Flasked seeds and cells,- NOW WHAT?

[Florida_guy_26]

Right again Philip. I also have some bap or bezylaminopurine that came in a few weeks ago. I have not used it yet, but I saw posts from someone on another site who used just about 200ppm in each flask and had amazing results with new buds coming off every available part of the rhizome. of some orchids. I do not know how wells it works with some orchids, but I think for catts it works well. Thank you for the info, and I will see what happens or if things change.

[Florida_guy_26]

Ok guys, I looked at phyto-tech labs and while the prices for the actual medium are good, they wanted $2.50 for 2 containers of replication medium but $13+ for shipping and what they call a handling/processing fee. That is ridiculous for what should only cost 6$ at the most for priority mail and they offer no usps only ups and fed ex shipping. That is the most retarded thing I have ever heard of and a way to swindle people into paying more for unnecessary services and postage. I was hoping there are some other suggestions for places that choose not to rip off our hard earned money or are at least a bit more reasonable on shipping and processing fees. I also emailed some people to ask them if they have any kind of replication medium that will work. I guess in time I will figure it all out.

[gixrj18]

Originally Posted by Florida_guy_26

Ok guys, I looked at phyto-tech labs and while the prices for the actual medium are good, they wanted $2.50 for 2 containers of replication medium but $13+ for shipping and what they call a handling/processing fee. That is ridiculous for what should only cost 6$ at the most for priority mail and they offer no usps only ups and fed ex shipping. That is the most retarded thing I have ever heard of and a way to swindle people into paying more for unnecessary services and postage. I was hoping there are some other suggestions for places that choose not to rip off our hard earned money or are at least a bit more reasonable on shipping and processing fees. I also emailed some people to ask them if they have any kind of replication medium that will work. I guess in time I will figure it all out.

I use Phytotech medias, the trick is to place a big order, and stock up...that kind of helps. I'm not familiar with replication medium, but if multiplying is what you are trying to do, this can be acheived simply by adding a little coconut water to your medium (I use P-723, and replate with P-785). This will cause the protocorms to put out a lot more plants.....the more coconut water, the more proliferation. Not sure if that is what you are looking for, but maybe that'll help. Good luck!

[Florida_guy_26]

Thank you very much Jasen, that does help. I will try sterilizing everything and adding a tiny bit of coconut water to see if that helps. At the stage I am at, I doubt anything will as I do not have any growth whatsoever in the flasks. I will try some bap when I collect more cell samples and I will also try different formulas and combinations but as I said, I do not think there are many totipotent cells in orchids that will grow no matter how young or old the growth is. I have yet to see any cell sample that even shows cell replication or multiplication in any of the flasks I have to this point. No green, no bumps, no irregular spots on the medium - nothing. I will try again with different cells from different areas of the plants and hope for the best. Thank you again though.

[gixrj18]

Originally Posted by Florida_guy_26

Thank you very much Jasen, that does help. I will try sterilizing everything and adding a tiny bit of coconut water to see if that helps. At the stage I am at, I doubt anything will as I do not have any growth whatsoever in the flasks. I will try some bap when I collect more cell samples and I will also try different formulas and combinations but as I said, I do not think there are many totipotent cells in orchids that will grow no matter how young or old the growth is. I have yet to see any cell sample that even shows cell replication or multiplication in any of the flasks I have to this point. No green, no bumps, no irregular spots on the medium - nothing. I will try again with different cells from different areas of the plants and hope for the best. Thank you again though.

It's really hit-or-miss. I have a closet full of flasks that haven't germinated, I would say only about 30% of the flasks that I have sown have germinated. So don't get discouraged, it's normal....you just have to flask as many as you can, and hope for the best.

[King_of_orchid_growing:)]

Cloning is no different.

Not many orchids have been successfully cloned.

[Royal]

I've already weighed in on this, but I have to jump in here and disagree for a second. My success, like Jasen's, has been hit or miss. BUT, neither of us are pros. We both have had to "make do" with the materials we have at hand. Plus, our environments are hardly laboratories. I work in my kitchen and have a dog and a cat. Germinating seed is an involved process and a bold undertaking, but we have some success without taking out a second mortgage, and that's pretty good for a hobbyist. However, commercial laboratories that can sink lots of cash into facilities and equipment can probably germinate 95% of the viable seed that comes through the door.

Cloning is the same, but even more bold of an undertaking. It's possible in the home, and labs around the world are successful in making meristem clones of many types of orchids. Some genera elude them, but cloning is big business. But again, these labs are good at what they do and have all the tools required. I know some growers that are cloning in their home labs, but they start by taking advantage of all the research that preceded them. They all have LF hoods, dissecting scopes, rotators, shakers, etc. None of them are making PLB's from root tips, leaf tissue, diced up new growths, or cells extracted from anywhere. They all use established protocol for meristem culture.

