Cloning after the first flush

[RedOctober]

I was wondering if there are any drawbacks to cloning a mushroom from the second or third flush i.e. tired growth, senescence, loss of potency etc…

Additionally if one does find a good clone, how many generations can one clone before these aging factors become noticeable and problematic?

[Sideshow]

I don't think there would be any drawback from cloning a 2nd or 3rd flush fruit. I always clone 1st flush, but that's just me. I think the wise choice in picking a clone specimen is to choose one which pins early; not coincidentally, these are usually the same ones that grow largest.

I have not cloned clones before, so I don't know what to tell you there. Common advice here, I think, is not to go past 1 or 2 clone generations.
It's easy enough to store the mother culture in the fridge and expand it again on agar, LC, etc when ready to use the culture again.

[BuckarooBanzai]

Senescence generally sets in when a culture has been actively growing for 2-4 months. That doesn't mean the culture is useless - but it will be notably slower/weaker than it was initially.

Every fragment of the culture retains an "awareness" of how old it is.

It doesn't matter what flush it is - it matters how long the culture has been growing since the spores first germinated.

Keeping a culture cold (no freezing - freezing kills 'em) slows down it's metabolism and slows the clock. Keeping a master culture in the fridge and expanding from it can stave off senescence for 6-12 months.

But all cultures will eventually get old and slow down.

[RedOctober]

Interesting, thanks guys! Buckaroo, do you store the fruit itself in the fridge or the LC that you make from the fruit?

I think I once stored a monster fruit in the fridge but when I went to make a culture from it a month later, it never grew. I had to chuck it unfortunately. It was like a 130g fruit too...sux.

It's a good tip too Buckaroo that it's not how much you expand a culture but how chronologically old it is. I never thought of it that way.

I think someone on here once told me that he clones not necessarily the largest fruit but the best cluster. He said this is the secret to getting dense and even pinsets.

[Hippie3]

i do like clusters.

[larshroom]

Originally Posted by BuckarooBanzai

Keeping a culture cold (no freezing - freezing kills 'em) slows down it's metabolism and slows the clock. Keeping a master culture in the fridge and expanding from it can stave of senescence for 6-12 months.

But all cultures will eventually get old and slow down.

I wonder if that would work for colonized grain...To use as a master for transfers.

[Hippie3]

it helps but it dries out over time too.

[BuckarooBanzai]

Storing colonized grain works better than storing LCs. Grain is a much more complete nutrient source than sugar water. But any form of long term storage of living tissue will eventually cause the sample to slow down and result in decreased performance.

You can stop all metabolic processes by storing in liquid nitrogen and samples should (theoretically) be good almost forever.

But you can store tissue samples much easier than using liquid nitrogen.

If you want to store a strain - just dry it. Completely dessicated mushroom material can be revived. I have successfully re-animated and fruited 3 year old stem that was rock hard using agar. There are documented cases in the literature of 30 and 40 year old dry samples being reanimated. LC would probably work as well as agar, but I've never tried it.

[larshroom]

Well, the strain I have now is BPK. It's good but it's not the strain I'm interested in preserving but more of just having colonized grain for a g2g transfer since from what I understand, it will slow eventually and has a limited lifetime. I just did my 2nd g2g transfer so I figure I have a couple left before I need fresh spores.

[BuckarooBanzai]

If you keep one master grain jar from the original spores in the fridge, it will stay healthy and usable for at least 3-4 months.

Chances are good it will last longer, but after 5-6 months, lots of variables creep in.

Murphy's law applies to planning for culture storage. If you trust that a single grain jar will be usable for 6 months and don't do anything else to save the strain - the jar will fail.

If you forget about it and leave it the back of the fridge for a year without needing it, it will be perfect when you finally test it.

[larshroom]

Okay, so I just did a g2g transfer which would be the second off the original spore. If, when one of the jars gets colonized, I put it in the fridge. Then in 3 or 4 weeks, after I pc some more quart jars, could I grab the one in the fridge, get a teaspoon for each jar I just pc'd, then put it back in the fridge and keep grabbing spoonfuls til it's gone.

Right now I'm just trying to get a flow going and inoculating seems to take alot longer. Once I have a personal supply built up, this probably wouldn't be such an issue.

I am buzzed so I apologize if this is inarticulate.

[larshroom]

Also sorry for hijacking this thread.

[BuckarooBanzai]

Originally Posted by larshroom

Okay, so I just did a g2g transfer which would be the second off the original spore. If, when one of the jars gets colonized, I put it in the fridge. Then in 3 or 4 weeks, after I pc some more quart jars, could I grab the one in the fridge, get a teaspoon for each jar I just pc'd, then put it back in the fridge and keep grabbing spoonfuls til it's gone.

That is correct. It would be good to start one or two more jars to store as fail safe masters. A grain jar can contaminate during transfers before it is empty.

Once fully colonized, keep them in the fridge at all times. DON'T freeze them.

[larshroom]

Thanks again Buckaroo.

As a side note, I was just thinking how bitchin the internet and forums like this is. I'm kinda old so I didn't have this when I was young. For example. I bought a new car a couple of years ago and felt stupid when I googled How to wash a car. It opened a whole new world of car detailing that I never knew about...Same as this a couple of months ago when my daughter bought a kit from a headshop.

[BuckarooBanzai]

Don't forget to thank AlGore for inventing The Internets in the first place.

Tis a magical age of easily accessed information. A shame that more people don't take advantage of it to educate themselves about the world around them...