View Full Version : Message from me
eaperez
04-16-2009, 06:40 PM
Hello to everyone,
My name is Eden A. Perez. I am from the Philippines and I'm here at the University of Hawaii since 1999. I graduated in 2006 with Ph. D. in Plant Pathology (Virology). I am currently working on my project entitled "Rehabilitation of BBTV infected areas in Hawaii" thus my work involves indexing bananas for use as starting material of in vitro cultures and micropropagation of virus-free bananas. Hope to meet all of the members through chat here. I welcome questions regarding tissue culture and genetic improvement of bananas. Aloha......Eden
xavierdlc61887
04-16-2009, 06:45 PM
u should meet "GABE" hes in hawaii too and does what u do!!!!
mskitty38583
04-16-2009, 06:53 PM
welcomoe to the org....ok question number 1: in laymans terms please....why cant we tc an aeae and get a varigated species??????????????
eaperez
04-16-2009, 07:39 PM
Hi to all,
Thanks so much for welcoming me.......eden
eaperez
04-16-2009, 07:44 PM
sorry, i think i don't know an aeae. is this a varigated banana? i think i have an idea if it is.....eden
eaperez
04-16-2009, 07:47 PM
welcomoe to the org....ok question number 1: in laymans terms please....why cant we tc an aeae and get a varigated species?????????????? know what an aeae is.
sorry, i replied to all. now i discovered how to send a reply to each of the emails. i do not know what an aeae is. if it is varigated banana, i think i know why it cannot be tc to produce more varigated plantlets.....
eaperez
04-16-2009, 07:49 PM
u should meet "GABE" hes in hawaii too and does what u do!!!!
i know one Gabe. i think he is one and the same Gabe who is one of my lab assistants.....
xavierdlc61887
04-16-2009, 08:11 PM
look for him here maybe it is!!!
Caloosamusa
04-16-2009, 08:27 PM
Welcome to Bananas dot org Dr. Perez,
If we can help you in any way just ask if you can't find it on the threads. I will be asking you many questions about plant pathology, especially mangos, and some other plants. I know your focus may be viruses, but I know you know more than we do about plant pathogens.
Best wishes and welcome to the organization!!:2239:
Taylor
04-16-2009, 08:51 PM
Welcome Eden.
His username is Gabe15. And yes, he is a lab assistant. I'd gather that it is the same person!
Here he is:
Gabe's Member Profile (http://www.bananas.org/member-gabe15.html)
Thanks for all the work you are doing to produce virus-free crops in Hawaii.
eaperez
04-16-2009, 09:08 PM
Welcome Eden.
His username is Gabe15. And yes, he is a lab assistant. I'd gather that it is the same person!
Here he is:
Gabe's Member Profile (http://www.bananas.org/member-gabe15.html)
Thanks for all the work you are doing to produce virus-free crops in Hawaii.
I looked at Gabe15's profile and found out that he's my lab assistant. i will tell him about this tomorrow.
thanks,
eden
Taylor
04-16-2009, 09:10 PM
It's a small world!
Any information you will share with us here in your visits will be greatly appreciated. :)
eaperez
04-16-2009, 09:11 PM
Welcome to Bananas dot org Dr. Perez,
If we can help you in any way just ask if you can't find it on the threads. I will be asking you many questions about plant pathology, especially mangos, and some other plants. I know your focus may be viruses, but I know you know more than we do about plant pathogens.
Best wishes and welcome to the organization!!:2239:
Thanks. I'll try to give you the best answer on all your questions. If I didn't answer today, you'll get them the next day. it is now 4:10 pm here (HI time) and will continue my work till 6:00 pm.
buzzwinder
04-16-2009, 09:20 PM
Welcome to the Org. Eden, Glad you joined us, and looking forward to your insites on some of our members questions, this is and has been a GREAT place to learn about Banana culture, as well as alot of humor tossed in for good measure! :bananas_b
conejov
04-16-2009, 09:25 PM
Welcome!
eaperez
04-16-2009, 10:11 PM
Welcome!
