Simple method for separating and preserving ustilaginoidea virens
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A preservation method and technology for Aspergillus oryzae are applied in the field of separation and preservation of Aspergillus oryzae, which can solve the problems of pathogenic degradation of strains, time-consuming and space-consuming of Aspergillus oryzae, and achieve simple steps, strong repeatability, The effect of taking up less space
Active Publication Date: 2015-12-16
INST OF PLANT PROTECTION SICHUAN ACAD OF AGRI SCI
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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
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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 O2g, Ca(NO 3 ) 2 4H 2 O0.5g, peptone 5g, agar 10g, and distilled water were dissolved to 1000mL, sterilized at 121°C for 30min by high temperature and damp heat, 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 the rice fields where the disease occurred in Sichuan, and dried at room temperature for 7-10 days for later use. From the collec...
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 Chloramphenicol 100μgml for the growth of miscellaneous bacteria -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 412H 2 O2g, Ca(NO 3 ) 2 4H 2 O0.5g, peptone 5g, agar 10g, and distilled water were dissolved to 1000mL, sterilized at 121°C for 30min by high temperature and damp heat, 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 ...
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Abstract
The invention relates to the technical field of separating and preserving ustilaginoidea virens, in particular to a simple method for separating and preserving the ustilaginoidea virens. The separation method comprises the following steps that fresh yellow, green and black rice false smut ball samples are collected from an attacked rice field, drying is conducted at the room temperature for 7-10 d for standby application; rice false smut balls of which the covering depth of chlamydospore powder is larger than 1 mm are selected in the collected samples and placed on an ultra-clean working table, and sterilization is conducted through an ultraviolet lamp for 30-40 min; single-grain rice false smut balls are taken through tweezers under the aseptic condition, and a small amount of chlamydospore powder on the surfaces of the rice false smut balls is shaken off slightly on a PSA culture medium; 150-200 microliters of sterile water is taken through a pipette and drips on a PSA flat plate containing chlamydospore. By means of the simple method for separating and preserving the ustilaginoidea virens, the ustilaginoidea virens can be separated to the objective strains rapidly and preserved, the problems that time is consumed, and the efficiency is low during separation of the ustilaginoidea virens are solved, and meanwhile the problems that in the preservation process of the strains, the pathogenicity is deteriorated, and the space is occupied and consumed are solved.
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 (Ricefalsesmut) is caused by Aspergillus oryzae ( Villosiclavavirens (Nakata) E.Tanaka&C.Tanaka) is an important fungal disease caused by the fungus infecting the ear and forming a rice bent ball several times larger than the grain, and its surface is covered with a large number of powdery yellow or dark green chlamydospores . Rice balls 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; Lietal., 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 ...
Claims
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Application Information
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Patent Type & Authority Applications(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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