Tissue Culture - Umami Style

it is very challenging and takes a lot of practice and patience. I got to try this hands on and the learning curve is steep. Even the transition from working with your eyes directly on an object vs working on a screen was a challenge. But like anything else practice makes perfect.

It does seem like most advertise/show off shoot tc vs meristematic. Never got that.

lastly, why hasnt anyone brought sour d back to life yet!?!?
 
Rico D said:
it is very challenging and takes a lot of practice and patience. I got to try this hands on and the learning curve is steep. Even the transition from working with your eyes directly on an object vs working on a screen was a challenge. But like anything else practice makes perfect.

It does seem like most advertise/show off shoot tc vs meristematic. Never got that.

lastly, why hasnt anyone brought sour d back to life yet!?!?
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Sour is very much alive and well in my greenhouse. I just dusted her with zeclair and jelly zonuts!
 
Really good info in this thread, much appreciated. Looking forward to popping your beans and hopefully finding some stock to breed with in the future! TC is quite a ways down the line for me but I think it will come in handy when I have the bandwidth to do so. Any suggested reading material for 101 level TC knowledge? I got the book Plants In Test Tubes and my dog ate it up before I could comb it over :ROFLMAO:
 
Jive Genetics said:
Really good info in this thread, much appreciated. Looking forward to popping your beans and hopefully finding some stock to breed with in the future! TC is quite a ways down the line for me but I think it will come in handy when I have the bandwidth to do so. Any suggested reading material for 101 level TC knowledge? I got the book Plants In Test Tubes and my dog ate it up before I could comb it over :ROFLMAO:
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read this: https://summit.sfu.ca/_flysystem/fedora/2022-11/etd21287.pdf

and watch a bunch of youtube videos.
 
Its great to see threads of this nature...finally. What experience do you have using turmeric and benzoin(sambrani) resin-based fumigation methods for microbial contamination surface sterilization of explants as opposed to using chemicals that become toxic to explants and surfaces as doses increase?
 
Rumpelstiltskin said:
Its great to see threads of this nature...finally. What experience do you have using turmeric and benzoin(sambrani) resin-based fumigation methods for microbial contamination surface sterilization of explants as opposed to using chemicals that become toxic to explants and surfaces as doses increase?
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Honestly, none. I have had great success with iodine + soap and then varying levels of chlorine for keeping explants sterile. The biggest issue that I have faced is endophyte contamination. I seem to have alleviated that for the most part by running a sterile media and fertilizer. Also, I don't recommend using PPM (plant preservation mixture) for general use, but it does work pretty well for endophyte issues.
 
ZODA did 6 nodes in 20 days. With an average of 10 fresh explants per "micro mom" per month, the growth rate is exponential and your only limitation is shelf space and how long you want to live in front of a flow hood....


20240206_121711.jpg
 
So lets check out what we got after another week of lab work. Zeclair is looking great for one weeks worth of growth!

20240213_085753.jpg


Payzos is also looking nice and healthy in stage 1.

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This far, my contamination rate is less than 1%. Hopefully that will continue but endophytes have a habit of popping up when you least expect them... Sometimes as far as a few months into the process. Meristem cultures (as shown here) are usually immune, but It does happen resulting in a loss of individual explants.

When the contamination does occur, I remove the other healthy explants and transplant them into fresh media and hope for the best.

20240214_093345.jpg
 
Umamiseedco said:
Honestly, none. I have had great success with iodine + soap and then varying levels of chlorine for keeping explants sterile. The biggest issue that I have faced is endophyte contamination. I seem to have alleviated that for the most part by running a sterile media and fertilizer. Also, I don't recommend using PPM (plant preservation mixture) for general use, but it does work pretty well for endophyte issues.
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for sterilization how about HOCL?

my lettuce raft has never looked better......
 
Any free available chlorine (FAC) will work just fine. I make mine from calcium hypochlorite at 5000ppm chlorine, and the active chemical from that mix is HOCL. You can use bleach as well, hypochlorite (OCl-). Both are effective at disinfection, but chemically speaking – hypochlorous acid is most effective for this purpose.
 
Umamiseedco said:
Any free available chlorine (FAC) will work just fine. I make mine from calcium hypochlorite at 5000ppm chlorine, and the active chemical from that mix is HOCL. You can use bleach as well, hypochlorite (OCl-). Both are effective at disinfection, but chemically speaking – hypochlorous acid is most effective for this purpose.
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5000..<--interesting.

the hocl plant Info of I have read says they start at under 50 ppms

and that 5 ppms is good enough.[.yeh right..maybe for a reservoir].[.jk]

..of course we like to push things as far as we can....[for science]
 
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