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04-17-2009, 08:13 AM | #21 (permalink) |
Orang Puteh
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Re: Message from me
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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04-17-2009, 12:48 PM | #22 (permalink) | |
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04-17-2009, 01:10 PM | #23 (permalink) |
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Re: Message from me
Hi,
thanks. eden |
04-17-2009, 01:17 PM | #24 (permalink) |
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Re: Message from me
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 |
04-17-2009, 01:20 PM | #25 (permalink) |
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04-17-2009, 01:21 PM | #26 (permalink) |
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04-17-2009, 01:33 PM | #27 (permalink) | |
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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 |
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04-17-2009, 01:42 PM | #28 (permalink) | |
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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 |
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04-17-2009, 02:12 PM | #29 (permalink) | |
The causasian Asian!
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04-17-2009, 03:44 PM | #30 (permalink) | |
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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 |
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04-17-2009, 05:37 PM | #31 (permalink) |
The causasian Asian!
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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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04-17-2009, 05:50 PM | #32 (permalink) | |
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04-17-2009, 06:52 PM | #33 (permalink) | |
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04-17-2009, 06:55 PM | #34 (permalink) | |
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Re: Message from me
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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 |
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04-17-2009, 07:07 PM | #35 (permalink) |
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Re: Message from me
I'd like to place my order for an aeae plantlet.
thanks Michael |
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04-17-2009, 07:10 PM | #36 (permalink) |
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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.
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04-17-2009, 07:49 PM | #37 (permalink) | |
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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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04-17-2009, 08:05 PM | #38 (permalink) |
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04-17-2009, 08:08 PM | #39 (permalink) |
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04-17-2009, 08:10 PM | #40 (permalink) |
The causasian Asian!
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Re: Message from me
Thank you Eden, I will be trying that. Now all I need is some AeAe pups.
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