Thanks!!!!
eden
Chironex
04-17-2009, 12:33 AM
Hi Eden! I am particularly glad that you joined us here. As an amateur tissue culturer I am happy to be able to learn more about new studies and sharing of techniques.
I have been experiencing several challenges in embryo rescue, finding a good medium to promote growth. I would also like to find out a technique for some of the Ensete varieties such as ventricosum Maurelii and perrieri. Apparently there has been little success with traditional M&S medium with Ensete. I may also have access to E. v. 'Montebeliardii' which is far more rare than 'Maurelii'. I have read some research on a newer medium, but wondered about any experience that you may have.
Welcome to Bananas.org!
eaperez
04-17-2009, 01:13 AM
Hi Eden! I am particularly glad that you joined us here. As an amateur tissue culturer I am happy to be able to learn more about new studies and sharing of techniques.
I have been experiencing several challenges in embryo rescue, finding a good medium to promote growth. I would also like to find out a technique for some of the Ensete varieties such as ventricosum Maurelii and perrieri. Apparently there has been little success with traditional M&S medium with Ensete. I may also have access to E. v. 'Montebeliardii' which is far more rare than 'Maurelii'. I have read some research on a newer medium, but wondered about any experience that you may have.
Welcome to Bananas.org!
Aloha! thanks to your message. i have not done any embryo rescue on bananas. and i would like to direct your inquiries to Gabe15 (also a member and one of my lab assistants) whom i know has been doing a lot of embryo rescues in our lab for his own research. i only do micropropagation of bananas right now and i assume embryo rescue should not be difficult unless the seeds are not viable. are you excising the embryos from the seed? are those seeds mature or immature? by the way, are you doing research? i have been doing tissue culture for more than 30 years on a lot of different crops. i am willing to give you some advice on your next mail. thanks,
eden
Chironex
04-17-2009, 01:33 AM
Thanks Eden, yes I have talked at length with Gabe about the struggles. In particular we have been trying to rescue embryos from M. ingens. They seem to have an endogenous contamination issue. I was going to attempt surface sterilization of the seeds, prior to excision, using silver nitrate as described by Dr Frederic Bakry. But I believe that this still will not cure the issue of endogenous contamination. I have used standard surface sterilization using bleach solutions, physan 20 and hydrogen peroxide. Every time, I either get contamination, or no growth. I have also tried modified N&N medium with the additions of kinetin and also ethrel. Still nothing. I am wondering if I should increase the percentage of PPM in the medium? I was also curious whether you thought it would help by placing the embryos directly into a PPM soak prior to placing them onto the medium? Would this tend to kill the embryo? I don't have much luck with PPM so far.
I did a lot of research, particularly with sharks and cubomedusae when I was at the U of Miami in 1976, but not much since then. I worked with Dr Samuel Gruber. I was pre-med with a Biology major. Began working on my thesis, but due to family medical issues, had to withdraw from continuing my education. I miss research as you can probably sense! Quite a bit rusty, but nonetheless, the fire still burns.
Tog Tan
04-17-2009, 07:41 AM
Hiya Eden and welcome to the board from the other side of the world! :02: It's great to have an expert to lead this bunch of crazy 'naner nuts.
Looking forward to learn more from you. Have a good day! :ha:
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.
Scot Nelson
04-17-2009, 12:48 PM
Hello to everyone,
My name is Eden A. Perez. I am from the Philippines and I'm here at the University of Hawaii since 1999. I graduated in 2006 with Ph. D. in Plant Pathology (Virology). I am currently working on my project entitled "Rehabilitation of BBTV infected areas in Hawaii" thus my work involves indexing bananas for use as starting material of in vitro cultures and micropropagation of virus-free bananas. Hope to meet all of the members through chat here. I welcome questions regarding tissue culture and genetic improvement of bananas. Aloha......Eden
Hi Eden, it's nice to see you here. :goteam:
Scot N.
eaperez
04-17-2009, 01:10 PM
Hi,
thanks.
eden
eaperez
04-17-2009, 01:17 PM
Hi Eden, it's nice to see you here. :goteam:
Scot N.
