ArchAngeL01

what exactly is tissue culture? and can i do it at home?

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Brad

ron_mcb

plant tissue culture is basically a method used for propagating more organisms from the original organism using cells...the cells are fed using a growth medium.

the main purpose of plant tissue culture is to produce disease free plants and,or quickly produce lots of cloned plants with little material.

it seems to be expensive and time consuming..you will have to worry about contamination.you you will probably need to keep your work environment as sterile as a hospital.


i dont believe you can just jump into it. you have to read about it and then you may be able to get yout feet wet..i think there are books on the subject of performing micropropagation at home. maybe the experts will tell us all about it,i can only supply general info. hope this helps.

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naw mek smaddi ditate fi yuh. naw wan gwan a pawty yuh naggo.

Lagniappe

It's not as expensive or complicated as one might think. There have been several discussions on this forum . Use the search and read the older threads.

ArchAngeL01

thank you ,i'll search it, thank you both

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Brad

Taylor

We have our own resources! Take advantage of them! Here:

Info:Tissue Culturing - Bananas Wiki

Michael_Andrew

Isn't that what you have when you blow you nose in a kleenex. I'll post a picture later.

Michael

ArchAngeL01

trying to be funny huh?

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Brad

Frankco

Hi ArchAngeL01,

Plant tissue culture isn't really all that hard and tissue culturing banana, at least from what I have found, is a lot easier than most plants. I tissue culture plants in a spare bedroom, just to keep the cats out. You could set up a small area in your living room, kitchen or were ever you have enough space for a small desk and some shelves to place your jars. In a short order you can have more banana plants than you know what to do with. In just 2 and ½ months from one Musa Basjoo I now have 80 small explants. In another 2 months I should have 256. I should have in 4 months, if account for 10% not making it, somewhere around 4000, that is if I can get enough jars:-)

My room is definitely not sterile as a hospital! I use a 30 gal aquarium set on its side to give me the sterile area in which to work with the plants. It’s cheap, easy to setup and works as well as an expensive laminar flow hood.
Most of the stuff you need you can get at a thrift store or garage sales. The most expensive thing you need is a pressure cooker to sterilize your media/jars. You can do it in a microwave but it can be a little tricky and can be messy if you are not careful. If you use a microwave you should add a couple of jars extra to act as heat sinks. This keeps your media from bubbling over too quickly.

As for bananas, I found you need very little lighting but need warmth (76-82 deg F). Actually at the start, when you stick the first banana in the jar, keeping it under very low light helps so, having lights setup isn’t necessary at the beginning. When you first start a Banana, it has a tendency to turn black under strong lights because of the phenolic extrudes. Low light helps with this problem. You will need to take the little banana explant out of the first jar and switch it to a new jar about once a week until the phenolic extrudes stop (about 4 or 5 times). After that it’s clear sailing.

Almost a year ago I started to make some videos on how I got started with micropropagation. I have changed and improved the way I do things since then but, the videos will give you an idea on how to do some of the things. The videos don't cover bananas but an African violet. I'll try to do one on bananas in the future. If you want to see the videos you can do a search on YouTube for micropropagation. There are 11 videos, I think:-)

Frank

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Frankco
Zone 5
N.W. Indiana

ArchAngeL01

Thank You for helping me!.......i'll whatch the vids and i already have a pressure pot ! YAY!! but im confused of what part of the banana to put in the jars!? is there any chemicals ? thnxx

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Brad

Frankco

Hi ArchAngeL01,

You have a pressure cooker, that's great!

The resident banana tissue culture expert, Gabe, put up a nice video on YouTube that shows how to trim the banana. You can watch it here.


As far as the part to use take a look at this page.
Micropropagation of Musa

As for the plant growth regulators, you will need these two, BAP and NAA. You can wait to get the NAA if you want. You won't need it until you are done multiplying the bananas. Depending on how many you want, you won't need it for at least a couple of months.

6-BENZYLAMINOPURINE SOLUTION or BAP (100 ml)
6-BENZYLAMINOPURINE SOLUTION (1.0 mg/mL) - Plant Growth Regulators - B130

NAPHTHALENEACETIC ACID SOLUTION NAA (100 ml)
NAPHTHALENEACETIC ACID SOLUTION (1.0 mg/mL) - Plant Growth Regulators - N605

You can get a smaller quantity of NAA from Carol's web site. You will not need as much of NAA as you will BAP. Carol sells it in 10 ml bottles. That’s enough for 20 liters of media as you will use .5ml per liter. Her shipping prices are better than Phytotech's too.
Home Tissue Culture* Group Catalog

You have to keep these plant growth regulars refrigerated. It is probably best to divide some in small plastic bottles and freeze it. They will last longer that way, about a year. Be careful that anyone who has access to your frig knows what they are. If there are children in your house, don’t keep them in the frig. It would be best to keep them in a cool, secure place and just buy fresh when you need more.

For Musa Basjoo you will need to use 5mg or 5 ml, ( 5mg=5ml with Phyototech’s BAP solution), per liter of media. This will give you the most multiplication. You should get at least 10+ little sprouts with the first explant. Cut each little sprout and put those in the same media and you should get 10+ from each one of those. The sprouts can be very small but each will grow. I got 16 from my first explant. The second round is proving to be just as good. So, on the second round you should get at least 10 x 10 or a hundred little explants in about 2 months. You should get better than this though.

