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A strain of cadmium removes rhizobia kg2, a bacterial agent containing said rhizobia and uses thereof

A kind of rhizobia, the technology of use, applied in the field of biology, can solve the problem of undiscovered leguminous crops and so on

Active Publication Date: 2020-06-09
SICHUAN AGRI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In summary, the prior art has not yet found a rhizobia that can improve the quality of leguminous crops in a cadmium-stressed environment and can be applied to the microbial remediation and immobilization of cadmium-contaminated soil in farmland.

Method used

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  • A strain of cadmium removes rhizobia kg2, a bacterial agent containing said rhizobia and uses thereof
  • A strain of cadmium removes rhizobia kg2, a bacterial agent containing said rhizobia and uses thereof
  • A strain of cadmium removes rhizobia kg2, a bacterial agent containing said rhizobia and uses thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Screening and Identification of Example 1 Bacterial Strains

[0031] Cadmium-removing Rhizobium pusense (Rhizobium) KG2 was isolated from soybean nodules planted on the edge of Longmang tailings dam in Xinjiuxiang, Panzhihua.

[0032]Separation adopts the plate streaking method, and the surface sterilized soybean root nodule tissue liquid is streaked on the YEM solid medium, and the purified rhizobium KG2 is obtained by observing the morphological characteristics of the colony and the morphological characteristics of the bacteria cells under the microscope. DNA was extracted according to the GUTC method, and it was identified as Rhizobium pusense by 16S rRNA, three housekeeping genes (atpD-recA-glnII), nitrogen fixation genes nifH gene and nodA gene sequencing and phylogenetic tree analysis. Among them, the sequence numbers of the 16S rRNA, atpD, recA, glnII, nifH and nodA gene sequences used to detect Rhizobium KG2 in the GeneBank database are: MF141000, MF141003, MF14...

Embodiment 2

[0033] Embodiment 2 contains the preparation of the bacterial agent of Rhizobium KG2

[0034] (1) Preparation of sterile YEM solid and TY liquid medium: 1.5 g of tryptone, 1.0 g of mannitol, K 2 HPO 4 0.5g, MgSO 4 ·7H 2 O 0.2g, NaCl 0.1g, and dilute to 1.0L with water; then use inorganic acid or inorganic base to adjust the pH value of the solution to 7.2, add agar powder (18.0g / L) after subpackaging, 121-125°C Sterilize under high pressure steam for 30 minutes to obtain sterile YEM medium, and pour the sterile YEM solid medium plate when the temperature drops to about 50°C; mix 5.0 g of tryptone, 3.0 g of yeast powder, and CaCl 2 2H 2 O 0.7g, dilute to 1.0L with water, adjust the pH value of the solution to 7.2 with inorganic acid or alkali, and sterilize with high pressure steam at 121-125°C for 30min to obtain sterile TY liquid medium.

[0035] (2) Activation of strains and preparation of strains: Streak and activate the KG2 strains preserved in glycerol tubes or slope...

Embodiment 3

[0036] The tolerance analysis of embodiment 3 Rhizobium KG2

[0037] The heavy metal tolerance analysis of rhizobia KG2 was carried out.

[0038] Get the rhizobia KG2 that embodiment 1 obtains, with 10 8 Bacteria cells were inoculated to heavy metal Cd 2+ Concentrations were 0, 20, 40, 60, 80, 100, 120, 150 and 200mg / L in TY liquid medium. Cultivate at 150r / min and 28°C for 48h, then measure the colony OD value in each medium at 600nm, the results show that the heavy metal Cd 2+ Concentration is the OD of rhizobia KG2 in the TY liquid medium of 120mg / L 600 The value is 0, that is, the heavy metal Cd 2+ The minimum lethal concentration (MLC) for Rhizobium KG2 is 120mg / L.

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Abstract

The invention provides a cadmium (Cd)-removing Rhizobium (Rhizobium pusense) KG2, a bacterial agent containing the Rhizobium and its ability to fix or remove Cd in soil or water. 2+ Applications. The invention proves that rhizobia KG2 has high-efficiency Cd fixation ability and strong nitrogen fixation function, which can promote the growth of leguminous crops, increase the stress resistance of leguminous crops, and reduce the enrichment and absorption of soil Cd by plants. The rhizobia KG2 of the present invention is applied in the bioremediation of farmland cadmium-contaminated soil in the form of bacterial agent or bacterial fertilizer, which reduces the enrichment and absorption of cadmium by crops in farming soil, promotes the growth of crops in heavy metal-contaminated soil, and improves the yield of crops. output and quality.

Description

【Technical field】 [0001] The invention belongs to the field of biotechnology. More specifically, the present invention relates to a kind of rhizobia, and also relates to the use of said rhizobia. 【Background technique】 [0002] With the massive development and utilization of mineral resources, the rapid development of industrial production and the widespread use of various chemical products, pesticides and fertilizers, pollutants containing heavy metals enter the environment through various channels, causing soil, especially farmland soil, to become increasingly polluted by heavy metals . my country's annual food production reduction due to heavy metal pollution exceeds 1×10 7 t, grains contaminated by heavy metals up to 1.2×10 7 t, the total economic loss and death is 20 billion yuan. Heavy metal pollution has very little mobility in the soil, is not easy to leach with water, and is not degraded by microorganisms. After entering the human body through the food chain, th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C02F3/34B09C1/10C05F11/08C12R1/41C02F101/22
CPCB09C1/10C02F3/34C02F2101/22C05F11/08C12N1/20C12N1/205C12R2001/41
Inventor 余秀梅李艳梅崔永亮涂卫国陈强闫敏沈甜王琼瑶辜运富赵珂陈香归
Owner SICHUAN AGRI UNIV
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