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
04-17-2009, 01:20 PM
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.
Hi Bob,
thanks. i'm enjoying and i'm learning a lot too.....
eden
eaperez
04-17-2009, 01:21 PM
Hiya Eden and welcome to the board from the other side of the world! :02: It's great to have an expert to lead this bunch of crazy 'naner nuts.
Looking forward to learn more from you. Have a good day! :ha:
Hi Tog,
thanks.
eaperez
04-17-2009, 01:33 PM
Hi Eden! I am particularly glad that you joined us here. As an amateur tissue culturer I am happy to be able to learn more about new studies and sharing of techniques.
I have been experiencing several challenges in embryo rescue, finding a good medium to promote growth. I would also like to find out a technique for some of the Ensete varieties such as ventricosum Maurelii and perrieri. Apparently there has been little success with traditional M&S medium with Ensete. I may also have access to E. v. 'Montebeliardii' which is far more rare than 'Maurelii'. I have read some research on a newer medium, but wondered about any experience that you may have.
Welcome to Bananas.org!
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
04-17-2009, 01:42 PM
Thanks Eden, yes I have talked at length with Gabe about the struggles. In particular we have been trying to rescue embryos from M. ingens. They seem to have an endogenous contamination issue. I was going to attempt surface sterilization of the seeds, prior to excision, using silver nitrate as described by Dr Frederic Bakry. But I believe that this still will not cure the issue of endogenous contamination. I have used standard surface sterilization using bleach solutions, physan 20 and hydrogen peroxide. Every time, I either get contamination, or no growth. I have also tried modified N&N medium with the additions of kinetin and also ethrel. Still nothing. I am wondering if I should increase the percentage of PPM in the medium? I was also curious whether you thought it would help by placing the embryos directly into a PPM soak prior to placing them onto the medium? Would this tend to kill the embryo? I don't have much luck with PPM so far.
I did a lot of research, particularly with sharks and cubomedusae when I was at the U of Miami in 1976, but not much since then. I worked with Dr Samuel Gruber. I was pre-med with a Biology major. Began working on my thesis, but due to family medical issues, had to withdraw from continuing my education. I miss research as you can probably sense! Quite a bit rusty, but nonetheless, the fire still burns.
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
04-17-2009, 02:12 PM
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
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.
eaperez
04-17-2009, 03:44 PM
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.
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
04-17-2009, 05:37 PM
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?
Scot Nelson
04-17-2009, 05:50 PM
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
Eden, there are AeAe plants growing on the Big Island if you want some. You can e-mail me.
eaperez
04-17-2009, 06:52 PM
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?
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
04-17-2009, 06:55 PM
Eden, there are AeAe plants growing on the Big Island if you want some. You can e-mail me.
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
04-17-2009, 07:07 PM
I'd like to place my order for an aeae plantlet.
thanks
Michael
Michael_Andrew
04-17-2009, 07:10 PM
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
04-17-2009, 07:49 PM
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.....
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:
http://www.bananas.org/gallery/watermark.php?file=14068 (http://www.bananas.org/gallery/showphoto.php?photo=13746&ppuser=2761)
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.
eaperez
04-17-2009, 08:05 PM
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:
http://www.bananas.org/gallery/watermark.php?file=14068 (http://www.bananas.org/gallery/showphoto.php?photo=13746&ppuser=2761)
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.
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
04-17-2009, 08:08 PM
I'd like to place my order for an aeae plantlet.
thanks
Michael
i'll let you know when we've got keikis (sister plants or baby plants) from them.
eden
Chironex
04-17-2009, 08:10 PM
Thank you Eden, I will be trying that. Now all I need is some AeAe pups.
eaperez
04-17-2009, 08:16 PM
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.
hey Michael,
thanks. i am getting some aeae plants from Scot Nelson next week.
vBulletin® v3.6.8, Copyright ©2000-2020, Jelsoft Enterprises Ltd.