Bacillus subtilis for preventing and treating phytophthora blight of pepper
A technology of Bacillus subtilis and Pepper Phytophthora, applied in the direction of chemicals for biological control, methods based on microorganisms, bacteria, etc., can solve the problems of difficult to effectively control Pepper Phytophthora, adverse effects on health, and destruction of agricultural ecological balance, etc. problem, to achieve good suppression effect, short cycle and high safety effect
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Embodiment 1
[0031] Embodiment 1: Isolation, screening and identification of Bacillus subtilis BS04
[0032] (1) Separation and screening: Soil samples were collected from the rhizosphere of healthy peppers using the five-point sampling method from fields with severe epidemics in pepper planting areas such as Wuhan, Jingzhou, Yichang, and Jingmen, Hubei. When sampling, shovel off the top 5cm soil layer, take 500g of fresh soil samples, and bring them back with fresh-keeping bags.
[0033] Weigh 10 g of the soil sample and add it to a conical flask filled with 90 mL of sterile water. The conical flask is filled with a number of glass beads, vibrate at 28 °C and 120 r / min for 30 min, and obtain a soil suspension after standing still.
[0034] Dilute the soil suspension to 10 -5 、10 -6 、10 -7 100 μL of soil suspension with different dilutions was drawn and spread on NA medium, cultured at 28°C for 48 hours, and single colonies of bacteria with different colors and shapes were picked and pu...
Embodiment 2
[0047] Sequence comparison of the sequencing results on NCBI Genebank (comparison number: lcl|28017), found that its similarity with the 16S rDNA sequence of Bacillus subtilis was as high as 99%, and the antagonistic strain BS04 was preliminarily determined to be Bacillus subtilis (Bacillus subtilis) . Cluster analysis was performed using MEGA4.1 software, and a phylogenetic tree was constructed. The results showed that B. subtilis (KF725636) was in a small branch with B. subtilis (KC506778.1), and was in a large branch with the other two Bacillus subtilis. Thus, combined with the morphological and physiological and biochemical characteristics, it was further demonstrated that the BS04 strain was Bacillus subtilis. Embodiment 2: the antibacterial spectrum determination of bacillus subtilis BS04 bacterial strain
[0048] Bacillus subtilis BS04 strain against pepper blight (Phytophthora capsici), tobacco black shank (P.parasitica var.nicotiana), cotton Verticillium dahliae (Vert...
Embodiment 3
[0054] Embodiment 3: the disease prevention mechanism of bacillus subtilis BS04 bacterial strain to capsicum blight
[0055] (1) Effect of BS04 strain on the mycelial growth of Phytophthora capsici
[0056] After inoculating and culturing Phytophthora capsici on PDA medium for 3 days, a 5 mm diameter bacterial block was punched out and placed in a sterile petri dish filled with 20 mL PD medium, and cultured at 28°C for 2 days. Treat the cultured Phytophthora capsici bacterial block with the sterile culture filtrate and bacterial suspension of the BS04 strain respectively for 24 hours, and use sterile water treatment as the control, pick the hyphae and observe its morphology under an inverted microscope, and repeat each treatment. 3 times.
[0057] The test results showed that after the Phytophthora capsici was treated with the bacterial suspension of strain BS04 and the sterile filtrate, the mycelium branches increased, the top appeared deformed, the growth was slow, and the ...
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