Production method and application of a salt-alkali-tolerant bacillus siamese and its live bacterial preparation
A technology of bacillus and live bacteria preparations, applied in the field of agricultural biology, can solve problems such as limited control effects, achieve scientific production methods, improve rice yield and quality, and reduce production costs
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Embodiment 1
[0030] Example 1: Screening of Bacillus siamese PAPM-15
[0031] (1) Screening of saline-alkali-tolerant strains
[0032] Bacillus siamese PAPM-15 was isolated from rice rhizosphere soil. The soil was sampled in the moderately saline-alkali soil of Bincheng District, Binzhou City, Shandong Province in September 2018. The specific separation method is as follows: mix the soil sample uniformly, weigh 5 g, put it into a triangular flask containing 95 mL of sterile water and 10 glass beads, and shake at 37° C. and 180 rpm for 30 min. Take 1mL of soil suspension for 10 -1 -10 -7 Serial concentration gradient dilution, then take 10 -5 , 10 -6 , 10 -7 Three dilutions were spread on plates containing selection medium and incubated upside down at 37°C for 2 d. A single colony was picked and streaked onto a plate containing selective medium, and cultured upside down at 37°C for 2 days. Pick a single colony and transfer it to the slanted surface of the preservation medium test tu...
Embodiment 2
[0048] Example 2 Identification of Bacillus siamese PAPM-15
[0049] (1) Morphological and physiological and biochemical characteristics
[0050] The morphological characteristics of the Bacillus siamese PAPM-15 strain are: cultured on NA medium at 37° C. for 48 hours, the colony diameter is about 0.5-0.8 mm, white and opaque, round, with irregular edges, rough surface, raised, folded ; the bacteria are rods, such as figure 1 shown. The NA medium is a nutrient agar medium, and its formula is: beef extract 3 g, peptone 10 g, NaCl 5 g, agar 20 g, water 1000 mL, pH 7.0.
[0051] Physiological and biochemical characteristics of the Bacillus siamese PAPM-15 strain: Gram-positive bacilli; positive in catalase test, positive in starch hydrolysis test, positive in nitrate reduction test, positive in malonic acid utilization test, and positive in gelatin liquefaction test , the methyl red test was negative, the D-glucose fermentation test was positive, the D-mannitol fermentation te...
Embodiment 3
[0055] Example 3 Preparation of Bacillus siamese PAPM-15 inoculum
[0056] The preparation method of described Bacillus siamese PAPM-15 strain microbial inoculum comprises the following steps:
[0057] 1) Activation of strains: The cryopreserved strain of Bacillus siamese PAPM-15 was transferred to the slant of a NA medium test tube, and cultured at 32-37° C. for 24-36 hours for activation. The NA medium is a nutrient agar medium, and its formula is: beef extract 3 g, peptone 10 g, NaCl 5 g, agar 20 g, water 1000 mL, pH 7.0.
[0058] 2) Preparation of seeds in a triangular flask: scrape the activated Bacillus siamese PAPM-15 bacterial lawn with an inoculating loop, inoculate it in NB medium, and cultivate at 32-37° C. for 24-36 hours. The NB medium is a nutrient broth medium, and its formula is: beef extract 3 g, peptone 10 g, NaCl 5 g, water 1000 mL, pH 7.0.
[0059] 3) Seed tank strain preparation: transfer the triangular flask seeds into a 10L seed tank containing 6L NB m...
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