Re: Going to buy the Tissue Culture kit
 

OK, it took some time, but now me & a friend are ready to TC. We didn't go for the TC-kit, but bought the stuff we needed from Phytotech-Lab. They shipped fast and cheap, and answered our questions the best they could. (Even though they probably got tired of hearing from 2 crazy Norwegians!) They have 2 different culture mediums specifically for bananas: Basal medium and Multiplication medium, the latter you have to add sugar. You also need agar, and they reccomended special lids for the containers, wich in our case are baby-food jars.

Apparently, it's possible to TC very young seedlings. I am still confused about wich part of the banana-plant you sterilize and use for your very first step-culture. I have read you can use almost any part of the plant with meristomatic cells, even the flower-stem(?) In that case you could theoretically TC grocery-store bananas if you take tissue from where they "hang together", but as I said, I am very confused. Some people talk about chopping the plant-material into a fine paste, and TC this. It seems some people also go inside the stem, just above the corm, and take tissue from here. If anyone can verify and/or explain further, I would be very grateful.

Erlend

Re: Going to buy the Tissue Culture kit
 

For over 5 years now (I think), I've been engaged in email exchange with this forum started by Carol M. Stiff, PhD, who also owns "Kitchen Culture Kits" (www.kitchenculturekit.com ): hometissueculture@yahoogroups.com . This forum has wide following around the world, although most of them are here in the US and Canada. At the end of last year, she had a sale on her equipment and kits. I just don't know if the sale is still on.

Keith Benson is a consistent contributor in this forum and has discussed his home banana micropropagation techniques here. I thought I saved a copy of his article on banana TC culture on the web, but I couldn't find it. He made it look easy.

Most of the members of that forum will disagree with Joe's statement that "you don't need a sterile environment". I realize that he was refering to the culture of mushrooms. But in the case of bananas, which is akin to orchids, sterility is of outmost importance. Gabe has already decribed his experience on this.

As someone who has designed mechanical systems for clean rooms in the high-tech/bio-tech/pharmaceutical industries, I am very much impressed by the resourcefulness of many of the members in designing/making their own equipment, e.g., hoods/clean box, etc. Using anywhere from melamine laminate lined particle boards to rubbermaid plastic storage boxes for their hoods/clean box. Of course, they can't make your own HEPA filters, but there are other sources of HEPA filters than the high tech industry suppliers. Even home depot sells them. Then, if they have the correct size fan, depending on the level of cleanliness you want to achieve (the amount of cfm determines the Class), access gloves, and closures, they're all set.

PPM may be expensive, but if you consider that you can use ordinary materials with it, like distilled water from Walmart, compared to chlorinating/h. peroxide treatment, ozonating, UV irradiation (which if the water processors are supposed to do anyway) by yourself, it might be a trade-off. And PPM stays with the material longer than those other chemicals.

As a friend of mine would say, "My 2 cents."

Re: Going to buy the Tissue Culture kit
 

Hi Erlend,

Do you need to get both media or just one? Which did they suggest? What are you going to use as a hood? I didn't see any medium for bamboo on the website, but I haven't called them yet to ask.

I also am very close to doing some TC. I have a 2'x2'x4.5" 99.99% .3 filter and a motor from some old computer equipment (400-450cfm). I'm not sure what I'm going to use to build my flow hood yet. Somebody may suggest sizes of cabinet to build here (Chong?). I've got everything I need except the cabinet built, phyto-lids and some media. I only have around 200 small jars and 10 larger jars.

On the subject of where to get tissue. I've never heard of using bananas from the store but it really wouldn't cost much to try it. Have you ever read 'Plants From Test Tubes' by Lydiane Ryte? This and help from a friend who has done TC for years is what I'm using.

Good Luck!

Hi Chong,

Any help and guidance you can give me on materials and size to build would be greatly appreciated! If you can help me to get this set up right, it will just prove to the world you are the "King Of Clean!". Thanx!

Re: Going to buy the Tissue Culture kit
 

OK, I tried my first tissue culture now. As explants I utilized the tissue located just above the corm, inside the stem. Common sense made me believe this is where meristomatic cells are located. Can anyone verify if this is right or wrong? Can you utilize other plant-parts in other techniques? Anyone? :2140:

Re: Going to buy the Tissue Culture kit
 

Erlend, 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.

Re: Going to buy the Tissue Culture kit
 

Thankyou Gabe!
How far up in the pseudostem can you go? Just above the corm or even further up?

Erlend

Re: Going to buy the Tissue Culture kit
 

Erlend, imagine if 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. This way, you don't sacrifice an entire plant. Good luck with it! I just had my first TC lab at school yesterday. African Violets and Crotons :( . Oh well! Bananas are next...

Re: Going to buy the Tissue Culture kit
 

Thanks bigdog, I am starting to get it now. I believe I may have left a little too much leaf-bases on the ones I have in culture at the moment, but next time I will be certain about the explant!
Offcourse, I will let everyone know how easy or hard I found the whole process of TC.

Erlend

Re: Going to buy the Tissue Culture kit
 

Erland do you know how long you will have to wait to find out if its started to grow is it days weeks or months ?

Are there any step by step (WITH PHOTOS ) on how to do it.

Re: Going to buy the Tissue Culture kit
 

Erlend, did you try making a paste out of any of the tissue? Which medium did you use? Keep us posted.

Re: Going to buy the Tissue Culture kit
 

Erlend,

I am so interested in hearing about your progress on this!
Good luck, and I hope you have great success!!!

Re: Going to buy the Tissue Culture kit
 

Mark: I found it very difficult to find beginners info on the net. There is Keith Bensons' page, but that one is down for the moment. I have been searching inibap.org for articles. None of wich have photos and all of wich use professor-language! I have heard 2 weeks is when you should start to see small shoots forming.

DAndra: No, did not make a paste. Just used tips of what I believe (hope) to be the atypical meristem. Used the banana basal medium from phytotechlab.com. They also have a multiplication-medium wich I will use for my first subculture. They said you don't need to add sugar to the basal medium, while the multiplication-medium needs sugar added. Offcourse, I also used PPM wich has a psycho price of diamonds.

AnnaJW: Thankyou! I will keep everyone posted on the progress. But I am afraid I will have to try a few more times. Right now we are "benchmarking", checking for things that have not gone optimally, and searching for "best practice"!

Erlend

Here's a do-it-yourself site...
 

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, here's the new site...
 

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.

Still sorry, change hor to hort...n/m
 

n/m