Preparation and application of a strain of Alcaligenes faecalis that antagonizes Fusarium graminearum and efficiently degrades don
A technology of Fusarium graminearum and Alcaligenes faecalis, applied in the field of microorganisms, to achieve strong degradation ability, good application prospects, and the effect of reducing disease index
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Embodiment 1
[0033] The screening of embodiment 1 bacterial strain
[0034] Bioaccumulation of antagonistic bacteria of Fusarium graminearum: Weigh 5 g of field rhizosphere soil (collected in the rice-wheat rotation area of Shipai Town, Zhongxiang, Hubei) into 250 mL of 1 / 5 beef extract peptone medium mL Erlenmeyer flask at 28°C, 180 rpm for 1 day.
[0035] Plate primary screening: Use the soil solution in the above steps to spread gradient dilution on LB medium, select single colonies with obvious differences in colony shape and color from the plate, and put them into PA bottles containing LB medium After overnight activation, the single colonies were streaked again to obtain purified single colonies. A hole punch with a diameter of 7 mm was used to punch out the bacterial block at the edge of the Fusarium graminearum colony, and then the bacterial block was inoculated in the center of the PDA plate. Take two pieces of sterilized filter paper (0.7 cm in diameter) and stick them symmet...
Embodiment 2
[0038] Morphological observation and molecular biology identification of embodiment 2 bacterial strains
[0039] Morphological characteristics: The antibacterial NF037 was serially diluted and spread on the LB plate, cultured upside down at 28°C for 2 days, the colony of the antibacterial NF037 was yellow and rounded. Bacteria are spherical in shape and do not produce spores.
[0040] 16S rDNA molecular identification: First, streak the LB medium with the antagonistic bacteria NF037, and then select a single colony from the plate, put them into a PA bottle for activation, shake at 180 rpm at 28°C for 1 day until a large amount of bacterial liquid is produced. Referring to the literature, the bacterial solution was used as a template. Using the universal primer 27F (5 ， -AGAGTTTGATCCTGGCTCAG-3 ， ) and 1492R (5 ， -TACGGCTACCTTGTTACGACTT-3 ， ) for amplification. The PCR reaction amplification system is 50 μL: 1 μL each of upstream and downstream primers 27F and 1492R, 1 uL ...
Embodiment 3
[0042] Example 3 Inhibition of Fusarium graminearum spore germination by Alcaligenes faecalis NF037
[0043] Preparation of conidia of Fusarium graminearum: Fusarium graminearum was mixed with 100 ml of CMC medium in a 250 ml flask, shaken at 180 rpm at 21°C-25°C for 3 to 5 days. Filter the culture with sterile filter paper to remove the mycelia to prepare the macroconidia suspension, collect the spores after centrifugation at 10000 rpm for 20 min at 4 °C, wash the spores twice with distilled water, and dissolve them in 0.1% (v / v) Tween 20 solution to adjust the concentration of the suspension to 4.0 x 10 5 Conidia / mL.
[0044] Alcaligenes faecalis NF037 and 4×10 5 The Fg spores of Fusarium graminearum were mixed at a ratio of 1:1 and cultured on a shaker at 180 rpm for 12 h. Take a 7mm piece of filter paper and place it in the center of the PDA plate, then take 10 μL of the mixture and spot it on the filter paper. Each treatment was repeated 3 times, and photographed and ...
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