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51st state
06-06-2008, 10:36 AM
following on from the Ensete Perrieri thread... Is there sufficient demand from .org members to jointly put some of the more unusual species into TC?

I seem to remember a number did this with Siam Ruby a while back. maybe we could create 'wishlists' and see what level of demand is out there.
I know that there are specialists out there doing this but it would be a great way for .org to spread some species out more widely across the globe.
any thoughts? :2722:

Gabe15
06-06-2008, 11:02 AM
Iʻm working on it right now, I have a whole setup in my house ready to go, Iʻm just waiting for my media to come in and hopefully I can start producing some. I already have much of my collection in Hawaii in TC already (including some very unusual species, and many plants not yet available on the market), when I get back there I plan to start selling plants by January hopefully.

Chironex
06-06-2008, 11:03 AM
Kev, I couldn't agree more. I am reading up on TC technique and will be able to do it in a few months or less. I was discussing this with Gabe too. It will definitely help the banana world and I want to be part of that.
Once I get set-up to where I know what I am doing, I will be offering this service to our membership. I am not sure just yet, but I think that taking the tissue from the explant can be done without causing too much injury. From what I have read, tissue is taken from the meristem, which is either growing root tips or in shoots. So, it shouldn't be that invasive as to cause unrecoverable injury to the source plant (explant).
Just need to get a comfort level on my own stock first. Then Dr Frankenstein will be ready to party!!!!

AltadenaGarden
06-06-2008, 11:51 AM
Would anyone like to post a good link about tissue culture for bananas. I am not looking to do it but am interested in how it works.

chong
06-06-2008, 12:38 PM
Would anyone like to post a good link about tissue culture for bananas. I am not looking to do it but am interested in how it works.

Here's one from one of our members, kgbenson:
Micropropagation of Musa (http://www.geocities.com/k2benson/TCoverview/TCban/TCban.html)

Richard
06-06-2008, 01:42 PM
It turns out that different generas of plants require different kinds of tissue culture. In other words, some plants are more troublesome than others. Here's a paper I came across while researching the origins of some pineapple guava varieties:

ScienceDirect - Scientia Horticulturae : Improvements in somatic embryogenesis protocol in Feijoa (Acca sellowiana (Berg) Burret): Induction, conversion and synthetic seeds (http://dx.doi.org/10.1016/j.scienta.2006.10.030)

Mark Hall
06-07-2008, 03:11 AM
Chong, Thanks for the link. Very interesting reading. I did think there would be more pictures after the meristem was removed showing various stages of growth.

damaclese
06-07-2008, 09:07 AM
Iʻm working on it right now, I have a whole setup in my house ready to go, Iʻm just waiting for my media to come in and hopefully I can start producing some. I already have much of my collection in Hawaii in TC already (including some very unusual species, and many plants not yet available on the market), when I get back there I plan to start selling plants by January hopefully.

Gabe Scot and i are looking for a Banana called Mysor its appears to be vary drought and heat tolerant if we got one do you think any one on the org would be interested in Tc of them?

Chironex
06-07-2008, 09:16 AM
Pauly, I forgot to mention to you that I have made arrangements for 2 Mysore and have another contact for more if needed. PM me with your address if you want and I can have them sent to you before I get there.

Richard
06-07-2008, 10:08 AM
I have heard from other Musa (fruiting) growers and read a few articles indicating that Tc of Musa usually produces a new cultivar of the plant and less often a plant that is completely true to form of the parent. What are your experiences with this?

Chironex
06-07-2008, 10:50 AM
I am not an expert yet, but this contradicts what I have read. The clone produced through micropropagation is an identical copy of the parent (as are all other plantlets multiplied from the same explant.) The only possible way that this can be deviated is through contamination or mutagens deliberately applied to the culture.

Gabe15
06-07-2008, 11:45 AM
Gabe Scot and i are looking for a Banana called Mysor its appears to be vary drought and heat tolerant if we got one do you think any one on the org would be interested in Tc of them?
Mysore is a very common banana, it shouldnʻt be too hard to find. Going Bananas has them. Other common names for it is Pisang Ceylon and Misi Luki, different accessions may vary slightly but they are all the same basic cultivar. If theres that much interest in it, I could look into TCʻn it later on.

I am not an expert yet, but this contradicts what I have read. The clone produced through micropropagation is an identical copy of the parent (as are all other plantlets multiplied from the same explant.) The only possible way that this can be deviated is through contamination or mutagens deliberately applied to the culture.
I have heard from other Musa (fruiting) growers and read a few articles indicating that Tc of Musa usually produces a new cultivar of the plant and less often a plant that is completely true to form of the parent. What are your experiences with this?

This is not true, though there is some truth in it. Bananas can and do indeed mutate during micropropagation, however precautions can be taken to lower the rate of mutation. In the field, bananas mutate naturally, this is the reason we have so many cultivars and different varieties in the same groups. In micropropagation, you are generally dealing with many plants at a time (I produce about 300-400 explants in 3hrs, you can see it adds up), and forcing them to grow and proliferate rather quickly. Usually, in a very small number of your explants, there is some mutation. To reduce this, in our lab, we will only subculture a plant 5 times after initiation (this results in about 4,000 explants per original plant). If we run out of explants for that variety, and have tissue subcultured all of the lines 5 times, we will then go collect a new plant from the field, preferably one that was not grown from tissue culture or if not available, has been in the ground for at least 2 years. If a lab continues to reuse the same material over and over, eventually the mutation rate would be unreasonably high and you will see many "off-types". Some mutations can be beneficial (such as dwarfing or increased yield), but most often they are not. Even with these precautions, from time to time a farmer will find an off-type in their field, but tissue culture still has many advantages over traditional propagation.