If I were to endeavor to do some "cloning" on my own, I would try to replicate what successful labs are doing. Learn more about the process and set achievable goals. If you get 30% success - awesome, that is success. You could master your own technique that you invented, but wind up with 0% success. That would be discouraging.

Look, all I'm saying is don't give up. You seem like a pretty persistent guy and I admire that. Just make sure to eliminate as many variables as you can. Read up, do a little more research on the front end. There's a TC listserv to which Phillip and I both subscribe. There are lots of papers on this topic out there, probably a ton at your local college. And maybe start with some seed or some stem props. The technique is similar, and practice makes perfect. Having some measure of success as you work toward your goal may help.

[King_of_orchid_growing:)]

Originally Posted by RoyalOrchids

There are lots of papers on this topic out there, probably a ton at your local college.

Scientific articles are everywhere on the net, you just gotta find them and figure out what they're saying. It can be very technical sometimes.

Many are inaccessible to the general public for free.

If you're affiliated with a lab or a university then you can access them for free.

On Google Scholar you'll find the articles regarding Phalaenopsis root tip meristem cloning and leaf tip Phalaenopsis and Laelia meristem cloning. It's been done already. I have the scientifically published articles. There are more than one regarding this kind of meristem cloning, they're not by the same authors either. Some come complete with photo plates.

It doesn't have to be just apical meristems or adventitious meristems anymore.

Originally Posted by RoyalOrchids

There's a TC listserv to which Philip and I both subscribe.

Really...you're on it too? What name do you go by?

I forgot about this resource. The Home Tissue Culture Listserv, is run by Carol Stiff, it's an awesome resource for those interested in tissue culture. It covers the gamut, it doesn't just deal with orchids. Everything from bananas to woody plants, etc. is talked about. It's a worldwide network btw, not just people from the US are on it.

[Royal]

Originally Posted by King_of_orchid_growing:)

Scientific articles are everywhere on the net, you just gotta find them and figure out what they're saying. It can be very technical sometimes.

Many are inaccessible to the general public for free.

If you're affiliated with a lab or a university then you can access them for free.

Sometimes all you need is a public library card and you can get things from universities on inter-library loan. I've been out of school for a while, but I can still access anything at the local college though my public library.

Originally Posted by King_of_orchid_growing:)

It doesn't have to be just apical meristems or adventitious meristems anymore.

I understand, but this all cutting edge. We probably won't stumble upon the same success in our homes that large research institutions have achieved after decades of work. If it was as easy as chopping up some plant tissue and sticking it in a test tube, we wouldn't be having this conversation.

[Florida_guy_26]

I have read through so many articles online- some from college students doing research on cloning in college labs, some grad students writing reports on cloning success and the use of different medium for each separate tissue sample. Yes, I have been unsuccessful as of yet, but I will not give up. It would help if someone had the way/means to develop a workable cloning medium just for cloning orchid tissue. I have read there is no medium to date that will work for cloning all genera or an all purpose cloning medium period. I have read hours worth of work by doctors, pharmacists, grad students, professors, lab study reports, cell studies etc that my mind could explode. Almost everything you read now can contradict another article and you all know that to be true. I am going by theory right now and flying by the seem of my pants so to speak. I am not discouraged because I finally have sterile flasks and that for me was one challenge, but now I am out to prove that some how mother nature can reproduce without the benefit of sperm and ova or ovum. I know snails and slugs can do it and so can many species of arthropods- and because of that, I know eventually I will prove that no matter how hard I must work, I can produce new orchids without the process of hacking the living hell out of plants. I know that I can get cells from a plant some how or some way that will grow into new plants on their own. So far I just do not have the proper medium to actually get anything growing. I have read articles that say certain medium is required yet there is nothing available that is really any different than what I already have. I also read the proper hormones, nutrients, and essential vitamins and amino acids can cause cell growth. I do believe that no matter what medium you use, there are cells that will grow on it, you just have to find them. Call me crazy- that is my opinion on this whole mess and I will keep trying until i prove myself right or everyone else wrong about how to clone. It has been done by many people true, but I am sure there are tissues that will grow with less stimulation than what everyone else thinks is needed. I have read protoplasts, basal stem cells, root tip cells and various others are totipotent and maybe the ones I have just need time to grow. I will find cells that will grow on just normal motherflasking medium as it has all the right micro and macro nutrients, proper salt content, ph, vitamins and hormones to initiate growth from any cells. If a seed which has a protective outer cover can germinate in it, then I believe that a cell from other parts will eventually become a new plant. Maybe I have not given enough time to these cells to actually grow, but in any case, I am out to change the world one flask at a time.