Bacillus subtilis Pro6A5, microbial inoculum and preparation method thereof, and application of bacillus subtilis Pro6A5 in muskmelon cultivation
A technology of Bacillus subtilis and melon, which is applied in the field of biological control, can solve the problems of narrow genetic background of germplasm resources, difficulty in popularization and implementation, and destruction of ecological balance, and achieve the effects of improving the ecological environment, overcoming the stress obstacles of saline-alkali land, and high-efficiency control
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Embodiment 1
[0027] Embodiment 1: Isolation and screening of Bacillus subtilis
[0028] (1) Isolation and purification of Agrobacterium
[0029] Collect melon fields (collected by Liang Shen in Duzhai Village, Putaojia Township, Lankao County on September 20, 2016) 300g of moist soil in the 5-20cm soil layer, weigh 10g after crushing, and place in a container filled with 100ml of sterile water In the Erlenmeyer flask, shake for 3-5 minutes, and heat in a water bath at 80°C for 10 minutes. Measure the upper soil suspension and dilute with sterile water to prepare 10 -2 、10 -3 、10 -4 and 10 -5 Soil suspension; pipette the above 10 -3 、10 -4 、10 -5 Spread 50 μl of soil suspension on the plate of NA medium (3g of beef extract, 1g of yeast powder, 5g of peptone, 10g of glucose, 12g of agar, 1L of water, pH6.8~7.0, sterilized at 121℃ for 20min) Coat slabs on bench and allow to dry. Place them upside down in a biochemical incubator and incubate at 37 °C for 48 h. After monospore colonie...
Embodiment 2
[0042] Example 2: Identification of Bacillus subtilis Pro6A5
[0043] (1) Analysis of physical and chemical properties
[0044] 1) Colony characteristics and bacterial morphology observation
[0045] Bacillus Pro6A5 was inoculated on NA medium and cultured in a biochemical incubator at 30 °C for 48 h. Observe the colony shape, color and growth of the strains on the culture medium. Bacillus colonies cultured for 48 h were picked and stained with Gram on glass slides to observe the size and shape of the bacteria.
[0046] 2) Physiological and biochemical test determination of bacterial strains
[0047] The physiological and biochemical experiments of Pro6A5 strains were carried out using the culture medium and experimental methods recommended by "Common Bacteria System Identification Manual" and "Bacillus". Physiological and biochemical tests mainly include: methyl red reaction, contact enzyme test, starch hydrolysis test, hydrogen peroxide test, gelatin liquefaction test, c...
Embodiment 3
[0062] Embodiment 3: the preparation of microbial bacterial agent
[0063] The preparation steps of Pro6A5 bacterial agent are as follows:
[0064] (1) Strain cultivation and preparation
[0065] Inoculate Bacillus subtilis Pro6A5 into test tube slant soybean casein solid medium (tryptone 17g, soybean peptone 3g, NaCl 5g, KH 2 PO 4 2.5g, glucose 2.5g, agar powder 12g, add water to volume 1000ml, pH7.0, sterilize at 121℃ for 20min), culture at 28~35℃ for 24~36 hours to obtain bacteria;
[0066] (2) Preparation of seed solution
[0067] Insert the test tube bacteria prepared in step (1) into soybean casein liquid medium (tryptone 17g, soybean peptone 3g, NaCl 5g, KH 2 PO 4 2.5g, glucose 2.5g, add water to volume 1000ml, pH7.0, sterilize at 121°C for 20min), shake culture at 28-35°C for 12-16 hours to make seed solution;
[0068] (3) Preparation of fermentation medium
[0069] Glucose, corn flour, soybean cake powder, and dipotassium hydrogen phosphate were mixed with wate...
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