Richard
06-07-2008, 11:52 AM
Thanks Gabe for that excellent clarification.

D_&_T
06-07-2008, 05:39 PM
Here is a link another member posted a while back.

Kitchen Culture Kits - Intro Page (http://www.kitchenculturekit.com/Index.htm)

Gabe15
06-07-2008, 07:57 PM
Chong, Thanks for the link. Very interesting reading. I did think there would be more pictures after the meristem was removed showing various stages of growth.

Iʻll try to find some pictures, but in the meantime, I made up this diagram showing basically what happens after the initiation (when tissue culture becomes micropropagation). You will come across inconsistencies in actual practice, such as dead explants, contamination, varying degrees of proliferation, size of regenerated meristems and duration between meristem regenerations. But its generally what happens, or at least the way I do it is.

NOTE: The plants do not actually get smaller and smaller, I just only had a finite working space to show the examples.
http://www.bananas.org/gallery/watermark.php?file=10126&size=1 (http://www.bananas.org/gallery/showphoto.php?photo=10126&ppuser=5)

mm4birds
06-07-2008, 08:47 PM
good luck on this project. hopefully it will make it easier to obtain great plants for our own collections and prevent some of the rarer plants becoming extinct.:nanablowskisses:

Richard
06-07-2008, 08:49 PM
I received the copy of "Plants From Test Tubes" by Kyte and Kleyn today and started reading it. So far the writing style and information content is excellent.

Chironex
06-09-2008, 10:54 PM
Richard I got the same book from Amazon last week and I'm reading it too. I have also made contact with the Doctor that runs the Kitchen Culture website. She has sent me some more info as well as given my email to her associate who specializes in bananas. I must have downloaded a gazillion pages of info on TC already. Now if i could just find the time to read all of it. Burned through a fresh ink cartridge already.

Richard
06-09-2008, 11:34 PM
Richard I got the same book from Amazon last week and I'm reading it too. I have also made contact with the Doctor that runs the Kitchen Culture website. She has sent me some more info as well as given my email to her associate who specializes in bananas. I must have downloaded a gazillion pages of info on TC already. Now if i could just find the time to read all of it. Burned through a fresh ink cartridge already.

:) I would take all those pages you printed from the internet and throw them into the paper recycling. :) No really, some of them probably are worthwhile, but for any research in just about any subject: first read a thorough, well-rated book like the one we have. 2nd, if the book does not contain what is referred to as "survey of the literature", then find one and read further on topics as needed.

Also I would caution that reading the "how to" of Tiissue Culture is a bit like reading how to be a master cabinet maker. I expect it is going to take me a lot of patience and practice, including going through the amateur, apprentice, and journeyman phases.

sandy0225
06-10-2008, 06:07 AM
Yes, I've been on the home tissueculture webgroup for quite some time now >2 years, and I've learned a lot just from what others are posting. It seems the biggest issue is getting the right chemical blend for propagation of particular plants, they do recommend certain chemicals for bananas, but getting them "dialed in" can be a challenge. Also the biggest problem is contamination. Everything has to be sterile and kept that way. Even the smallest particle of dust contains thousands of different contaminants from bacteria to fungus.
I haven't actually tried tc yet because of lack of time issues, maybe this winter. I said that last year too, though.
I did get a cloning machine though, that sounded very do-able to me. I'm supposed to get it today.

Chironex
06-10-2008, 06:29 AM
I said that last year too, though.
I did get a cloning machine though, that sounded very do-able to me. I'm supposed to get it today.

A cloning "machine"? Do you have a link to it? I would be interested to see it. What does it do?

musa_monkey
06-10-2008, 06:59 AM
This maybe ?

Hydroponics Cloning Machines - Plantlighting Hydroponics & Grow Lights (http://www.plantlightinghydroponics.com/hydroponics-clone-machines-c-37.html)

MediaHound
06-10-2008, 09:38 AM
Looks like this thread is really blowing wind into some sails!!
I'm elated to see all the interest everyone has into tissue culturing and bringing some of the more rare bananas into production. I have always had an interest in TC'ing but have never taken the initiative. Perhaps watching the success of our members will help convince me to find the time and space and set it up properly.

Iʻll try to find some pictures, but in the meantime, I made up this diagram showing basically what happens after the initiation (when tissue culture becomes micropropagation). You will come across inconsistencies in actual practice, such as dead explants, contamination, varying degrees of proliferation, size of regenerated meristems and duration between meristem regenerations. But its generally what happens, or at least the way I do it is.

NOTE: The plants do not actually get smaller and smaller, I just only had a finite working space to show the examples.
http://www.bananas.org/gallery/watermark.php?file=10126&size=1 (http://www.bananas.org/gallery/showphoto.php?photo=10126&ppuser=5)

This is great! Thanks for taking the time to create it.
BTW, I copied it over into the banana diagrams (http://www.bananas.org/gallery/showgallery.php?cat=593) category in the gallery.

