Cadmium-tolerant lysinibacillus sp. for promoting growth of rhizosphere of solanum nigrum L. and application thereof
A technology of Bacillus lysinus and nightshade, applied in the direction of bacteria, microorganisms, microorganisms, etc., can solve the problems of few effective strains and lack of pertinence, and achieve the goal of promoting growth and development, improving extraction efficiency, and good application prospects Effect
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Embodiment 1
[0023] Isolation and Purification of Lysinibacillus sp. NT1
[0024] This embodiment includes the following steps:
[0025] Step 1. Take the rhizosphere soil of Solanum nigrum, collect the plants with their roots, cut off the aboveground part, remove the large soil around the roots, use a brush to brush the rhizosphere soil within 1cm of the root system into a sterile fresh-keeping bag, and bring it back to the experiment Store in a refrigerator at 4°C, and screen within 2 days. Weigh 5g of soil sample, put it into a conical flask filled with glass beads and 50mL of sterile water, shake and cultivate on a shaker at 37°C and 200rpm for 30min, and keep the above culture solution in a water bath at 80°C for 10min; 10 mL of the suspension was transferred to an Erlenmeyer flask containing 100 mL of LB-Cd (Cd content 10 mg / L) liquid medium, and enriched at 37°C and 200 rpm for 24 hours. Repeat this operation 2 times for enrichment culture.
[0026] Step 2, take the above-mentione...
Embodiment 2
[0030] Isolation and identification of growth-promoting ability of Lysinibacillus sp. NT1
[0031] This embodiment includes the following steps:
[0032] 1) Identification of IAA-producing ability: using Salkowski's reagent microplate colorimetric method, referring to Loper's method, the isolated strains were inoculated in 20 mL of TSB liquid medium containing tryptophan (100 mg / L), at 28 ° C, Shake culture at 200r / min for 48h, then centrifuge at 8000r / min for 10min. Take 2 mL of the centrifuged supernatant, add 50 μL of orthophosphoric acid with a volume fraction of 83% and 4 mL of Salkowski reagent, and the solution turns pink, indicating that IAA is produced. Take the above-mentioned color developing solution, develop color at 25°C for 30 min in the dark, and measure the absorbance value at a wavelength of 530 nm. Replace the culture medium with distilled water and use the same reaction as a control to adjust to zero, prepare standard IAA gradient solutions of 10, 20, 30,...
Embodiment 3
[0037] Enhanced Extraction of Cadmium from Solanum nigrum by Lysinibacillus sp. NT1
[0038]This embodiment comprises the following steps: take plump nightshade seeds, disinfect and soak them, pre-cultivate them for 4 weeks, select robust and consistent nightshade seedlings in a hydroponic tank, use Hoagland nutrient solution as the hydroponic solution, and add CdCl from an external source 2 , the treatment concentration was 4mg / L, the growth-promoting bacteria were cultured in LB liquid medium for 24 hours, and added to the hydroponic solution (the concentration of bacteria in the hydroponic solution was about 1×10 7 CFU / mL), the hydroponic medium and bacterial solution were replaced every 5 days, and the culture was continued for 8 weeks in a glass greenhouse. Set up a blank control without cadmium and no bacteria (CK), and a control with cadmium and no bacteria (CK-Cd).
[0039] The growth index measurement of described Solanum nigrum: take out whole plant Solanum nigrum g...
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