Bananas.org

Welcome to the Bananas.org forums.

You're currently viewing our message boards as a guest which gives you limited access to participate in discussions and access our other features such as our wiki and photo gallery. By joining our community, you'll have access to post topics, communicate privately with other members (PM), respond to polls, upload photos, and access many other special features. Registration is fast and simple, so please join our community today!

If you have any problems with the registration process or your account login, please contact us.

Go Back   Bananas.org > Banana Forum > Tissue Culturing & Other Propagation Techniques of Banana Plants
The Facebook Platform
Register Photo Gallery Classifieds Wiki Chat Map Search Today's Posts Mark Forums Read

Tissue Culturing & Other Propagation Techniques of Banana Plants This forum is for discussing propagation techniques of banana plants. Tissue culturing is the popular process of creating clones from a source plant. There are other techniques to propagate banana plants however, such as nicking corms or dividing corms. Learn more inside.


Members currently in the chatroom: 0
The most chatters online in one day was 17, 09-06-2009.
No one is currently using the chat.

Reply   Email this Page Email this Page
 
LinkBack Thread Tools
Old 11-27-2009, 07:49 PM   #1 (permalink)
 
Location: CAracas, Venezuela, South America
Zone: Tropics
Name: Alberto Albert
Join Date: Jul 2009
Posts: 62
BananaBucks : 21,023
Feedback: 0 / 0%
Said "Thanks" 24 Times
Was Thanked 13 Times in 8 Posts
Said "Welcome to Bananas" 0 Times
Default making extracts before trying explants

Most of you might know what I´m talking about. This is a debt to Jack Daw..
with whom I got confused with Dave? Rohsen...

I very well agreed with his considertion of some less technical discussionj of TCíng for newbies. I went through all , well almost all the threads on the forum and extracted what I considered more imporant and useful.
Please excuse me if I didn´t mention some of you.
There´s still some more reading to go through but it can´t wait ´till next year. Thanks everybody, Alberto.
alah is offline   Reply With Quote Send A Private Message To alah

Join Bananas.org Today!

Are you a banana plant enthusiast? Then we hope you will join the community. You will gain access to post, create threads, private message, upload images, join groups and more.

Bananas.org is owned and operated by fellow banana plant enthusiasts. We strive to offer a non-commercial community to learn and share information. Receive all three issues from Volume 1 of Bananas Magazine with your membership:
   

Join Bananas.org Today! - Click Here


Sponsors

Old 11-27-2009, 08:27 PM   #2 (permalink)
 
Location: CAracas, Venezuela, South America
Zone: Tropics
Name: Alberto Albert
Join Date: Jul 2009
Posts: 62
BananaBucks : 21,023
Feedback: 0 / 0%
Said "Thanks" 24 Times
Was Thanked 13 Times in 8 Posts
Said "Welcome to Bananas" 0 Times
Default Re: making extracts before trying explants

Quote:
Originally Posted by alah View Post
Most of you might know what I´m talking about. This is a debt to Jack Daw..
with whom I got confused with Dave? Rohsen...

I very well agreed with his considertion of some less technical discussionj of TCíng for newbies. I went through all , well almost all the threads on the forum and extracted what I considered more imporant and useful.
Please excuse me if I didn´t mention some of you.
There´s still some more reading to go through but it can´t wait ´till next year. Thanks everybody, Alberto.
The only thing nice about responding to your own thread is that you earn a few bucks
I tried to attach a word doc but must have done something wrong.... again

So I´ll try to paste it here:

