A kind of culture method of high-purity Nomura reyesi yeast cell

A technology of Nomura reinhardtii and a culturing method is applied in the field of high-purity Nomura reinhardtii yeast-type cell culture, and can solve the problem that yeast-type colony boundaries are not obvious, conidia dispersion effect is poor, yeast-type cell purity is not high, etc. problem, to achieve the effect of long duration, obvious boundary and high purity

Inactive Publication Date: 2017-03-15
齐肇然
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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 conversion of Nomura reyesi from yeast-type cells to hyphae-type is affected by the population density. Yeast-type cell colonies can only appear in the end area of ​​the streak, and the purity of the obtained yeast-type cells is not high. In the microscope field of view 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, it is not convenient to determine the time to scrape the yeast-type cells, so it is urgent to explore the cultivation method to obtain high-purity Nomura yeast cells

Method used

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  • A kind of culture method of high-purity Nomura reyesi yeast cell

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Embodiment 1

[0049] A method for cultivating high-purity Nomura yeast cells comprising the following steps:

[0050] ① Add 20g of maltose, 8g of peptone and 8g of yeast extract to 900ml of distilled water to dissolve, then add 8g of agar powder and stir evenly to obtain a mixed solution, sterilize the mixed solution at 120°C for 20 minutes, and then evenly pour it on the plate to 2 The thickness of millimeters, the culture medium is obtained after cooling;

[0051] ② Use a sterilized inoculation loop to pick up the conidia of Nomura fungi reyesi, place them 5 cm above the surface of the culture medium obtained in step ①, tap the inoculation loop, and sprinkle the conidia of Nomura fungi reyesi evenly on the surface of the culture medium obtained in step ①. On the substratum, obtain the substratum inoculated with the conidia of Nomura Reidi, the distribution density of the Nomura conidia inoculated substratum is 0.5 / cm 2 ;

[0052] ③ Place the culture medium inoculated with conidia of Nom...

Embodiment 2

[0055] A method for cultivating high-purity Nomura yeast cells comprising the following steps:

[0056] ① Add 50g of maltose, 30g of peptone and 30g of yeast extract to 950ml distilled water to dissolve, then add 15g of agar powder and stir evenly to obtain a mixed solution, sterilize the mixed solution at 125°C for 30 minutes, and then evenly pour it on the plate until 3 The thickness of millimeters, the culture medium is obtained after cooling;

[0057] ② Use a sterilized glass rod to dip the conidia of Nomura fungi reyesi, place it 7 cm above the surface of the culture medium obtained in step ①, tap the glass rod, and sprinkle the conidia of Nomura fungus reyesi evenly on the surface of the culture medium obtained in step ①. On the substratum, obtain the substratum inoculated with the conidia of Nomura Reidi, the distribution density of the Nomura conidia inoculated substratum is 3 / cm 2 ;

[0058] ③ Place the culture medium inoculated with conidia of Nomura reyesi obtaine...

Embodiment 3

[0061] A method for cultivating high-purity Nomura yeast cells comprising the following steps:

[0062] ① Add 40g of maltose, 20g of peptone and 15g of yeast extract to 920ml of distilled water to dissolve, then add 12g of agar powder and stir evenly to obtain a mixed solution, sterilize the mixed solution at 122°C for 22 minutes, and then evenly pour it on the plate to 2.4 The thickness of millimeters, the culture medium is obtained after cooling;

[0063] ② Use a sterilized inoculator to pick up the conidia of Nomura fungi reyesi, place it 6 cm above the surface of the culture medium obtained in step ①, tap the inoculator, and evenly sprinkle the conidia of Nomura fungus reyesi on the surface of the culture medium obtained in step ①. On the substratum, obtain the substratum inoculated with the conidia of Nomura Reidi, the distribution density of the Nomura conidia inoculated substratum is 1 / cm 2 ;

[0064] ③ Place the culture medium inoculated with conidia of Nomura reyesi...

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Abstract

The invention discloses a culture method for high-purity Nomuraea rileyi yeast type cells. The method comprises the following steps that a culture medium is prepared, Nomuraea rileyi conidiospores are evenly dispersed on the newly-prepared culture medium to obtain the culture medium inoculated with the Nomuraea rileyi conidiospores, the distribution density of the culture medium inoculated with the Nomuraea rileyi conidiospores is 0.5-3 / cm<2>, the inoculated culture medium is placed in the illumination environment under the temperature of 24-30 DEG C and the humidity of 75-90%, the semitransparent liquid drop shaped Nomuraea rileyi yeast type cells are obtained on the surface of the culture medium after 96-120 h, and then the Nomuraea rileyi yeast type cells are amplified and cultured. According to the culture method for the high-purity Nomuraea rileyi yeast type cells, the duration time of the growing period of the yeast type cells is long, and the conversion time of the yeast type cells to the hypha state is correspondingly prolonged, so that the yield of the yeast type cells is improved, no mycelium is detected in the microscope examination, and 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. 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 yeast cells. [0003...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/16
CPCC12N1/16
Inventor 齐肇然殷幼平王中康邵长文李富宽董芳邱奉同
Owner 齐肇然
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