Thread stuck by the way, everyone, so we can use it as a general reference and a place to point people to.

We do have a page in the wiki about tissue culturing, it's here:
Info:Tissue Culturing - Bananas Wiki (http://www.bananas.org/wiki/Info:Tissue_Culturing)
It's fleshing out, but these things take time. Feel free to add to it, we can use both this thread and the page in the wiki as a springboard for information.

Cheers

Dean W.
06-10-2008, 11:15 AM
I want to keep track of this thread. I have an Ensete perreri that might be TC material.

Chironex
06-28-2008, 02:06 PM
Dean, keep in mind that TC'ing a banana requires the original pseudostem to be destroyed. We dissect the apical meristem from the base of the pseudostem. You might be able to re-grow from the corm.
There is always a risk of contamination in TC work, however small the percentage (Less than 2% hopefully), so always keep a back-up, just in case. I have several Ensete perrieri seeds in germination, so if I get a decent germination rate, I plan to TC one of them.

Richard
06-28-2008, 03:24 PM
I thought that banana TC has been successfully initiated from growth tips and cambium material in rhizomes. Is this true and just not a preferred practice, or ?

Chironex
07-03-2008, 04:57 PM
I thought all might be interested to know that I have been emailing the guy who created Little Prince. It is a sport that was created in TC from 4 mutants. It is patented as a variety, so TC'ing it will require permission. I will invite him too join our forum.

mskitty38583
07-04-2008, 12:23 AM
that would be awesome scot!

Chironex
07-04-2008, 10:43 AM
Update: He just emailed me that he will be joining as soon as he gets his computer back online. Apparently a Microsoft security update has locked him out of his desktops and he is having it repaired.
Bottom line is he will be joining us soon. His name is Randy, but I don't have his last name.
It will be exciting to include him among our membership. It will also be helpful to anyone who develops their own sport and seeks to have it patented, from someone who knows the ropes.

Micronana
08-07-2008, 03:26 AM
Hi New member here,
The diagram from Gabe 15 was brilliant, I am microing musa basjoo, dwarf cavendish and ensete v. maurelii (although the last 2 are new to me, and only just initiated).
I have never tried being quite so brutal with them, I usually just cut them down the middle. Might try quartering some in future.
By the way, I've recently read that the ensete don't produce pups spontaniously. Mine in culture have already started sprouting! I'll try damaging a few anyway to see the results. Has any one got any experience of them?

Thanks Dawn

Gabe15
08-07-2008, 07:35 AM
Hi New member here,
The diagram from Gabe 15 was brilliant, I am microing musa basjoo, dwarf cavendish and ensete v. maurelii (although the last 2 are new to me, and only just initiated).
I have never tried being quite so brutal with them, I usually just cut them down the middle. Might try quartering some in future.
By the way, I've recently read that the ensete don't produce pups spontaniously. Mine in culture have already started sprouting! I'll try damaging a few anyway to see the results. Has any one got any experience of them?

Thanks Dawn

Hello and welcome Dawn,
Ensete do not naturally produce pups in the ground, but in culture they will proliferate like normal with the benzyladenine in the media.

51st state
08-10-2008, 04:15 PM
Hi Dawn
welcome aboard the best banana site in the world!!!!!!!!!!!!!

great to see you trying TC'ing some nana's. If you can pull it off I've got a few species you might want to have a go at

Mark Hall
08-10-2008, 05:14 PM
Me too! Welcome by the way:waving: Where are you in the UK?

Micronana
08-19-2008, 04:48 AM
Hi 51st State
I'd be interested to hear what varieties you've got going

and Mark Hall
I'm located near Bognor Regis on the south coast.

Work to do - ho hum

Dawn:bed:

51st state
08-20-2008, 11:46 AM
Hi Dawn
see my profile, which lists the species/varieties I'm growing

kgbenson
08-24-2008, 04:28 PM
Chong, Thanks for the link. Very interesting reading. I did think there would be more pictures after the meristem was removed showing various stages of growth.


Yeah - I just never got around to taking many or to posting them. Lemmie look back into some of my files and if I can find them I will try to get them on there.
Keith

griphuz
08-28-2008, 09:58 AM
Maybe this article is usefull for the Ensete's?
http://www.barc.ernet.in/webpages/letter/2002/200210-24.pdf
Regards,
Remko.

Chironex
09-01-2008, 12:35 AM
I know that there are some of us who are doing micropropagation. I would like to see if there is any interest in combining our orders from various suppliers to buy in larger quantities. As an alternative, we could approach vendors for a Bananas.org membership discount and buy at wholesale pricing.

We could do the same with Agri-Starts, Phytotech Labs, etc. All they could do is say no. I just ordered more TC culture jars and noticed a huge discount if ordering by the hundred lot rather than 25 lot. Savings is almost 20% in just this example. Larger quantities could be greater obviously. M&S media is 12-20% lower in quantity.

Anyone interested? Let's talk about it and see if there is enough interest. We have numbers of members into this or interested in TC. Let's make use of it - a nice membership benefit in my opinion.

damaclese
09-01-2008, 10:40 AM
wow! the institute for advanced banana research funded and supported by bananas.org. Interesting concept! Scot every time I read one of your threads the possibilities just explode in my mind.!!!

