Analytical method for strain group structure of arbuscular mycorrhizal fungi of phellodendren amurense rupr.

A technology for arbuscular mycorrhizal fungi and mycorrhizal fungi, which is applied in biochemical equipment and methods, and the determination/inspection of microorganisms, can solve the problems of large fluctuations in analysis results and poor accuracy, and achieve high accuracy and stability High and repeatable results

Inactive Publication Date: 2008-12-31
HEILONGJIANG UNIV
View PDF0 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The object of the present invention is to provide a method for analyzing the structure of the arbuscular mycorrhizal fungi of the barkberry in order to solve the problems of large fluctuations and poor accuracy of the analytical results of the current analytical method for the arbuscular mycorrhizal fungi of the barkberry

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Analytical method for strain group structure of arbuscular mycorrhizal fungi of phellodendren amurense rupr.
  • Analytical method for strain group structure of arbuscular mycorrhizal fungi of phellodendren amurense rupr.

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment approach 1

[0012] Specific embodiment one: present embodiment Phellodendron amurense arbuscular mycorrhizal fungi flora structure is analyzed according to the following steps: one, Phellodendron amurense arbuscular mycorrhiza and Phellodendron amurense rhizosphere soil arbuscular mycorrhizal fungus total DNA extraction; Two, Nested-PCR amplification Fungal 18S rDNA NS31 / Glo1 region: Perform three PCR amplifications; 3. Use the third PCR amplification product in step 2 to perform denaturing gradient gel electrophoresis: use denaturing polyacrylamide gel, and the denaturing gradient range of the gel is 30%~ 60%, the electrophoresis voltage is 130V, the electrophoresis time is 8 hours, and the electrophoresis temperature is 60°C; 4. Using DNA sequencing technology, the flora structure of the arbuscular mycorrhizal fungi of Phellodendron amurense can be analyzed;

[0013] The primers for the first PCR amplification in step 2 are GeoA2: 5′-CCAGTAGTCATATGCTTGTCTC-3′ and Geo11: 5′-ACCTTGTTACGACT...

specific Embodiment approach 2

[0022] Specific embodiment two: the difference between this embodiment and specific embodiment one is: in the step one, extract the Phellodendron amurense arbuscular mycorrhiza total DNA according to the following steps: a, the length is 1~1.5cm with double distilled water After washing for 3 to 5 times, put it into a centrifuge tube, add 40 μL of TE buffer solution, and crush the mycorrhizae with a sterile pipette tip, then add 10 μL of Chelex-100 solution with a mass concentration of 20% chemical integration resin, and then boil water Bath for 5 minutes, ice bath for 5 minutes, centrifuge at 15,000 r / min for 5 minutes, and take the supernatant to obtain the total DNA of the arbuscular mycorrhiza of Phellodendron amurense. Other steps and parameters are the same as those in Embodiment 1.

specific Embodiment approach 3

[0023] Specific embodiment three: the difference between this embodiment and specific embodiment one is: in step one, the total DNA of arbuscular mycorrhizal fungi in the rhizosphere soil of Phellodendron amurense is extracted by the modified Bead-Beating method. Other steps and parameters are the same as those in Embodiment 1.

[0024] Heilongjiang University

[0025] A method for analyzing the structure of arbuscular mycorrhizal fungi of Phellodendron amurense

[0026] 5

[0027] 1

[0028] 22

[0029] DNA

[0030] Artificial sequence

[0031]

[0032] Primer GeoA2 for PCR amplification of arbuscular mycorrhizal fungal flora of Phellodendron amurense.

[0033] 1

[0034] ccagtagtca tatgcttgtc tc 22

[0035] 2

[0036] 23

[0037] DNA

[0038] Artificial sequence

[0039]

[0040] Primer Geo11 for PCR amplification of arbuscular mycorrhizal fungal flora of Phellodendron amurense.

[0041] 2

[0042] accttgttac gacttttact tcc 23

[0043] 3

[0044] 60 ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

Disclosed is an analysis method of a corktree arbuscule mycorrhizal fungi community structure, which relates to an analysis method of a fungal community structure. The method solves the problems of big fluctuation of analytic results and poor accuracy of the current corktree arbuscule mycorrhizal fungi community structure analysis method. The analysis steps comprise: firstly, extracting DNA; secondly, Nested-PCR; thirdly, denaturing gradient gel electrophoresis; and fourthly, adopting the DNA sequencing technique, so the corktree arbuscule mycorrhizal fungi community structure can be analyzed. The method implements the detection and analysis to an arbuscule mycorrhizal fungi heredity conserved region 18S rDNA NS31 / Glo1, the copy number of corktree arbuscule mycorrhizal fungi DNA is high through three times of PCR amplification, thereby acquiring mass target products of arbuscule fungi 18S rDNA NS31 / Glo1 region segments, and facilitating the execution of the third step of denaturing gradient gel electrophoresis; therefore, the method has the advantages of high accuracy, good repeatability and high stability.

Description

technical field [0001] The invention relates to a method for analyzing the structure of arbuscular mycorrhizal fungi. Background technique [0002] The arbuscular mycorrhizal fungi of Phellodendron amurense may be composed of A.scrobiculata, A.spinosa, G.aggregatum, Rubus (G. rubiforme), S. calospora, G. mosseae, G. etunicatum, G. versiforme and G. versiforme Composed of various arbuscular mycorrhizal fungi such as G.diaphanum, the flora structure is complex. The existing analysis methods have the problems of large fluctuations in analysis results and poor accuracy. Contents of the invention [0003] The object of the present invention is to provide an analysis method for the structure of the arbuscular mycorrhizal fungi of the amurberry in order to solve the problems of large fluctuations and poor accuracy in the analysis results of the current analysis method for the arbuscular mycorrhizal fungi of the amurberry. [0004] The structure of the arbuscular mycorrhizal fu...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/68C12Q1/02
Inventor 蔡柏岩接伟光葛菁萍王雪阎秀峰
Owner HEILONGJIANG UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap