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Method for inducing rice blast bacteria to produce pathogeneic protein

A rice blast, pathogenic technology, applied in bacteria, fermentation and other directions, can solve the problem of unable to produce pathogenic protein and so on

Inactive Publication Date: 2008-01-30
YUNNAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The main purpose of the present invention is to overcome the deficiency that rice blast bacteria cannot produce pathogenic proteins under artificial culture conditions, and provide a method that can induce rice blast bacteria to produce pathogenic proteins

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0009] The weak pathogenic bacterial strain 93-12-1a, 95-8-3c, 93-1-2a, 95-22-1a of the rice blast bacterium preserved on the slant is inoculated in glucose (15g / L), yeast powder (5g / L), fat powder (12g / L) and H 2 On ordinary solid medium composed of O (1L), culture at 28°C. After forming larger colonies, use an inoculation needle to pick 3 pieces of about 0.5cm 2 The mycelium block was transferred to 50mL by Na 3 C 6 h 5 o 7 -2H 2 O (2.5g / L), KH 2 PO 4 (5.6g / L), NH 4 NO 3 (2.0g / L), MgSO 4 -7H 2 O(0.2g / L), CaCl 2 -H 2 O(0.1g / L), Biotin(5.0μg / L), Thiamine(10.0mg / L), Trace elements(0.1mL), Sucrose(15g / L), Citric acid(5.00g / L), ZnSO 4 -7H 2 O(5.00g / L), Fe(NH 4 ) 2 (SO 4 ) 2 -6H 2 O(1.00g / L), CuSO 4 -5H 2 O(0.25g / L), MnSO 4 -H 2 O(0.05g / L), H 3 BO 3 (0.05g / L), Na 2 MoO 4 -2H 2 O(0.05g / L) and H 2 O (1L) ordinary liquid culture medium, 28 ° C 150rpm shaking culture, 3 days later, in a sterile state, filter with 2 layers of gauze, and the filtered mycel...

Embodiment 2

[0011]The medium pathogenic bacterial strain 93-2-1a, 95-7-1d, 94-006-1a, 94-12-2a of the rice blast bacterium preserved on the slant is inoculated in glucose (15g / L), yeast powder (5g / L), fat powder (12g / L) and H 2 On ordinary solid medium composed of O (1L), culture at 28°C. After forming larger colonies, use an inoculation needle to pick 3 pieces of about 0.5cm 2 The mycelium block was transferred to 50mL by Na 3 C 6 h 5 o 7 -2H 2 O (2.5g / L), KH 2 PO 4 (5.6g / L), NH 4 NO 3 (2.0g / L), MgSO 4 -7H 2 O(0.2g / L), CaCl 2 -H 2 O (0.1g / L), Biotin (5.0μg / L), Thiamine (10.0mg / L), Trace elements (0.1mL), Sucrose (15g / L) and H 2 O (1L) ordinary liquid culture medium, 28 ° C 150rpm shaking culture, 3 days later, in a sterile state, filter with 2 layers of gauze, and the filtered mycelium pieces are washed with sterile water until colorless. Transfer the washed mycelium block to 50mL by Na 3 C 6 h 5 o 7 -2H 2 O (2.5g / L), KH 2 PO 4 (5.6g / L), MgSO 4 -7H 2 O(0.2g / L), C...

Embodiment 3

[0013] The strong pathogenic strains 94-014-1a, 95-11-1a, 95-19-1a, 95-23-4a of the rice blast bacterium preserved on the slant were inoculated in glucose (15g / L), yeast powder ( 5g / L), fat powder (12g / L) and H 2 On ordinary solid medium composed of O (1L), culture at 28°C. After forming larger colonies, use an inoculation needle to pick 3 pieces of about 0.5cm 2 The mycelium block was transferred to 50mL by Na 3 C 6 h 5 o 7 -2H 2 O (2.5g / L), KH 2 PO 4 (5.6g / L), NH 4 NO 3 (2.0g / L), MgSO 4 -7H 2 O(0.2g / L), CaCl 2 -H 2 O(0.1g / L), Biotin(5.0μg / L), Thiamine(10.0mg / L), Trace elements(0.1mL), Sucrose(15g / L), Citric acid(5.00g / L), ZnSO 4 -7H 2 O(5.00g / L), Fe(NH 4 ) 2 (SO 4 ) 2 -6H 2 O(1.00g / L), CuSO 4 -5H 2 O(0.25g / L), MnSO 4 -H 2 O(0.05g / L), H 3 BO 3 (0.05g / L), Na 2 MoO 4 -2H 2 O(0.05g / L) and H 2 O (1L) ordinary liquid culture medium, 28 ° C 150rpm shaking culture, 3 days later, in a sterile state, filter with 2 layers of gauze, and the filtered myceliu...

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Abstract

This is a method for inducting rice blast bacterium to generate sick-causing protein, which is belonged to plant-protecting field, whose technical project are: inoculate the bevelly conserved rice blast strain in common solid medium, and extendly propagte at 28deg C; after big strain is formed, commutate the strain to 50mL common liquor medium, and oscillation culture for 3 days at 28deg C 150rpm; flush the culture liquid with asepsis water until the hyphal block is colorless; commutate the flushed hyphal block to 50mL sick-causing protein induction medium, and oscillation culture for 3 days at 28deg C 150rpm; filter hyphal liquor, and get the extract of rice blast bacterium excreting protein after freezing out; reserve at -20deg C. This method can induct rice blast bacterium to generate lots of sick-causing proteins, and this is signality because sick-causing protein could not be generated in rice blast bacterium sick-causing mechanism research neither at home nor abroad.

Description

Technical field: [0001] The invention relates to a method for inducing the rice blast fungus to produce pathogenic protein, which belongs to the field of plant protection. Background technique: [0002] Rice blast caused by Magnaporthe grisea is a devastating disease of rice production, which occurs in all rice producing areas in the world, causing 11-30% yield loss every year, and economic losses amounting to 157 million US dollars, which is enough to provide a year for 60 million people. Therefore, it is widely concerned by rice breeders, plant pathologists and geneticists all over the world. Magnaporthe grisea is also a good test material for classical genetics and molecular genetics, and the disease system is a model system for the study of fungi-plant interactions. Research in recent years has obtained a lot of genetic resources and information on morphological differentiation and pathogenic factors related to infection, which has laid a good foundation for the genomic...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P21/02
Inventor 李成云朱有勇周江鸿杨静苏源周晓罡李进斌王云月刘林业艳芬叶敏
Owner YUNNAN AGRICULTURAL UNIVERSITY
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