The first leaves have appeared!

Well, my baby orchids have been busy. In general, the protocorms have gotten significantly larger. Some have even gotten their first leaves. In this first photo, you can see one nice leaf a little right and up from center. A few other protocorms in this picture show a little green spike on top, which will become their first leaf in a week or two.

Also interesting are the white protocorms in the lower right of the photo. These were clearly alive at one time, because they are much larger than the seeds I initially planted. However, this jar is the "mother flask", and I removed a number of the protocorms to "replate flasks". I must have dripped a drop of the calcium hypochlorite sanitizer into the mother flask, and it apparently killed these few protocorms. Notice that while they turned white, they did not rot or decompose in any way. That is because there is no fungus or bacteria in the flask, so there is nothing to consume the corpses.

Ordinarily, I might have transferred all the protocorms from the mother flask to the replate flasks, but I had too many protocorms, and I am a little unsure of my technique, since this is my first attempt at flasking. So, I decided to thin out the mother flask, and keep it going, just in case.


In the next photo you can see the contents of a replate flask. Ideally, I should have separated each protocorm, and spaced them a little bit apart. However, I was afraid of handling them too much, because of the risk of contamination, so I instead put little clumps of protocorms spaced apart. They'll just have to work it out for themselves. The big protocorm on the left is getting ready to put out its first leaf. The plants in the mother flask seem to be further along. I don't know if I replated too soon, or if the plants are not crazy about the replate medium, or if this is just normal, persnickety orchid behavior. But they are green and growing, so they cannot be too unhappy.


In this last picture, you can see the result of an interesting experiment. As I said above, I had more protocorms than I needed. So, what the heck... Do you suppose they might be willing to live in some nice sphagnum moss? I really didn't think they would survive. If you look back to my previous post, you can see how tiny they were - just little green dots smaller than the head of a pin.

But live they did. I dampened the sphagnum with deionized water, with some KLN rooting hormone, sprinkled on the protocorms, covered the pot loosely with plastic wrap, and crossed my fingers. Maybe twice a week, I've been spraying them with deionized water with a tiny bit of MSU fertilizer added, and sure enough, they are growing well.

In the center of the photo, you can see one nice leaf. I guess my fluorescent lights are a little too strong, because the leave has a tinge of purple. But that doesn't seem to be a problem. Also, in the upper right, you can clearly se the faintest beginnings of a leaf on that protocorm.

So as I had hoped, life wants to live.

The little orchids continue to grow. Today I decided to "replate" the protocorms. They originally started in a sealed jar containing a germinating medium. This medium has a fairly low level of nutrients, and the idea is to move the developing plants to something a little stronger when they are able to tolerate it.

Also, because the jar was sealed, eventually the air in the jar would become too unbalanced for the plants to survive. The new jars have a lid with an 1/8" hole. The hole is plugged with cotton, and the cotton is saturated with copper sulfate. This should be sufficiently germicidal to keep the interior of the jars sterile. We shall see.

One last reason to do the replate: as the protocorms grow, the jar becomes too crowded. Even though I thought I started with a reasonable amount of seed, I had way more seeds germinate than could continue to grow in a single jar.

So, I once again set up a fish tank on its side, sprayed calcium hypochlorite, and moved the protocorms to the new media. Even after spreading the protocorms over 5 jars, I had a bunch left over. So, here are a few photos of the protocorms. Sorry to say that they don't look like much at this stage.

However, they are photographed on a piece of tissue, so you can get an idea of how small they are by how magnified the tissue looks.

A few years ago, I started testing my orchids for viruses. One phal tested positive, so I decided to try setting seed, because I've read that viruses are not transmitted via mature seeds.

So now I have orchid seeds, and a bunch of chemicals to germinate them. And so, here are a few thousand of my new orchids.

This first photo shows some of the smallest orchid plants you will ever see. The round green ones are embryos that have just germinated. The white ones are seeds that did not, for one reason or another, survive.


These seeds are growing on a sterile agar medium. I mixed up the medium, and put it in a pressure cooker for 1/2 hour to sterilize. The seed surfaces also have to be sterilized, to remove fungus and bacteria on the exterior of the seeds, but without killing the seeds themselves.

This is done by soaking the seeds in calcium hypochlorite. The seeds then must be rinsed in sterile water. This is tricky, because they are much smaller and lighter than grains of sand.

The seeds are then added to the sterile agar, without re-contaminating anything. I did this in a fish-tank turned on its side, liberally sprayed with more of the calcium hypochlorite. My work bench was covered with industrial polyethylene - also liberally sprayed. And yes, the basement reeked of chlorine for a little while.

This next photo shows the mason jars containing the seeds and agar. You obviously cannot see the seeds at this scale. I did a total of four jars. Two grew something altogether unlike orchids. Maintaining perfect sterility is hard...


And lastly, this photo shows a few of the trendy new chemicals I bought to make up the agar. I've spared you pictures of the beakers, flasks, hypodermics, etc. used in the process.


After the embryos spend a month or two in these jars, they will become "protocorms". These must then be thinned out, and transferred to fresh sterile "replate medium".

Several months after that, they should have actual roots and leaves, and should be strong enough to be grown in regular pots, in the open. At least, that is the plan...

Practical microbiology. Fun, fun, fun.