Gabe15

M. velutina, about 15 days old.


M. balbisiana, about the same age, seem to be slower to germinate and grow in the same medium.


Ensete glaucum subcultured from a single embryo, 25 explants.

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microfarmer

Looks good! Can you give us the Cliff's Notes version of the procedure? What tools would I need?

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bikoro child

wow!!! that's great !!! they are in agar agar?

Las Palmas Norte

I've never been able to germinate any of the store bought banana seeds (Thompson & Morgan etc). Is there a trick beyond soaking and a warm growing medium? Have these store bought seeds lost their viablity with age? If so, how fresh do the seeds have to be, and where would one source them out?
Sorry for all the questions ... thanks in advance. Cheers, Barrie.

51st state

Wow, Gabe thats some awesome photos, loving the Glaucum.

Can you germinate from seed?

I may have a source for some 'special' plants which it might be fun to propagate.

Let me know and I'll PM you.

Kev

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bigdog

That's cool, Gabe. Now do that with some Musa ingens, and you're in business!

Gabe15

I should have written more down, but I was in a bit of a hurry at the time. The pictures I posted above are of embryos germinating on a tissue culture medium. Each banana seed contains an embryo which is the actual plant that grows from the seed, the rest of the seed is there to feed and protect the embryo as it germinates and grows. The technique is called embryo rescue germination, the embryos are removed from the seed and (if all goes well) forced to germinate. This is a common method for germinating seeds from hybrid plants which may not germinate normally (usually due to lack of a fully developed and/or functional seed), it is also good for plants that are normally difficult to germinate as well.

The 3 species I cultured were just for practice, they are not rare or necessarily difficult to germinate normally. The Ensete glaucum was originally a single germinated embryo which I induced shoot proliferation on (made many meristems, instead of just one), and then subcultured (divide and re-culture) once to produce 5 new plants, and each of those was then subcultured later to produce many new plants each, I had so many (about 50) that I had to throw half of them away, I kept 25 of them which are now growing. They are genetically identical and suitable for subculture an additional 3 more times if I wanted to create more. They are very young right now, which is why they just look like a mass of green tissue instead of full plants.

I have also put into culture various other Musa and Ensete seeds, but have had limited success with some of them. I will be trying out new and different culture mediums and methods in my trials to come.

I am lucky to have training in tissue culture and full access to a tissue culture lab and all of the equipment and materials necessary. However, I think it could be done with those kitchen culture kits for home tissue culture, or if you do your research on basic tissue culture you can probably rig something cheap and easy. The materials are not very expensive themselves, but the equipment like an autoclave and hood is where you would need a real labs help, but like I said earlier, with enough experimentation it can probably be done at home.

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Gabe15

I actually did! Unfortunately I only had one seed left and it seemed to be contaminated on the inside (it had been in soil at one point), so the culture was over taken by mold within a few days. But if/when I get more I will try it again. The same thing happened with E. perrieri. Embryos of M. beccarii var. hottana, M. ornata, M. laterita, Ensete sp. 'Thai Superbum' and Musella lasiocarpa are all in culture as well but have not shown any signs of life yet, but that could be due to a number of different variables.

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inkcube

embryo rescue itself might be difficult in a kitchen lab but with a pressure cooker for an autoclave, fruit jars/babyfood jars, and a fish tank on its side as your hood plus a little ingenuity seed sowing can easily be done at home. i started back in the early '80's with orchid seed in a kitchen lab.

getting chemicals may be tough since they are usually sold in quantities that would be large for hobbyist plus some are quite expensive and others can be difficult. also buying a good scale would be necessary and that might put you on a DEA watch list, good precision scales are considered drug paraphernalia.

mrbungalow

Try this one for chemicals and equipment: Home Page - PhytoTechnology

For bananas, go for the "Banana multiplication media". Basically, just add (sterile) water, meristem, and a drop of PPM (wich you buy somewhere else.)

I have not been sucessfull yet, but I also lack a sterilizer. (Microwave or pressure cooker)

The race is on to produce TC-plants!!

