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Pseudomonas sp., microbial biocontrol agent containing Pseudomonas sp. and application of microbial biocontrol agent

A technology of pseudomonas and biocontrol agents, applied in the field of microbial technology and biological control, can solve the problems of low utilization rate of fertilizers, decline in agricultural benefits, damage to the physical structure of soil, etc., achieve good disease control effect, and promote growth. And the effect of increasing production and simple preparation process

Active Publication Date: 2021-08-13
INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] In recent decades of agricultural production, problems such as the unreasonable use of agricultural inputs such as chemical fertilizers, pesticides, and herbicides have caused the accumulation of various toxic and harmful substances, destroyed the physical structure of the soil, and caused increasingly serious soil acidification and organic matter. The decline of the soil has caused the imbalance of functional microorganisms in the soil and the decline of soil fertility. The utilization rate of fertilizers is not high, crop diseases occur frequently, and agricultural benefits decline. To solve the obstacles to agricultural production in our country and achieve sustainable agricultural development, it happens to be in harmony with microbial bacteria. The function of the agent is consistent with the characteristics and expertise of the microbial agent

Method used

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  • Pseudomonas sp., microbial biocontrol agent containing Pseudomonas sp. and application of microbial biocontrol agent
  • Pseudomonas sp., microbial biocontrol agent containing Pseudomonas sp. and application of microbial biocontrol agent
  • Pseudomonas sp., microbial biocontrol agent containing Pseudomonas sp. and application of microbial biocontrol agent

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Experimental program
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Effect test

Embodiment 1

[0042] Embodiment 1, the separation and screening of biocontrol strain ZL8

[0043] Specific steps include:

[0044] Step 1, isolation and preservation of Pseudomonas

[0045] The biocontrol bacterial strain ZL8 provided by the present invention is to be isolated in the sclerotium of healthy polyporus (polyporus, non-Pleurophytes Polyporaceae Arboria medicinal fungus) sclerotia collected in Ningshan County, Shaanxi Province, and the separation process is as follows: first, with flowing water The material is rinsed repeatedly to remove the soil attached to the surface and most of the soil microorganisms. Then wash with 70% ethanol for 3 minutes, and soak in 2% hypochlorous acid for 2, 5, and 7 minutes respectively. The results show that some bacteria still grow in the plate when soaked in sodium hypochlorite for 2 minutes, and the materials soaked for 5 and 7 minutes can achieve surface disinfection. Thorough effect, but there is no growth of endophytic bacteria when soaking ...

Embodiment 2

[0059] Embodiment 2, identification of biocontrol strain ZL8

[0060] Genomic DNA of strain ZL8 was extracted using a bacterial genomic DNA extraction kit, and identified by 16S rRNA gene. The specific steps were as follows:

[0061] Use the aforementioned bacterial 16S rRNA gene universal primer 27F / 27R, and use the strain ZL8 genomic DNA as a template; carry out PCR reaction according to the following reaction system (25 μL): 2×M5 supermix 25 μl, upstream primer (10 μM) 2 μl, downstream primer (10 μM) 2μl, bacterial solution 1μl, the balance is ddH 2 O. PCR reaction program: pre-denaturation at 94°C for 10 minutes, (denaturation at 94°C for 30 seconds, annealing temperature at 55°C for 30 seconds, extension temperature at 72°C for 1 minute) 35 cycles, 72°C for 5 minutes, 4°C to terminate the reaction.

[0062] The PCR product is about 1500bp. The PCR product was sent to Beijing Nuosai Genome Research Center Co., Ltd. for sequencing. The sequencing result is shown in SEQ I...

Embodiment 3

[0068] The preparation of embodiment 3, ZL8 biocontrol agent

[0069] The specific steps are: insert the biocontrol strain ZL8 into the LB liquid medium, cultivate at 30° C. and 200 rpm for 12 hours, and make a seed liquid; inoculate the seed liquid into the fermentation medium (using LB liquid medium) with an inoculum size of 1%. medium, 30°C, 200rpm shaking culture for 48h, to obtain a fermentation broth, the total concentration of viable bacteria was 1 × 10 9 CFU / mL or more is the prepared biocontrol agent.

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Abstract

The invention discloses Pseudomonas sp. ZL8 which is preserved in China General Microbiological Culture Collection Center, and the preservation number is CGMCC NO. 21874. The invention further discloses a microbial biocontrol agent containing the Pseudomonas sp. and a preparation method of the microbial biocontrol agent, and application of the ZL8 and the biocontrol agent thereof in inhibiting plant pathogenic fungi and promoting plant growth. The biocontrol strain ZL8 provided by the invention has a relatively strong inhibiting effect on cucumber brown spot bacteria, sclerotinia sclerotiorum, wheat scab, potato verticillium wilt or salvia miltiorrhiza pathogenic fungi, and has a good disease control effect and growth promoting and yield increasing effects in cucumber and panax notoginseng plants.

Description

technical field [0001] The invention belongs to the field of microbial technology and biological control technology, and in particular relates to a strain of pseudomonas, a microbial biocontrol agent containing the bacterium and an application thereof. Background technique [0002] In recent decades of agricultural production, problems such as the unreasonable use of agricultural inputs such as chemical fertilizers, pesticides, and herbicides have caused the accumulation of various toxic and harmful substances, destroyed the physical structure of the soil, and caused increasingly serious soil acidification and organic matter. The decline of the soil has caused the imbalance of functional microorganisms in the soil and the decline of soil fertility. The utilization rate of fertilizers is not high, crop diseases occur frequently, and agricultural benefits decline. To solve the obstacles to agricultural production in our country and achieve sustainable agricultural development, ...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01N63/27A01P3/00A01P21/00A01G7/06A01G22/05A01G22/25C12R1/38
CPCC12N1/20A01N63/27A01G7/06A01G22/05A01G22/25Y02A40/10
Inventor 袁媛南铁贵刘天睿韩鹏杰郑媛黄璐琦
Owner INST OF CHINESE MATERIA MEDICA CHINA ACAD OF CHINESE MEDICAL SCI
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