51st state
09-02-2008, 04:33 PM
agree with all that. got to be a way of having members subscribe to wanting a particular species/variety before you TC it so that you can gauge demand and have customers ready and waiting before you start. Not sure how people like agristarts would react to 'hobbyists' clubbing together though... I'm in

damaclese
09-03-2008, 09:22 AM
well market demand i say if agrastarts wants to keep are business then they should cut down the minimum order to accommodate hobbyist but i really don't think they are all that interested in small orders or they would have adapted already? besides i think we are all capable of taking care of each others Banana needs. I'm sure if we took and inventory of all the plants and supply's that every one on the org had we would find that theres more then adequate stocks to keep every body happy for years!

rohsen
11-09-2008, 06:29 PM
hello there you all im from amsterdam i make aliving tissue culturing plants and would like to share the following files with you, for anyone trying to micropropagate banana it is essential information!!!

here goes...

Chironex
11-09-2008, 08:57 PM
rohsen, thank you for sending these protocols and propagation media formulations. Do you have a recommendation list of which medium to use for best results on different types of banana plants? Do you have a method for preparing the medium?

rohsen
11-10-2008, 02:32 AM
as im just getting started on musa spp im not sure yet, i do know some cultivars respond well to different formulations, banana is known as an easy species! what i usually do and i advice you the same is check the references given with the media, you can google this reference and see what comes up. if i cant find info i just set up trials using different media, i make my choices depending on what ingredients i have available what growth regulators (hormones or plant growth regulators PGR's for short such as 6-BAP GA3 IBA IAA NAA) required and the price of growth regulators. as far as media preparation goes i usualy buy pre-formulated media such as MS , MS gamborg, Rugini olive medium etc. only rarely do i formulate the media myself. (its a hastle) I only use agar in the initiation of cultures to screen for contaminants, after initiation i use liquid media in a temporary immersion system. any advice on media for musa basjoo ? im looking into propagating this species im also interested in ensete ventricosum

regards
Dave

in my gallerie pictures of my modest lab

Chironex
11-10-2008, 03:31 AM
Dave, I am also just beginning to tissue culture bananas. I use pre-measured M&S medium and add BAP for initiation and shoot proliferation. For rooting, NAA is commonly used. I have not yet tried Basjoo or E. ventricosum. I have 3 varieties in culture now as well as trials of 3 different bananas in embryo rescue. For these I want to make a modified N&N medium which is reportedly more robust for embryo rescue.
Photos of my laminar flow hood are in my gallery, too.

Gabe15
11-10-2008, 04:51 AM
Some banana varieties are very slow in the normal media (such as Lakatan, Fe'i bananas, some Callimusa and Australimusa species), so to help them grow I use full strength MS (instead of the normal 1/2 strength), and its also been recommended to use 30g/L sucrose, currently we are using only 20g/L for normal micropropagation. There is also an issue with different varieties reacting adversely to BA, some varieties proliferate too much and go through some type of dedifferentiation to the point where I cannot regenerate usable explants from the tissue (perhaps it is possible with somatic embryogenesis, but that would be a whole different project...), so for these bananas that have reacted poorly to the BA (E. glaucum and Fe'i varieties), I will experiment with using little or no PGRs, especially upon initiation.

rohsen
11-10-2008, 07:24 AM
First of all nice to meet you guys!! Gabe, for species proliferating too fast you may want to consider using a growth retardant such as Paclobutrazol. im attaching a publication on use of this and other substances with musa. you'll find it an interesting read! as for embryogenesis .. not so much a different project as you think, very do-able especially if you have a protocol and media formulations.....! which are , naturally , attached!

I dont see to much here on temporary immersion systems, and if there ever was one technique for making loads of musa ....
I have used it succesfully with papaya pineapple and other species.

regards

Dave

Gabe15
11-10-2008, 12:22 PM
First of all nice to meet you guys!! Gabe, for species proliferating too fast you may want to consider using a growth retardant such as Paclobutrazol. im attaching a publication on use of this and other substances with musa. you'll find it an interesting read! as for embryogenesis .. not so much a different project as you think, very do-able especially if you have a protocol and media formulations.....! which are , naturally , attached!

I dont see to much here on temporary immersion systems, and if there ever was one technique for making loads of musa ....
I have used it succesfully with papaya pineapple and other species.

regards

Dave
Thanks for the papers Dave. I would love to try all of these things out one day, but I've been so busy that I don't have time to be making different media for side project this semester, I just use what our lab produces for normal micropropagation for the time being. Hopefully I will have more free time next semester and can experiment more with different media for things like embryo rescue (I use the normal microprop. media which works sometimes, but not always great), somatic embryogenesis, and these hard to culture Musa.

