Dendrobium Dehiscence

A while back, in this post, I self pollinated Dendrobium Hamana Smile.  Almost exactly two months later, the seed pods were mature and began to split.  For an orchid, that's fast!

D. Hamana Smile Orchid Seeds
©2012 MadOrchidist.com

As you can see, I was ready for the seeds with a clean coffee filter.  Once these opened more fully, a gentle tap knocked out all the seed I'll ever need.  I haven't gotten them flasked up yet, so I will update this post when I do . . .

A Flask 6 Months in the Making

Back in July 2011 I attempted my first orchid pollination.  Since then I have been pollinating other flowers and learning/testing laboratory flasking procedures with seeds obtained elsewhere.  Now the seed capsule resulting from that first pollination attempt is mature, and it is finally time to put the lab to use with my own seeds!

Ripe Seed Pod with Coffee Filter Safety Net
©2012 MadOrchidist.com

I brought the pod-parent plant inside recently so that I could keep an eye on the ripening pod.  I have been propping a clean coffee filter under it to catch seeds in case it should burst when I'm not looking.  Well, today I caught it in the act.  I walked into my office and noticed that the end had suddenly turned yellow; closer examination revealed that it had begun to split open.  Time to harvest those seeds!

Dried LC Schilleriana Orchid, Cut From Ripe Seed Pod
©2012 MadOrchidist.com

Above is a photo of what's left of the original flower I pollinated - I'd say it's a bit less attractive now than it was nearly 6 months ago.  I snipped the pod from the plant and cut away the stem and dried flower remnant.

Preparing the Pod for Seed Harvesting
©2012 MadOrchidist.com

Since the capsule had only just begun to dehisce (dry and open), I helped it along a bit with a scalpel.  Here is what it looks like split open:

Splitting an Orchid Seed Pod
©2012 MadOrchidist.com

Since I didn't do anything new with cleaning and flasking the seeds, and all my glove-box photos look pretty much alike, I'll leave that part out of this post.  Instead, here are some more photos of the seeds to give a sense of scale to both their size (very small) and number (very large).

 Here is a low-power micrograph of the seeds still in a section of the capsule:

Orchid Seed Micrograph
©2012 MadOrchidist.com

 You can see some individually along the edge, but in the middle it is just a dense fluff of innumerable seeds.  I tapped and scraped out a bunch onto a clean sheet of white printer paper.  There are more seeds there than I could ever possibly need or want!  I sowed a portion of them into 4 mother flasks tonight and put a bunch more into cold storage.  That way, if these flasks become contaminated, I can try it again.

Thousands and Thousands . . . and Thousands of seeds
©2012 MadOrchidist.com

Below is my favorite picture of the day.  That is the razor sharp edge of a surgical scalpel, and each of those white flecks is a tiny seed!

Tiny Tiny Seeds on a Scalpel
©2012 MadOrchidist.com

It's one thing to know that Orchids have very small seeds; it's another entirely to look through the 'scope and really "feel" how small they are - or exhale while opening the capsule and see a cloud of them rise into the air . . . both experiences I feel richer for having had.

With my limited magnification power and light sources, it's hard to determine if there are viable embryos in there.  I didn't see anything that positively confirmed viability, but that doesn't necessarily rule it out either.  Time will tell . . .

Little Victories

Progress!  The tiny green dots in the blurry photo below are actually what this blog madness is all about:

Epidendrum Orchid Protocorms In Vitro
©2012 MadOrchidist.com

These Epidendrum seeds have germinated; at this stage, the little green balls of undifferentiated and dividing cells are called protocorms.  Most importantly, there are no obvious signs of contamination! All three of the flasks I made up with these seeds (dry and then sterilized) are looking good; here is another one:

Succesful Orchid Germination
©2012 MadOrchidist.com

In this case, the orchids themselves (reed-stem Epidendrums with no ID) aren't that exciting, but that fact that I was able to sterilize, flask, and germinate them successfully at home is.  In fact, of the more than a dozen mother flasks I have sown with various seeds so far, none are showing any contamination (yet) - a good sign that my home-grown laboratory and procedures are viable.

Further progress toward my eventual goal of actually breeding my own new orchids is also being made.  Today I noticed that the seed capsule of the first cross I made back in July 2011 has matured.  It is turning yellow and has just begun to split at the end where the flower was.  This eliminates any chance of sowing these with the "green pod" method, since I must presume the seeds are contaminated now that they have been exposed to the outside world.  I will definitely get on to flasking them up soon.

LaleoCattleya Seed Capsule Maturing
©2012 MadOrchidist.com

These will hopefully be the fist new orchids that are completely my own, from pollination to germination.  This was just the shot of encouragement I needed to stay excited about this orchid odyssey . . . 

New Year's Flasking

Happy New Year!  2011 was the beginning of a new chapter in my orchid hobby, and I am looking forward to continuing the madness in 2012.  I started off right, with some ambitious orchid seed flasking.

A while back I ordered dry seeds of a few orchid species from Thailand on eBay.  They arrived in tidy little packets, and I popped them in the refrigerator for safe keeping until I could get around to them.  I actually only ordered the first two in the picture below; the seller threw in a couple of others.  What a nice surprise!

Gastrochilus bellinus, Ascocentrum ampullaceum,Dendrobium pachyphyllum, and Gastrochilus patinatus
©2012 MadOrchidist.com

Here's an open packet showing the bright yellow seeds of Dendrobium pachyphyllum:

Dendrobium pachyphyllum Seeds
©2012 MadOrchidist.com

And here are is a closeup of some of the Ascocentrum seeds using my iPhone and an antique dissecting microscope - what a cool combo!  If you look closely, you can see the fibers of the white printer paper to give you a sense of how small they are:

Ascocentrum ampullaceum seeds magnified under my stereo dissecting scope
©2012 MadOrchidist.com

I got a little ambitious and decided that I would try to sow all four species in one flasking session in the glove box.

