Plant endogenetic bacterium with heavy metal tolerance and application thereof
A technology of endogenous bacteria and heavy metals in plants, applied in the fields of agriculture and environmental pollution control, can solve the problems of long cycle, slow growth, small biomass, etc., and achieve the effects of improving stress resistance, promoting plant growth, and improving repair efficiency
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Embodiment 1
[0026] The heavy metal-tolerant plant endophytic bacteria Y1-3-9 strain of the present invention is isolated and purified from Mosla chinensis leaves, and the isolation and identification method is as follows:
[0027] First clean the surface of the leaves of Elsholtzia vulgaris with tap water, then wash it with sterile water, then soak and disinfect with 75% ethanol for 5 minutes, rinse with sterile water and then immerse in 2.5% sodium hypochlorite solution for 5 minutes, and rinse with sterile water several times. Place the sterilized leaves in a mortar and grind them to homogenize. Take 0.1ml of the homogenate and spread it on an LB plate (10g peptone, 5g yeast extract, 10g NaCl, 1000mL water, 20g agar, pH 7.0), and incubate at 28°C for 3 days , Pick a single colony, streak it on the LB plate and save it after purification. The strains were inoculated into LB culture broth, shake culture at 30°C for 18h, take 1.0mL bacterial broth in Eppendorf centrifuge tube, centrifuge at 1...
Embodiment 2
[0029] Y1-3-9 (CCTCC NO: M 2011251) was streaked and inoculated into LB medium containing different heavy metal concentrations, and cultured at 30°C for 3 days to observe whether it could grow and its growth. The results are shown in Table 1. The Y1-3-9 strain can tolerate a variety of heavy metals (the minimum inhibitory concentrations of Cu, Pb, Cd, and Ni are 3.1, 4.8, 4.5, 1.7 mM, respectively).
[0030] Table 1 Growth of strains on LB medium with different concentrations of heavy metals
[0031]
[0032] In the above table, "+" means growth, and "-" means no growth.
Embodiment 3
[0034] Inoculate Y1-3-9 (CCTCC NO: M 2011251) in LB culture broth, shake culture for 20 hours, centrifuge at 4℃ to collect the bacteria, and use SM broth (glucose 1.0g, sucrose 1.0g, sodium citrate 1.0g) , Malic acid 1.0g, mannitol 1.0g, sodium acetate 1.0g, KH 2 PO 4 0.4g, K 2 HPO 4 2.0g, MgSO 4 0.2g, CaCl 2 0.1g, CuSO 4 1mg, NiSO 4 1mg, ZnSO 4 5mg, FeSO 4 5mg, MnSO 4 3mg, CoSO 4 1mg, Na 2 MoO 4 1mg, H 3 BO 3 2mg, biotin 2 (VB 6 ) 10mg, Thiamine (VB 1 ) 2mg, cyanocobalamin vitamin (VB 12 )0.1mg, pantothenic acid (VB 3 ) 5mg, folic acid 2mg, riboflavin 5mg, niacin 5mg, distilled water 1000mL, pH 6.4. Add the vitamin after filtration and sterilization. ) Wash and centrifuge twice, suspend the bacteria in SM culture medium, and inoculate SMA culture medium according to 5% of the inoculum [Add filter sterilized ACC (1-aminocyclopropane-1-carboxylic acid deaminase) to the sterile SM medium ), make the final concentration 3mM], culture for 48h under the condition of 28℃1...
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