You can get this agar to start out with. It’s $4.20 and will make a lot of media for you. Use 2 ¼ teaspoons per liter.
Amazon.com: Agar Powder - 2 Ounces: Health & Personal Care

Get some packets of MS nutrients. They are a $1 each. Each one will make 1 liter of media.
MURASHIGE & SKOOG (MS) BASAL SALT MIXTURE - Media, Basal Salts, & Vitamins - M524

Just pour the MS nutrients into 1 liter of distilled water. Add 4 teaspoons of sugar, 5 ml of BAP (use a 1 milliliter syringe to measure).

You will have to adjust the pH as it will be a little low. You want to get the pH close to 5.5 You can ballpark this with some pH papers. Just add the baking soda very slowly to raise the pH, a very tiny pinch at a time. Add 2 ¼ teaspoons of agar, heat to a boil and pour enough media into baby food or canning jars to cover ½ inch of the bottom and then put that in your pressure cooker for 15 minutes. You’re done with the media!

The web pages and video that I listed, I hope, will guide you the rest of the way.
Good luck and let me know how you do.
Frank

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Frankco
Zone 5
N.W. Indiana

ArchAngeL01

it dosn't seem as complicated as i thought ,i cant wait to try it...im moving soon so im going to wait to start it so i won't have to move everything.....thank you for the info! the only thing is what next step do you take from taking the plant out of the jar and how do you know when it's ready? thnxx you so much!

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Brad

Frankco

Hi ArchAngeL01,

Ah, that's where the NAA comes in. With the your last round of multiplication, let the plants get a little bigger. Let them will grow into tall single shoots, 3 to 4 inches. Divide the shoots keeping the base on them and put them in media with .5mg of NAA.

Different strenghts of auxins and types of auxins, such as NAA or IBA will work differently for different species of banana. Reports of up to 2mg per liter of NAA have been found to promote rooting with some bananas. NAA at .5mg per liter will work fine for Musa Basjoo.

The NAA promotes the little banana plants to develop roots. When the bananas get to be about 6 to 7 inches tall, (you'll need tall jars), and have nice roots, take them out of the jars, wash all of the agar off under the sink and plant them in a covered container with good potting soil. Keep them well watered and out of direct sun light.

Let them grow some and slowly increase the amount of light they get and slowly allow more air to circulate into the covered container. You can allow more air in the container by increasing the gap of the cover. Do this slowly, they need time to adjust.

When they get to be about 20 inches tall they should be out in the sunlight and getting fresh air. At that point they are ready to brave the world on their own.

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Frankco
Zone 5
N.W. Indiana

ArchAngeL01

sounds good! how long does the proccess take? i just got my ice cream pup in the mail today! can't wait to get 'em in the ground! and start my lab work hehehe do you do it?

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Brad

Frankco

I'm not sure with your ice cream banana. You're going to need a pup to sacrifice first I wouldn't try it with your only ice cream banana.

For Musa Basjoo, if I wanted to just do two rounds of multiplication, I would say about 6 months.

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Frankco
Zone 5
N.W. Indiana

ArchAngeL01

ok i cant wait! i get so excited about my naners i can't sleep at night!! hahhaahha what kinds do u grow?

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Brad

Frankco

Right now just Musa Basjoo. Our -18 below zero this winter did in the ones I had outside. It may have been that I left them covered with hay too long though. When I uncovered them last weekend the temp inside the cage filled with hay was pretty warm. Maybe I cooked them.

I brought in 10 little pups so I have pleny to plant ouside. The ones in the jars are still too tiny and only on their second round of multiplication.

Ya, I like bananas too. I do want to get some different kinds. They'll have to keep them in the house. It gets too cold here.

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Frankco
Zone 5
N.W. Indiana

ArchAngeL01

my basjoo is doing great ,its beggining to pup

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Brad

alah

Hi, Bradley. TAKE IT EASY. While I was reading this thread it crossed my mind the amount of sleep I´m losing each night!, BEFORE I read your quote!
This isn´t really difficult, even though I haven´t even tried it yet!
But as some say, measure twice, cut once. Read some more although sometimos you get more confused so ( take extracts NOT explants ), as I´m doing; e.g. from this thread I copied and pasted on another document the follwing info:

“As Gabe mentioned before, we generally use 1/2 strength M&S medium, with BA or BAP during initiation.
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.

No, 1 mg/ml in water. If you are dissolving it in DMSO use about 25 mg/ml. You will use about 5 mg of this BAP solution in your agar.
Note: It's quite alkaline, so be sure to adjust your pH.

When you first start a Banana, it has a tendency to turn black under strong lights because of the phenolic extrudes. Low light helps with this problem. You will need to take the little banana explant out of the first jar and switch it to a new jar about once a week until the phenolic extrudes stop (about 4 or 5 times). After that it’s clear sailing.”

One other point. When I started reading I had no idea about IAA, NAA, 2,4-D not even MS! If you don´t, anybody can post some info. So, take it easy, Alberto.

alah

Sorry Bradley, I forgot something very important. I HOPE that very soon somebody extracts sleeping potion, a very potent one, from bananas!
I´m going to try to sleep.

ArchAngeL01

thanx for the info alah do u think home remedies would work ? eg sugar ...vitamin pills

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Brad