Smellyhobbit said:

Ok I have a real question:

How is your growth on grains so fucking fast? I would never see that much growth after 8 days. If I didn’t shake my jars after they reached maybe a 1-2” diameter it would take probably 2-3 weeks to see growth like that.

What am I doing wrong?

---

Was that directed at me, Smelly?

If so, yes about a full plate sliced like this per jar. Also, those are pint jars rather than quarts, so that could be a bit deceptive.

Everything inside the blue circle goes in one jar.
Well... everything that doesn't miss the jar, anyway.

ReverendMyc said:

Smellyhobbit said:

Ok I have a real question:

How is your growth on grains so fucking fast? I would never see that much growth after 8 days. If I didn’t shake my jars after they reached maybe a 1-2” diameter it would take probably 2-3 weeks to see growth like that.

What am I doing wrong?

---

Was that directed at me, Smelly?

If so, yes about a full plate sliced like this per jar. Also, those are pint jars rather than quarts, so that could be a bit deceptive.

Everything inside the blue circle goes in one jar.
Well... everything that doesn't miss the jar, anyway.

---

ReverendMyc said:

Ya. Small hands make your "jars/myc" look huge.

Always picking on my tiny hands. You guys are just mean.





EDIT: Lemon for scale. Didn't have a banana.

---

Traveled said:

It's classic LMEA this a touch of yeast.

I m a bit disappointed by this batch because I didn't achieve totally clear agar.
Maybe I put too much of agar and or lme ?

---

covertjoy said:

Update on original Ps. Cyanescens dilution plates.

No visible germination on any of the original Ps. Cyanescens dilution plates I did which were on water agar. However I was scanning a few under the microscope and found this.



There are no nearby spores so I think this one looks like a fairly safe monokaryon.

I was able to spot it using the 10x microscope objective which is nice since that allows me to cover a lot of surface area fairly quickly. I think the technique for catching the germinations early is not searching for single spores which is labour intensive but a few days after inoculation to start to scan the plates daily or twice daily looking for for hyphae using the 10x objective.

Going to transfer this straight away. Planning to do as small a transfer as possible. I think I am ok with doing the transfer face down if I am able to so the hyphae comes into contact with nutrient agar immediately. From what I have seen they don't get starved of oxygen too quickly for them to find the edge although I may be wrong for a mycelium of such small size.

It looks like hyphae grew from both sides of the spore. Cool.

Here is the completed transfer. It is face down.





I'll keep an eye on it under the microscope. For all I know this spore might have germinated days ago and has terminated.

---

0mar said:

Day 6 check up on this ghost plate. Some nasty shit is starting to grow in between the dishes but the slow growth is starting to turn  white now. Waiting on some new dishes to try a different recipe to see if that helps.

---

0mar said:

So the transparent part is mold or the white? Or both? Man I wish I knew where I’m going wrong

---

0mar said:

So the transparent part is mold or the white? Or both? Man I wish I knew where I’m going wrong

---

covertjoy said:

On my other dilution plates which were on LME agar I have had a number of visible germinations from Ps. Natalensis #2 which I have transferred and are growing well. I didn't observe their germination under the microscope so I can only suspect that they are monokaryotic at this point and need to do further work to prove them.

No germinations on any of the others (Ps. Natalensis #1, Ps. Hispanica, Ps. Allennii). I inoculated regular germination plates for all of these also and of them only the Ps. Natalenis #2 had prolific germination. Ps. Natalensis #1 had very sparse germination and the other two had no germination at all. So I conclude that these spores are in a bad state or don't like the conditions. It has been hot lately, over 30 Celcius and the plates have been sitting out with temperature swings as well so perhaps this has been a factor. I used dishwasher rinse aid to help unclump the spores so perhaps this was a factor too. It is interesting that the Ps. Natalensis #2 germinated well if this was the case though.

---

Ps. Natalensis #2 regular germinations:


Ps. Natalensis #1 regular germination:


Ps. Hispanica regular germination:

0mar said:

RIP thanks boys.

---

could try some more transfers from the germ plate. definitely don't toss any of those plates though. maybe one sprout a plin in a couple weeks