A strain of Pseudomonas syringae and its application in the control of apple ring spot
A technology of Pseudomonas and cloves, applied in the field of microorganisms, to achieve the effects of environmental friendliness, low production cost, and stable control effect
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[0029] The preparation method of biocontrol agent provided by the present invention comprises the following steps:
[0030] (1) Streak inoculation of strain B-1 in NA medium, and activate and cultivate at 30°C for 24 hours to obtain a single colony A of activated strain B-1;
[0031] (2) Inoculate the single colony A of the activated bacterial strain B-1 in the NB medium with a bottled volume of 80mL / 250mL, 180r·min -1 , 30 ° C constant temperature shaking culture for 12 hours, to obtain the strain B-1 fermentation seed liquid B;
[0032] (3) The fermented seed liquid B was inoculated into NB culture medium at an inoculum size of 1:100 for expanded cultivation, and incubated at a constant temperature at 30°C for 24 hours to obtain 10 9 cfu mL -1 Cell culture medium C of B-1 strain;
[0033] (4) put 10 9 cfu mL -1 Cell culture medium C of strain B-1, at 4°C, 12000r·min -1 Centrifuge for 15 minutes, collect the supernatant and filter it through a 0.22 μm microporous membra...
Embodiment 1
[0038] Isolation Screening and Strain Identification of Pseudomonas syringae B-1
[0039] 1. Isolation and screening of strains
[0040]Mature healthy fruits were collected from the commercialized Fuji apple orchard in Yantai, Shandong. After surface disinfection, the fruit tissues were ground, added sterile water, and separated by conventional gradient dilution coating. They were cultured with NA medium at 28°C and colonies were picked. Colonies with obvious morphological differences were purified and preserved on NA medium, and the primary screening and multiple re-screening of antagonistic bacteria were carried out with Apple ringworm as the target pathogen, and finally a bacterial strain with strong antibacterial activity was obtained, named B -1.
[0041] 2. Identification of strains
[0042] (1) Morphological characteristics: After the strain B-1 was cultured on NA medium for 24 hours, the colony was milky white, round or nearly round, with a smooth surface and a raise...
Embodiment 2
[0048] Effect of Pseudomonas syringae B-1 on Mycelia Growth of Apple Ringworm
[0049] Configure PDA medium, add bacterial suspension of bacterial strain B-1 to it, make its final concentration be 10 5 、10 6 、10 7 and 10 8 cfu mL -1 , with the PDA without adding bacterial strain B-1 as a control, the activated good apple ringworm was inoculated in the above-mentioned PDA medium. Incubate in dark at constant temperature at 25°C for 5 days, and measure the colony diameter.
[0050] The above-mentioned PDA medium used has the following components and formula: weigh 200g after peeling the potatoes, cut into small pieces and boil in water for 15-20min, add 20g of glucose and 15g of agar powder after filtering through four layers of gauze, and dilute to 1000mL. The pH value is natural, and it is sterilized by high-pressure steam at 121°C for 20 minutes.
[0051] The results showed that adding different concentrations of bacteria strain B-1 had a significant inhibitory effect o...
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