Question
QUESTION: I pollinated denrobium orchid on 15th september and i have a seed pod now. I wanted to sow this by myself and wants to see the tiny plants. No matter about the time. I am not much convenient about the lab techniques. Any idea can i even see few plants. please help.

ANSWER: Hemali, before sterile techniques came into use people would sow the seeds on top of the orchid mix in which the mother plant was growing.  It was learned that a tiny fungus specific to that type of orchid must penetrate the seed and converts cellulose to sugars the seed needs to germinate.  Unfortunately, the germination was very poor and very few, if any, seedlings developed.  Many formulas were developed for growing the seeds in bottles under sterile conditions.  There are now standard media in powder form (much like baby formula) that need to be mixed with a specific amout of sterile water in small bottles and the bottles with the mix sterilized in a pressure container, cooled and stoppered before then sterilizing the seed and sowing it on the culture medium in the bottles.  The main expense is the culture medium which is quite modest.

So, your choices are to try the sterile technique or sow the mature seeds on the surface of the existing mix in which you are growing the plant.  It sounds as if you favor the latter.  If unsuccessful, you will need to try the sterile tecnnique.  If the latter, I can provide you with more detail for supplies, etc..

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QUESTION: Can you please describe me the sterile tecnnique. But I don't know anything about chemistry.
thanks
hemali

Answer
If you wish to attempt this by yourself, you will need to get a number of supplies.  You will need glass bottles and rubber stoppers that fit them, a pressure cooker (takes about 15 minutes at 15 lbs of pressure to sterilize).

The first step is to acquire the orchid culture media.  Usually this comes supplied with instructions for adding water and other materials.  Google the "Orchid Mall" and go to "Service, Supplies, and Software" for a location to acquire the growth media.  Be sure to get a seed germination medium suitable for dendrobiums.

Once you have prepared the germination medium, add it to you bottles and stoppered them, insert the bottles into your sterilizer and sterilie at 15 pounds of pressure for 15 minutes.  Then let cool for an hour or more and remove the bottles.  If stoppers have come off from any, place the stoppers back onto the bottles while they are still in the sterilizer.  The bottles so prepared should  be set aside until the culture medium has solidified into a gel.

Next, you sterilize the seed in a solution of 1 part bleach in 10 - 15 parts of water (add a few drops of mild dish detergent to the bleach solution and shake it up gently to expose all of the seeds to the bleach solution.  The soap will help to reduce the surface tension on the seeds, increasing exposure to the bleach.

I use a low tech way of seed transfer to the bottles of culture media.  I turn on an oven and open the door so hot air emerges from the open and creates a semisterile condition near the open door.  I then take a bottle of sterile culture media, in one hand and suck some seeds/bleach solution with a syringe or eye dropper and slightly unstopper the bottle and squirt the seed solution into the bottle and quickly replace the stopper on the bottle.  This is all done as near to the opening of the oven as you can tolerate.  You only need enough culture media in each bottle to cover the bottom an inch or two.
Place the incubated bottles in a fairly low light, but warm, environment.  It is possible that some of the bottles will have been contaminated by mold spores and any bottles with mold growth need to be discarded.  Successfully incubated bottles will have green grasslike growth on the culture media in about a month.

Normally, this is a two step process.  The step above is called the "mother flask" as it is where the seeds germinate.  If the seeds germinate, then, the next step is called "replate" in which seeds are transferred to bottles of a special growth medium.  This step reuires a special sterile transfer box and provides both increased seedling growth, but also increased risk of contamination.  To avoid this risk, when I prepare the mother flask media, I add small pieces of apple or banana before sterilizing.  This provides some growth inducing properties without using a transfer box and the risk of contamination.  The latter, howver, is not as effective as preparing or using a replate medium.