Growing Orchids From Seed Printed: 9/25/2010

For the 1’st agar medium you need:

- hydrogen peroxide 3% to sterilize the seed

- 6 gr. agar-agar
- 1 ltr distilled water
- 2 gr. fertilizer 20-20-20
- 5 gr. sugar

Preparation
Boil the distilled water with the sugar and fertilizer. Stir the solution till the powder is
completely dissolved.
Then stir the agar powder in the solution. Let the media boil for ± 2 min. and keep stirring.
Now you can fill the flasks and sterilize them.
Concerning the sterilization you can find full explanation on the site of Lotte and Thomas.
It can take several months before you will see protocorns.

Don't put too much seed in one flask so that they can grow as long as possible before you
have to reflask.

Affordable Plant Tissue Culture:

Orchids
 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

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 Growing Orchids From Seed Printed: 9/25/2010

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. 

o See this for a refresher:  http://www.kitchenculturekit.com/sivbposter.htm

o 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.

o Specific media recommendations can be found at:  www.phytotechlab.com
o Type "orchid" in the search window, or contact garys@phytotechlab.com

Disinfect your seeds.  There are several methods available including a "new" one I
was told about on the new listserv

o homemade filter unit

o coffee filter method
o sucrose and peroxide (new and easiest method!!)
o green pod method

 WRAP your containers to prevent contamination and at room temperature.  Some

species "like" a dark incubation while others will tolerate light.

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 Growing Orchids From Seed Printed: 9/25/2010

 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). 

Knudson C media or MS media needed

for orchid seed culture can be purchased www.phytotechlab.com
from these companies which do sell to www.caissonlabs.com
hobbyists.  I am not sure if there is a www.austratec.com/au
homemade media for orchid seeds but will www.kitchenculturekit.com
look into it.
www.ppm4plant-tc.com
PPM, Plant Preservative Mixture, is not
www.phytotechlab.com
typically used in orchid seed culture but I
www.austratec.com/au
recommend for hobby tissue culture
www.kitchenculturekit.com (only as part of a kit;
because it helps to control contamination.
cannot sell as individual item)
Table sugar is sucrose Your kitchen
health food stores

gelatin (use 1/2 tsp per 3 tablespoons liquid

Agar media; process in microwave; culture at cooler
temperatures so the "jello" does not melt.

All companies mentioned above.

Seeds are available, sometimes, for free for
those who are rookies and just testing the
system.  Email me and I can probably find you
Seeds or a seed pod
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

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 Growing Orchids From Seed Printed: 9/25/2010

_____ 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.

Disinfection (or Disinfestation) of Orchid Seeds

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.

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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 Growing Orchids From Seed Printed: 9/25/2010

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.
   

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.

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.

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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 Growing Orchids From Seed Printed: 9/25/2010

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.

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.

3.   Using sucrose and peroxide:

This setup requires a 5% solution of sugar, small test

tube or small container, hydrogen peroxide (3%), and
a transfer pipette.

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 Growing Orchids From Seed Printed: 9/25/2010

Seeds  are placed in a small test tube or container and  5% sucrose solution (with a few
drops detergent)  is added.   Five percent sucrose is 5 grams of sugar ( a gently rounded
teaspoon)  in 100 ml water.  This is allowed to "incubate" for 12 hours.   The sucrose aids in
germination of fungal spores.

The sucrose is then removed with a transfer pipet.  Hydrogen peroxide (3 %) is added and
the seeds are allowed to disinfect in the hydrogen peroxide for 30 minutes.   This should kill
the bacteria and germinating fungal spores.

1.  Mix the solution with the pipet and add a few drops to
each media bottle. 

2.   Tilt the bottle back and forth to distribute the seeds over
the top of the agar medium.   Cap and seal.

This is fully described by Aaron Hicks at his site.  See: 

www.orchidseed.com

1.

 
3. Peroxide and seeds           4.  Seeds spread on surface

5.  Peroxide absorbed             6.  Seeds germinating

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 Growing Orchids From Seed Printed: 9/25/2010

2.

