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View Full Version : Musella Lasiocarpa tissue culture help


jarceleste
09-10-2010, 09:41 AM
Hello ppl.:bananas_b

Three weeks ago, I've started my Musella lasiocarpa tissue culture.

In the first 4 days it starts to increase the central part, but after that and till the momment, I don't see any changes in the explant.
I don't have contamination in the media.
I've used MS media with vitamines, and no plant hormones.

Can anybody tell me what's wrong?

I have some photos since the beginning <br>http://www.pikipimp.com/pp/pimped_photo/s/image/54/434/18/IMG00128-20100823-2242-compiled.jpg?ts=1284128064685<br>http://www.pikipimp.com/pp/pimped_photo/s/image/54/434/28/IMG00129-20100823-2242-compiled.jpg?ts=1284128295427<br>http://www.pikipimp.com/pp/pimped_photo/s/image/54/434/53/IMG00133-20100826-1951-compiled.jpg?ts=1284128340116<br>http://www.pikipimp.com/pp/pimped_photo/s/image/54/434/148/IMG00137-20100826-1953-compiled.jpg?ts=1284128740671<br>http://www.pikipimp.com/pp/pimped_photo/s/image/54/434/68/IMG00141-20100827-2147-compiled.jpg?ts=1284128416285<br>http://www.pikipimp.com/pp/pimped_photo/s/image/54/434/73/IMG00150-20100831-1732-compiled.jpg?ts=1284128456034<br>http://www.pikipimp.com/pp/pimped_photo/s/image/54/434/83/IMG00151-20100831-1732-compiled.jpg?ts=1284128495455<br>http://www.pikipimp.com/pp/pimped_photo/s/image/54/434/103/IMG00152-20100831-1733-compiled.jpg?ts=1284128536298

Gabe15
09-10-2010, 12:08 PM
It looks like you put the wrong part of the plant in. You just have a piece of pseudostem which has no capacity to continue growth or regenerate plants. You need to have a the meristem in there, which in bananas is a layer that lies between the corm and the pseudostem. In order to get a piece of meristem, you just include a chunk of tissue which has corm and pseudostem, that way you know there is some meristematic tissue in there as well.

I started to make a guide video but have not had the chance to work on it more for a while, but I do have this one small section on YouTube which shows how to actually cut the plant during initiation (there is no commentary on it, that is to come later but people were bugging me to put up whatever I had so this is it for now). If/when I start the video guide again, I will likely even make a new clip. This process is simple once you learn it, but can be difficult to conceptualize unless you see it done.

It is even difficult to see whats going on in the video perhaps, but basically I had a plant which I trimmed down to a small section of corm of and pseudostem, put it in a mild bleach solution for a few minutes and trimmed it down to a small a piece of tissue that includes the a small section of meristem. Again, you know that there will be meristematic tissue somewhere in your piece if you preserve some pseudostem and some corm attached to the explant. When you have that last small piece, you can either put it in the medium as is, or you can quarter it vertically (top to bottom/pseudostem to corm)to induce more shoot proliferation (which is a normal practice). It is also good to put each of those 1/4 pieces into its own vessel as this helps to ensure that you can keep the line going in case of contamination...don't put all your explants in one vessel!

When you do it right next time, you may also want to change your medium. I have never TC'd Musella, and it may do better with a modified medium, but standard banana initiation medium is 1/2 strength MS with 5 micrograms/L BA. If you do not have the BA (hormone), then there will be little to no shoot proliferation (depends on the variety though) and thus you will not be multiplying your plants very rapidly. See the details of the premade multiplication medium below. This is a good cheap one to buy (for initiation and subculturing only), or you can copy the recipe and make your own. If you make your own, you can start from scratch or you can get the " Banana AGS Basal Medium", and then add sugar, a gelling agent and adjust how much hormone which is important when switching from subculturing to rooting.


MUSA (BANANA) MULTIPLICATION MEDIUM - Media, Basal Salts, & Vitamins - M462 (http://www.phytotechlab.com/detail.aspx?ID=429)
BANANA AGS BASAL MEDIUM - Media, Basal Salts, & Vitamins - B144 (http://www.phytotechlab.com/detail.aspx?ID=124)

YouTube - banana tissue culture (http://www.youtube.com/watch?v=p4DUwSGJYNE)