How to Tissue Culture Bananas

As explants I utilized the tissue located just above the corm, inside the stem.
yes that is where the meristem tissue is located. It is also located in whats called the infloresence-tip (where bracts are produced during flowering) and also for some varieties they have successfully used parts of the male flower. Gabe15
imagine you were peeling away leaves and leaf bases from the pseudostem. Eventually, you would get down to a small nub in the center, just above the corm. That will be your apical meristem. On a more mature corm, you can use the lateral meristems that have formed, but haven't yet started to grow. BigDog
Look at www.une.edu.au/agronomy/AgSSrHortTCinfo.html. There is a lot of infor about creating your own medium and sterilization. I also remember reading that you can transfer sterilized plant material into a sterilized container of media on a cutting board over a pot of boiling water. The steam is supposed to kill any microbes in the air. Plant material is sterilized in a 10% bleach solution and the agar and container are sterilized in a pressure cooker.
Sorry, it's changed to http://www.une.edu.au/agss/hor/horti...al-science.php and is the same article I was referring to.
the meristem are equivalent to the "eyes" on potatoes, but instead on potatoes, these are located around the banana corms, you can easily spot them after you clean up the corm of roots, soil and dirt.
I also read that you can use NaDCC. I'd imagine you can get this at a pool and spa store since it's used as a disinfectant for swimming pools. It may be sold as sodium dichloro-s-triazinetrione.
Tissue Culture Col 1 Be sure to click on "next page" tab.
Error Page | Aggie Horticulture

http://www.bananas.org/gallery/water...le=4809&size=1

http://www.bananas.org/gallery/water...le=4810&size=1

http://www.bananas.org/gallery/water...le=4811&size=1

http://www.bananas.org/gallery/water...le=4812&size=1

http://www.bananas.org/gallery/water...le=4813&size=1

Israeli banana flag??
http://www.bananas.org/gallery/watermark.php?file=4789


Israeli net house
http://www.bananas.org/gallery/water...le=4790&size=1


Israeli net house
http://www.bananas.org/gallery/watermark.php?file=4794


Israeli net house construction
http://www.bananas.org/gallery/watermark.php?file=4793


Israeli net house
http://www.bananas.org/gallery/watermark.php?file=4798


Israeli net houses of various colors
http://www.bananas.org/gallery/watermark.php?file=4791


Israeli bagged bananas
http://www.bananas.org/gallery/watermark.php?file=4795


Israeli bananas with grey bags
http://www.bananas.org/gallery/watermark.php?file=4796


Israeli bananas being readied for transport
http://www.bananas.org/gallery/watermark.php?file=4797


Israeli bananas of various kinds
http://www.bananas.org/gallery/watermark.php?file=4799


For comparison, bananas being shipped to market in India
http://www.bananas.org/gallery/water...le=4800&size=1
In the premade media from PhytoTech Labs, it has BA in it which is the chemical responsible for shoot multiplication, however it also inhibits shoot elongation and root formation (which is ideal for subculturing when multiplying your stock). So what we do is cycle the explants through the medium with the BA in it, and when the time comes to root the explants, they are switched to a similar medium which has no BA in it, this allows the shoots to elongate, roots to form and no longer induces extra shoot proliferation.
Shawn, i found this info on bananas wiki:
PROPAGATION Back To: Menu Bar
Bananas are propagated from offshoots (suckers or keikis) or corms (bullheads). If enough buds are present, large bullheads can be halved or quartered.
Planting material should be treated for nematodes:

(1) Cut off bottom half of corm and, if discolored, trim off up to 2/3 of the bottom of the corm until only clean white tissue remains.
(2) Trim off about 1/2 inch of tissue around the sides of the corm.
(3) If bullheads are used, cut off the pseudostem 3-4 inches above the top of the corm.
(4) Either,
(a)Immerse the trimmed corms in a hot water bath at 50 - 52 degrees C (122 - 126 degrees F) for 15 - 20 minutes. Before planting, place the corms in a transparent plastic bag at room temperature until new roots begin to appear.
Or,
(b) Coat the corms with parafilm wax prior to shipment or storage.