Chironex
11-10-2008, 02:40 PM
Dave, I have read some about liquid media, but from everything I have read, it seems more expensive in terms of equipment, for the hobbyist like me. Is this the same process as developing callus culture and so on? Is there an inexpensive way to set-up this process for a home lab?
Thanks for the research articles.

rohsen
11-10-2008, 06:53 PM
Scot, indeed the initial cost will be higher than when using babyjars plastic jars or other disposables, however the results are amazing, and after the initial layout the returns wil justify the expense, in some species the multiplication rates will improve by 500% !!! for commercial labs these numbers are .... well you can imagine. added to this are less handling, no use of agar, and especialy in temporary immersion systems you get partially aclimatized and sturdier plants, every time you immerse the plants in liquid media you also renew the air in the culture vessel wich has considerable benefits. I distribute a TIS system in europ and south america wich includes an airpump culture vessels, the works. for 40 1 liter vessels including pump airfilters and all you need. would set you back at least 2500 euros, you would than be able to produce 10s of thousands of plants. so this is for pros, however it is possible to build a set yourself using lowcost options.
attached a paper listing lowcostoptions for developing countrys. and since a home TC lab, and indeed many commercial labs, needs to watch spending it will have some interesting options.

basicly if you can micropropagate conventionally...you can use bioreactors. you would first propagate the old way, and then seed your bioreactor with the product after you established your cultures in agar. see it as the next step towards lots and lots of plants

hope you enjoy the papers,

regards

Dave (who has no university education whatsoever started out as a hobyist)

ps. in this publication you will find in the chapter BIOREACTORS AS A LOW COST OPTION FOR TISSUE CULTURE a discription on how to build a very cheap aparatus that wil absolutly do the trick. ofcourse if anyone cares to try i will happily assist!!

rohsen
11-10-2008, 08:07 PM
I love to share, i have many interesting books on micropropagation tissue culture etc.
if there is a special section of this site to do so please let me know.
for now a very good book for all people interested. Its a real knowledge booster!!!
regards

dave

please if im breaking any rules by sharing this let me know asap!!

rohsen
11-17-2008, 09:20 PM
For those interested,

here's another paper i found in one of my books, it has some info on optimal media formulations for specific cultivars. interesting.

regards

griphuz
11-18-2008, 05:36 AM
If I'm correct all TC from bananas is done from meristem tissue right? So you would have to sacrifice a sucker to go TC right?

Did anyone ever try to TC starting with just a piece of leaf?
I've heared it might be possible to re-differenciate the cells to differenciete them later on into leaf, root, or whatever...
Kind regards,
Remko.

Chironex
11-18-2008, 09:16 AM
I have heard of being able to use male bud tissue, but have not tried it. It seems to me that I have read about someone being able to develop callous tissue from other parts of the plant, yet I have not read any research using thismeans. The article that rohsen attached "bananas" above, included some charts showing propagation results from other means - as best I can recall. If there are other places on the plant where adventitious tissue is formed, I would think that it could also be used.

Gabe15
11-18-2008, 01:03 PM
If I'm correct all TC from bananas is done from meristem tissue right? So you would have to sacrifice a sucker to go TC right?

Did anyone ever try to TC starting with just a piece of leaf?
I've heared it might be possible to re-differenciate the cells to differenciete them later on into leaf, root, or whatever...
Kind regards,
Remko.

You do have to "sacrifice" a sucker, but if successful, you won't actually kill it and will end up with exponentially more identical plants.

Creating meristematic tissue from leaf tissue is not done, and has not been done with bananas as far as I know (one method of somatic embryogenesis). However, pieces of corm and the very bottom portions of sheath have been used during somatic embryogensis, but this is not done to micropropagate due to unacceptably high mutation rates, it is often done when trying to transform a plant through gene transfer however. Also, immature flowers (usually male, sometimes female) are used in the same fashion. Only shoot tip cultures (meristem) is normally used for mass producing banana plants.

Jack Daw
01-16-2009, 06:55 PM
Once the cell has been assigned a role in the organism, it cannot be "reprogrammed, booted and reinitialized", if it were possible in such a manner, we would use that on people, to kill cancer cells for instance and force them to reinitialize properly and get a new role, which wouldn't harm the organism.

BTW - we would be talking here also about the possibility of reinitializing old cells - literally immortality key.

It would not only be the key to bioprogramming, but also to mutation processes beyond our technological levels and the only way we must go to truly understand the ways of the G :).

griphuz
01-16-2009, 07:45 PM
Ehm, Jack,
From what I read, differenciated cells (in animals and humans) can indeed not go back to stemmcells. But in plants tissue-cells (differenciated cells) can sometimes indeed go back to meristematic cells to later form other tissues...or I must have read it wrong meaning the differenciated cells go directly to OTHER differenciated cells.
This trait is called 'totipotenci' and occurs naturally in plants when propagated vegetatively (e.g. through cuttings or layering).

This is invoked in TC, end in SOME plant species leaves, roots, or other differenciated tissue can indeed be used for TC and the mass production of plants.
Experts, please correct me if I'm wrong...
Kind regards,
Remko.

Chironex
01-16-2009, 08:13 PM
These links should help to explain it. It will also spare you my lengthy repy.

