Multi-heavy-metal-resistant strain for producing indoleacetic acid and application of multi-heavy-metal-resistant strain
A technology of indole acetic acid and heavy metals, applied in the field of agricultural microorganisms, can solve problems such as plant poisoning
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Embodiment 1
[0028] Example 1: Isolation, purification and identification of resistant strains
[0029] 1. Medium ① LB medium: peptone 10g, yeast extract 5g, sodium chloride 10g, agar 20g, distilled water 1000mL, pH 7.0-7.2, sterilized at 121°C for 20min. LB liquid medium: without agar, other conditions are the same as above. ②DF salt medium: every liter of medium contains Na 2 HPO 4 6.0g, KH 2 PO 4 4.0g, MgSO 4 .7H 2 O 0.2g, glucose 2g, gluconic acid 2g, citric acid 2g, (NH 4 ) 2 SO 4 2g, 0.1ml each of component 1 and component 2 solutions, H2O 1L, pH 7.0-7.2. Sterilize at 121°C for 20min. Component 1: H 3 BO 3 10mg, MnSO 4 .H 2 O 11.19mg, ZnSO 4 .7H 2 O124.6mg, CuSO 4 .5H 2 O 78.22mg, MoO 3 10mg dissolved in 100ml sterilized distilled water; component 2: FeSO 4 .7H 2 O100mg dissolved in 10ml sterile distilled water. ③ DFa liquid medium: After the prepared 0.5mol / LACC liquid was filtered through a 0.2μm membrane to remove bacteria, ACC was added to the medium wit...
Embodiment 2
[0039] Embodiment 2: Growth and IAA-producing characteristics of bacterial strains under the stress of different concentrations of heavy metals
[0040] Put 100mL of LB liquid medium containing L-tryptophan (200mg / L) in a 500mL Erlenmeyer flask, adjust the pH of the liquid medium to 7.0, sterilize at 121°C, and after 20min, press 1% (v / v ) Inoculation amount After inoculating CDL29 in the logarithmic growth phase, place at 28°C, 200r·min -1 In the shaker culture, in the culture stage, the culture solution is taken from time to time, and the amount of IAA produced is measured by the method of quantitative determination. The result is as figure 2 Shown, show that the growth of 22.5h thalline reaches the steady phase, and the IAA concentration that thalline secretes reaches the maximum in 45h culture fluid and is 107.74mg / L, and thalline is in steady growth at this moment, and OD value is maximum.
[0041] Add different concentrations of heavy metals to the above medium, 2.5mg...
Embodiment 3
[0042] Example 3: Different heavy metal polluted soils, the growth-promoting effect of bacterial strains on wheat seedlings
[0043] After pre-alcohol sterilization, the wheat seeds were put into a petri dish filled with a little distilled water and placed in an incubator for germination treatment. After the seeds germinated, they were planted in a 50ml centrifuge tube filled with 40g of soil for cultivation. The cultivation time was 20 days. The selected soil is composed of multiple heavy metals, and the two soils polluted by cadmium in different degrees are soil A and soil B. The specific soil properties are shown in Table 1:
[0044] Table 1 Basic properties and heavy metal element contents of two kinds of soils
[0045]
[0046] Set up 4 treatments and three repetitions, namely: A soil without inoculation treatment (ML), A soil with inoculation treatment (ML+B), B soil with no inoculation treatment (JN), and B soil with inoculation treatment (JN +B). Inoculate 5ml (1....
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