Meri-clone technique

[a4jarl]

Dear All,

I've been flasking for a little over a year with a good level of success. I've exclusively used home made formulas without the addition of fruit pulp.
Would anyone happen to have a simple meri-cloning protocol worth trying? I'm looking for something rather easy to carry out in a home-made lab.

Thanks in advance

[margo]

Welcome, a4jarl! Glad you joined us!
Sorry I can't help with any idea, but I wonder if you can share your media and home flasking technique? I tryied once but... got nothing... Would love to try again!

[Brutal_Dreamer]

Welcome, a4jarl. There are a few great articles here on flasking, but I don't know if anyone has posted about home meri-cloning. I would also be interested in learning more about that too.

Cheers,
BD

[dudleyleroux]

Hey a4jarl,

The idea with a meristem is to initiate callus or undifferentiated cell growth from a meristem. This is usually done by using an equal quantity of auxin and cytokinins. it is very likely that the media you make for replating would work if you added 1mg/l Benzylaminopurine (BAP) and 1mg/l Napthaleneacetic acid (NAA). Once you have suitable callus growth you cut off a suitable section and place it in a media that has more cytokinin (eg: BAP) which would initiate cell elongation or shoot formation. Once again, as soon as you have enough shoot growth, you switch to a media with more auxins (eg: NAA), which will induce root formation. You should then be able to deflask.

Hope this helps.

[Frankco]

Hi a4jarl,

You can do it from the orchids you have in flasks fairly easily as long as they are very young, just developing. It might depend on the orchid you are germinating too. I am able to do this with Phal, Dendrobium and Cattleya. After germinating, and after the orchids have 3 or 4 tiny leaves on them, you can cut the top off and place them in the same media but with 1mg of 6-benzylaminopurine (BAP). The tiny shoots will develop into plants plus they will develop many more shoots. Each shoot then can be used to continue the process on and on.

I now skip the step of germinating them first and put 1mg of BAP directly in the gemination media. They start off normally but then produce many shoots.

[Frankco]

Here are what the orchids shoots of Cattleya look like after being in media with BAP. Each of these shoots can be cut and placed into other jars.



When you are done multiplying them, place them in media without BAP.

Here are some grown from shoot tips.

[Brutal_Dreamer]

Thanks Frankco! Welcome to the site!

cheers,
BD

[dudleyleroux]

That's excellent Franko, I never thought of that. I will be giving that a go as soon as i have some seedlings big enough. Thanks for sharing.

right! i now have some seeds in media with BAP, lets see what wickedness happens *evil grin*

[Louis_C]

Hi a4jarl,

You can do it from the orchids you have in flasks fairly easily as long as they are very young, just developing. It might depend on the orchid you are germinating too. I am able to do this with Phal, Dendrobium and Cattleya. After germinating, and after the orchids have 3 or 4 tiny leaves on them, you can cut the top off and place them in the same media but with 1mg of 6-benzylaminopurine (BAP). The tiny shoots will develop into plants plus they will develop many more shoots. Each shoot then can be used to continue the process on and on.

I now skip the step of germinating them first and put 1mg of BAP directly in the gemination media. They start off normally but then produce many shoots.

Where I can get BAP n NAA?

[Ron-NY]

There are many lab supply companies that will carry growth regulators. Also many Biological supply houses will carry these as well. I have purchased from a lab supply co in North Logan, Utah.

Also..please note that these chemicals need extra care using..they are mutagenic and may be carcinogenic.