A method for cultivating high-purity Nomura reyesi yeast-type cells in large quantities

Abstract

The invention discloses a method for culturing high-purity nomuraea rileyi yeast cells on a large scale. The method comprises the following steps: preparing a culture medium, uniformly scattering nomuraea rileyi conidia on the culture medium with pH value of 6.5-8.5 to obtain a nomuraea rileyi conidium inoculated culture medium, putting the inoculated culture medium in a light environment at a temperature of 24-30 DEG C and with humidity of 75-90%, obtaining semitransparent liquid drop shaped nomuraea rileyi yeast cells on the surface of the culture medium after 96-120 hours and then carrying out enlarge culture on the nomuraea rileyi yeast cells, wherein the distribution density of the nomuraea rileyi conidium inoculated culture medium is 0.5-3 / cm<2>. The method for culturing high-purity nomuraea rileyi yeast cells on a large scale has the advantages that the growing period duration of the yeast cells is long and the state switching time from the yeast cells to hyphae is correspondingly delayed, so that the yield of the yeast cells can be increased; mycelia can not be detected in microscopic examination, so that the purity is high.

Description

technical field [0001] The invention relates to the field of insecticidal fungal cells, in particular to a method for cultivating high-purity Nomura reyesi yeast cells in large quantities. Background technique [0002] As an entomogenous fungus, Nomura lyseri has become a hotspot in theoretical research and production application in the field of biological control. Nomura lyseri has two types of growth phenomena on the surface of solid medium, that is, the transition from yeast-type cell state to hyphae growth state. On the one hand, , the yeast-type cells of Nomura reyesi are similar to the worms in the worm body, and the blastospores produced in liquid fermentation grow rapidly and are suitable for large-scale industrial production; on the other hand, in the ATMT system developed by Shao Changwen, the yeast-type cells As a recipient cell, whether it is industrial production or laboratory research, it has put forward a realistic demand for the cultivation of Nomura reyesi y...

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Problems solved by technology

[0003] At present, there are two methods for obtaining the yeast-type cells of Nomura reyesi: one is to use an inoculator to draw a line on the surface of the solid medium for preparation. This method has a small amount of preparation and is not effective in dispersing conidia. The transformation of Nomura reyesi from yeast-type cells to hyphae is affected by population density and pH, and yeast-type cell colonies can only appear at the end of the line, and with the growth of Nomura reyesi, its metabolism The product will cause the pH value of the medium to drop, and when the pH value of the medium drops below 6.5, the conversion of yeast-type cells to hyphae-type growth will become more and more rapid, so the purity of the obtained yeast-type cells is not high. There is a large amount of mycelium in it, which requires multiple transfers and purifications to obtain high-purity yeast-type cells; the second method is to use the method of preparing conidia suspension. Although this method has a better dispersion effect of spores, it is easy to pollution, and the boundary of yeast-type colonies is not obvious when they grow out, and it is not convenient to determine the time to scrape yeast-type cells. Therefore, it is urgent to explore the method of cultivating high-purity Nomura yeast-type cells in large quantities.

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