Phalaenopsis propagation on commercial scale

[Metallising]

Dear all,

I have been wondering for a while how Phal are propagated on commercial scale, to make it happen fast and in numbers how is it done?

Is stem propagation viable on a commercial scale? From the papers I have been reading, a single node in the multiplication medium (case study used P793) will produce about 10 new plants over a period of 30-60 days. After that period, the new plants are cut from the node and transferred to maintenance medium. The old node is transferred to new multiplication medium and continue to produce new plants for an indefinite period of time.

The papers stated this as the best method so far, I mean, the last paper dates back to 2004. In the meanwhile what kind of procedure have been adopted by the industry?

I have a reasonable sized greenhouse and would like to propagate some of my phalaenopsis to see how it does in the climate here. Of course the idea of doing it in large scale is very appealing for the commercial potential, so, here I am for reality check

Thanks for your time.

[Metallising]

To my initial post I would like to add some information I have been gathering from papers I found.

So, it seems commercial propagation of Phals start with node propagation. Once protocorm like bodys (PLB) start showing up from the node, they are cut and put in a bioreactor where they will multiply. Once there are protocorms in sufficient number, these are taken out of the bioreactor and placed in growing medium so they can develop as new plants.

The whole process of multiplying PLB in a bioreactor is completely new for me. Any info on this?

[Leafmite]

Check this out!

[Metallising]

Hey. I have seen it before. My question is more related to how Phals are propagated commercially, the video is more focused on how Phals are grown since the seedlings are deflasked.

Thanks for your suggestion, but I doubt they will share that information :P

[theloyalplum]

I'm no expert, but meristem propagation is probably how clones are done. It takes advantage of the totipotency of a plant cell where a single undifferentiated cell (or mass of cells) can be propagated into a larger mass of cells, separated as many times as necessary to get protocorms that can then be grown into individual plants. Many many growth hormones and nutrients are involved and it's very time consuming! I have not done anything with orchids, but I was able to play around in a lab a year ago with Arabidopsis, or the lab mouse of the plant world. From these undifferentiated meristem cells genetic modification can be done using agrobacterium (again don't know about orchids, but it might be interesting looking into it). This is probably where mutations occur, when DNA replication or cell division goes awry as more and more "copies" of the cells are made.

tl;dr: Get undifferentiated plant cells. Place in flasks with controlled nutrient and hormones. Ctrl+C and Ctrl+V. Place protocorms in flasks with different hormones to encourage differentiation. Grow. Replate into new flasks if necessary. Grow... Profit?

This is probably terribly inaccurate, so take it with a grain of salt. I've only ever done anything with arabidopsis, but from what my professor has stood by is that almost any plant can be propagated, genetically modified and grown in this way with a single piece of meristem tissue.

*goes off to do accounting homework*

Also, interesting approach to animal biology, where scientists are try to dedifferentiate animal cells into pseudo-embryonic cells that can then be manipulated to grow into specific cells and maybe even used to seed 3D printed organs tailored to a person to reduce rejection rates... Future. Okay, I should probably stop procrastinating...

[Metallising]

tl;dr: Get undifferentiated plant cells. Place in flasks with controlled nutrient and hormones. Ctrl+C and Ctrl+V. Place protocorms in flasks with different hormones to encourage differentiation. Grow. Replate into new flasks if necessary. Grow... Profit?

Once the protocorms are placed in flasks they will develop in new plants, is that what you are saying?
The idea behind the bioreactor is to multiply protocorms in a liquid medium, this requires a specific hormone called thidiazuron otherwise the protocorms will develop in new plants instead of multiplying.
This method is much more efficient than flasking protocorms with the propose of multiplication. The bioreactor is automated and doesn't require any labor, protocorms multiply faster because there's oxygenation inside the bioreactor. Also, since there's more oxygen crowding is less of a problem and higher densities are acceptable.
I found a protocol for mass multiplication of protocorm-like bodies using bioreactor publish at Plant Cell, by Park So Young. Still digesting

[theloyalplum]

Originally Posted by Metallising

Once the protocorms are placed in flasks they will develop in new plants, is that what you are saying?
The idea behind the bioreactor is to multiply protocorms in a liquid medium, this requires a specific hormone called thidiazuron otherwise the protocorms will develop in new plants instead of multiplying.
This method is much more efficient than flasking protocorms with the propose of multiplication. The bioreactor is automated and doesn't require any labor, protocorms multiply faster because there's oxygenation inside the bioreactor. Also, since there's more oxygen crowding is less of a problem and higher densities are acceptable.
I found a protocol for mass multiplication of protocorm-like bodies using bioreactor publish at Plant Cell, by Park So Young. Still digesting

Essentially. Given a right mix of hormones, the protocorms will remain as a mass of undifferentiated cells until you want them not to be. It's the magic of a lab. Since you can completely control temperature and nutrients and all that, you can create the best environment to multiply the mass of cells to your heart's desire. The shaker tables are used to constantly mix the solution so everything is as even as possible and the protocorms are only put onto solid medium after they're ready to grow into plants!

[Metallising]

What you wrote is in line with what I have been reading.
The paper I referred before reports a multiplication by 10 of PLB from which protocorm when the flasks are put in a shaker table, while in the bioreactor method the multiplication factor is 17. So, if you start culturing 100 protocorms you will end with 1700 after 8 weeks.
They report 83% rooting success once the protocorms are cultured in this Hyponex medium. There are more references to this medium in other papers but so far haven't been able to find it. Already asked to Phytotechlab, they don't have it.

[theloyalplum]

Are you looking to try? or just to propagate your own? There's been numerous studies using different hormonal and nutrient concentrations and medias for meristem propagation for plants in general and for orchids specifically. It's really not that hard and general use media packages are available out there. A sanitary setup is required though, so it might be a little much. If it's just for curiosity's sake, very little has been done with orchids in terms of experimentation because of how long they take to mature; not exactly the ideal model or test subject for genetic engineering.

[Metallising]

I want to try propagate my own, but I want to take the challenge a bit further and do it in a bioreactor like the pros do. The materials are not super expensive, even the laminar flow hood can be built for less than 400$, the blower and the HEPA filter cost 300$, the rest is up to your imagination.
The type of bioreactor (RITA) for this purpose is low-tech, is cheap as it doesn't require sophisticated mechanisms for pH and temperature control/adjustments.

More likely I will fail a few times until something actually multiply in my homemade lab. To keep me motivated I will flask in more "traditional" ways too, so I can get some experience and see some results faster.

I'm not prepared yet to start ordering parts, as I still have some questions about the process. I'm very enthusiastic about this I will do my best to make it worth. Fortunately I have a phD in microbiology at home, might be of some use

I'm not really into GMO, I just want to propagate them! I will follow a protocol established and proven to work, less see what happens.

Unfortunately the media packages available are not exactly what I'm looking for. This Hyponex media is want I need, but I still have some doubts about it, the formula used is 6.5N – 4.5P – 19K 1g/l + 20N – 20P – 20K 1 g/l + 1% potato homogenate. There's no reference to micronutrients, vitamins or hormones, which confuses me. Should I assume all those things missing are present in the 1% potato? Or is definitely something missing?