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Equipment and method for determining field rhizosphere soil in-situ zymogram

A technology of rhizosphere soil and zymography, applied in measurement devices, instruments, fluorescence/phosphorescence, etc., can solve the problem of blurred focus in "hot spots", and achieve the effect of advanced equipment, easy operation and high precision

Inactive Publication Date: 2018-10-23
INST OF GEOGRAPHICAL SCI & NATURAL RESOURCE RES CAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is a problem because in zymography using the gel as the substrate medium, the focus of the "hot spot" is blurred due to the diffusion of the enzyme or substrate into the gel
[0006] In summary, in situ zymography can successfully distinguish the spatial heterogeneity of enzyme activity in rhizosphere and biopore soil, but currently there is no instrument suitable for in situ zymography monitoring in the field in the world, only a simple indoor darkroom- Fluorescence-camera components have yet to be developed into a set of monitoring instruments and technologies that are convenient for field applications, realizing the comprehensive functions of field in-situ monitoring and digitalization of enzyme spectrum images

Method used

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  • Equipment and method for determining field rhizosphere soil in-situ zymogram

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Experimental program
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Effect test

Embodiment 1

[0041] Embodiment 1: The equipment for measuring field rhizosphere soil in situ zymogram

[0042] Before the detailed description of the method for measuring the in situ zymogram of field rhizosphere soil according to the present invention, the structure of the device for measuring the field rhizosphere soil in situ zymogram of the present invention will be described first, so as to understand the present invention more clearly.

[0043] figure 1 A specific embodiment of the device for measuring in situ zymogram of field rhizosphere soil according to the present invention is shown. It can be seen from the figure that the equipment for measuring the in situ zymogram of field rhizosphere soil in the present invention includes a fiber membrane, a reaction device, a soil root culture device, a fluorescence reaction darkroom-zymogram imaging device and an image processor.

[0044] The fiber membrane 1 may be a polyester fiber membrane with a pore size of 0.45 μm, but is not limite...

Embodiment 2

[0054] Example 2. In situ monitoring of rhizosphere soil β-1,4-glucosidase enzyme activity zymogram

[0055] 1. Preparation of β-1,4-glucosidase standard substance-polyester fiber membrane medium: choose methyl 7-hydroxycoumarin as fluorescent standard substance, set 0, 0.5, 1, 2, 4, 6, 8, 8 concentration gradients of 10 μM, put 0.45 μm polyester fiber membranes of 2cm×2cm into standard substance solutions with different concentration gradients in a graduated cylinder, soak for 1 minute, and prepare standard substances with different concentration gradients-polyester Fiber membrane medium, and accurately record the volume of fluorescent standard substance absorbed by each standard substance-polyester fiber membrane medium.

[0056]Make the zymogram standard curve of β-1,4-glucosidase: open the fluorescence reaction darkroom door 8, put the prepared standard substance-polyester fiber membrane medium on the movable supporting plate 9 of the fluorescence reaction darkroom 7, clos...

Embodiment 3

[0061] Example 3: In situ monitoring of rhizosphere soil β-1,4-N-acetylglucosaminidase (NAG) activity zymogram

[0062] 1. Preparation of β-1,4-N-acetylglucosaminidase standard substance-polyester fiber membrane medium: choose methyl 7-hydroxycoumarin as fluorescent standard substance, set 0, 0.5, 1, 2, 4 , 6, 8, and 10 μM with 8 concentration gradients, put a 0.45 μm polyester fiber membrane of 2cm×2cm into the standard substance solution with different concentration gradients in a graduated cylinder, and soak for 1 minute to prepare different concentration gradients. Standard substance-polyester fiber membrane medium, and accurately record the volume of fluorescent standard substance absorbed by each standard substance-polyester fiber membrane medium.

[0063] Make the zymogram standard curve of β-1,4-N-acetylglucosaminidase: open the fluorescence reaction darkroom door 8, put the prepared standard substance-polyester fiber membrane medium on the removable holder of the fluo...

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Abstract

The invention relates to the multidisciplinary fields such as soil microbiology, soil chemistry, ecology, molecular biology, environmental science, physical geography, global climate change and the like, and specifically provides equipment for determining field rhizosphere soil in-situ zymogram, and a method for determining field rhizosphere soil in-situ zymogram by using the equipment. With the equipment and the method of the present invention, the field rhizosphere soil in-situ zymogram can be successfully determined, the determining is rapid and convenient, and the equipment and the methodcan be used for the research on the influence of roots, litter falls, soil animals, biological pores and the like on the activity of extracellular enzymes related to soil carbon, nitrogen and phosphorus cycles.

Description

technical field [0001] The invention relates to soil microbiology, soil chemistry, ecology, molecular biology, environmental science, physical geography, global climate change and other multidisciplinary fields, and specifically provides a device for measuring the in situ zymogram of field rhizosphere soil and methods. Background technique [0002] Soil extracellular enzymes are involved in numerous biochemical processes and are the drivers of soil organic matter turnover and nutrient cycling in terrestrial ecosystems. Most extracellular enzymes are thought to be derived from microorganisms, however, plant roots and soil animals have a strong influence on the abundance of extracellular enzymes by either directly producing enzymes or indirectly releasing organic substrates that stimulate microbial growth. Soil extracellular enzymes exist in highly heterogeneous environments, so many soil processes driven by them exhibit high spatial heterogeneity. The rhizosphere and biolog...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6428
Inventor 张心昱孙晓敏温学发
Owner INST OF GEOGRAPHICAL SCI & NATURAL RESOURCE RES CAS
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