Biocontrol strain ZHR0 for prevention and control of sugarcane smut and application thereof
A technology of smut and biocontrol fungicide, applied to the biocontrol strain ZHR0 for the prevention and treatment of sugarcane smut and its application field, which can solve environmental pollution, difficult to overcome disease resistance and yield and quality traits, difficult to control chemical drugs, etc. problem, to achieve good control effect
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Embodiment 1
[0017] The Bacillus amyloliquefaciens ZH0 (Bacillus amyloliquefaciens ZH0) strain is a strain of Bacillus amyloliquefaciens isolated from sugarcane leaves in September 2013.
[0018] A kind of screening method of bacterial strain, described bacterial strain is Bacillus amyloliquefaciens ZHRO (bacillus amyloliquefaciens ZHRO) bacterial strain, and the method comprises the following steps:
[0019] (1) Take several sugarcane leaves and put them into a sterilized plastic bag and bring them back to the laboratory quickly. Take 5g of sugarcane leaves and cut them into pieces after being sterilized and put them into a sterilized triangular flask, add 50ml of sterile water, and sterilize Glass beads 5g, shake on a shaker at 150-180rpm for 30min, then let stand for 10min;
[0020] (2) Draw the supernatant and dilute to 10 -4 、10 -5 、10 -6 For three gradients, draw 0.1mL liquid from each of the three gradient dilutions and inoculate them on LB solid medium, culture at a constant tem...
Embodiment 2
[0025] Described Bacillus amyloliquefaciens ZHR0 (bacillus amyloliquefaciens ZHR0) and the first group of confrontation culture test of sugarcane smut, concrete method is as follows:
[0026] First draw two mutually perpendicular straight lines at the center of the plate bottom containing the PDA medium, draw 5ul of the described Bacillus amyloliquefaciens ZHR0 (bacillus amyloliquefaciens ZHR0) bacterial strain cultivated overnight on the LB liquid medium with a sterilized gun tip to inoculate in plate center; draw 5ul sugarcane smut bacteria solution (1X10 6 CFU / ml, positive burden spores 1:1) were dropped along two vertical lines at a distance of 2 cm from the center, and placed in an incubator at 28°C for 3 days. The colony in the center of the plate is the Bacillus amyloliquefaciens ZH0 (Bacillus amyloliquefaciens ZH0) strain, surrounded by four colonies of S. amyloliquefaciens.
[0027] It can be seen that the growth of smut is significantly inhibited near the ZHR0 side ...
Embodiment 3
[0030] The second group confrontation culture test of described Bacillus amyloliquefaciens ZHR0 (bacillus amyloliquefaciens ZHR0) and sugarcane smut, concrete method is as follows:
[0031] (1) will be divided into 1, 2, 3 and 4 four regions equally on the plate containing PDA medium;
[0032] (2) The first test group: draw two mutually perpendicular straight lines at the center of area 1, draw 5ul of the described Bacillus amyloliquefaciens ZHR0 (Bacillus amyloliquefaciens ZHR0) bacterial strain cultivated overnight on the LB liquid medium with a sterilized pipette tip Inoculate at the intersection of two mutually perpendicular straight lines; draw 5ul sugarcane smut bacteria liquid (1X10 6 CFU / ml, positive burden spores 1:1) drop along two vertical lines at a distance of 1cm from the intersection point;
[0033] (3) The first control group: draw two mutually perpendicular straight lines at the center of the area 3, inoculate 5ul sterile water at the intersection of the two ...
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