Bacillus pumilus rp01 and its application
A technology of Bacillus pumilus and bacterial agents, applied in the field of bio-agricultural science and technology, can solve the problems of poor ecological adaptability and inability to achieve crop growth-promoting effects, and achieve the goals of improving phosphorus nutrition, high inorganic phosphorus solubility, and promoting plant growth Effect
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Embodiment 1
[0022] Screening and Identification of Bacillus pumilus RP01
[0023] 1. Screening of Bacillus pumilus RP01
[0024] 1. Material preparation
[0025] (1) Test soil
[0026] The soil was collected from the vegetable rotation (lettuce-pepper-soybean-pakchoi) at the National Purple Soil Fertility and Fertilizer Effect Monitoring Base of Southwest University in Beibei District, Chongqing City. High planting area. Select 3 pieces of soil with a range of 1 × 1m, randomly collect 10-20cm soil according to the 5-point sampling method, mix the soil samples evenly, put them in an ice bucket, and bring them back to the laboratory and store them in a refrigerator at 4°C for later use. The basic physical and chemical properties of the soil are listed in Table 1.
[0027] Table 1 Basic physical and chemical properties of the tested soil
[0028]
[0029] (2) Configure culture medium
[0030] Phospholytic bacteria selection medium: NaCl 0.2g; MgSO 4 ·7H 2 O 0.1g; MnSO 4 4H 2 O 0...
Embodiment 2
[0058] Determination of Phosphorus Solubilizing Ability of Bacillus pumilus RP01
[0059] Referring to the method in the literature (Yu Weibin, 2010), prepare LB liquid culture medium, take 100mL and divide into 250mL Erlenmeyer flasks. After autoclaving, inoculate RP01, culture at 28°C and 120r / min for 24 hours, and dilute the bacterial concentration of the fermentation liquid to 10 8 cfu / mL, spare.
[0060] Prepare phosphate-dissolving bacteria selective liquid medium, take 100mL and divide it into 250mL Erlenmeyer flasks, and after autoclaving at 121°C for 30min, the test group was inoculated with 2mL of the above bacteria with a concentration of 10 8 cfu / mL fermentation broth, the control group was added 2 mL of high-temperature sterilized LB liquid medium, and each treatment was repeated three times. All were cultured at 28°C and 120r / min for 7 days. Use the diluted plate colony counting method (phosphorus-dissolving bacteria selective medium, cultivated at 28°C for 3 ...
Embodiment 3
[0066] Determination of Growth-promoting Ability of Bacillus pumilus RP01 on Tobacco Seedlings
[0067] After the petri dish is sterilized, cover the filter paper, moisten it with distilled water, evenly spread the K326 tobacco seeds on the filter paper, and accelerate germination at 25±1°C for 3-4d (the filter paper of the petri dish should be kept moist during this period). The tobacco seeds after the germination are transferred to the seedling trays equipped with substrate (60% humus soil+40% perlite), and 2 tobacco seeds through the germination are put in each hole, which are covered with substrate, and each hole is reserved after emergence. A tobacco seedling.
[0068] When the tobacco grows to 1 heart of 3-4 leaves, start to inoculate the RP01 bacterial liquid (the bacterial liquid is prepared according to the method in Example 2). Set 3 treatments: control, bacterial solution concentration 10 3 cfu / mL and 10 6 cfu / mL two test groups. Repeat four times, inoculate 1mL...
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