Detection method of living cell biomass of Phanerochaete chrysosporium under heavy metal stress

Abstract

The invention discloses a method for detection of the living cell biomass of phanerochaete chrysosporium under the heavy metal stress. The method comprises the steps as follows: 1, pellet culturing: adding the phanerochaete chrysosporium into a liquid culture medium for constant-temperature culturing, and adding heavy metal solution for stress treatment; 2, chromogenic reaction: adding the phanerochaete chrysosporium pellets obtained after heavy metal stress into sterile MTT solution and performing chromogenic reaction for 0.5-4 h at the temperature of 30-50 DEG C, and then quickly adding hydrochloric acid solution to stop the reaction, so as to obtain the mixed liquor after reaction; 3, centrifugal treatment: performing centrifugal treatment on the mixed liquor to remove the liquid supernatant and obtain the lower sediment; 4, extracting detection: adding an extracting agent into the sediment for extraction, determining the absorbance of the extract liquor, and calculating the living cell biomass of the phanerochaete chrysosporium under the heavy metal stress. According to the invention, the detection method is simple and convenient, wide in application and strong in practicability.

Description

technical field [0001] The invention relates to the field of microbial technology and environmental toxicology, in particular to a detection method for the biomass and activity of Phanerochaete chrysosporium live cells under heavy metal stress. Background technique [0002] At present, heavy metal pollution in the environment has become a global problem. Heavy metals in the environment accumulate and persist in organisms through the food chain effect, which increasingly threatens the ecological environment and human health. The use of microorganisms to treat heavy metals in industrial wastewater has the advantages of low cost, little impact on the environment, and high efficiency, and has attracted more and more attention from scientific and technological personnel. In recent years, white rot fungi have been widely studied for their extensive and unique degradation ability to various xenobiotics and good adsorption effect on heavy metal ions in wastewater. Most of the stud...

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Problems solved by technology

[0004] Some traditional detection methods, such as wet and dry weight method, blood count method, plate colony count method and fluorescence microscope count method, etc. have been widely used to detect the activity of microorganisms, but the traditional methods have many shortcomings, such as: blood count method 1. The plate colony counting method is time-consuming and laborious, and often has technical inaccuracies; the fluorescent microscope counting method uses various dyes to stain the cells, and uses a microscope to observe the different colors of the dead / living cells to count, which often occurs The color difference between dead and living cells after staining is not obvious, which is easy to confuse

In addition, Phanerochaete chrysosporium is a filamentous fungus. In the process of liquid culture, its hyphae are easily entangled to form mycelial balls with a diameter of about 3 mm. Form large blocks or large sheets together, mycelia and culture medium cannot form a homogeneous system, so traditional methods are not suitable for determining the biomass of living cells

The simple cell weight (dry weight or wet weight) cannot reflect the physiological state and aging degree of the bacteria in the treatment system, and cannot even distinguish between cell death and life.

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