BTW, Gabe, could you elaborate on the differences between TC and embryo-rescue?

Cheers
Erlend

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Gabe15

Your best bet is to probably use a pressure cooker to sterilize the media containers, with the media inside of it (before it gels), the PPM is supposed to reduce contamination but in my trials with it, it cannot handle the mold alone, it needs extra sterilization.

You can get fancy with the medium, but what I am using is basic MS media, sugar, thiamin HCl and gelrite, with minute amounts of hydrochloric acid or sodium hydroxide to achieve a pH of 5.6-5.8. I am experimenting also with levels of shoot proliferation hormones, this essentially makes the embryo either germinate into one or many plants at a time. If anyone is serious about trying and wants more detailed info, PM or email me.

Tissue culture is just a basic term for the culturing of tissue in a controlled environment, in vitro. Embryo rescue means you use the embryo from the seed as your source of tissue, in "regular" tissue culture, you would likely be using the meristematic tissue taken from a shoot which has not flowered yet, although other parts can be used for different reasons.

I also do normal micropropagation tissue culture (standard, as what Agri-Starts and such do), and will hopefully be able to send some of the interesting banana plants here back to the mainland so people can grow them.

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inkcube

if you go to this link:
germination

and down to post #3 you will see my post on the basics of embryo rescue. it was basically developed to TC difficult to germinate hybrids and perfected in the day lily labs.

use a pressure cooker, it works on the same principle as an autoclave. ppm is a waste of money. when working in a home lab the main detail is to pay attention to sterility.

mrbungalow

PPM is almost more expensive than gold by weight! As a novice to TC, I bought some just to make sure. Is it possible to be sterile enough as a home TC-experimenter? Or would you reccomend other antimicrobials?

Also, do you have any tips on using explants etc.?

Erlend

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inkcube

a pressure cooker will adequately sterilize, they use the same principles of pressure plus heat as an autoclave - pressure cookers will reach a temperature of 250F/121C, @ 15lbs of pressure you need 20-30 minutes to be effectively sterilized. also longer time do not equal more sterile - 15lbs of steam pressure for 15 minutes will kill all bacteria, viruses, spores, and other living things. the key with a pressure cooker is to monitor its temp by watching the release valve (the weight you set on the nozzle) on top which rocks as the steam is let off. be sure set the weight on 15, the weight is designed so that it will lift and hiss at the desired pressure. you want it to rock slowly, about 10 rock per minute. put about 3cm (~8 oz) of water in the cooker to generate your steam. always let the cooker get cold before opening it if it is hot. the hot steam inside containers condenses to a liquid and will suck in one volume of non-sterile air. if you let the cooker get cold, the dust in the room air collects on the surfaces it strikes and no non-sterile air gets sucked inside the containers. you also do not want to tighten the lids of the containers you sterilize (screw them on loosely) - the pressure change will make it difficult to open.

when i used to do this in the kitchen PPM did not exist and i never had problems with contamination - good technique is always the ket to success with TC.

sandy0225

I'm going to have to try that next winter when I get bored. This time of the year is already starting to get busy. That really looks like fun!

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Northern Tropics Greenhouse
1501 East Fuson Road
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specializing in bananas, heirloom tomatoes and water gardening plants~
check out our new online store at our website!

microfarmer

That's the same setup I used for a mushroom culture experiment. I sterilized the grain, with water, in the jars, in the pressure cooker. Worked like a charm. Very low tech. I might just might dig out the equipment and try it again. Maybe some Chanterelles this time...

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mrbungalow

Gabe, How did you sterilize the embryos before putting them in the media?

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Gabe15

I sterilized the seeds in bleach for a few minutes, then cut them open on a sterile surface. The seeds are sterile on the inside if unplanted, I have had mixed results with seeds that have been in soil prior to culture, some showed instant contamination and others appeared to be clean (but have yet to germinate).

Alternatively, if you are working with fresh fruit, you can simply sterilize the fruit in bleach and then cut in open on a sterile surface and everything inside will be sterile, I have had good results with this and you also get the freshest embryos.

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