Bacillus subtilis Y13 for effectively preventing and treating camellia anthracnose and use thereof
A technology for Bacillus subtilis, Camellia oleifera anthracnose, applied in the application, bacteria, fungicides and other directions, can solve the problems of weak or even blank technology, no biological pesticides for preventing Camellia oleifera anthracnose, etc., achieves good development and application prospects, easy industrial production, The effect of simple culture conditions
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Embodiment 1
[0030] Isolation of Bacillus subtilis Y13
[0031] The healthy leaves of common camellia oleifera are taken, separated, purified and preserved by conventional methods.
[0032] After the camellia oleifera was rinsed with water, an appropriate amount of healthy tissue was selected, weighed separately, cut into 5mm×5mm tissue pieces, soaked in 75% ethanol for 1min, then soaked in 0.1% mercuric solution for 40s-70s, and then placed Soak in 75% ethanol for 30s, take it out and rinse it with sterile water 4 times. Add 10ml of sterile water and a little sterilized quartz sand into a sterile mortar to grind into a homogenate, cover and let stand for 30 minutes, serially dilute the homogenate 10 times, and pipette 0.1ml to prepare After smearing evenly on a good NA plate, put it into an incubator for cultivation, record the amount of bacteria after 48 hours, and select according to the shape, color and size of the colony, purify and store on the plate, and separate 5 batches in total...
Embodiment 2
[0034] Screening of Bacillus subtilis Y13
[0035] Preliminary screening: use the confrontation culture method. Screening of endophyte antagonistic pathogenic fungi. Put the 6mm-diameter bacterium cake of Camellia anthracnose bacteria in the center of the plate, and then use the inoculation ring to pick up the suspension of the bacteria to be tested and inoculate 2 to 3 kinds of endophytic bacteria equidistantly (3cm from the center of the plate). Incubate in the dark at 25°C in an incubator, observe and record the presence and size of the bacteriostatic zone after 4 days, and repeat 3 times in total. Select 8 bacterial strains with a bacteriostatic zone width greater than or equal to 4mm for re-screening, followed by Y5, Y12, Y13, Y19, Y21, Y29, Y31, and Y35 for re-screening (see Table 1).
[0036] Table 1 Antibacterial effect of endophytic bacteria on Camellia oleifera anthracnose
[0037]
[0038] Re-screening: use fermentation method. After the 8 initially screened ...
Embodiment 3
[0042] Inhibitory Effect of Bacillus subtilis Y13 on Mycelia of Camellia oleifera Anthracnose
[0043] Put the 6mm-diameter bacterium cake of Camellia anthracnose bacteria in the center of the potato sucrose medium PDA plate, and then spot the antagonistic bacteria Y13 at an equal distance of 3cm, repeat three times, culture at 25-28°C, and measure the diameter of the inhibition zone after 4 days. Depend on figure 1 It can be seen that Bacillus subtilis Y13 has a strong antibacterial effect on Camellia oleifera anthracnose.
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