The present disclosure discloses a visualized screening method for an Aspergillus recombinant strain with multigene editing and belongs to the technical field of gene engineering. CRISPR-Cas9 is used in the disclosure to cleave spore color change-related genes and a target gene in Aspergillus at the same time, such that editing of the target gene is visualized and an Aspergillus niger strain with multigene editing can be rapidly and efficiently screened out through spore phenotypes. Through different combinations of visualized genes and non-phenotypic change genes, rapid screening of the strain with multigene editing and simultaneous screening of multiple visualized genes are realized, and use of resistance genes in industrial strains is reduced.

A method for determining a level of one or more live microbes in a sample of a Cannabis plant includes (a) introducing a DNase enzyme into a sample including (i) Cannabis plant matrix derived from lysed cells of a Cannabis plant and (ii) live microbial cells including microbial genomic DNA, (b) introducing an anionic detergent into the sample, where the anionic detergent is in an amount effective to inactivate the DNase enzyme in the sample and lyse the live microbial cells in the sample, (c) introducing a known quantity of control DNA into the sample, (d) contacting the sample with a plurality of polynucleotide capture agents, (e) isolating the plurality of polynucleotide capture agents, thereby isolating polynucleotides from the microbial genomic DNA and/or control DNA in the sample, and (f) amplifying at least a portion of polynucleotides from the microbial genomic DNA isolated in step (e).

The invention relates to methods for controlling mycotoxin production in field populations of heterothallic and homothallic filamentous fungi through application of biocontrol compositions comprising selected strains of the heterothallic and homothallic filamentous fungi, the selected strains having a lineage of low mycotoxin production or no mycotoxin production and having opposite mating types (heterothallic) or both mating types (homothallic) (e.g., MAT1-1, MAT1-2).

A method of producing a microbial fermentation broth includes fermenting at least two of plum, dandelion, pine needle, and thistle to prepare each fermentation stock solution, mixing the fermentation stock solutions and fermenting the mixed fermentation stock solutions a to prepare a fermentation mixture, mixing the fermentation mixture with coffee meal and fermenting the mixed fermentation mixture with the coffee meal to prepare a coffee meal fermented solution, and aging the coffee meal fermented solution.

The invention relates to methods for controlling mycotoxin production in field populations of heterothallic and homothallic filamentous fungi through application of biocontrol compositions comprising selected strains of the heterothallic and homothallic filamentous fungi, the selected strains having a lineage of low mycotoxin production or no mycotoxin production and having opposite mating types (heterothallic) or both mating types (homothallic) (e.g., MAT1-1, MATT-2).

The disclosure provides a method of reducing aflatoxin contamination in one or more nuts and/or reducing the amount of Aspergillus flavus and/or Aspergillus parasiticus resulting in the presence of aflatoxin contamination in one or more nuts. In an embodiment, nuts are treated with ultraviolet (UV) light, ozone, and peroxide, or combinations thereof. In an embodiment, nuts are treated with UV light and ozone. In an embodiment, nuts are treated with UV light and peroxide. In an embodiment, nuts are treated with ozone and peroxide.

The disclosure provides methods of preparing aflatoxin-contaminated nuts.

The invention relates to methods for controlling mycotoxin production in field populations of heterothallic and homothallic filamentous fungi through application of biocontrol compositions comprising selected strains of the heterothallic and homothallic filamentous fungi, the selected strains having a lineage of low mycotoxin production or no mycotoxin production and having opposite mating types (heterothallic) or both mating types (homothallic) (e.g., MAT1-1, MAT1-2).

The present disclosure belongs to the technical field of microorganisms, and specifically relates to a non-toxin-producing Aspergillus flavus strain, a microbial inoculant, a preparation, and use. The present disclosure provides a non-toxin-producing Aspergillus flavus strain EXY1A109 with a deposit number of CCTCC No: M20221465. The strain can effectively inhibit the production of aflatoxin B.sub.1, aflatoxin B.sub.2, aflatoxin G.sub.1, and aflatoxin G.sub.2 by a toxin-producing Aspergillus flavus strain. The results of examples show that when an Aspergillus flavus strain EXY1A109 spore suspension and a strain CGMCC 3.4408 spore suspension are mixed in an equal volume, the Aspergillus flavus strain EXY1A109 spore suspension and the strain CGMCC 3.4408 spore suspension each have a concentration of 110.sup.6, and the strain EXY1A109 has a toxin-producing inhibition rate of 99.71% against the strain CGMCC 3.4408. This Aspergillus flavus strain shows a desirable application effect.

A method for determining a level of one or more live microbes in a sample of a Cannabis plant includes (a) introducing a DNase enzyme into a sample including (i) Cannabis plant matrix derived from lysed cells of a Cannabis plant and (ii) live microbial cells including microbial genomic DNA, (b) introducing an anionic detergent into the sample, where the anionic detergent is in an amount effective to inactivate the DNase enzyme in the sample and lyse the live microbial cells in the sample, (c) introducing a known quantity of control DNA into the sample, (d) contacting the sample with a plurality of polynucleotide capture agents, (e) isolating the plurality of polynucleotide capture agents, thereby isolating polynucleotides from the microbial genomic DNA and/or control DNA in the sample, and (f) amplifying at least a portion of polynucleotides from the microbial genomic DNA isolated in step (e).