Hope this helps! Chironex
The most common thing used is the apical meristem. Basically this is a small piece we cut out of the p-stem where it joins the corm. It is about the size of a small bean. Usually, it is one of the last areas to die in a banana plant. If that part of yours is still intact, we might be able to carve it out and TC it. It's worth a shot. Chances are, if the corm and p-stem are both mush, then the meristem may also be gone. But, it wouldn't cost much to send the top 1/2 inch of the corm and maybe an inch of p-stem. Chironex
I would recommend cutting down the plants and making a horizontal cut deep into the meristem, this will destroy the apical dominance and encourage the growth of new pups. I've done this with good success on even very small plants. And although I've never tried it, I have heard lawn starter fertilizer works well, it must have some kind of shoot proliferation hormone in it which would make sense for starting off newly planted lawns. Gabe15
If you want a plant to pup fast, you can cut off the corm to under the meristem or close to it (pretty much leveling it with the ground or soil in your pot), basically killing the plant, but it'll jumpstart the pup process. THEN wait til the new plants are around 12" (which could take as little as 2-3 weeks). Each time you remove a pup, a new one (if another hasn't already) should start sprouting out within a few weeks. Keep doing this process until the original corm just seems to be completely choped up... that's when it's been expended. You might even give up after a bunch of pups have already came up and been removed, then pull up the rhizome to put with your yard waste, and find that there's MORE eyes or pups that were under the surface. I'm still not highly experianced as I've only done this once, so I can't tell you how long this may go on. It could potentially in my eyes go on for 6 months or more. Give it a shot and see what happens!!!! Jpfloors
The meristem is the actual area of cell division in the plant. It is located inside pseudostem on top of the corm at the base of the plant (except when flowering, it travels up the pseudostem and out of the top, producing the flowers and the rest of inflorescence, it ends up in the male bud where it eventually terminates). Other meristems are also located at various points around the corm, when these sprout they form their own pseudostems and these are what become suckers. In many plants, the apical meristem produces a hormone which discourages the growth of too many extra shoots (called "apical dominance"), if you destroy the apical meristem it will break this dominance and allow other shoots to grow out. Many of the buds for new suckers are already formed on the plant, they are just being inhibited until the plant gives them permission to grow.

Here is a diagram taken from Simmonds book, what he labels as "gp" (growth point) is the meristem, as well as the same corresponding area on the sucker.
http://www.bananas.org/gallery/water...le=9418&size=1 Gabe15
Level 10 Propagation Questions
either u can watch the videos here or go to the website i got them from

Super Starts Plant Tissue Culture - Learn


Re: inexpensive vials, racks, etc. ?
________________________________________
It turns out that in the container industry, a "vial" is usually has a volume of 1 fluid ounce (30 mL) or less -- often more like 1 fluid dram (1/8 of a fluid ounce). What I'm looking for has a volume of about 4 oz (115 mL) which the container industry call a "jar". Today I found some reasonably priced suppliers for just about every kind of vial, jar, and what-not you can imagine:
SKS Bottle & Packaging, Inc.

Yankee Containers
In particular I found the 4 oz size of "clear plastic straight-sided jars" with lids for $0.44 each in quantities of 24, $0.39 each in quantities of 280, and $0.33 each in quantities of 1100. Richard
reference article from India
________________________________________
some interesting stuff in there about using tap water and sucrose solutions instead of expensive media and sterile water. I don't have the patience required to do this myself but for those of you that do, excellent info
Attached Images
200210-24.pdf


He did say that they typically will not subculture a line more than four times to reduce the risk of mutations and then resort back to collecting material from an old established plant.
Hydroponics | Advanced Nutrients | Hydroponic Gardening
However, even if you get really really tiny TC plants, right out of the test tube, they are actually quite easy to grow if you know how to care for them.
Also, TC plants have been shown to grow faster, fruit earlier and produce larger bunches than suckers grown in the same (optimal) conditions Gabe15
TC plants can be tested easily for diseases, all you need to do is test an explant early on in the micropropagation Gabe15
Micropropagation of bananas
________________________________________
Produce your own pups by the thousands.

for the laboratory
Banana Micropropagation by Ian Maguire



for the home
http://www.geocities.com/RainForest/...ropropBAN.html

The plant is orginally from Indonesia and was found in the wild. I was first showed a picture of it almost 3 years ago, and now its finally becoming somewhat available. The orginal one was sold for $2000, then they were wholesale for $500 a peice, then I heard of them being $100 at some markets in Thailand.
Yet another way to micropropagate plants, this is all about activating the plants own ability to generate all necesary PGRs and let it grow in sugar-hormone free media under high levels of Co2 strong illumination and a low relative humidity. no sugar is needed because the plant is stimulated by the raised illumination to photosynthezise and use the Co2 as a carbon source. by eliminating sugar you eliminate most contamination problems. the explants will be single nodes with 1 or 2 leaves attached to allow photosythesis. All this happens in a closed large vessel with hundreds of plants in regular trays using fibrous substrates such as cellulose vermiculite mixtures, the sytems stimulates root growth and plants need not aclimatize!! they come out fully hardened and rooted !! the most important factor is the C02 level, values of 1500 to 3000 ppm are discussed, the vessel needs to be force ventilated to keep these levels at a constant.

