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PCR identification method for entomopathogenic fungi in orange habenaria

An identification method, a fungal technology, applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., to achieve the effects of reducing surface tension, enhancing activity, and increasing dissolved oxygen content

Inactive Publication Date: 2019-03-01
HUIZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

And do not see the report of the PCR identification method of endophytic fungus of orange-yellow phoenix orchid in the prior art

Method used

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  • PCR identification method for entomopathogenic fungi in orange habenaria
  • PCR identification method for entomopathogenic fungi in orange habenaria
  • PCR identification method for entomopathogenic fungi in orange habenaria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 Isolation and purification of endophytic fungi

[0051] A method for isolating and cultivating endophytic fungi of orange-yellow phoenix orchid, comprising the steps of:

[0052] (1) Take the orange-yellow Yufenglan plant, rinse off the sand on the surface of the plant, blot the water on the surface of the plant with filter paper, place it in a clean petri dish with filter paper to dry naturally, and then transfer it to an ultra-clean workbench;

[0053] (2) Decompose the air-dried plants into main stems, leaves, roots and tubers, and cut them into small pieces with a length of 3 to 4 cm;

[0054] (3) Soak the cut section in ethanol with a mass fraction of 70% for 1 to 3 minutes;

[0055] (4) Take out the small section soaked in ethanol, and then soak it in sodium hypochlorite with a mass fraction of 2.5% for 1 to 2 minutes;

[0056] (5) Take out the small section soaked in sodium hypochlorite, and place it on sterile absorbent paper to absorb the water;

...

Embodiment 2

[0060] Example 2 Morphological identification of endophytic fungi

[0061] After culturing the isolated and purified strains on the separation medium for several days, observe the colony morphology with the naked eye and record the fungal colony morphology; then use the direct picking method to observe the fungal morphology under a microscope Characteristics and structure, spore insertion site and shape, size, arrangement, and microscopic features with or without septum, for preliminary identification.

[0062] Endophytic fungi were classified according to their colony morphology and microstructural characteristics. The results of the first photograph and microscopic examination are as follows: figure 1 , 2 As shown, the results of the second photograph and microscopic examination are as follows image 3 , 4 shown. Among them, the first and second times were on April 20, 2018 and June 11, 2018, respectively. The growth stages of the original plants in the two microscopic ...

Embodiment 3

[0065] Example 3 Molecular biology identification

[0066] A PCR identification method for endophytic fungi of orange yellow phoenix orchid, comprising the steps of:

[0067] A, the cultivation of bacterial strain: get the mycelium of endophytic fungus of orange-yellow phoenix orchid and cultivate;

[0068] B. DNA extraction: the bacterial DNA was extracted by the CTAB method;

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Abstract

The invention relates to the technical field of fungi identification, in particular to a PCR identification method for endophytic fungi of orange habenaria. The PCR identification method comprises thefollowing steps: A, the mycelia of the endophytic fungi of the orange habenaria is fetched to be cultivated; B, the DNA of the mycelia is extracted through a CTAB method; C, the extracted sample DNAis subjected to electrophoresis detection; D, the extracted sample DNA is subjected to PCR amplification; E, the product of PCR amplification is subjected to electrophoresis detection. According to the PCR identification method, the species of the endophytic fungi in the orange habenaria can be identified through the PCR identification method, and the species: trichoderma asperellum, the trichoderma virens and the fusarium solani of parts of endophytic fungi existing in the orange habenaria can be finally identified, and therefore the further studying on the symbiotic relationship of the orange habenaria and the endophytic fungi of the orange habenaria is possible.

Description

technical field [0001] The invention relates to the technical field of fungal identification, in particular to a PCR identification method for endophytic fungi of orange-yellow phoenix orchid. Background technique [0002] Orchids are the most diverse family of angiosperms, including many important medicinal and ornamental flowers. Under natural conditions, orchids can only grow normally if they form a symbiotic relationship with certain fungi. Due to its economic value and its special biological relationship with fungi, in recent years, orchids have become important materials for studying the intercropping relationship between plants and fungi, and the diversity and specificity of fungi. Endophytic fungi can form symbiotic relationships with plants, becoming a mutualistic symbiotic organism. Recently, some scholars put forward an evolutionary hypothesis: the relationship between the diversity of symbiotic microbial groups formed by plant and animal hosts plays an importan...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/686C12Q1/00
CPCC12Q1/686C12Q1/6895
Inventor 陈兆贵郑静敏陈双庆邓文言连琼华
Owner HUIZHOU UNIV
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