OT---Embryo Rescue Technique

Kenneth Hixson khixson@nu-world.com
Mon, 10 Aug 2009 16:09:15 PDT
Hi, Anita:

the plant breeding side
> of it.
> Where do you go to learn the lab techniques used in such cutting edge stuff 
> as embryo rescue?
> How much lab equipment does one person need to do this sort of stuff any way?

First, if all you want to do is learn plant breeding, you don't need to
use tissue culture/embryo rescue.  Mankind has been breeding plants for
thousands of years without it.  It wasn't until about 1900 that Mendel's
laws became generally known and people started using scientific
principles--before that, open pollinated seedlings were raised, and
crops and plants improved without even knowledge of genetics.  And yet,
crops were improved, roses, carnations, primroses etc were produced that
were unlike those that preceded them.  Embryo rescue/tissue culture has
become common in the last forty years or so, but even now, a great deal
is done with simple crossing and selection of seedlings.  A knowledge of
genetics and statistics are probably more helpful than how to use embryo

If you do wish to make "wide" crosses, embryo rescue is about as hard as
making jelly.  I've done it (yeah, both--embryo rescue-and made jelly).
If you cultured bacteria or fungi on petri dishes, you've already used
many of the techniques necessary--and you probably already know how to
do that.  An internet search on embryo rescue, or tissue culture, should
turn up leads. There are at least a couple groups that help beginners,
and they'll probably turn up in a web search.  Judith Freeman
(TheLilyGarden) used to teach embryo rescue to groups at lily
conventions, sometimes fifty people at a time--no scientists, just
gardeners, and provided a handout.  If I still have the one I was given,
  it may be possible to provide a copy.  If you wish to email me, I'll
provide some URLs also.  A couple to start:

Try U. of Texas--	http://aggie-horticulture.tamu.edu/tisscult/…

Equipment and supplies--
> http://www.hometissueculture.org/ 
	There is also a link to a listserv, which I haven't visited.

At one time Carolina Biological Supply offered supplies, including Agar
and Murashiga and Skoog mixes (mineral/hormone additives), but I don't
know if they still do so.  I used an agar mix, but newer things are on
the market, and are probably easier to use.

In a pinch, you can use unflavored Gelatin--which is mostly agar anyway.
Jelly jars work as well as test tubes, if larger, a little harder to
cap and sterilize, but that's what you are doing when you process jelly-
sterilizing the mixture in the jelly jars (before you put the embryos
in the jars please).  You probably should add some sugar to the agar as
an immediate energy source for the embryos.  Adding a mineral salt
mixture (fertilizer/Murashiga and Skoog mixture) is needed if the plants
are to remain in the container for a long period of time--a few months.
As a transfer hood, I used a ten-gallon aquarium on its side with a
cling-wrap plastic closure, but I've seen cardboard boxes, with the
opening closed with clear plastic.  To sterilize the interior of the
"transfer hood", I used rubbing alcohol, but 10% chlorine solution
works, it's just harder on skin, instruments, etc.  I used a spray
bottle after liberating the window cleaner solution from it.  A laundry
"spritzer" bottle works.

You do need patience, it helps if you have a place where the temperature
doesn't change much (because air pulling in and out of the containers
when the temperature warms and cools will introduce fungi/bacteria).
One lady I know does very well in the second (upstairs) bathroom.
Flourescent light setups help, and a knowledge of how to transfer the
plantlets from the culture containers to the potting mix--which may be
the hardest thing of all.

In other words, it isn't hard, "proper" equipment and supplies are nice,
but you can make do, and you don't need to know everything to be
successful, anymore than you need to be an auto mechanic to be able to
drive a car.  It does take practice.  And no, my first batch of jelly
was not a success either.


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