Orderly Preparation - don't want to mix them up!
©2012 MadOrchidist.com

After numbering and labeling syringes to correspond to the right species and flasks, I got to sterilizing the seeds. 

Orchid Seed Solution
©2012 MadOrchidist.com

Since my first attempt hasn't yet shown any contamination, I used the same procedure for these seeds.  The biggest difference is that I did four syringes instead of one, and I only used the "filter" cap I made on one of them.  The others I just let settle a bit longer after every rinse to avoid squirting out too many seeds.

Once more into the box
©2012 MadOrchidist.com

Four at once turned out to be a bit more than was comfortable.  It required me to have a lot more stuff in the glove box (multiply empty flasks and sterile water jars by four), and the possibility for cross-contamination or a mix-up seemed high.

Crowded and Messy in the glove box
©2012 MadOrchidist.com

Finally, mother flasks of the four species are sealed and out of the box.  This took a while, and it made me realize how non-ergonomic my glove box is.  I have to hunch over it to get the best view through the acrylic window on top while working, and it gets to my neck eventually.

New Orchid Flasks
©2012 MadOrchidist.com

The whitish particles below are a thin layer of seeds and water in a finished flask.

Orchid seeds in vitro
©2012 MadOrchidist.com

Here are some representative photos of these species from the Internet Orchid Species Photo Encyclopedia (click on the name below a photo for more info).

Gastrochilus patinatus

Ascocentrum ampullaceum

Dendrobium pachyphyllum

Gastrochilus bellinus

Flasking My First Orchid Seeds

This is what it's all about; sowing some Orchid seeds in flasks!  A couple of my own crosses capsules' are nearing maturity, but I couldn't pass up the opportunity to practice on some that just came along.

Epidendrum seed capsules ready for flasking
©2011 MadOrchidist.com

The other night was the Orchid Society Holiday Party.  Our host had some Reed-stem Epidendrums out in the yard that he says set seed all the time.  As it happened there was a dry pod that had split open and a green one still sealed, both on the same plant.  I have been considering whether I should go with the "green pod" method or wait for my pods to spit naturally and then sterilize the seed, and here was an opportunity to try both and compare!  He let me take them both to sacrifice to the learning process in my lab (thanks Van!).

Pod Parent Tag
©2011 MadOrchidist.com

Given that the Epidendrum was probably pollinated by an insect, I have no way of knowing what the pollen parent was.  All I know for sure is that the pod parent is pink; its tag is pictured above, plus I saw a few of the flowers on another spike.  Since this is sort of an experiment, I am not really worried about the pedigree.  If I end up growing anything from these they will be good for landscaping; Reed-stem Epis are practically bulletproof outdoors in this climate.  Let's hope the seeds are hardy too:

Epidendrum Orchid seed capsule dried and split open (dehisced), fine seeds visible
©2011 MadOrchidist.com

I decided to deal with the open capsule first.  Once an orchid seed pod dries, it splits open to release the thousands upon thousands (sometimes even millions) of tiny seeds inside.  After this happens, the seeds must be considered contaminated and their surfaces sterilized before sowing.

Epidendrum Orchid Seeds being sterilized in a syringe
©2011 MadOrchidist.com

I opted to use household bleach in a ratio of one part bleach to nine parts distilled water as my sterilizing solution.  Several online tutorials and discussions recommended using a syringe to clean the seeds.  I bought dispensing syringes with caps on Amazon.com and made one cap into a filter by melting a couple of holes in it with a heated pin and then placing a small piece of coffee filter inside.

Basically, I drew up about 10mL of the sterilizing solution through the filter cap and then banged the syringe on my hand and the table for at least ten minutes to try and eliminate all of the tiny air pockets that cause the seeds to float and prevent the solution from doing its job.  Below is a close up photo of the tiny orchid seeds in the syringe of sterilizing bleach solution:

Tiny Epidendrum Orchid seeds floating in sterile solution
©2011 MadOrchidist.com

 I put the syringe, several jars of autoclave sterilized distilled water (for rinsing seeds), a few pre-made flasks with seed sowing media, and a jar of stronger bleach solution (to clean the outside of the syringe and any tools I used) into my Glove Box and sealed it up.

Tools and materials in the glove box
©2011 MadOrchidist.com

It is unfortunately impossible for me to have my hands in the gloves working and take pictures at the same time.  At some point a small tripod and/or an intrepid volunteer will enable me to post more photos of what goes on inside the sterile glove box.

Briefly, I squirted out the bleach solution, drew up a rinse of distilled water a couple of times, and then squirted a small remaining amount of water with the seeds into a few flasks.  The result is pictured below:

Tiny Orchid Seeds in Flask
©2011 MadOrchidist.com

I got some seeds on the sides of the flask, but there are plenty in there on the agar media too.  I also ended up with more water than I would have liked in the flasks, so we'll see if that hurts things any.

I'll try to get to the green pod tonight or tomorrow so that I can make a fair comparison of the results.  I also received some seeds today in the mail that I ordered from Thailand on eBay.  I think I will give today's flasks a few days to see if contamination develops, so I can change my methods if necessary before trying again.  For now, those seeds will go into my pollen storage unit in the refrigerator.

As for these flasks, now we wait . . . it's a theme with orchids . . .