A medium for screening and enumerating amylase-producing fungi from complex environmental samples

A complex environment, amylase technology, applied in the field of microbial engineering, can solve problems such as heavy workload, long culture time, and difficult to determine the boundaries of fungi

Inactive Publication Date: 2018-07-24
HUBEI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method mainly faces three problems during the cultivation process: the environmental sample has complex components and many bacteria, which is easy to cause pollution; the fungus grows too fast and quickly occupies the entire plate medium, so the boundaries between fungi are difficult to determine. Unable to count accurately; there are many types of fungi, and the target microorganisms cannot be intuitively selected, such as amylase-producing fungi
However, this method needs to prepare a large amount of medium, and the total culture time is long and the workload is large.
There are few reports on the count of amylase-producing fungi

Method used

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  • A medium for screening and enumerating amylase-producing fungi from complex environmental samples
  • A medium for screening and enumerating amylase-producing fungi from complex environmental samples
  • A medium for screening and enumerating amylase-producing fungi from complex environmental samples

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Embodiment 1 preparation contains bacterium and fungal antibiotic mixed culture medium SK3N1

[0053] (1) S medium (basic medium) composition: NaNO 3 2g,K 2 HP0 4 1g, KCl 0.5g, MgSO 4 0.5g, FeSO 4 0.01g, 10g of soluble starch, 15g of agar, 1000mL of water, 0.1g of Triben blue.

[0054] When preparing, first use a small amount of cold water to make starch into a paste, pour it into boiling water, heat it on the fire, add other ingredients while stirring, and make up water to 1000mL after melting. Sterilize at 121°C for 30 minutes.

[0055] (2) Add kanamycin to a final concentration of 54 mg / L, then add nystatin solution to a final concentration of 1.8 μg / mL to prepare a mixed culture medium containing bacterial and fungal antibiotics SK3N2, and store it for future use.

Embodiment 2

[0057] Select complex environmental samples--Baiyunbian medium-temperature Daqu samples and process them

[0058] Take 10 g of the pulverized Baiyunbian medium-temperature Daqu sample, put it in a conical flask filled with 90 mL of sterile water and glass beads, and shake it for 30 min in a shaker at 37 °C (120 r / min). The processed samples were serially diluted, and 10 -2 、10 -3 and 10 -4 is the final coating plate concentration.

Embodiment 3

[0060] Culture and enumeration of fungi

[0061] Take the activated Baiyunbian mesophilic Dakoji suspension 10 -2 、10 -3 and 10 -4 Each of the three gradients was 100 μL, and the plate was spread on the SK3N1 medium to do three replicates. Incubate at 28°C. The colony morphology was observed and counted every 12 hours.

[0062] Colonies appeared after 1 day on the SK3N2 medium plate, the colonies were round or oval, and the hyphae adhered to the surface of the medium. After 5 days of culture, the number of colonies on the plate of SK3N2 medium remained basically unchanged, the shape of the colonies was round or oval, and individual colonies were easy to identify. The fungal colonies are small, the hyphae are short, and there is little overlap between colonies and colonies. It is very convenient to count amylase-producing fungi in complex environmental samples, so as to facilitate the analysis of the quantitative and structural characteristics of amylase-producing fungal c...

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Abstract

The present invention proposes a culture medium for screening amylase-producing fungi from complex environmental samples, which consists of: NaNO3 2g, K2HP04 1g, KCl0.5g, MgSO4 0.5g, FeSO4 0.01g, soluble starch 10g, agar 15g, Tribyl blue 0.1g, water 1000mL, final concentration of kanamycin 54μg / mL, final concentration of nystatin 0.9-1.8μg / mL. The medium is a medium containing a mixture of bacterial and fungal antibiotics. It can completely inhibit the growth of bacteria. The fungal colonies are smaller, the hyphae are shorter, and there is less overlap between colonies. Amylase fungi: the culture medium can cultivate, separate and count amylase-producing fungi in a complex environment at one time, and the preparation is simple, the counting operation is simple and easy, no purification is required, and the screening cycle is short.

Description

technical field [0001] The invention belongs to the technical field of microbial engineering, and in particular relates to a culture medium for screening and counting amylase-producing fungi from complex environmental samples. Background technique [0002] Complex environmental samples such as soil samples, koji samples, etc. contain a large number and types of microorganisms, and various microbial aggregates with the same or similar functions constitute different microbial functional groups, such as the starch-degrading microbial functional groups concerned in the present invention. The study of microbial functional groups has important practical significance in industry, agriculture, microbial management, medicine and genetic engineering. [0003] One of the important methods to study microbial functional groups includes the application of specific selection medium for isolation, culture and counting. However, this method mainly faces three problems during the cultivation...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/04C12Q1/06
Inventor 杜剑晖熊小毛杨团园张明春马向东
Owner HUBEI UNIV
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