(B.massiliogorillae)YC9L and application thereof
A technology of bacillus and gorilla, applied in the field of microbial resources, can solve the problems that the functional genes of active substances have not been excavated and researched, and achieve the effect of increasing root length, germination rate and plant height
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Embodiment 1
[0037] Example 1 Screening of YC9L bacterial strain
[0038] (1) Separation
[0039] Taking the rhizosphere soil of the vegetation in the Tianhe Green Belt in Guangzhou as the screening soil sample, weigh 10g of the soil sample and put it into a 250mL triangular flask (with glass beads) filled with 90mL of sterile water, and shake it on a shaker for 30min. Fully disperse the bacteria and let it stand for 20-30s. Then in a 90°C water bath for 10 min, the supernatant was taken for 10-fold incremental dilution, using 10 -3 -10 -4 For dilution, pipette 100 μL of each gradient and spread on LB solid plate, and incubate upside down at 37°C for 24 hours. Observe and according to the growth characteristics of the colony shape, size, color, etc., pick the colonies and purify them on the LB solid medium, and then save the isolated bacteria for future use.
[0040] (2) screening
[0041] The ability of the isolated strains to produce IAA was determined by the Salkowski colorimetric ...
Embodiment 2
[0042] Embodiment 2 YC9L bacteria belong to Gram-positive bacteria
[0043] Pick a single colony of YC9L and culture it in liquid LB medium for 24 hours, pipette 4 μL of sterile water onto a glass slide, pipette 1 μL of bacterial liquid into the added sterile water, mix well, and then dry naturally. Fix with flame 2-3 times; first dye with crystal violet for 1 min, wash with water, and dry naturally; mordant with iodine solution for 1 min, wash with water, and dry; decolorize with 95% ethanol for 20 seconds, wash with water, and dry; counterstain with safranin for 1 min, wash with water , dried, and observed under a 100-fold oil immersion microscope, the thalline was blue-purple ( figure 2 A), YC9L is a Gram-positive bacterium.
Embodiment 3
[0044] Example 3 YC9L bacteria can produce spores
[0045] Pick a single colony of YC9L and culture it in liquid LB medium for 24 hours, pipette 4 μL of sterile water onto a glass slide, pipette 1 μL of bacterial liquid into the added sterile water, mix well, and then dry naturally. Pass the flame 2-3 times to fix. Add malachite green dye solution to the fixed smear, clamp the glass slide with tweezers and heat it on the flame of alcohol lamp, so that the dye solution will steam for 3-4 times within 30 seconds; wash with water and dry; reconstitute with safranin aqueous solution Dye for 1min, wash with water, and dry. Carry out microscopic examination with 100 times oil lens, spore is in green, and thalline is in red ( figure 2 B), YC9L bacteria can produce spores.
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