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Cd (cadmium) removal rhizobium pusense KG2, strain containing rhizobium pusense and purpose of Cd removal rhizobium pusense KG2

A rhizobia, KG2 technology, applied in the biological field, can solve problems such as undiscovered legume crops

Active Publication Date: 2018-03-20
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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  • Cd (cadmium) removal rhizobium pusense KG2, strain containing rhizobium pusense and purpose of Cd removal rhizobium pusense KG2
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  • Cd (cadmium) removal rhizobium pusense KG2, strain containing rhizobium pusense and purpose of Cd removal rhizobium pusense KG2

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Experimental program
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Effect test

Embodiment 1

[0030] Screening and Identification of Example 1 Bacterial Strains

[0031] The cadmium-removing rhizobium KG2 (Rhizobiumpusense) 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 Rhizobiumpusense 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 16SrRNA, atpD, recA, glnII, nifH and nodA gene sequences used to detect Rhizobium KG2 in the GeneBank database are: MF141000, MF141003, MF1...

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 0.7g, dilute to 1.0L with water, then 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 a sterile TY liquid medium.

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

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 Cd removal rhizobium pusense KG2, a strain containing the rhizobium pusense and application of Cd removal rhizobium pusense KG2 to fixation or removal of Cd<2+> in soil or a water body. The invention proves that the rhizobium pusense KG2 has the efficient Cd fixation capability and stronger nitrogen fixation functions; the leguminous crop growth can be promoted; the stress resistance of the leguminous crop can be increased; the enriching adsorption quantity of the plants on the Cd in the soil can be reduced. The rhizobium pusense KG2 is applied to the biological restoration of the farmland Cd pollution soil in a form of a strain or a bacterial fertilizer; the enriching adsorption of the crops on the Cd in the farmland soil can be reduced; the growth of the crops in the metal pollution soil is promoted, so that the yield and the quality of the crops are improved.

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 Applications(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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