A liquid sporulation method is described. A liquid culture is prepared by adding bacterial cells to a sporulation broth. The liquid culture is incubated. Spores are harvested from the incubated liquid culture by separating the spores from a culture supernatant. In some embodiments, the harvested spores are suspended in an aqueous medium to form a spore suspension. At least a portion of the culture supernatant is combined with the spore suspension.

A liquid sporulation method is described. A first liquid culture is prepared by adding bacterial cells to a sporulation broth, wherein an optical density (OD.sub.600) of the first liquid culture is in a range of 0.001 to 0.01. The first liquid culture is incubated and an optical density (OD.sub.600) thereof is increased to be in a range of 0.2 to 2.0. Second liquid culture is prepared by adding the incubated first liquid culture to a predetermined amount of additional sporulation broth, wherein an optical density (OD.sub.600) of the second liquid culture is in the range of 0.001 to 0.1, and a ratio of a volume of the second liquid culture to a volume of the first liquid culture is in a range of 10:1 to 150:1. The second liquid culture is incubated so an optical density (OD.sub.600) thereof is increased to be in a range of 1.0 to 4.0.

A liquid sporulation method is described. A first liquid culture is prepared by adding bacterial cells to a sporulation broth, wherein an optical density (OD.sub.600) of the first liquid culture is in a range of 0.001 to 0.01. The first liquid culture is incubated and an optical density (OD.sub.600) thereof is increased to be in a range of 0.2 to 2.0. Second liquid culture is prepared by adding the incubated first liquid culture to a predetermined amount of additional sporulation broth, wherein an optical density (OD.sub.600) of the second liquid culture is in the range of 0.001 to 0.1, and a ratio of a volume of the second liquid culture to a volume of the first liquid culture is in a range of 10:1 to 150:1. The second liquid culture is incubated so an optical density (OD.sub.600) thereof is increased to be in a range of 1.0 to 4.0.

The present invention provides a preparation method of Beauveria bassiana (Bals.) Vuill microsclerotium and a formulation thereof, and an application of the formulation thereof, belonging to the technical field of microsclerotium formulation preparation; the Beauveria bassiana microsclerotium formulation is prepared by the following steps of: 1) mixing conidia of Beauveria bassiana with Tween 20 aqueous solution to obtain spore suspension; 2) inoculating the prepared spore suspension into a liquid induction medium for induction culture to obtain microsclerotium; 3) mixing the prepared microsclerotium with a padding for drying to obtain dried microsclerotium; 4) mixing the prepared dried microsclerotium with an auxiliary to obtain a microsclerotium formulation. The Beauveria bassiana microsclerotium formulation produced by the method has low production cost, strong product tolerance and good insecticidal activity, moreover, after rehydrated, the formulation may rapidly germinate hyphae and produce spores to infect pests.

The present invention provides a preparation method of Beauveria bassiana (Bals.) Vuill microsclerotium and a formulation thereof, and an application of the formulation thereof, belonging to the technical field of microsclerotium formulation preparation; the Beauveria bassiana microsclerotium formulation is prepared by the following steps of: 1) mixing conidia of Beauveria bassiana with Tween 20 aqueous solution to obtain spore suspension; 2) inoculating the prepared spore suspension into a liquid induction medium for induction culture to obtain microsclerotium; 3) mixing the prepared microsclerotium with a padding for drying to obtain dried microsclerotium; 4) mixing the prepared dried microsclerotium with an auxiliary to obtain a microsclerotium formulation. The Beauveria bassiana microsclerotium formulation produced by the method has low production cost, strong product tolerance and good insecticidal activity, moreover, after rehydrated, the formulation may rapidly germinate hyphae and produce spores to infect pests.

Fusion proteins containing a targeting sequence, an exosporium protein, or an exosporium protein fragment that targets the fusion protein to the exosporium of a Bacillus cereus family member are provided. Recombinant Bacillus cereus family members expressing such fusion proteins are also provided. Genetically inactivated Bacillus cereus family members and recombinant Bacillus cereus family members that overexpress exosporium proteins are also provided. Seeds coated with the recombinant Bacillus cereus family members and methods for using the recombinant Bacillus cereus family members (e.g., for stimulating plant growth) are also provided. Various modifications of the recombinant Bacillus cereus family members that express the fusion proteins are further provided. Fusion proteins comprising a spore coat protein and a protein or peptide of interest, recombinant bacteria that express such fusion proteins, seeds coated with such recombinant bacteria, and methods for using such recombinant bacteria (e.g., for stimulating plant growth) are also provided.

Fusion proteins containing a targeting sequence, an exosporium protein, or an exosporium protein fragment that targets the fusion protein to the exosporium of a Bacillus cereus family member are provided. Recombinant Bacillus cereus family members expressing such fusion proteins are also provided. Genetically inactivated Bacillus cereus family members and recombinant Bacillus cereus family members that overexpress exosporium proteins are also provided. Seeds coated with the recombinant Bacillus cereus family members and methods for using the recombinant Bacillus cereus family members (e.g., for stimulating plant growth) are also provided. Various modifications of the recombinant Bacillus cereus family members that express the fusion proteins are further provided. Fusion proteins comprising a spore coat protein and a protein or peptide of interest, recombinant bacteria that express such fusion proteins, seeds coated with such recombinant bacteria, and methods for using such recombinant bacteria (e.g., for stimulating plant growth) are also provided.

The present invention relates to modified bacterial spores, and particularly, although not exclusively, to modified bacterial spores which comprise one or more heterologous genes, but without the introduction of antibiotic resistance genes. The invention extends to methods for producing such modified bacterial spores, vectors and kits for introducing heterologous genes in a spore.

The present invention relates to modified bacterial spores, and particularly, although not exclusively, to modified bacterial spores which comprise one or more heterologous genes, but without the introduction of antibiotic resistance genes. The invention extends to methods for producing such modified bacterial spores, vectors and kits for introducing heterologous genes in a spore.

The present disclosure provides inoculant compositions and methods for enhancing the survival and/or stability of microbial spores in an inoculant composition. In some embodiments, inoculant compositions of the present disclosure comprise microbial spores in a carrier consisting essentially of one or more water-miscible anhydrous liquids, such as cyclohexanol, cyclohexanone, N-methyldiethanolamine, and/or N-methyl-2-pyrrolidone.