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A Simple Isolation and Preservation Method of Aspergillus Oryzae

A preservation method and technology of rice koji, which is applied in the field of rice koji isolation and preservation, can solve the problems of strain pathogenicity degradation, time-consuming and space-consuming rice koji, and achieve simple steps, strong repeatability, The effect of taking up less space

Active Publication Date: 2018-03-06
INST OF PLANT PROTECTION SICHUAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the above technical problems, the present invention provides a simple method for isolating and preserving rice koji, using the method of the invention, the target strain can be quickly isolated and preserved, which solves the time-consuming and low-efficiency problem of oryza koji isolation. At the same time, it also solves the problems of pathogenicity degradation and space consumption of strains during storage

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Supply medium:

[0023] PSA medium for isolating Aspergillus oryzae: 300g of potato, 15g of sucrose, 10g of agar, distilled water to 1000mL, sterilized at 121℃ for 40min, add inhibitor when the medium is cooled to a temperature of 45±3℃ which is not hot Miscellaneous bacteria growth Chloramphenicol 100μgml -1 Mix well and pour into a Petri dish for later use.

[0024] XBZ medium for preserving Aspergillus oryzae: 300g potato, 20g sucrose, Na 2 HPO 4 12H 2 O 2g, Ca(NO 3 ) 2 4H 2 O 0.5g, peptone 5g, agar 10g, and distilled water were dissolved to 1000mL, sterilized at 121°C for 30min, and cooled for later use.

[0025] 1) Isolation and purification of Aspergillus oryzae strains

[0026] (1) Isolation of Aspergillus oryzae strains: In mid-August 2012, 228 fresh yellow rice ball samples were collected from rice fields where the disease occurred in Sichuan, and dried at room temperature for 7-10 days for later use. From the collected samples, select rice balls cover...

Embodiment 2

[0034] Supply medium:

[0035] PSA medium for isolating Aspergillus oryzae: 300g of potato, 15g of sucrose, 10g of agar, distilled water to 1000mL, sterilized at 121℃ for 40min, add inhibitor when the medium is cooled to a temperature of 45±3℃ which is not hot Miscellaneous bacteria growth Chloramphenicol 100μgml -1 Mix well and pour into a Petri dish for later use.

[0036] XBZ medium for preserving Aspergillus oryzae: 300g potato, 20g sucrose, Na 2 HPO 4 12H 2 O 2g, Ca(NO3 ) 2 4H 2 O 0.5g, peptone 5g, agar 10g, and distilled water were dissolved to 1000mL, sterilized at 121°C for 30min, and cooled for later use.

[0037] 1) Isolation and purification of Aspergillus oryzae strains

[0038] (1) Isolation of Aspergillus oryzae strains: In mid-August 2013, 230 fresh yellow rice ball samples were collected from the rice fields where the disease occurred in Sichuan, and dried at room temperature for 7-10 days for later use. From the collected samples, select rice balls cov...

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Abstract

The invention relates to the technical field of aspergillus oryzae isolation and preservation, in particular to a simple method for isolating and preserving aspergillus oryzae. Its separation method comprises the following steps: collect fresh yellow, green and black rice bent ball samples from diseased paddy fields, and dry at room temperature for 7-10d for subsequent use; select chlamydospore powder from the collected samples to cover rice bent rice with a thickness greater than 1mm Put the ball on the ultra-clean workbench and sterilize it with ultraviolet light for 30-40 minutes; take a single rice ball with tweezers under sterile conditions, and gently shake off a small amount of chlamydospore powder on the surface of the rice ball and put it on the PSA medium On; use a pipette gun to take 150-200ul sterile water drops on the PSA plate containing chlamydospores and so on. The method of the present invention can be used to quickly isolate and store the target bacterial strain, which solves the problems of time-consuming and low efficiency in the isolation of Aspergillus oryzae, and also solves the problems of pathogenicity degradation and space consumption of the bacterial strain during storage. .

Description

technical field [0001] The invention relates to the technical field of aspergillus oryzae isolation and preservation, in particular to a simple method for isolating and preserving aspergillus oryzae. Background technique [0002] Rice false smut is caused by Aspergillus oryzae ( Villosiclavavirens (Nakata) E.Tanaka & C. Tanaka) is an important fungal disease caused by the fungus infecting the panicle and forming a rice ball several times larger than the grain, and its surface is covered with a large amount of powdery yellow or dark green thick Cystospores. Rice curlers not only seriously threaten the yield and quality of rice, but also produce mycotoxins that are highly toxic to humans and livestock (Koiso et al., 1994; Li et al., 1995). In recent years, with the increase in the use of N fertilizer in rice and the promotion of hybrid rice in a large area, the severity of rice false smut is increasing. In severe years, the resulting yield loss can be as high as 50%. [00...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12N1/04C12R1/645
Inventor 伏荣桃卢代华王剑龚学书
Owner INST OF PLANT PROTECTION SICHUAN ACAD OF AGRI SCI
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