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Affordable Plant Tissue
Culture:
Orchids |
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Carol M. Stiff, Kitchen Culture Kits, Inc., Lufkin, TX
www.kitchenculturekit.com
kck@turbonet.com
| Orchids are beautiful, mysterious
plants which can be easily cultured in your home "lab" using materials and
instructions described in my last article. Seeds shown here were germinated under
sterile conditions at my home (left photo). The photo in the middle is from
seeds I germinated about 3 years ago and are about ready to be planted in the
greenhouse. My plants are not old enough to produce a flower as shown on the right
but this is an example of what you might be able to produce. How do you do
this? From the last article, I described simple tissue culture procedures that can
be done by the hobbyist. Using those procedures, slightly modified for the small
seed of the orchid, you can become an "orchid grower".
Note that I'm getting pretty good on growing these in sterile conditions but once they
leave the "bottle", I'm in trouble. If you are new to orchid growing,
consult the experts and see how they do it. Do not be afraid to ask for help and
join a listserv if needed. There is a new one now specific to orchid tissue culture
- go to my website and click on "Listservs" ( www.kitchenculturekit.com ) or email me at kck@turbonet.com
Here are the steps involved: Get materials needed for culture: Knudson C medium or MS medium, table sugar,
PPM, agar, seeds or a seed pod plus items mentioned in my last paper on home tissue
culture.
- See this for a refresher: http://www.kitchenculturekit.com/sivbposter.htm
- Seeds are available, sometimes, for free for those who are rookies and are just
testing the system. Email me and I can probably find you some free non-valuable seed
to play with.
Make your media and set up your clean box.
Disinfect your seeds. There are several methods available including a
"new" one I was told about on the new listserv
- homemade filter unit
- coffee filter method
- sucrose and peroxide (new and easiest method!!)
- green pod method
WRAP your containers to prevent contamination and at room
temperature. Some species "like" a dark incubation while others will
tolerate light.
WAIT. Seeds can take several weeks to several months
to germinate. Watch for contamination and discard those cultures.
Subculture to fresh media. There is much information
on the web about the specifics of this so do some reseach while you are waiting. It
is good to ask the source of your seeds what they recommend.
ENJOY. Be prepared for the plants to take over your
house. When I started this a few years ago, the cultures took up a small space on my
plant shelf in the kitchen nook. They have since taken over my guest room for
growing, not to mention other areas of the house where I prepare media, store media,
process media, etc. (I'll discuss more about this in the future).
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| Knudson C media or MS media needed for orchid
seed culture can be purchased from these companies which do sell to
hobbyists. I am not sure if there is a homemade media for orchid seeds but will look
into it. |
www.phytotechlab.com
www.caissonlabs.com
www.austratec.com/au
www.kitchenculturekit.com |
| PPM, Plant Preservative Mixture, is not
typically used in orchid seed culture but I recommend for hobby tissue culture because it
helps to control contamination. |
www.ppm4plant-tc.com
www.phytotechlab.com
www.austratec.com/au
www.kitchenculturekit.com
(only as part of a kit; cannot sell as individual item) |
| Table sugar is sucrose |
Your kitchen |
| Agar |
health food stores gelatin
(use 1/2 tsp per 3 tablespoons liquid media; process in microwave; culture at cooler
temperatures so the "jello" does not melt.
All companies mentioned above. |
| Seeds or a seed pod |
Seeds are available, sometimes, for free for
those who are rookies and just testing the system. Email me and I can probably find
you some free non-valuable seed to play with Purchase
from Aaron Hicks: ahicks51@cox.net |
|
Knudson C Medium with
Sucrose:
_____ 1 packet Knudson C medium
_____ 1 ml PPM
_____ 5 teaspoons sugar IF MEDIA DOES NOT INCLUDE SUCROSE
Prepare media according to instructions in manual: Add ingredients
to 1 quart or 1 liter of filtered or distilled water. Mix well. Adjust pH to 5.5. Measure
out 3 tablespoons to a baby food jar. Add agar to each jar before processing. Sterilize
media as described in manual. |
1/2 MS Medium with Sucrose:
_____ 1 packet MS basal medium with vitamins (no sucrose)
_____ 1 ml PPM
_____ 4 tablespoons sugar
Prepare media according to instructions in manual: Add ingredients
to 2 quarts or 2 liters of filtered or distilled water. Mix well. Adjust pH to 5.5.
Measure out 3 tablespoons to a baby food jar. Add agar to each jar before processing.
Sterilize media as described in manual. |
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Disinfection (or Disinfestation) of Orchid Seeds
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1. Using a homemade filter
unit:
This setup includes the filter unit, 70% isopropyl alcohol,
10% bleach solution and sterile water plus sterile plate and forceps |
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Pour dry seeds into filter unit (go to http://www.kitchenculturekit.com/filter.htm
for instructions on making this).Dip unit up and
down into 70% alcohol until all seeds are wet.
Spray down inside of filter with 70% alcohol such that
interior is disinfected completely. |
 
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| Disinfect forceps in 70% alcohol. Use forceps
to lift filter unit out of alcohol and transfer to solution of 10% bleach + detergent. Dip unit up and down again to wet all the seeds with the bleach
solution. Continue this for 10 minutes. |
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| Using a sterile forceps, lift the filter unit
out of the bleach solution and allow to drain slightly Place on a sterile plate.
Using a sterile spatula, small spoon, or rounded butter
knife, transfer some seeds to a sterile jar of medium.
Spread the seed over the surface of the medium. Add some
sterile water with the sterile spoon to the medium surface to help spread the seeds (if
necessary). Cap and seal with tape. |
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2. Using a paper coffee filter:
This setup includes the paper coffee filter, a
funnel, 10% bleach solution, sterile water plus sterile plate or paper towel, and forceps
or spatula/knife |
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| Seed are placed in a test tube or small
jar. A solution of 10% bleach + few drops detergent are added and the seeds and
solution are mixed for 10 minutes. The mixture is
poured into a funnel lined with a paper coffee filter which has been first wetted down
with more bleach solution.
Sterile water is poured over the seeds several times to rid
them of bleach. |
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Using a sterile spatula, small spoon, or
rounded butter knife, transfer some seeds to a sterile jar of medium. Liquid medium
can also be used if you have a shaker unit. The vessel shown here is a Combiness
unit which works great for the in vitro life of the plants.
Spread the seed over the surface of the medium. Add some
sterile water with the sterile spoon to the medium surface to help spread the seeds (if
necessary). Cap and seal with tape. |
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| Happy
culturing.............................carol |
