, centrifuge at 8000rpm / min at 4°C After 20 min, the supernatant was collected to detect PAL enzym...

Embodiment 3

[0087] Detection of Activation Protein EPTP's Control Effect on Tobacco Mosaic Virus

[0088] (1) Indoor control experiment of tobacco mosaic virus disease

[0089] Preparation of TMV virus juice: Weigh 1g of fresh tobacco leaves with obvious virus symptoms, put them into a mortar, add 40mL of 0.05M PBS (pH 7.0) and 2g of quartz sand, grind into a homogenate, and put 2 layers of gauze Filtration, the obtained virus filtrate (30 virus particles / ml) can be used for friction inoculation of tobacco (every strain of tobacco virus inoculum is advisable to evenly cover the top two new leaves of this strain of tobacco), for the following experiments or examples.

[0090] Select tobacco with uniform growth and robustness at the 7-8 leaf stage, and use pot experiments to determine the control effect of activator protein EPTP on tobacco mosaic virus disease. The test is divided into two modes of action: protection and treatment:

[0091] Protection group: administer once, inoculate wit...

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Abstract

The invention relates to penicillium protein elicitor EPTP and application thereof in improving plant disease resistance. The protein sequence of the EPTP protein elicitor is shown as SEQ ID NO.2, and the molecular weight is 19247 Da. EPTP induces significant increase of activity of phenylalanine ammonialyase, peroxidase and polyphenol oxidase, and when the elicitor induces tobacco anaphylactic reaction, an anaphylactic reaction marker gene hin1 is induced to express; it is indicated through semiquantitative RT-PCR results that EPTP can induce tobacco to obtain systemic acquired resistance, cause up-regulation of the tobacco blade resistance gene transcription level, and induce expression of resistance gene related to the salicylic acid path and the jasmonic acid path. After being applied, EPTP can improve resistance to viruses and pathogenic bacteria by plants, takes effects efficiently, does not pollute environment, reduces dosage of chemical pesticide and has wide application prospects as a plant immunity activator.

Description

technical field [0001] The invention belongs to the technical fields of agricultural microbiology and biochemistry and molecular biology, and specifically relates to a penicillin elicitor EPTP, and also relates to its application in improving plant disease resistance. Background technique [0002] Protein elicitors can induce plant resistance, enhance plant defense against various pathogenic bacteria, and do not produce toxic side effects and do not cause resistance characteristics of pathogenic bacteria. The role of protein elicitors in the biological control of plant diseases has attracted more and more attention. At present, protein elicitors have shown good application prospects in inducing plant resistance. However, most of the reported protein elicitors are derived from bacterial hapin protein, and few protein elicitors derived from fungi have been found. [0003] Fungi have abundant resources of antiviral active substances, and protein peptides are one of the most imp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/385C12N15/31A01N47/44A01P1/00
Inventor 赵秀云祁高富李娜薛守聪任加庆
Owner HUAZHONG AGRI UNIV
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