OT---Embryo Rescue Technique

Adam Fikso adam14113@ameritech.net
Mon, 10 Aug 2009 18:03:15 PDT
Carolina Biological Supply st5ill offered the items noted as of three years 
ago. ----- Original Message ----- 
From: "Kenneth Hixson" <khixson@nu-world.com>
To: "Pacific Bulb Society" <pbs@lists.ibiblio.org>
Sent: Monday, August 10, 2009 6:09 PM
Subject: Re: [pbs] OT---Embryo Rescue Technique

> 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
> rescue.
> 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.
> Ken
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