4.   Using a green seed pod:

This setup requires 70% isopropyl alcohol, 10%

bleach solution and sterile water plus sterile plate
and forceps

Green pods can be cleaned with a tooth brush

and then subjected to 70% isoproyl alcohol for a
few seconds followed by 10% bleach solution for
about 20 minutes.

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 Growing Orchids From Seed Printed: 9/25/2010

The pod is removed and placed on a sterile

surface.

It is then cut in half and the seeds are gently

removed with a forceps or knife blade and put on
media as described above.

Here are some of my orchids.  They are Oncidium sphaecelatum varieties that I have been culturing for a
friend and we are now looking for buyers.  Contact me if interested.  I've never sold plants before so I'm
new to all of this.   We are not sure of the varieties due to a mixup  (I gave them the names indicated below)
but have 3 distinct phenotypes developing.

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 Growing Orchids From Seed Printed: 9/25/2010

       
"OS 2" series                                 "lou thomas" series                                 "G" series

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 Growing Orchids From Seed Printed: 9/25/2010

"lou thomas" (my favorite)

   
"G" series (has a nice pseudo-bulb on most)

baby pictures

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 Growing Orchids From Seed Printed: 9/25/2010

For more information, see these sites:

Orchids Orchid Seed Sowing Manual (Barry Cohen)

InvitroLabs  (Peter W Spear) Specialising in Ericacious Plants and Orchids; Specializing in Australian Cool  
   Growing Orchids.. Phone , invitro@ihug.co.nz
   Tissue Quick Plant Laboratories containers, filter discs, and more   TQPLlab@awinthrop.freeserve.co.uk

   Akerne Orchids
   Blue Pagoda Orchids (includes sales to hobbyists)
   Alan Bickell's Orchids
   Orchids of La Joya
   Peter and Harriet Spear's InVitro Labs
   Rescue and Propagation of Orchids (A Non-Sterile Method)
   Vengers' Orchid Flasking
  G & B Orchids (Plants, seeds, chemicals, media, lab equipment)
Barry Gardner's "Growing Orchids in Australia" including "Simple Orchid Flasking", Advanced Orchid Flasking" and growing the
  Catasetinae family  with lots of photos.   Includes green pod maturation times
http://members.iinet.net.au/~barryg/
  The Orchid Seedbank Project  (Aaron Hicks)  -   www.orchidseed.com
  www.easyorchids.com
  www.growmorebiotech.com
  Troy Meyer's Conservatory - Orchid Flasking Service
  Orchid Web
  Lottie and Thomas Orchideen (Great Resources!!)
  ftp://ftp.nmt.edu/pub/orchids/flasking
   http://www.orchidsource.com/GBSeedManual.html
  http://www.ceiba.org/elpahuma/propagation.htm (there is a good manual to download)
  http://www.med-rz.uni-sb.de/med_fak/physiol2/disa/orchid.htm
  http://www.orchid-guide.com/propagation/Growing_from_seed.htm
  http://www.rbgkew.org.uk:80/ksheets/orchid-prop.html
  http://instruct1.cit.cornell.edu/courses/hort400/orchid/orchid1.html
  Colchicine and Orchids
Thomas Ederer:  info about parts of the flowers that can help you to pollinate Paphios.  "Seed germination"
  and then "How can I get some seeds?"   http://www.8ung.at/ederer_thomas/orchideen/
  Easy Orchids     www.easyorchids.com
  Seed Culture Recommendation:    www.phytotechlab.com - contact garys@phytotechlab.com
 

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 Growing Orchids From Seed Printed: 9/25/2010

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

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.
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.
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

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 Growing Orchids From Seed Printed: 9/25/2010

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.

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
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.

Contact Information
Carol M. Stiff, PhD
Kitchen Culture Kits, Inc.
905 Champions Drive
Lufkin, Texas 75901-7235
Resume (pdf)
936-699-3551  FAX 936-699-3553
kck@turbonet.com

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