Chironex

I have to say, I am having a blast doing this so far. I keep a journal to log all of the details, so if I make a mistake, it is noted. (Like roasting the 2 culture jars in the pressure cooker!!! LOL)

It is amazing to me that that tiny piece of apical meristem can create so many banana plants. After I feel like i have it down pat, I am going to attempt to induce some mutant variegates.

Thank you for all of your kind words and support.

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Scot

bigdog

Scot, I've found the best way to rescue embryos from Musa seeds is with a small vice, mounted on a table. You just slowly tighten the vice until you hear a crack, then remove the seed from the vice. Find the crack, and pry it open with a scalpel, knife, something sharp. I haven't yet mangled the first embryo with this method.

Regarding contamination...it really is ridicuously easy to grow mold, lol. We did some cultures the other day, and 3 out of our 9 cultures got some contamination. I didn't feel so bad though, seeing the next group get every single one of theirs contaminated! This was with a professional setup, but most of the students had very little or no tc experience. Even with a constant flame, wiping down with ethyl alcohol, 10% bleach soak, rinsing with distilled water many times, and working as close to the hood as possible, contamination is still possible. It only takes one little particle! Good luck with your setup there.

Frank

Chironex

Thanks! I think I will buy a C - clamp to do the same thing. That way I can sterilze it in the pressure cooker, wipe it with alcohol and put in in the hood to crack the seeds. I try to do as much under the hood as I can.
So far, only one contaminated out of 7 cultures. It was one of the Ingens embryo rescues, and we know that they seem to be likely to have endocarpic contaminates, no matter how much I sterilize them. But, I will say that out of the 5 I did, 1 or 2 look like there is some promise. The other 2 have no contamination, but are doing nothing, so I may have injured the embryo upon removal. The 5th one was the contaminated.

The Manzano and the California Cold hardy banana cultures appear to be aseptic so far. I will be doing 2 more cultures tomorrow or this weekend.

I appreciate your encouragement.

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Scot

Richard

Go Scot, go! I see from your avatar that your cortex stem has gone banana.

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Back in business at plantsthatproduce.com

Chironex

The way I see it, if Jimmy Buffett can have Parrotheads, I can have BananaHeads, right?

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Scot

Chironex

Here is one of 3 pics posted in my gallery of a Manzano TC. It is beginning to shoot now.

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Scot

Chironex

Here is one that was just put into TC today for one of our members. The surgery went well, and the prognosis is excellent.

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Scot

damaclese

Scot you are so cool dude thanks for sharing this with us congrats looking forwerd to more info and pics keep them coming! from the pics they defanatly look good to me! how are the Engen seeds doing? can't wate till you can Tc those im sure you will have no problims selling them to the Org rember im in line to LOL

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harveyc

Scot, thanks for all the photos, comments, etc.

Whey you refer to "California Cold Hardy", are you referring to the one sold by Shawn in Lodi, CA? Or is there something else referred to as that?

Harvey

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Chironex

Yes, I believe that is the one. Thanks for your kind remarks.

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Scot

Bob

Ditto for me.

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ursula

with all the experience you all have you make me feel very insecure!
cant you discuss technical theatre for a while?
ursula

MediaHound

Good work Scot!

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Apologies in advance if I am slow to reply to your PM. I suggest posting in the forums for support if you need something urgent.

Chironex

Today put 6 E. perrieri embryos into culture along with 1 laterita and 1 itinerans embryo. Also took one ingens seed from germination tray and rescued the embryo onto culture media. Gabe was very helpful while I was doing this and Frank's idea of using a small vise to crack the seed hull worked like a dream! It's the only way to go from now on. Look how small an itinerans seed is and imagine trying to split it without damaging an embryo the size of a pin head! Using the vise, the embryos just fell out, or were easily removed if they were stubborn. Note: this method works best on dry seeds, if soaked, they just split open slightly. Embryos were easier to extract from the dry seeds.

We will see how they look as time passes. No pic's yet, my camera resolution is not that great, so they would be blurred, if visible at all.
Seeds were first sterilized in Physan 20 with some Tween 20 as a surfactant. Then they were rinsed for 20 minutes in 10% bleach water solution. The seeds were then cracked, as aseptically as possible, and put into a quick rinse in 10% bleach solution, rinsed 3 times in sterile water, then placed upright onto M&S media.
I hope to get my hands on some better media for embryo rescue at some point, but this was all that was available for now.
Now we wait...

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Scot

Chironex

Technical theatre - ummm, project your voice from the diaphram, that's the extent of my knowledge - you got me there. lol

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Scot

51st state

Looking good there Scot. keep it up.

Once you've got it nailed I'll try get some unusual stuff over to you

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Chironex

Kev, that would be fun. Wait until I get more trials in with different media. Some variances occur with different banana varieties. Everything is being logged so that the formulas that work on certain bananas will already be known. Need to get the medium for embryo rescue, as the normal M&S is not as robust.

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Scot

edd82

Wow that's so cool :-) Good luck with it.

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Chironex

Yesterday 8 more ingens embryos were dissected and placed into embryo rescue. Still waiting for chemicals to use with the new medium using modified Nitsch & Nitsch.

The Viente Cohol was sub-divided again into 12 plantlets. The California Cold-hardy banana has not done anything since induction. Can't figure out why, as there does not appear to be any contamination. It just seems to be happy taking its time.

The Manzano culture is also taking its sweet time to do anything. Next cultures I do I will split up using charcoal in the medium and citrate. I have also been doing some reading about alternative media such as straight peat with some hormones. Others are cotton balls, perlite, vermiculite and gellan gum. I got some of each to test the differences in cultures.

Phytotech Labs has a Banana culture medium that also shows some promise, so I am going to set-up some trials with it, too.

The hope is to aggregate banana TC data and construct a "best culture environment" database for each banana type, including Ensete and Musella. Sometimes the strength of the medium including the additives can vary the results dramatically.

There is also an antibiotic that has been shown to induce variegation that remains true to type. This may be a key to use in AeAe culturing.

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Scot

harveyc

Wow, Scot, you've really got into this! What are your objectives in producing plants? Are these for your own use, to give away to friends and family, or do you plan on selling plants?

While I have no plans to for commercial applications, I thought I might like to learn TC so that I could produce more plants for my own use. For instance, a fast maturing variety like Viente Cohol might work well being grown as an annual so I wouldn't have to worry about cold hardiness. I'm thinking I could TC some plants in the fall or winter and grow them in pots in my greenhouse until spring and hope for ripe fruit by the end of summer or early fall from several plants. I have lots of relatives and friends to share the fruit with.

I'm not sure I have the discipline necessary to carry out the TC process, though. I just need to focus my attention, perhaps.

Thanks for sharing!

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