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A Phytophthora polysaccharide elicitor gep1 and its application in improving plant disease resistance

A technology of elicitors and polysaccharides, applied in the fields of application, plant growth regulators, plant growth regulators, etc., can solve the problems of isolated glucan elicitors, etc., and achieve the effect of alleviating symptoms and inhibiting proliferation

Inactive Publication Date: 2017-09-26
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the isolation of glucan elicitors from Phytophthora fungus

Method used

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  • A Phytophthora polysaccharide elicitor gep1 and its application in improving plant disease resistance
  • A Phytophthora polysaccharide elicitor gep1 and its application in improving plant disease resistance
  • A Phytophthora polysaccharide elicitor gep1 and its application in improving plant disease resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] A Phytophthora polysaccharide elicitor Gep1 is prepared by the following steps:

[0054] 1. Cultivation of Phytophthora tobacco (P.parasitica) fermentation broth and extraction of crude polysaccharides:

[0055] Inoculate the activated Phytophthora tabacum (P.parasitica) (China Agricultural Microorganism Culture Collection and Management Center, strain preservation number ACCC36285) mycelium block into (picking the mycelium block with a diameter of 1 cm at the edge of the colony) potato sucrose culture solution In 250mL (600g of potatoes, 20g of glucose, dilute to 1000mL with water, the final concentration of kanamycin is 50μg / mL), cultivate in the dark at 26°C and 200r / min for 24 days, and obtain the fermentation broth (the cell concentration is 1×10 7 CFU / mL).

[0056] The fermentation broth was first filtered with 4 layers of gauze, the filtrate was centrifuged at 3000r / min for 10min, and the supernatant was collected. The resulting supernatant was concentrated to ...

Embodiment 2

[0066] Monosaccharide composition analysis of polysaccharide elicitor Gep1:

[0067] Weigh 10 mg of the polysaccharide sample into an ampoule, add 2 mL of 2M trifluoroacetic acid solution, seal with an alcohol blowtorch, and hydrolyze at 120°C for 3.5 hours. After cooling, remove the insolubles by filter membrane filtration, remove trifluoroacetic acid by rotary evaporation under reduced pressure, wash with a small amount of methanol, continue rotary evaporation, repeat 3 times, remove residual trifluoroacetic acid, add 0.5mL distilled water to dissolve. Use a 20×20cm silica gel plate to separate, the developer is ethyl acetate-pyridine-absolute ethanol-water (volume ratio=8:1:1:2), and the color developer is aniline-diphenylamine-phosphoric acid (2% Aniline acetone solution, 2% aniline acetone solution, and 85% phosphoric acid are mixed in a ratio of 5:5:1). Take the standard solution (10mg / mL) of each monosaccharide (glucose, fructose, ribose, mannose, galactose, xylose, L-ar...

Embodiment 3

[0070] Detection of physicochemical properties of polysaccharide elicitor Gep1:

[0071] Thermal stability detection of polysaccharide elicitor Gep1: Take 100 μL of polysaccharide elicitor Gep1 (1 mg / mL), incubate in water baths at 40, 60, 70, 80, 90 and 100 °C for 5 min, and then quickly place them on ice, After cooling, centrifuge at 4°C and 12000r / min for 5min, and take the supernatant. After the solution returned to room temperature, its biological activity was tested according to the biological activity identification method described in Example 2. Distilled water was used as a control, and each treatment was repeated 3 times. In addition, 100 μL of polysaccharide elicitor (1 mg / mL) was taken, boiled water bath for 20, 30, 40, 50 and 60 min, and sterilized by high-pressure damp heat at 121 °C for 20 min, and its biological activity was detected by the same method as above.

[0072] The result is as Figure 5 As shown in middle A, a1-a6 in the figure are Gep1 after 20, ...

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Abstract

The invention discloses a phytophthora polysaccharide exciton Gep1 and an application thereof to the improvement of plant disease resistance. The phytophthora polysaccharide exciton Gep1 is separated and purified from tobacco phytophthora; and thin layer chromatography confirms that the monosaccharide composition of polysaccharide is glucose, and Gep1 has thermal stability, can tolerate acid and alkaline, still has activity after being treated at high temperature of 100 DEG C for 60min and under the condition that the pH value is 2-12 and can induce tobaccos to generate resistance when sprayed on leaf surfaces of plants so as to have relatively good preventing effect for tobacco mosaic viruses. Six grams of polysaccharide exciton can be separated out of each liter of phytophthora fermentation solution, so that the yield is far higher than that of protein exciton. After being applied to a farmland, the polysaccharide exciton is easy to decompose in plants, soil and water bodies, free of residues, free of pollution to the environment and relatively safe to non-target organisms such as people and livestock and hardly causes drug resistance. Therefore, the phytophthora polysaccharide exciton Gep1 has very wide application and market prospects.

Description

technical field [0001] The invention belongs to the technical fields of agricultural microbiology and biochemistry and molecular biology, and specifically relates to a Phytophthora polysaccharide elicitor Gep1, and also relates to its application in improving plant disease resistance. Background technique [0002] Sugar elicitors refer to sugar substances that can activate plant autoimmunity and improve plant resistance to disease and stress, and belong to biological elicitors (Yin H et al.2010). Sugars have always been considered as a class of inert compounds, which are only used as metabolic energy sources in organisms or as structural protection materials (such as plant cell walls). Ayers et al. discovered in 1976 that oligosaccharide fragments of the cell wall could induce soybeans to produce phytoalexin, and since then people have begun a new understanding of the function of sugars. At present, it is generally believed that oligosaccharides are a class of elicitors tha...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P19/04C08B37/00A01N43/16A01P1/00A01P21/00C12R1/645
Inventor 赵秀云祁高富霍瑞李锡宏许汝冰王瑞谭军郭利
Owner HUAZHONG AGRI UNIV
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