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Epicoccum nigrum, microbial agent as well as preparation method and application of microbial agent

A microbial inoculant, the technology of E. nigricans, applied in the field of microorganisms, can solve the problems of agricultural production and human health threats, bacterial resistance, and the application of Fusarium graminearum has not yet been disclosed.

Active Publication Date: 2022-04-29
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, Fusarium graminearum can also produce mycotoxins, such as B-type chorphene, deoxynivalenol (DON) and zearalenone (ZEN), which pose a threat to agricultural production and human health
The excessive use of fungicides has caused serious environmental pollution and bacterial strain resistance problems. The disease problem caused by Fusarium graminearum urgently needs a green and environmentally friendly solution
In agricultural production, the application of plant endophyte E. niger to the prevention and control of Fusarium graminearum has not been published

Method used

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  • Epicoccum nigrum, microbial agent as well as preparation method and application of microbial agent
  • Epicoccum nigrum, microbial agent as well as preparation method and application of microbial agent
  • Epicoccum nigrum, microbial agent as well as preparation method and application of microbial agent

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] The strains were isolated as follows:

[0069] Leaves of maize plants collected in a maize-producing area in Beijing, China. Surface disinfection was first performed by soaking the samples in 70% alcohol for 1 minute, then transferred to 2% sodium hypochlorite for 2 minutes, and finally soaked in 70% alcohol for 30 seconds. After this treatment, they were washed three times with sterile distilled water and placed on sterile filter paper for further drying. The sample was cut into small fragments (about 5 mm) and placed on the medium surface of Potato Dextrose Agar (PDA), and ampicillin (50 μg / ml) was added to the medium to inhibit the growth of bacteria in the PDA medium. Take 100 μl of water eluted in the last step of surface disinfection and also add it to the surface of PDA medium to evaluate the efficiency of surface sterilization. All media were cultured in a dark incubator at 25°C. After the hyphae grew out, the mycelium at the tip of the colony was taken and t...

Embodiment 2

[0071] The morphological identification and molecular biological identification of strain 38L1 are as follows:

[0072] ① 38L1 morphology identification

[0073] The preserved 38L1 was activated and cultured in PDA medium, and the culture condition was dark culture at 25°C for 5 days. Afterwards, a bacterium cake with a diameter of 5 mm was punched out at the edge of the activated culture colony, placed in a freshly configured PDA medium, and cultivated under the same culture conditions for 5 days. Observe the front and back of the photographed colony (such as figure 1 shown in A).

[0074] ② DNA extraction and PCR amplification

[0075] After the strain was cultured in PDA medium for 7 days, the genomic DNA was extracted with DNeasy Plant Mini kit (Qingdao Branch Biotechnology Co., Ltd.). The concentration of extracted DNA was measured by NanoDrop spectrometer. PCR specifically amplified the internal transcriptional spacer (ITS), large subunit (LSU) and β-tubulin (TUB) g...

Embodiment 3

[0079] The inhibitory effect of strain 38L1 on fungi such as Fusarium graminearum was determined by plate confrontation, as follows:

[0080] Get a bacterium cake with a diameter of 5 mm from the edge of the pathogenic fungal colony that has been cultured for 5 days and inoculate it in the center of the PDA plate. The 38L1 bacterium cake with a diameter of 5 mm was taken from the edge of the 38L1 colony that was subjected to the same activation treatment and inoculated on both sides near the edge of the plate, and the inoculated 38L1 bacterium cake and the phytopathogenic fungus bacterium cake were in a straight line. All culture dishes were placed in a 25°C incubator for 7 days in the dark. The experiment was performed 3 times, and each treatment was repeated 3 times. By observing and counting the growth of pathogenic fungi, the inhibitory effect of endophytes on them can be judged. Such as figure 2 As shown, on the potato dextrose agar (PDA) medium, the endophytic fungus...

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Abstract

The invention discloses Epicoccum nigrum, a microbial agent and a preparation method and application of the Epicoccum nigrum and the microbial agent. The collection number of the Epicoccum nigrum 38L1 in the China General Microbiological Culture Collection Center (CGMCC) is CGMCC (China General Microbiological Culture Collection Center) No.40003. The invention further discloses a preparation method of the Epicoccum nigrum 38L1. The strain has an excellent inhibition effect on various pathogenic bacteria such as fusarium graminearum, botrytis cinerea, colletotrichum gloeosporioides, botrytis cinerea and sclerotinia sclerotiorum, and has a good application prospect.

Description

technical field [0001] The application relates to Epicoccus niger, a microbial agent, a preparation method and application thereof, and belongs to the technical field of microorganisms. Background technique [0002] Among plant endophytic fungi, fungi of the genera Epicoccus, Penicillium and Trichoderma enhance host tolerance to biotic and abiotic stresses. Among them, studies have shown that Epicoccus can antagonize the growth of various plant pathogens by producing antibiotics. Among them, Epicoccus niger is more valuable for research, because the secondary metabolites it produces contain antifungal substances and can exhibit biological control activities against various plant diseases. The antioxidants and antibiotics isolated therefrom have antagonistic activity against various fungi such as anthracnose and Botrytis cinerea. [0003] Fusarium graminearum is one of the most common and damaging fungal diseases in wheat production, known for causing devastating ear infect...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N3/00A01N63/30A01P3/00C12R1/645
CPCC12N1/14C12N3/00A01N63/30Y02A40/10
Inventor 王双超郭立华可莱
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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