Alfalfa growthpromoting rhizobacteria MJM-11 and application thereof
A technology of MJM-11, strain, applied in the field of microorganisms, can solve problems such as salt tolerance improvement
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Embodiment 1
[0059] Embodiment 1, screening and identification of alfalfa rhizosphere growth-promoting bacteria
[0060] 1. Screening
[0061] (1) Take 1g of rhizosphere soil sample in 50mL of PAF medium, shake culture at 28°C. The PAF medium contains peptone, casein hydrolyzate, MgSO 4 , K 2 HPO 4 ,glycerin.
[0062] (2) Take 1 mL of shaken PAF culture medium, place it in 50 mL of PAF medium, and culture with shaking at 28°C.
[0063] (3) Take 1mL of the obtained PAF culture solution, place it in 50mL of DF salt culture solution, and incubate with shaking at 28°C.
[0064] The DF salt broth contains KH 2 PO 4 , Na 2 HPO 4 , MgSO 4 , FeSO 4 , glucose, gluconic acid, citric acid, (NH 4 ) 2 SO 4 , H 3 BO 3 , MnSO 4 , ZnSO 4 , CuSO 4 , MoO 3 .
[0065] (4) Take 1 mL of the obtained DF salt culture solution, and add it to 50 mL without (NH 4 ) 2 SO 4 , but containing 3mM 1-aminocyclopropane-1-carboxylic acid (ACC) in the above-mentioned DF salt culture solution, culture...
Embodiment 2
[0075] Embodiment 2, the application of strain MJM-11 (Enterobacter ludwigii) in the preparation of ACC deaminase
[0076] 1. Preparation of ACC deaminase
[0077] TSB medium: tryptone 17g, soytone 3g, NaCl 5g, glucose 2.5g, K 2 HPO 4 Dissolve 2.5g in 1000ml of distilled water and adjust the pH value to pH=7.5.
[0078] ADF medium: KH 2 PO 4 4.0g, Na 2 HPO 4 6.0g, MgSO 4 ·7H 2 O 0.2g, FeSO 4 ·7H 2 O 0.1g, glucose 2.0g, gluconic acid 2.0g, citric acid 2.0g, trace elements 0.1mL, add in distilled water to dissolve, adjust the pH value to pH=7.5, set the volume to 1000ml, add 1-amino Cyclopropane-1-carboxylic acid (ACC), so that the final concentration is 3.0mmol / L (you can first prepare the mother liquor 0.5mol / L); among them, trace elements (100mL): H 3 BO 3 10mg, MnSO 4 11.2mg, ZnSO 4 124.6mg, CuSO 4 78.2mg, MoO 3 10mg.
[0079] None (NH 4 ) 2 SO 4 DF medium: KH 2 PO 4 4.0g, Na 2 HPO4 6.0g, MgSO 4 ·7H 2 O 0.2g, FeSO 4 ·7H 2 O 0.1g, glucose 2.0g, gluc...
Embodiment 3
[0087] Embodiment 3, the application of bacterial strain MJM-11 (Enterobacter ludwigii) in preparing IAA synthesis
[0088] 1. Preparation of IAA
[0089] DF medium: KH 2 PO 4 4.0g, Na 2 HPO 4 6.0g, MgSO 4 ·7H 2 O 0.2g, FeSO 4 ·7H 2 O 0.1g, glucose 2.0g, gluconic acid 2.0g, citric acid 2.0g, (NH 4 ) 2 SO 4 2.0g, 0.1mL of trace elements, dissolved in distilled water, adjust the pH value to pH=7.5, and dilute to 1000ml. The preparation of trace elements is the same as above.
[0090] The test strain MJM-11 (Enterobacter ludwigii) was cultured in DF medium at 28°C and 180rpm for 2 days, and then transferred to different concentrations (0, 50, 100, 200, 500 μg L-Trp·mL -1 ) tryptophan (L-Trp) in fresh DF medium, cultured at 28° C., 180 rpm for 2 days, and collected the fermentation product. Sampling to measure the OD of the bacterial solution 600 value, and then put the rest of the fermentation product (bacteria liquid) at room temperature (25°C) at 8000rpm for 10min...
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