Biology: Totipotency in plants v. little totipotency in humans, mammal cells, totipotency in plants (http://en.allexperts.com/q/Biology-664/Totipotency-plants-v-little.htm)

Home > Publications > (http://www.eppc.org/publications/pubID.2374/pub_detail.asp)

griphuz
01-17-2009, 08:03 AM
Jack & I discussed this a little over PM and the links from Scot really helped;
It seems indeed that plant cells (most tissue) is not as far differencieted as animal/human cells are, and do not have the 'on-off' switched genes. Therefore they can return so to speack to other differentiations;

"A large portion of a plant consists of cells that are not highly differentiated. These are parenchyma cells which make up most of the soft tissues and are involved mostly in food manufacture and storage. They are capable of dedifferentiation so that they can form other highly specialized tissues, even meristemetic cells which are capable of forming shoots and roots under the right circumstances. "

(From the links Scot posted)

It seems we were both a bit right;
They can form meristematic cells again, but also other differencieted tissue, without meristem in between, right?

So indeed it should be possible to mass produce a banana into TC going from just a piece of leaf, if you have the right formula to reduce the unacceptably high mutation rates e.g.
For now, If I try, I'll just usa a pup, :)

Kind regards,
Remko.

griphuz
03-10-2009, 08:53 AM
Yeah, thanks dude, I think you need to be removed...

rohsen
05-30-2009, 02:09 PM
hi there here's an article on using growth retardants for cultivating Musa in temporary immersion systems, people interested in using these systems can contact me if they please

ps i just found i had already posted this paper....???!!! sorry for that

rohsen
05-30-2009, 02:36 PM
jack draw, griphuz, chironex, after reading your discussion heres some reading material for you all

rohsen
05-30-2009, 02:43 PM
and 2 more publications

rohsen
06-01-2009, 04:28 AM
some clarification on these texts
alginate encapsulation is where you drop a somatic embryo in a sodium alginate solution thus creating what is known as a synthetic seed. wonderfull though complex technique.
Soma means "coming from the self" genesis "to give birth" so soma-tic embryo-genesis stands for creating embryos from the plants own cells, be they leaf, flower, stem or any other cell. (with different results )
protoplasts are basically cells with their cell walls removed allowing for "easy" in vitro crossbreeding and all sorts of other "easy" tricks (note the cynical """ marks around easy)All these techniques are pretty advanced. but it never harms to read ...right?

ps its always cool (for posting members) to ask me for specific articles, i have access to most scientific databases

alah
07-29-2009, 02:12 AM
Yeah - I just never got around to taking many or to posting them. Lemmie look back into some of my files and if I can find them I will try to get them on there.
Keith

Hi, Keith. This is the kind of lecturing that I like, I need.. Please excuse me everybody else, you see, I´m areal novice. So keith, hope you´re still around and able to continue this with lecturing on the rest of the process.
Thanks, Alberto.

reenaevm
08-10-2009, 12:32 AM
hi all,

had a query regarding multiplication of plantlets in vitro.
i have successfully initiated and got around 3-4 shoots from each initiated corm. these hav now grown into lil plantlets.

i want to know if it is possible to further take them for multiplication and how to go about with the multiplication. i kno i need to cut off all the leaves and roots but i'm confused about whether i have to discared the original corm tissue or not, the original corm has started blackening and some of the leaves have also started yellowing.

I'm a first timer with banana tc, so i'v really no idea how to go about it.
the variety i'm culturing is grand naine bananas

thanks in advance

reena

rohsen
08-16-2009, 03:07 PM
as a rule i always reuse the parts that gave me shoots!! there may be many buds left in the part that already gave you some ... just cut of the browned bits and replant. nothing you can lose there...
regards dave

alah
09-07-2009, 12:57 PM
Here's one from one of our members, kgbenson:
Micropropagation of Musa (http://www.geocities.com/k2benson/TCoverview/TCban/TCban.html)

Help. This article seems very useful to me, but when I try to download it I get an error message saying " Address not valid". Why, Alberto.

chong
09-07-2009, 01:41 PM
Help. This article seems very useful to me, but when I try to download it I get an error message saying " Address not valid". Why, Alberto.

Check your computer's security settings. That may be preventing your computer from downloading this file. Or, you may not have enough RAM or HD memory. I just downloaded it again, just now, and I was able to save it as a *.html file. Another way that I saved it, just now also, was to highlight the entire article, and then copy and paste it on to a Word document. The latter procedure assumes that you are able to open the link to the article.

Hope that works for you.

Chong

jmoore
09-07-2009, 02:25 PM
1262 here's a pdf of the same article

alah
09-07-2009, 09:01 PM
Check your computer's security settings. That may be preventing your computer from downloading this file. Or, you may not have enough RAM or HD memory. I just downloaded it again, just now, and I was able to save it as a *.html file. Another way that I saved it, just now also, was to highlight the entire article, and then copy and paste it on to a Word document. The latter procedure assumes that you are able to open the link to the article.

Hope that works for you.

