Grain to grain transfers

[scruffy_hair86]

Iv'e been fiddling with kings oysters for a few months now and with each batch that I do im saving one jar to then inoculate the next ten.

I have done this probably 5 times so far and was wondering is there a point where this process will weaken the mycelium and alter the performance of the fungi.

Thanks for your help.

[El Presidente Hillbillios]

i aint familiar with king oysters. But i have done it for a year! just to see how far it would go. Was at least 30 transfers. It did start to lose its virility at the end though. Just not as strong myc and not as fast and vigorous

[MagicalMedic]

Random question: does mycelium grow faster in grain spawn than it does it liquid culture?

I've gathered from reading that the process of inoculating sterile liquid culture with a spore syringe, then drawing it off with a syringe via an injection port when ready and injecting this directly into substrate might be a relatively easy way to maintain sterility all the way through.. so why is grain spawn way more popular? If using a syringe & injection port with liquid culture you'd never even have to open the bottle

[El Presidente Hillbillios]

Grain spawn is a good bulky way of getting spawn into a bulk and its easy to evenly distribute. Lc adds alot of wetness that you have to also account for. mositure content is a big secret to doing any of it right really.

But to your point that is an easy way to keep a sterile culture. Then use that as a master to start gains or other LC's. Only touch it when you finished your working cultures.

[OPP]

The best route (arguably) is to go from spore print to agar (choosing the mycelium characteristics you want to use) to LC to bulk.

The agar ensures you don't have any contaminates and a fast colonising mycelium (although possible poor fruiter). The LC will "colonise" very quickly due to the high sugars which is then used to innoculate your Bird seed (or whatever you are using). this way you aren't waiting for the spores to "germinate". The mycelium is "alive" as soon as it goes into your spawn and takes off very quickly.

Some people do all or some of these steps. Whatever works for you.

Another option is to innoculate a small jar and when it is 100% colonised, with no signs of contamination, inject it with sterile water. Shake the jar and the mycelium with become suspended in the water. This mycelium water can then be used to innoculate other jars (similar to a LC).

Edited October 23, 2011 by OPP

[MagicalMedic]

Right yeah hadn't really thought about adding to much moisture. Using as a master sounds like it'd be a good idea though.

Like OPP said too, not waiting for germination (once you've got an established LC) would speed things up heaps. I was thinking though; LC's are much easier to make and safely inoculate than agar plates - so if you were to make syringes and inject into say ten prepped LC bottles, some might be contaminated but it took like five minutes to make them right? so you can just throw the contaminated cultures if there are any. The main advantage though is there is no transfer from agar to LC / grain which seems like the riskiest part of the adventure, the part that'd likely require a flowhood.

Is it harder to clearly observe mycelium growth in LC than it is on agar?

The agar in the bottom of a jar then adding water after colonisation is great - no flow hood / glow box needed at any stage in the process, and you still get the benefit of watching growth on agar and excluding contaminates. That is smart.

OPP I noticed you spell 'innoculate' with two n's as I was before the little red line starting irritating me.. is this correct australian / british spelling? All online dictionaries spell it with one n.

As you said anyway so many options; up to the individual. one might easily avoid needing a flowhood/glovebox at all with the liquid process and all transfers using syringes though. Appealing in its simplicity and apparently minimal risk.

[Hyphal]

The term used to describe a culture losing its vigour is 'Senescence' http://en.wikipedia.org/wiki/Senescence.

As stated above, a master or 'mother' culture can stop this to a point. Age is inevitable, at which point you need to start from a fresh print.

[tripsis]

Like OPP said too, not waiting for germination (once you've got an established LC) would speed things up heaps. I was thinking though; LC's are much easier to make and safely inoculate than agar plates - so if you were to make syringes and inject into say ten prepped LC bottles, some might be contaminated but it took like five minutes to make them right? so you can just throw the contaminated cultures if there are any. The main advantage though is there is no transfer from agar to LC / grain which seems like the riskiest part of the adventure, the part that'd likely require a flowhood.

You're giving LC far too much credit. It is a less desirable medium than agar to use and easy to contaminate, while simultaneously be far more difficult to identify it has been contaminated. Often the only way to tell if it is contaminated, is to waste time by injecting your grain jars and seeing what happens. A better method is to put a drop onto an agar plate and see what the resultant growth is like.

You never actually require a flowhood. A glovbox will suffice and even that is not strictly necessary. I spent my entire last trip in India cloning wild mushroom species to agar is open air situations, including on the train once. I just used peroxided agar to help reduce contamination, coupled with being quick and vigilant about transferring away from contamination.

Is it harder to clearly observe mycelium growth in LC than it is on agar?

It's not hard on either, but with agar you can clearly identify the type of mycelium that is growing. Moulds also tend to sporulate much faster on agar, making identification simple.

The agar in the bottom of a jar then adding water after colonisation is great - no flow hood / glow box needed at any stage in the process, and you still get the benefit of watching growth on agar and excluding contaminates. That is smart.

You still need to inoculate the agar in the jar, which would require opening the lid.

OPP I noticed you spell 'innoculate' with two n's as I was before the little red line starting irritating me.. is this correct australian / british spelling? All online dictionaries spell it with one n.

The correct spelling is with one 'n'.

As you said anyway so many options; up to the individual. one might easily avoid needing a flowhood/glovebox at all with the liquid process and all transfers using syringes though. Appealing in its simplicity and apparently minimal risk.

LC might seem appealing, but it's prone to problems, which often result is failure and wasted time/energy. Agar is actually a very simple and much more reliable.

[MagicalMedic]

Right well there's my answer at to why LC isn't so popular when it seemed appealing, thanks for such a comprehensive response. I guess all teks have their advantages/disadvantages for certain parts of the process, but that certainly puts LC in perspective.

Edited October 24, 2011 by MagicalMedic

[Heretic]

Just wondering if possible or feasible to inoculate grain or agar with myc from a PF tek jar? [ soory if its a dumb question ; I have very little mycology experience .]

[MagicalMedic]

Just wondering if possible or feasible to inoculate grain or agar with myc from a PF tek jar? [ soory if its a dumb question ; I have very little mycology experience .]

 

Yeah man of course - you only need to transfer a tiny little sample of the mycelium to inoculate either. The trick is doing so without contamination.