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Excition of inducing anti-disease ability of plant system

A technology for inducing plants and elicitors, applied in the field of biotechnology research, to achieve the effect of low concentration and strong biological activity

Inactive Publication Date: 2004-01-28
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, there are no literature and patent reports to isolate a new protein elicitor of this patent application from cotton blight

Method used

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  • Excition of inducing anti-disease ability of plant system
  • Excition of inducing anti-disease ability of plant system
  • Excition of inducing anti-disease ability of plant system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Embodiment 1: the preparation method of 90kDa protein elicitor PB90 of cotton blight:

[0016] Cotton blight was cultivated in synthetic culture medium for 10 days, filtered with double-layer filter paper, added ammonium sulfate to 80%, overnight at 4°C, centrifuged at 12000g for 10 minutes, collected protein precipitate, and washed with 1% volume of Tris.HCl (7.4 ) suspension.

[0017] Purified by fast protein liquid chromatography FPLC. Process: anion exchange chromatography, column SepharoseQFF, mobile phase: A, 20mmol / L Tris.HCl pH6.0; B, A+1mol / L NaCl; flow rate: 2.5ml / min, 30min gradient elution, collecting active peaks, The method for determining the activity peak is: after desalting the protein solutions in different collection tubes, dilute them 100 times, inject tobacco, and collect the components that can cause allergic necrosis of tobacco cells for further purification; hydrophobic interaction chromatography, Chromatographic column: Sepharose PHE HP; mobil...

Embodiment 2

[0018] Dilute the elicitor to 100pM, use a syringe with the needle removed, and inject it into the plant leaves along the back of the plant leaves. After 24 hours, the leaves at the injection site turn brown. After the elicitor treatment, until 15 days later, the plants are challenged to inoculate tobacco mosaic leaves Viruses, tobacco black shank bacteria, tobacco Ralstonia solanacearum, tobacco red spot bacteria, and cabbage anthracnose bacteria, etc., plants can show a systemic broad-spectrum disease resistance of up to 80% (induced resistance = number of control leaf lesions- The number of diseased spots on treated leaves / the number of diseased spots on control leaves×100%), the results showed that the elicitor-induced resistance effect obtained in this study could last for more than 15 days. image 3

Embodiment 3

[0020] Using immunogold labeling technique, it was proved that the elicitor was localized on the cell wall of Phytophthora. Figure 4

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Abstract

The invention is an protein exciton, able to induce the disease resistance of plant system, separated from phytophthora and the preparing method. It uses the deposition of ammonium sulfate to obtain a coarse protein exciton, and then makes purification by ion-exchange chromatography and hydrophobe-interaction churomatography to obtain a 90kDa protein excition. The excition can be applied to induce system broad-spectrum disease resistance of tobacco and little Chinese cabbage. The use concentration is 0.1-10nM. The effect of prevention and treatment is above 80%.

Description

(1) Technical field [0001] The invention discloses an elicitor for inducing plant system disease resistance, which belongs to the field of biotechnology research. It is specially used to prepare a protein elicitor from cotton blight, which is used to prevent and control various diseases of tobacco and Chinese cabbage. (2) Background technology [0002] Elicitors are substances that can induce defense responses in plants, such substances include proteins, oligosaccharides, and glycoproteins. There are two types of elicitors, biological sources and non-biological sources. Among them, biological sources include plant pathogenic fungi (such as Phytophthora spp.), plant pathogenic bacteria Erwinia amylovora, Pseudomonas syringae and plant Cell walls, etc., due to the characteristics of environmental safety, obvious effect, and abundant sources, biologically derived elicitors have broader application prospects in the prevention and control of agricultural plant diseases. [0003...

Claims

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Application Information

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IPC IPC(8): A01N63/00
Inventor 王源超张正光胡东维
Owner NANJING AGRICULTURAL UNIVERSITY
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