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Heavy-metal-resistant Cupriavdus sp. and preparation method and application thereof

A technology of copper greedy bacteria and heavy metals, applied in the field of preparation of copper greedy bacteria HXC-8 and its bacterial agent, can solve the problems of undisclosed copper greedy bacteria, and achieve the effects of improving stress resistance, slowing down stress, and increasing seed germination rate

Active Publication Date: 2019-12-31
HANSHAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the prior art does not disclose the remediation of Cu in soil 2+ Cuprovidus

Method used

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  • Heavy-metal-resistant Cupriavdus sp. and preparation method and application thereof
  • Heavy-metal-resistant Cupriavdus sp. and preparation method and application thereof
  • Heavy-metal-resistant Cupriavdus sp. and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment one, the isolation and cultivation of Mimosa rhizobia

[0033] Collect the whole plant of Mimosa, wash the collected Mimosa with clean water, remove the soil clods and stems, select a large, plump, fresh root nodule at the root, wash away impurities, soak in 95% ethanol for 5 minutes, and then rinse with 0.1 % HgCl 2 The surface was sterilized for 5 minutes, and then rinsed 10 times with sterile water under sterile conditions.

[0034] In the case of aseptic operation, shake the sterile water flowed down from the root nodule for the last wash, pour a small amount into the YMA medium to spread evenly, and culture it upside down at 28°C for 3 to 5 days. If no colonies appear, it proves that the copper greedy bacteria have been cleaned before inoculation, otherwise, the cause of the colonies should be analyzed, and it should be verified whether the colonies that appear in the culture medium inoculated with copper greedy bacteria are rhizobia colonies.

[0035]...

Embodiment 2

[0037] Embodiment two, the Gram stain identification of bacterial strain HXC-8

[0038] The purified strain HXC-8 was identified by Gram staining method, and the identification result showed that it was in the shape of a red short rod, so it was determined to be a Gram-negative short bacillus. The specific identification steps are as follows:

[0039] (1) Use an inoculation loop to pick a small amount of bacteria in a single colony of round translucent milky white nasal mucus, and evenly spread it on a glass slide dripped with sterile water; at the same time, pick a little Bacillus subtilis, and spread it on the same drop Sterile water, as a control; spread the bacteria evenly, after natural drying, burn 2-3 times on the alcohol lamp to fix;

[0040] (2) Add crystal violet dyeing solution dropwise for 1-2 minutes, wash away the dyeing solution with fine water, and absorb residual water with filter paper;

[0041] (3) Mordant with iodine solution for 1 min, and washed with wa...

Embodiment 3

[0045] Example 3, 16S rDNA sequencing identification of bacterial strain HXC-8

[0046] 1. Genomic DNA Extraction

[0047] 1) Take the strains on the medium in a mortar and grind with liquid nitrogen next to the flame of an alcohol lamp.

[0048] 2) Put the ground bacteria into a 1.5ml centrifuge tube, mark the name of the strain, add 0.6mL TE (pH 8.0), and pipette evenly with a pipette tip to fully suspend the bacteria.

[0049] 3) Add 250 μL of 10% SDS, and gently invert to mix.

[0050] 4) Add 3 μL of proteinase K (20 ng / μL), mix gently, and bathe in 37° C. water for 1 hour.

[0051] 5) Add 150 μL 5mol / L NaCl and mix gently.

[0052] 6) Add 150 μL of 2% CTAB, mix gently, and bathe in water at 65° C. for 20 minutes.

[0053] 7) Centrifuge at room temperature at 12000rpm for 20min.

[0054] 8) Carefully pipette the supernatant into a new 1.5ml centrifuge tube, add an equal volume of isopropanol, mix thoroughly, place at room temperature for 30min, centrifuge at 12000rpm,...

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Abstract

The invention belongs to the technical field of new strain screening technology and environmental microorganism technology, and particularly relates to heavy-metal-resistant Cupriavdus sp. HXC-8. TheCupriavdus sp. HXC-8 is separated from the roots of Mimosa pudica of Leguminosae, can resist heavy metal copper, improve the germination rate of seeds polluted by Cu<2+> and promote the growth of seedlings polluted by the Cu<2+>, can effectively remove copper ions in the environment, can be used as a preparation for remedying soil polluted by the Cu<2+>, and has wide application prospect.

Description

technical field [0001] The invention belongs to the field of new strain screening technology and environmental microorganism technology, and in particular relates to a heavy metal-resistant cupriavdus sp. HXC-8 and a preparation method and application thereof. Background technique [0002] In recent years, the heavy metal pollution of soil by mining, chemical industry and life has become increasingly serious. Heavy metals cannot be decomposed by themselves in the soil, remain for a long time, cause great harm, and are easily absorbed by crops to affect human health. Copper pollution is a common heavy metal pollution. Too much copper in the soil accumulates and stays in the roots after being absorbed by plants, which hinders the roots from absorbing other components, especially iron, leading to poor plant growth. Excessive copper entering the body will cause Nausea, vomiting, epigastric pain, acute hemolysis and renal tubular deformation and other poisoning phenomena. Biore...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01G31/00A01C1/00C09K17/14C09K101/00C12R1/01
CPCC12N1/20A01G31/00A01C1/00C09K17/14C09K2101/00C12R2001/01C12N1/205
Inventor 王桔红陈文
Owner HANSHAN NORMAL UNIV
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