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Method for reducing nitrate content of vegetables under protected cultivation conditions

A nitrate and vegetable technology is applied in the field of agricultural pollution prevention and control to achieve the effects of promoting plant growth, reducing nitrate concentration and reducing nitrate content

Inactive Publication Date: 2015-08-26
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] These methods have reduced the content of nitrate in planted vegetables to a certain extent, but also introduced new chemicals, and long-term treatment may bring new problems

Method used

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  • Method for reducing nitrate content of vegetables under protected cultivation conditions
  • Method for reducing nitrate content of vegetables under protected cultivation conditions
  • Method for reducing nitrate content of vegetables under protected cultivation conditions

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] The screening and separation of embodiment 1 target strain

[0037] (1) Culture medium preparation

[0038] Enrichment medium: KNO 3 10g, KCl 1g, MgSO 4 ·7H 2 O 0.5g, CaCl 2 1mg, FeSO 4 ·7H 2 O 10mg, KH 2 PO 4 0.5g glucose 7.5g, deionized water 1L, pH 7.0. Autoclave: 121°C, 20min.

[0039] Separation medium: KNO 3 10g, KCl 1g, MgSO 4 ·7H 2 O 0.5g, CaCl 2 1mg, FeSO 4 ·7H 2 O 10mg, KH 2 PO 4 0.5g, glucose 7.5g, agar 15g, deionized water 1L, pH 7.0. Autoclave: 121°C, 20min.

[0040] LB medium: tryptone 10g, yeast extract 5g, NaCl 10g, deionized water 1L, pH 7.0. Autoclave: 121°C, 20min.

[0041] (2) Enrichment and isolation of nitrate transforming bacteria

[0042] 1) Enrichment culture of transformed bacteria

[0043] Shake flask enrichment culture method was adopted. Potassium nitrate was the only nitrogen source. The specific steps are as follows: Take 10g of each soil sample, put it in 100mL enrichment medium, place it on a constant temperature sh...

Embodiment 2

[0049] The identification of embodiment 2 target bacterial species bacillus pumilus SrN7

[0050] (1) Molecular biological identification

[0051] Molecular biology identification of SrN7 strain

[0052] Inoculate the preserved SrN7 bacteria into LB liquid medium, cultivate overnight at 30°C on a constant temperature shaker at 180r / min, and extract the obtained bacterial liquid with Shanghai Sangon Biobacterial DNA Extraction Kit to amplify the 16S rDNA of the strain. After purifying the obtained PCR products, BGI was entrusted with sequencing.

[0053] PCR amplification primers are universal primers:

[0054] 27F: 5'-AGAGTTTGATCCTGGCTCAG-3' (SEQ ID NO: 2)

[0055] 1492R: 5'-GGTTACCTTGTTACGACTT-3' (SEQ ID NO: 3).

[0056] The PCR amplification system used a 25 μl reaction system, as shown in Table 1.

[0057] Table 1 PCR amplification system

[0058] Element Content (μl) DNA template 0.5 10×buffer 2.5

[0059] dNTP mixture (each 2.5...

Embodiment 3

[0064] The preparation of embodiment 3 microbial bacterial agents

[0065] (1) Activation and expanded culture of strains: inoculate 20 mL of LB medium with cryopreserved strains, and culture on a constant temperature shaker at 30° C. at 180 r / min for 24 hours. Then inoculate the activated bacterial solution into fresh 100mL LB culture solution with an inoculum amount of 1%, and cultivate it on a constant temperature shaker at 30°C for 24 hours at 180r / min.

[0066] The composition of the LB medium is: tryptone 10g L -1 , yeast extract 5g·L -1 , sodium chloride 10g·L -1 , Agar 15g·L -1 , pH 7.0.

[0067] (2) Fermentation culture:

[0068] Inoculate the expanded cultured bacterial solution into the fermentation medium with 10% inoculum amount, and culture it on a constant temperature shaker at 37° C. at 180 r / min for about 36 hours.

[0069] The number of live bacteria after fermentation is 2×10 9 , The sporulation rate is not less than 85%.

[0070] The composition of ...

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Abstract

The invention discloses a method for reducing nitrate content of vegetables under protected cultivation conditions. A strain of soil nitrate conversion bacteria is separated from greenhouse soil for protected planning for many years, and is called as Bacillus pumilus through classification, the number of the bacterial strain is SrN7, and the preservation number is CGMCC No. 9685. The microorganism strain is mixed with bran to prepare a microorganism bacterium agent after activation, enlarging cultivation and fermentation treatment, and the obtained microorganism bacterium agent is applied to soil at the roots of target vegetables. The obtained bacterium agent is a pure biological agent and is harmless for the body and the environmental health, the concentration of nitrate in protected soil can be reduced, so that the content of the nitrate in vegetable bodies is reduced, the plant growth can be promoted, and the basic physicochemical property of the soil is improved; compared with other methods for reducing the nitrate of the protected soil, under the precondition of not influencing normal production, the content of the nitrate in the soil can be reduced while production, and the effect of repairing while production is achieved.

Description

technical field [0001] The invention relates to the field of agricultural pollution prevention and control, in particular to a method for reducing the nitrate content of leafy vegetables under facility cultivation conditions by using microbial agents. Background technique [0002] Facility cultivation is one of the important modes of vegetable cultivation in my country. It has the advantages of small land occupation, high output, and little impact on the environment. It plays an important role in off-season and cross-regional cultivation. However, this high-input, high-yield planting model is carried out under strong human intervention. In order to ensure high yields of vegetables, a large number of agrochemical products, especially nitrogen fertilizers, need to be applied. The high input of fertilizers for many years has caused increasingly serious secondary salinization of the facility soil, especially the problem of excessive nitrate. Excessive nitrate in the soil will l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A01N63/00A01P21/00C09K17/00C12R1/07C09K101/00
Inventor 潘风山冯英杨肖娥孟茜罗莎陈宝凯兰唐琳
Owner ZHEJIANG UNIV
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