Bacteria d10 for improving the effectiveness of heavy metals and its application

A heavy metal and effective technology, applied in the fields of agricultural technology and environmental pollution control applications, can solve the problem of small total biomass, and achieve the effect of promoting plant growth and increasing the content of heavy metals in plants

Active Publication Date: 2020-06-23
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the biomass of herbaceous alfalfa is small, so it is not suitable for phytoextraction and restoration

Method used

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  • Bacteria d10 for improving the effectiveness of heavy metals and its application
  • Bacteria d10 for improving the effectiveness of heavy metals and its application
  • Bacteria d10 for improving the effectiveness of heavy metals and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The heavy metal-resistant plant growth-promoting bacteria D10 strain of the present invention is isolated and purified from the root nodules of alfalfa (Medicogo sativa L.) grown in heavy metal-contaminated soil, and the separation and identification method is as follows:

[0040] Gently pull the alfalfa from the soil, shake off the roots to loosen the soil and rinse with water. Select the plump, large-diameter pink nodules on the taproot and cut them off with scissors. Soak the cut nodules in deionized water for 4-5 minutes to remove the remaining impurities, soak and disinfect with 75% ethanol for 2-3 minutes, wash with water 2-3 times, soak in 3% NaClO solution for 5 minutes, and finally use sterile deionized Wash with water 5-7 times. Under sterile operating conditions, put the surface-sterilized nodules into YMA solid plates containing 0.3% Congo red [mannitol 10.0 g, yeast powder 1.0 g, MgSO 4 ·7H 2 O 0.2g, K 2 HPO 4 0.5g, NaCl 0.1g, anhydrous CaCl 2 0.05g,...

Embodiment 2

[0042] Example 2 Activation of strain D10 and preparation of bacterial suspension

[0043] The D10 (CCTCC NO: M 2016554) slant strain was inoculated into TY solid medium (tryptone 5.0 g, yeast extract 3.0 g, CaCl 2 0.3g, agar 20g, distilled water 1000ml, pH 7.0), cultured at 30°C for 3 days. Then select plump and viscous D10 colonies to inoculate in TY liquid medium (tryptone 5.0g, yeast extract 3.0g, CaCl 2 0.3g, distilled water 1000ml, pH 7.0), shaking at 150rpm at 30°C for 20-24h, so that the number of cells reaches more than 500 million CFU / ml.

Embodiment 3

[0044] Example 3 The ability of strain D10 to produce IAA

[0045] According to the determination method of Gordon and Weber (1951), TY liquid medium was divided into test tubes, each tube was 4 mL, and 1 mL of filter-sterilized 2.5 mg / mL tryptophan solution was added after high-temperature sterilization at 121 °C to make the medium in color. The final concentration of amino acid was 0.5 mg / mL. D10 (CCTCC NO: M 2016554) was inoculated into the above-mentioned medium and cultured with shaking at 30°C for 3 days. The fermentation broth was centrifuged at 12000 r / min for 5 min, 1 mL of the supernatant was taken, 50 μL of 10 mM orthophosphoric acid was added, and 2 mL of Sackowski's chromogenic reagent was added. Sterile medium was treated the same as above and zeroed as a control. The concentration of IAA in the fermentation broth was calculated by using IAA standard solution with concentrations of 0, 5, 10, 20, 40, and 60 mg / L as a standard curve in the same way. The results ...

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Abstract

The invention discloses a bacterium D10 for improving the effectiveness of heavy metals and an application thereof. A bacterium D10 that improves the effectiveness of heavy metals, named Sinorhizobium meliloti D10, is preserved in the China Center for Type Culture Collection, and the strain preservation number is CCTCC NO: M 2016554. The D10 strain was used as the raw material, and the bacterial suspension was prepared after being activated by the culture medium. In heavy metal polluted soil in mining areas, inoculation of D10 bacterial suspension can increase the growth of energy plants, increase the extractable copper content in plant rhizosphere soil, and increase the copper content and total copper absorption in plant roots. Alfalfa and ryegrass inoculated with D10 can be used for fixed remediation of heavy metal-contaminated soil, and sweet sorghum inoculated with D10 can be used for both fixed and extractive remediation. And after the D10 bacterial suspension was inoculated, the IAA content of the plant rhizosphere microorganisms increased, and the soil fertility increased.

Description

technical field [0001] The invention belongs to the application field of agricultural technology and environmental pollution control, and relates to a bacterium D10 for improving the effectiveness of heavy metals and its application. Background technique [0002] In recent years, heavy metal pollution in farmland due to mining activities has caused serious impacts on food safety and human health. Phytoremediation is an environment-friendly treatment measure that can realize the treatment of heavy metal pollution. However, most of the hyperaccumulators are difficult to grow in the wild due to their harsh agricultural production requirements, and the biomass yield is relatively small, which limits the large-scale farmland practical application of hyperaccumulators. It is feasible to use energy plants (such as sweet sorghum) with high biomass yield, fast growth rate and strong tolerance to heavy metals as plant extract materials. [0003] Rhizobium can not only symbiotically ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20B09C1/10A01G7/06A01B79/02C12R1/01
CPCA01B79/02A01G7/06B09C1/10C12N1/205C12R2001/01
Inventor 何琳燕盛下放王琪严警
Owner NANJING AGRICULTURAL UNIVERSITY
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