Bob

Welcome Eden, thanks for your responses to everyone's questions here. I don't have any but, enjoy learning any new info provided on my favorite fruit.

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Scot Nelson

Hi Eden, it's nice to see you here.

Scot N.

eaperez

Hi,
thanks.
eden

eaperez

yeah, nice to see you here too. i am enjoying this and i am learning too. imagine, i've been working on bananas for so long and the first question that showed me that i've got to learn more is the banana cultivar aeae. first time that i've seen this plant is in in Angela Keppler's yard. anyway. i google searched the image and voila. i think i have a good suggestion on how we can tc this plant though it is a slim chance of getting the same phenotype as the mother plant.
later.....
eden

eaperez

Hi Bob,
thanks. i'm enjoying and i'm learning a lot too.....
eden

eaperez

Hi Tog,
thanks.

eaperez

Hi Scot,
I have no experience in embryo rescue but i thought it is not so difficult as long as the seed is viable. i did immature embryo rescue on corn so this shouldn't be a problem. i know one media formulation for germination of banana embryos from somatic cells initiated through somatic embryogenesis. try this and tell me if it works: MS salts + Morel and Westmore Vitamins (100 mg/L myo-inositol, 1 mg/L thiamine-hydrochloride, 1 mg/L nicotinic acid, 1 mg/L calcium panthotenate, 0.01 mg/L biotin, and 10 mg/L folic acid) + 0.045 mgL BAP + 0.2 mg/L IAA + 30 g/L sucrose, pH 5.7.
aloha.....
eden

eaperez

Hi Scot,
I'm not certain about the endogenous contamination. I suggest you do the following: surface sterilize the whole fruit with 50% chlorox (commercial) for 20 - 30 minutes. open and collect the seeds in sterile condition (in laminar flow hood). either plate directly to the media or excise the embryo then plate onto the media. you'll be able to see if there is an endogenous contaminants or none at all depending on how good they were excised and transferred. I don't think PPM will harm the embryos when used supplemented onto the media or directly exposing the embryos at the concentrations that was suggested by the supplier. hope this works....
Eden

Chironex

Eden, I am all ears if you have a possible way to tc this chimera and have any chance of retaining phenotype of the mother. I believe it is a periclinal chimera from past discussions here. I am interested in trying to induce variegation on green AeAe to see what comes of it. I have some basjoo that I plan to use for variegation induction using streptomycin or spectinomycin. If this shows promise, then on to AeAe. There is some research done using this method that has produced variegates that remain true-to-type in subsequent generations. Heat and cold may also be able to be used to induce this trait as well. This is also in the same research article.

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Scot

eaperez

Hi Scot,
after reading past discussions on AeAe here, i think one way of propagating this and produce true-to-type to the mother plant is also through tissue culture, excising the apical dome and no cutting ( we do longitudinal cutting into quarters for initial establishment in micropropagation) and using very minimal hormone (BAP) so that only axillary shoots (suckers) will be initiated and not adventitious shoots that may have come from the sector of non-mutated part. the new shoots coming from the base of the explant should have the same phenotype as the initial explant. you may also put a wound to the apical dome to stop the apical dominance and redirect growth on producing axillary buds (suckers). in this way, micropropagation would be slow as compared to the usual protocol of micropropagation but way faster than in vivo propagation (waiting for suckers to grow out). if you want a lot of variations, then you can do the usual protocol of micropropagation. Chimeras can segregate in vitro and i think this depends on your technique of micropropagating them. i wish i can have a hand on doing this but i have no way of getting initial explants from here.
also, i think if the green AeAe still carry that mutated sector then you can induce separating or regenerating the varigated shoots/plants from them through the usual micropropagation system.
well, good luck......
eden

Chironex

I will try this once I find some AeAe pups. Thanks for confirmong what I was thinking. The hard part will be identifying just the apical dome tip. I imagine an investment in a stereoscope might prove useful.

But then, after initiation and getting some growth, do you think sub-culturing it by quartering it longitudinally would still work?

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Scot

Scot Nelson

Eden, there are AeAe plants growing on the Big Island if you want some. You can e-mail me.

eaperez

it isn't hard to identify the apical dome. that portion is solid white or cream in color. do not do cut the dome into quarters. you'll be getting regeneration of mixtures of adventitious shoots from different sectors of the dome and not only axillary buds at the base of the dome. as i've mentioned, inflicting wound to the dome would result on pushing the side shoots/suckers/axillary buds ("apical buds give rise to axillary buds in such a fashion that all three histogen layers are maintained") to come out.....

eaperez

Hi Scot,
Yeah, i would like to have some and try to either segregate the chimeras or retain the phenotype of the mother plant. Kooh-n-Hui is going to the Big Island next week. I'll send you email after talking to her. Thanks,
Eden

Michael_Andrew

I'd like to place my order for an aeae plantlet.

thanks
Michael

Michael_Andrew

But seriously I'm in total awe of your knowledge and theory. And if your successful at tc'ing an ae ae that would be awesome.

Chironex

I understand, but it is still like peeling the leaves from an artichoke except on a much smaller scale. You can see one of my photos of the a.m. here:


This one is sitting in a baby food jar Magenta cap. So, I would make horizontal slices off the top .5 mm as I understand it. Would you suggest making subsequent .5 mm cuts from the remaining a.m. to initiate separate cultures? I suppose it couldn't hurt. I just don't know if I can slice it thin enough without using a microtome.

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Scot

eaperez

don't make any cutting or slicing. you may want to peel off more of the leaves and expose the apical dome or meristematic region . poke the apex with scalpel blade to inflict wound and let the whole apex get established in the media. you don't need to separate cultures. if you want more cultures then excise apex from another suckers. the idea is to establish the aeae in in vitro culture and initiate axillary buds in a natural way but with the help of hormone to speed up the growth of the buds.

eaperez

i'll let you know when we've got keikis (sister plants or baby plants) from them.

eden

Chironex

Thank you Eden, I will be trying that. Now all I need is some AeAe pups.

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Scot