Rhizobium rosettiformans FN2, microbial inoculum thereof and application of Rhizobium rosettiformans FN2 and microbial inoculum in field of soil improvement
A rhizobia, FN2 technology, applied in applications, soil conditioning materials, bacteria, etc., can solve the problems of loss of microbial activity, inability of soil microorganisms to adapt, and change of soil microbial adaptability, etc., to improve stress resistance, increase colonization rate, The effect of good salinity resistance
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Embodiment 1
[0056] Example 1 Screening and identification of saline-alkali-tolerant nitrogen-fixing bacteria
[0057] 1. Configuration of culture medium
[0058] LB liquid medium: tryptone 5g / L, yeast extract 2.5g / L, sodium chloride 5g / L, deionized water 1000mL, pH=7.4.
[0059] LB solid medium: tryptone 5g / L, yeast extract 2.5g / L, sodium chloride 5g / L, agar 20g / L, deionized water 1000mL, pH=7.4.
[0060] Ashby nitrogen-fixing culture medium: potassium dihydrogen phosphate 0.2g, magnesium sulfate heptahydrate 0.2g, sodium chloride 0.2g, calcium carbonate 5g, mannitol 10g, calcium sulfate dihydrate 0.1g, deionized water 1000mL, pH=7- 7.5.
[0061] Ashby solid medium: potassium dihydrogen phosphate 0.2g, magnesium sulfate heptahydrate 0.2g, sodium chloride 0.2g, calcium carbonate 5g, mannitol 10g, calcium sulfate dihydrate 0.1g, agar 20g / L, deionized water 1000mL, pH=7-7.5.
[0062] 2. Isolation and screening of nitrogen-fixing bacteria
[0063] Weigh 10g of Suaeda salsa rhizosphere sa...
Embodiment 2
[0082] The saline-alkali tolerance experiment of embodiment 2 nitrogen-fixing bacteria FN2
[0083] 1. Solution configuration
[0084] Configuration of different gradient salinity solutions: set the salinity to 0.1g / L, 0.5g / L, 1.0g / L, 1.5g / L, 3.0g / L, set the alkalinity to 7.2, 7.6, 8.0, 8.4, A total of 20 treatments. Salinity is prepared with sodium chloride, and alkalinity is adjusted with hydrochloric acid and sodium hydroxide.
[0085] Configuration of non-salinity LB liquid medium: tryptone 5g / L, yeast extract 2.5g / L, deionized water 1000mL, pH natural.
[0086] 2. Determination of the growth curve of FN2 under different salinity
[0087] Inoculate the strain FN2 into the liquid LB medium and cultivate to OD=0.1 (bacteria density is about 10 8 cells / mL) and stored in a 4°C refrigerator for later use.
[0088] Inoculate the bacterium liquid of OD=1.0 with 5% inoculum amount in the culture medium of different salinities containing 10% non-salinity LB, measure the absorban...
Embodiment 3
[0090] Corn germination experiment under embodiment 3 nitrogen-fixing bacteria FN2 inoculation
[0091] Select large, round and plump corn seeds, soak them in 70% absolute ethanol for 2 minutes for disinfection, and rinse them with tap water. The salinity gradient is 0.1g / L, 0.5g / L, 1.0g / L, 1.5g / L, 3.0g / L, the alkalinity gradient is 7.2, 7.6, 8.0, 8.4, a total of 20 treatments. Cover each petri dish with filter paper, sow 10 corn seeds, inoculate 10 9 FN2 bacteria, and add the same volume of water with different salt-alkali gradients. Two parallels were set up for each saline-alkaline gradient. Azospirillum brasilense, which also has the ability to fix nitrogen, was purchased from the China General Microorganism Culture Collection Center (CGMCC) as a control. Observe how the corn seeds germinate after a week.
[0092] The results show that the germination rate of corn seeds inoculated with FN2 can reach more than 80% at any alkalinity and salinity of 3.0g / L, while the germ...
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