Bacillus velezensis and application thereof to relieving continuous cropping obstacles of apples
A technology of Bacillus velesi and Bacillus, applied in the field of agricultural microorganisms, can solve the problems of few Bacillus and continuous cropping obstacles of apples, and achieve the effect of promoting plant growth and reducing continuous cropping obstacles of apple trees.
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Embodiment 1
[0051] Example 1: Isolation and identification of bacterial strains
[0052] 1. Isolation and purification of strains:
[0053] The rhizosphere bacteria were separated by the improved dilution coating plate method, and fresh soil samples were taken from the rhizosphere soil of three continuous cropping apple orchards in Xicheng Town (XC), Wantou Village (WT) and Fengmaozhai (FM) in Yantai City. 3 ~ 4mm sieve to remove impurities in the soil. Weigh 5 g of rhizosphere soil, add it to a triangular flask (with glass beads) filled with 45 mL of sterile water, vibrate at 180 rpm for 30 min, let it stand for 5 min, and serially dilute to 1×10 with sterile water. -4 100 μL of the supernatant of the gradient dilution was spread on the LB plate, repeated three times, and cultured upside down in a constant temperature incubator at 28°C. After a single colony grows in the plate, pick a single colony of different shapes and streak on a new LB plate for purification. At the same time, th...
Embodiment 2
[0078] Example 2: Functional identification of Bacillus velezensis XC1
[0079] 1. Antibacterial effect:
[0080] The antibacterial effect of XC1 was evaluated by the filter paper method, and the XC1 strain was inoculated in LB liquid medium, 37°C, 180r min -1 Cultivate on a shaker for 24 hours, use a 5mm diameter puncher to punch out the bacterial mass at the edge of the pathogenic bacteria, inoculate it into the center of the PDA plate, put sterile filter paper at the 4 docking points 2cm away from the pathogenic bacterial mass, and inject 10 μL of XC1 bacterial solution, After 2 hours of pre-diffusion, culture at 28°C. Use the pathogenic bacteria not inoculated with the bacteria solution as a control, and observe at any time. After the control is full of the plate, use a vernier caliper to measure the diameter of the fungal colony using the cross method, and the diameter represents the antibacterial rate. Repeat the above operation 3 times.
[0081] Bacterial inhibition r...
Embodiment 3
[0092] Embodiment 3: pot test
[0093] 1. Experimental design:
[0094] The pot experiment was carried out at the National Apple Engineering Experimental Center and the Apple Replanting and Microbiology Laboratory of the South Campus of Shandong Agricultural University from March to October 2019. The sweet tea seeds are stratified at 4°C for about 30 days, and after the seeds turn white, they are sown in a cultivation pot equipped with a seedling substrate for seedling cultivation. When the seedlings grow to 6 true leaves, select plants with consistent growth and no pests and diseases, and transplant them on May 1 into clay pots with 6.5 kg of different treatment soil (the upper diameter is 25 cm, the lower diameter is 17 cm, and the height is 18 cm). , Sampling in mid-September to determine relevant experimental indicators.
[0095] The potting test soil was taken from a 32-year-old apple orchard in Qingwan, Manzhuangtan, Tai'an City, Shandong Province, from an area 80.00 c...
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