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A method of inducing Phytophthora capsici to produce toxic secreted protein

A technology of Phytophthora capsici and protein secretion, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve problems such as limiting in-depth research on pathogenic mechanisms

Active Publication Date: 2018-02-13
INST OF PLANT PROTECTION FAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current research shows that the lack of a method to induce Phytophthora capsici to secrete a large amount of toxic proteins limits our in-depth study of its pathogenic mechanism

Method used

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  • A method of inducing Phytophthora capsici to produce toxic secreted protein

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Embodiment 1

[0014] Scrape the surface of the stem base of the diseased pepper plant with a scalpel, and then rinse it with tap water. After natural air-drying, take a 0.5 cm tissue block from the disease-healthy junction and place it on V8 medium. Cultivate for 4 days under dark conditions at 25°C, 15% (final concentration) solid V8 medium is mainly composed of the following parts: V8 vegetable juice (purchased from Campbell Soup Company, USA) 150 mL, CaCO 3 0.2 g, 20 g agar, 1000 mL distilled water; inoculate the isolated weakly pathogenic strain of Phytophthora capsici FZ-1 (self-isolated) on the V8 medium, and cultivate for 4 days at 25°C in the dark until it forms Larger colonies; transfer 20 plates with a diameter of 5 mm to 150 mL of toxic protein liquid induction medium, culture at 25°C and 150 rpm with shaking for 18 days, the main component of the toxic secretion protein liquid induction medium: yeast extract 0.6 g, KH 2 PO 4 0.6 g, MgSO4.7H 2 O 0.25 g, VB1 0.001 g, glucose 25 g,...

Embodiment 2

[0016] Scrape the surface of the stem base of the diseased pepper plant with a scalpel, and then rinse it with tap water. After natural air-drying, take a 0.5 cm tissue block from the disease-healthy junction and place it on V8 medium. Cultivate for 3 days in the dark at 25℃, and inoculate the isolated strong pathogenic Phytophthora capsici strains ND-2, XM-3 into 15% solid V8 medium (V8 vegetable juice 150 mL, CaCO 3 0.2 g, 20 g agar and 1000 mL distilled water), cultured in the dark at 25°C for 3 days, until large colonies are formed; transfer 20 plates with a diameter of 5 mm to 150 mL toxic protein liquid for induction culture Base medium, shake culture at 25℃, 150 rpm for 20 days, toxic secretion protein liquid induction medium main components: yeast extract 0.8 g, KH 2 PO 4 0.7 g, MgSO4.7H 2 O0.3g, VB10.002g, glucose 30g, asparagine 1g, ß-sitosterol 0.02g, dilute to 1000ml; filter with Whatman filter paper, centrifuge the filtrate at 8000rpm for 30min, add 100% saturated ...

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Abstract

The invention relates to a method for inducing Phytophthora capsici to produce toxic secretory protein. The diseased tissue of pepper blight is brought back to the laboratory, the surface of the stem base of the diseased plant is first scraped off with a surgical blade, and then washed with tap water, and taken from the junction of disease and health. The 0.5 cm tissue pieces were placed in V8 medium, and cultured in the dark at 25°C for 3-4 days until large colonies were formed, then transferred the bacterial discs to the toxic protein liquid induction medium, and cultured with shaking for 15-20 days. Filter the mycelium with filter paper, centrifuge the filtrate at 8000rp for 30min, add saturated ammonium sulfate to the supernatant, centrifuge at 12000rpm for 30min, dialyze the precipitate, and obtain the toxic protein secreted by Phytophthora capsici after freeze-drying, and store at -70°C. The above method can induce Phytophthora capsici to produce a large amount of toxic secretory protein, which fills the gap that a large amount of toxic secretory protein cannot be obtained in the research of pathogenic mechanism of Phytophthora capsici at home and abroad, and is of great significance.

Description

Technical field [0001] The invention relates to the technical field of plant protection, in particular to a method for inducing Phytophthora capsici to produce toxic secreted proteins. Background technique [0002] Pepper blight is a devastating fungal disease that occurs worldwide. It was first discovered in New Mexico, USA in 1918. Phytophthora capsici that causes pepper blight Phytophthora capsici Leonian was named by LeonLeonian in 1922, and it belongs to the plant pathogenic oomycete. Phytophthora capsici survives the winter in the soil diseased body as oospores or chlamydospores, and can survive for several months or even longer. It has a wide host range and mainly infects many important vegetable crops such as peppers, tomatoes, eggplants, cucumbers, and pumpkins. Pepper blight can occur in both open and protected areas. The epidemic of the disease has caused serious loss of pepper. The general disease plant rate is 15-30%, and it can reach more than 80% in severe cases....

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P21/00C12R1/645
Inventor 刘裴清陈庆河李本金丁雪玲翁启勇
Owner INST OF PLANT PROTECTION FAAS
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