Group of aroma-producing fungi in Paphiopedilum barbigerum Tang etWang roots and application
A technology of Paphillium root and small leaves, applied in the field of microorganisms
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Embodiment 1
[0026] The separation of embodiment 1 endophytes
[0027] 1. Isolation of bacteria
[0028] Select 2-3-year-old root samples of robust Paphiopediphyllum microphylla, soak the rhizomes in soapy water for 2 hours, rinse them with running water for 3-5 hours, and gently scrape off the root hairs, root covers and other appendages with a scalpel on the ultra-clean workbench. objects, examined under a microscope, selected root segments with mycelium clusters, placed in a petri dish, rinsed with sterile water 3-5 times; first sterilized with 75% alcohol for 1 min, and then sterilized with 0.1% mercuric chloride solution 4-5min, wash 5 times with sterile water; cut the sterilized roots into 2-3cm small pieces, put them in a 6cm-diameter petri dish filled with sterile water, and gently use a scalpel and tweezers Scrape the root segment to free the mycelium cluster in the root segment cortex; culture at a constant temperature of 24°C, observe under the microscope after 24 hours, use a ...
Embodiment 2
[0031] The identification of embodiment 2 endophytes
[0032] (1) Morphological identification
[0033] 1. Identification method
[0034] a. Microstructural characteristics of colonies and hyphae
[0035] The bacterial strains XYDL040 and XYDL003 were inoculated in the middle of the PDA medium plate, a piece of sterile cover glass was picked up with tweezers, and inserted obliquely into the medium at an angle of 45 degrees at a certain distance from the inoculation point; when the hyphae When the cover glass grows to about 2 / 5, the cover glass is taken out, and the structural characteristics of the hyphae are observed under a microscope.
[0036] b. Diaphragm ultrastructure
[0037]1) Fixation of materials: Discard the culture medium and add electron microscope fixative solution to fix at 4°C for 2-4 hours, centrifuge the cells at low speed until mung bean-sized cell clumps can be seen at the bottom of the tube, wrap in 1% agarose, and use 0.1M phosphate buffer PB (pH7 .4)...
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