Chong

Thanks, Chong. Something´s gone wrong with my CPU.
It hangsup every once in a while. I´ll check tomorrow.

alah
09-07-2009, 11:51 PM
1262 here's a pdf of the same article
Thanks, James. I think I´ll stick to this protocol mainly because Phytotech has a distributor at Colombia. That means they´re right next to me and I can pay in our currency ( not US$ at 600% the official exchange!)

alah
09-08-2009, 12:41 AM
Yeah - I just never got around to taking many or to posting them. Lemmie look back into some of my files and if I can find them I will try to get them on there.
Keith
Excuse me Keith. I received this email, with use of Phyto Tech Lab products for banana MP. I have some doubts with their explanation, as well as your´s. Can you please check and clarify thanks, Alberto.

ligature
02-06-2010, 05:07 AM
kindly let me know about automisation of micropropogation by using bioreactors
madhu

kamayoi
02-08-2010, 12:43 AM
http://www.agriculture.gov.bn/DOA2009/pdf%20Publication/banana_TC3.pdf --> source : Brunei Agricultural research centre
banana TC..

ligature
02-08-2010, 10:24 PM
thanks for your information kindly give some company reference who will install bioreactors for our commercial tissue culture lab in India.
http://www.agriculture.gov.bn/DOA2009/pdf%20Publication/banana_TC3.pdf --> source : Brunei Agricultural research centre
banana TC..

ligature
02-08-2010, 10:31 PM
can any body give more info about bio-reactors technology for micro-propagation for commercial tissue culture units and even the company which supply and install equipments.

B S MADHUSUDHAN
info@ligaturebiotechnologies.com
ligature@live.in
+91 9449837234

kamayoi
02-09-2010, 03:44 AM
thanks for your information kindly give some company reference who will install bioreactors for our commercial tissue culture lab in India.

ur welcome fren..

dinhml
08-31-2012, 09:42 PM
Hi guys, Here's my 1 day reading


it's not that difficult as you think it is. try it. it should be a fun and also a challenge. different authors have different formulas, but it all boils down to.


find all necessary utilities, P.H. meter, pressure cooker, micropipettes, 250ml flash, electronic scales, forceps


steriles all of these in pressure cookers @ 250F, or with strong ethanol or isopropyl alcohol

ingredients:

1 L of de-ionized water

4.3 gram of MS (Murashige and Skoog salts)

5ml of BAP, sometimes called BATH for nutrients purposes(studies show 5ml of BAP is the minimum to get the maximum of plantlets out of the test tubes or whatever containers)

1ml PPM(plant preserve material) for antimicrobial purpose (i think imma experimenting skipping this PPM step) - BUY HERE (http://www.ppm4plant-tc.com/instructions.htm)


AGAR, which is jelly that comes in patri dishes in most labs, which i think the powder form can be obtain thru the supermarkets for making jelly, which then u can boil with water and make AGAR out of it

Add Sugar and coconut milk for experimenting, since all formulas are self-made by a guy in his lab, so can we too :) . i suspect u add sugar in this step if u gonna mix the powder agar with water by yourself. Coconut milk acts as a growth promotor so add it if u have any.
add all the stuffs above together in a sterile flash/container and measure the P.H., try to have it 5.8, if PH too high, add vinegar, if too low, add antacid tablets. PH is crucial, the exacter to 5.8, the better


then boil in pressure cooker or microwave it until everything boil

let the mixture cool

pour in jars (where explant gonna go)

_____________________________
FOR the Corm/or Explant

trim it to 1 inch if it is from a flower, bigger size for corms

dipped it in bleach solution with 2 drops of surfactant (dish soap) for 20 minutes, then wash 3 times with deionized water

remove again the outter tissues since it got damaged by the bleach, now u have a perfect living sterile explant

when trimming, remember to use sharp razors or knives to avoid dulling the tissue

transfer this explant to the prepared jars contained the mixture, close the lid or by parafilm if test tubes are used.

let it sit under flourescent light for 16 hours daily at room temperature 75-85F


Most important of all, KEEP EVERYTHING CLEAN FROM BEGINNING TO END

AND REMEMBER: Not all formulas are exact, try making your variation, u dont have to be exact


and lastly

report your success with formulas back on this page :)

have fun growing ALL

USEFUL LINKS

youtube 1 (http://www.youtube.com/watch?v=LACbr7keIEc&feature=plcp) : watch all of his series

watch this in vietnamese for using flowers as explant (http://www.youtube.com/watch?v=CsRaGG84aAs)this has 3 parts so find it

learn aseptic techniques from youtube

read tissue culture for home gardener (http://web.archive.org/web/20060502161631/http://www.une.edu.au/agronomy/hort/ptc_for_home_~1.htm) there ain't error, wait 1 minutes before page load

and read anything that is on tissue culture on bananas.org (http://www.bananas.org/f260/tissue-culture-4755.html)


and read Ian Mcguire (http://trec.ifas.ufl.edu/litzlab/Banana.htm)

LASTLY: any inputs to make the process shorten is always appreciated

bengtang
12-03-2012, 12:12 AM
Here's one from one of our members, kgbenson:
Micropropagation of Musa (http://www.geocities.com/k2benson/TCoverview/TCban/TCban.html)

Here is a cached copy
Micropropagation of Musa (http://web.archive.org/web/20091024223858/http://geocities.com/k2benson/TCoverview/TCban/TCban.html)

Thelazybanana
02-25-2013, 04:02 AM
A cloning machine? could you further explain how it works

lloydgibson1234@gmail.com
06-06-2016, 03:07 AM
Here's one from one of our members, kgbenson:
Micropropagation of Musa (http://www.geocities.com/k2benson/TCoverview/TCban/TCban.html)
Link sent me to a small business web page..nothing about bananas

Juicy Bananas
06-06-2016, 12:36 PM
Aloha,

I have recently been allowed to use a small facility for experimenting. This man is awesome, he wants to try new things out. He has an Orchid business so my first attempt will be my Vanilla. The specimen seems to be disease free. My other thought was native bananas... maybe Ma'oli to start with, or possibly just a Mysore. My main goal to to successfully TC the Fe'i. We plan on keeping our mushrooms in his lab for safe keeping as well.

