Diploid yeast cells induce meiosis and spore formation when they are starved of both nitrogen and carbon sources. At this point, their chromosomes replicate and undergo a secondary division to produce a haploid nucleus.

Operation method

basic program

Materials and Instruments

Yeast
YPD
Petri dish Incubator

Move

I. Spore formation on plates

1. Pick or selectively inoculate single colonies of yeast from a plate onto a spore forming plate.If there is no need for selective viability, cells can be pre-grown on YPD plates for a few days before being transferred to spore forming plates. Individual colonies can be grown on YPD plates for 3 to 4 days, while for small clumps of cells, they can be grown on the plate for 2 days.This pre-growth is not necessary, but it allows for a somewhat more efficient spore formation.When transferring yeast cells, a small number of cells should be used to coat a relatively large area of a spore-forming plate, and clumps of inoculum should not be seen.2. Incubate at 25°C for 4 days.3. Put a drop of water on the slide, pick a small number of cells diluted in water, and observe under a microscope with a magnification of 250×~400×, that is to say, you can see the tetrasporangium spores suspended in water.II. Spore formation in liquid medium1. In a suitable culture vessel, add YPD medium to culture, and wait for the spore-forming diploid cells to grow until the _OD600 is 2.5~3.0.

2. Transfer 1 ml of culture medium to a single-use sterile 15 ml polypropylene tube and centrifuge at 12 000 g for 5 min.

3. Pour off the supernatant, resuspend the cells with 5 ml of water, and centrifuge according to step 2.

4. Pour off the supernatant and resuspend the cells in 1 ml of spore-forming liquid medium supplemented with the nutrients required for the specific diploid cells.5. Incubate at 30°C and above 350 r/min for 2-3 days and examine spore formation under a microscope.

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