High-frequency protocorm-like body (PLB) formation directly from thin leaf sections

[Ki6bud]

Really interesting article: "High-frequency protocorm-like body (PLB) formation directly from thin leaf sections of Doritaenopsis hybrid was achieved in order to develop a mass-scale propagation system."

(PDF) An efficient direct induction of protocorm-like bodies from leaf subepidermal cells of Doritaenopsis hybrid using thin-section culture

[King_of_orchid_growing:)]

Were you interested in talking about this subject or were you just presenting this article for people who might be interested to read it?

[Orchid Whisperer]

Thanks for the link; interesting article.

[Ki6bud]

Hi Philip. This article looks to be a very good article in mass producing the hybrid phalaenopsis I own. I have successfully cloned this orchid by way of stem propagation in vitro. Now I would like to take the next step in my education by creating PLB's. My goal is when I retire in 4 years to produce 500 clones to raise. I have scoured the internet to find out how to mass propagate clones and this article looks to be the key. What do you think?

[dansyr]

I've read this and a couple other similar articles, and attempted a couple times according to a few different protocols. I got close once, but I think attempting regeneration starts getting over to the art side of tissue culture. I got callous formation, and started to get what I think were PLBs, but every attempt browned out ultimately. If you get it working, I'd love to hear some experienced wisdom. I found it much easier to just multiply by dividing the plantlets on BA-containing media. You can double or triple your plantlet numbers relatively easily every few weeks or months and so get up to large numbers relatively quickly without needing to go through the very finicky regeneration step.

[Ki6bud]

This article was written back in 2002 and I'm sure over the past 6 years, advances have been made. Because most companies do not want to give away their trade secrets, these advances are not published. So, then how do hobbyists find these trade secrets without reinventing the wheel? The answer may be out there with those who worked professionally in the industry who also would like to share information (as long as they did not sign a NDA).

[dansyr]

Originally Posted by Ki6bud

TBecause most companies do not want to give away their trade secrets, these advances are not published. So, then how do hobbyists find these trade secrets without reinventing the wheel?

As an dyed-in-the-wool academic believe me I'm generally skeptical, but I'd be cautious in assuming nefarious motives. While regeneration in a commercially-applicable species has obvious industrial applications, I know many academic-track researchers/friends who work in TC. Also, this isn't like protocols to CRISPR your orchids into glow-in-the-dark transgenics or whatever.

And there has been quite a bit of further research into induction of PLBs from various parts. I haven't done an exhaustive literature search but I've read several more recent papers, I think 2016 is the most recent. And I own a copy of an edited TC manual for a variety of plants, including Phals. The protocols are out there, and they're not that wildly different (hence why I don't think there's some industrial cabal holding out on us). It's just that when you get into trying to execute a finicky protocol, devils crop up in unexpected details. For example, my lab sometimes works on an anaerobic sulfur bacterium, and there are excellent cultivation protocols in existence around since forever, but successfully cultivating it reliably ends in failure until you're practiced. The growing parts of biology are often frustratingly experience-based and difficult to transcribe into a cookbook (why physicists get very annoyed at us).

Sorry that was kind of a ramble. I say all this to suggest, don't rush to assign ill intent here. And this is not to say that it's impossible - clearly many people can do it, and the directions are pretty accessible. I just spent a bit of time attempting it and don't care to sink any more time in given my goals, so I'm just warning you based on my experience. But, if you can get regeneration from leaf explants working, excellent, and I would love to question you for tips.

[Ki6bud]

Hi Dansyr. No ill will taken. I appreciate a good discussion on this subject. I worked in the semiconductor for most of my career and I have signed nondisclosure agreements to prevent competitors taking away market share. Is this nefarious? No. Is this smart business practice? Yes.

Most scientific papers we published were either older technology or did not have a significant impact on our proprietary processes. So, when I read papers from 2002 they are either early discoveries meant to inspire further development, or processes that have already been developed but have little consequences to the advance technology already obtained.

Having little knowledge of the larger commercial tissue culture labs, do they operate as the semiconductor industry in the sense of releasing valuable information? I would certainly like to know how the larger labs mass produce orchids faster, cheaper and better!

Bill Gates: “I choose a lazy person to do a hard job. Because a lazy person will find an easy way to do it.”