it is a very elegant way of producing plantlets, by lowering the humidity the plant will "sweat" away moisture and suck up nutrients, there is no longer a need to discover the perfect medium nor do you need to invest in PGRs or sugar, it will fix Co2 into plant matter along the way grow roots and be hardened by the forced ventilation, all you need to do is ventilate efficiently keep Co2 at the right level and pump up the PPF (light)

Bananas do great in this system pioneered by a japanese scientist named Toyoki Kozai.

I have very detailed discriptions on how to build these vessels, have sofar had no use for such a system but would wilingly suply info for people interested as long as experiences are shared. i will attach a few publications.
one on coffee and banana and one more general explaining it in more detail

regards Rohsen
Dave
Attached Images
photoautotrophic banana-coffee.pdf (188.6 KB, 37 views)

photoautotrophic microprop.pdf (332.5 KB, 32 views)


As Gabe mentioned before, we generally use 1/2 strength M&S medium, with BA or BAP during initiation. I have been experimenting with other media too, including kinetin and activated charcoal. Although, these are primarily to promote embryo rescue.

Then no hormones for rooting, or some use NAA, IBA or IAA (auxins) but I haven't tried them yet, as all of my cultures are still in stage 2 sub-culturing.
Generally, 1 mg/ml is adequate although I have seen much lower concentrations of BAP. I believe that this will vary based upon which banana cultivar is being cultured.

I do not use NAA until the rooting phase as it is a rooting auxin. Some add no NAA and just leave out the plant growth regulators altogether in phase 3 - rooting. Try using some in a few cultures and none in others until you find which one works best for your banana plant selection.
I use about 5-7 grams per vessel. I put about 3 tablespoons of medium per vessel and add 5-7 grams of agar. (This is about a third of a teaspoon of agar per vessel.) Gelrite is used instead of agar by some people. I like it more because it is colorless.
You can PM me, but you may have to wait for a reply. I have been getting lots of PM's and I have become very busy at work, so I don't always get to the PM's right away. Chironex
These are some photos of my first home grown tissue cultures, which have been growing for 6 days. You can see they have turned green and are starting to grow tiny leaves. When they grow large enough and start to form real leaves, pseudostems and pups (in a few more weeks), I can separate them and put them back into culture to form more plants.

http://www.bananas.org/gallery/water...e=10677&size=1
http://www.bananas.org/gallery/water...e=10675&size=1
http://www.bananas.org/gallery/water...e=10676&size=1
http://www.bananas.org/gallery/water...e=10678&size=1
Remko.
These cultures have in the media the hormone BA (benzyladenine), this suppresses root growth and shoot elongation but encourages shoot proliferation, so you can make multiple meristems on a single tissue mass. Eventually, I will use a very similar media except the BA will be absent, this will allow roots to grow, shoots to elongate, and no longer encourage multiplication, this will allow the growth of single rooted plants ready for transfer to soil.
alah is offline   Reply With Quote Send A Private Message To alah
Reply   Email this Page Email this Page






Currently Active Users Viewing This Thread: 1 (0 members and 1 guests)
 
Thread Tools


Similar Threads
Thread Thread Starter Forum Replies Last Post
making it hard cherokee_greg Banana Plant Soil, Additives, and Fertilizer 25 09-19-2009 06:35 PM
Making Pseudostems Into Paper Ohio'sBest Main Banana Discussion 7 08-10-2009 01:08 PM
Making Compost on the Cheap? BadPun Banana Plant Soil, Additives, and Fertilizer 13 07-19-2009 07:40 PM
Making potash momoese Main Banana Discussion 5 08-22-2008 10:54 PM
Making a greenhouse around bananas? wxman Main Banana Discussion 3 05-07-2008 09:22 PM


All times are GMT -5. The time now is 02:58 PM.





Follow us:
Twitter YouTube

All content © Bananas.org & the respective author.