I have a lot of learning and experimenting to do... but there is a lot of information out in the world, so it will just take time.



Mahalo

chong
06-06-2016, 08:18 PM
Quote:
Originally Posted by chong View Post
Here's one from one of our members, kgbenson:
Micropropagation of Musa

Link sent me to a small business web page..nothing about bananas

When I posted that link, it went to kgbenson's website and had a full presentation of banana tissue culture, including the selection and preparation of the meristem from the corm through various stages of division. I will try to find a copy of his presentation from that link. If I can find it, I will attach it if the copy is in word or pdf format.

Chong

greenfumb
03-22-2017, 07:34 AM
Do you have to use a Musa pup for the donor TC. Can a young clone plant be used. I am asking because I want to try TC and I dont have any adult plants, or pups. I only have some growing clones for tissue.

Gabe15
03-22-2017, 01:19 PM
Do you have to use a Musa pup for the donor TC. Can a young clone plant be used. I am asking because I want to try TC and I dont have any adult plants, or pups. I only have some growing clones for tissue.

If you are just experimenting, it is fine to use a young TC plant instead of a field grown sucker (if that's what you mean), just as long as it is a healthy growing plant.

If it were a serious professional project and you were producing them in high quantity, then it should be avoided. This is because there tends to be a small number of plants produced from TC which are "off-types", and the resulting plants are not true-to-type. If you use a young tissue cultured plant as starting stock, you have no guarantee the plant is true-to-type and you could be creating a bunch of off-type plants. For this reason, a well run banana tissue culture lab will only produce about 5 subculture cycles from a single sucker, and then discontinue that line and start over from a new field-grown sucker which has been verified true-to-type.

druss
03-23-2017, 12:00 AM
With regards to using the adult corm for tc, how long does the meristem last for after you cut it from the corm? Is it hrs? Days, can you preserve it for later use?

Gabe15
03-24-2017, 10:29 AM
With regards to using the adult corm for tc, how long does the meristem last for after you cut it from the corm? Is it hrs? Days, can you preserve it for later use?

There would be no reason to do this and really no way to. When you excise the tissue piece containing meristem, it needs to be put into culture immediately. You would only do this in a flow hood with everything set up and ready to go. There is maybe 5-10min of prep outside the hood, and then another 5-10min in the hood for initiation, but there should be no waiting on anything as this is the most critical time to avoid contamination and to get it right. If the initial culture is contaminated then nothing can be propagated from it. If you want to preserve meristem essentially, you either leave the plant intact and growing normally, or you put it into tissue culture, for all practical purposes there is no in-between.

Kegas76
03-24-2017, 01:13 PM
I think he might have been confused by the line, "For this reason, a well run banana tissue culture lab will only produce about 5 subculture cycles from a single sucker, and then discontinue that line and start over from a new field-grown sucker which has been verified true-to-type."

For those of us that have no experience with doing tissue cultures the "5 subculture cycles" sounds like 5 different batches of TCs (made from one plant) one after the other. And from that we lay-folk might assume there is a time gap between the batches.

Obviously after your last post we know this isn't the case.

druss
03-24-2017, 04:31 PM
Sweet, i was curious if you could store meristem for future use. But makes sense when you put it that way. Just asked about some ney poovan from the tc lab where I get my plants. They told me its is one of the more difficult varieties to tc so they only keep a few plantlets.
Which is why I wondered if the meristem could be stored.

Tytaylor77
03-25-2017, 12:21 AM
Sweet, i was curious if you could store meristem for future use. But makes sense when you put it that way. Just asked about some ney poovan from the tc lab where I get my plants. They told me its is one of the more difficult varieties to tc so they only keep a few plantlets.
Which is why I wondered if the meristem could be stored.

Chini chompa/ney poovan/new poovan/new pavan is an amazing grower. Great tasting tiny little bananas. Flesh is very white and harder than most. If you don't like mushy or slimey bananas you will love it. PM me and I can show some pictures.

greenfumb
03-25-2017, 02:05 AM
If you are just experimenting, it is fine to use a young TC plant instead of a field grown sucker (if that's what you mean), just as long as it is a healthy growing plant.

If it were a serious professional project and you were producing them in high quantity, then it should be avoided. This is because there tends to be a small number of plants produced from TC which are "off-types", and the resulting plants are not true-to-type. If you use a young tissue cultured plant as starting stock, you have no guarantee the plant is true-to-type and you could be creating a bunch of off-type plants. For this reason, a well run banana tissue culture lab will only produce about 5 subculture cycles from a single sucker, and then discontinue that line and start over from a new field-grown sucker which has been verified true-to-type.

That all makes a lot of sense. I am just experimenting trying to figure out if I want to get into TC, possibly as a business in the future. But for now I am just experimenting. I have a lot of lab glassware and lab equipment from when I had a lab 10 years back. Is there a publication dealing with the chemicals/hormones banana's need during the TC propagation process.