Plant Tissue Culture
Did you ever have a plant that was so unique or so beautiful that you
wished
you had
hundreds or thousands of them to enjoy or to sell? Plant
tissue culture (micropropagation) is a technique which will do just that
for
us. We
are going to discuss this tool which is used so extensively in the
nursery
business
and in plant biotechnology. It is a fascinating and useful tool which
allows the
rapid production of many genetically identical plants using relatively
small
amounts of
space, supplies and time.
Basically the technique consists of taking a piece of a plant (such as a
stem tip,
node, meristem, embryo, or even a seed) and placing it in a sterile,
(usually gel-based)
nutrient medium where it multiplies. The formulation of the growth
medium is
changed depending upon whether you are trying to get the plant to
produce
undifferentiated
callus tissue, multiply the number of plantlets, grow roots, or multiply
embryos
for "artificial seed".
For many who become superficially aware of the technique it seems
shrouded
in mystery
and is shrugged off as too technical to be of concern. Actually, it is
no
more of
a mystery than taking a cutting of your favorite house plant and growing
it
to share
with a friend. As for being technical, you can begin plant tissue
culture
with as little
as a cookbook approach and a feeling for sterile technique.
Some people have visions of scientists doing plant tissue cultures in
white
gowns
and masks in hospital-clean environments. Such conditions are excessive.
While it
is true that mold spores, bacteria, and other contaminants will grow and
overrun
a culture, air that is not moving has a minimum of contaminants. In
addition, disinfection of
implements, work surface and nearby areas helps eliminate
contaminants.
The guidelines for preparation and the laboratory protocol
provided here are given as a place to begin. Included
with is a limited discussion of some of the many options you have as
you
explore micropropagation. We can discuss these in more depth if you have
questions,
concerns or related experiences to share. I would be particularly
interested
in success and
challenges you may have had or are currently having in your
classroom.
Some suggestions are given for the following
(a) Selecting plant sources. Some species, or even clones are easier to
grow
in culture
than others. Some respond reluctantly to culture, some do not respond at
all, and
many plants have never been tried.
(b) Choosing a growth medium (price, convenience, type of plant and
purpose
of the
micropropagation all enter into this decision.) How important are the
kinds
of hormones
used? On limited scale, media ingredients are available at the grocery
and
health
food stores.
(c) Suggestions for media preparation and sterilization. There are
alternatives to
sterilization in a pressure cooker or an autoclave.
(d) Methods for cleaning, storing and manipulating explants (plant
pieces to
be cultured).
Given certain basics there are many options for procedure, equipment and
supplies
for plant tissue culture. Some of your decisions will be based upon the
amount of
time, money and space you have. Other decisions will be based upon why
you
are doing
plant tissue culture and what you expect as a result (more plants?) .
Catalogs, such as Sigma,
Carolina Biological, or Edmund Scientific are good reference and they are for
purchasing
needed materials..
I look forward to sharing tissue culture experiences with you.
References:
Debergh, P.C. and R.H. Zimmerman, eds. 1991. Micropropagation,
Technology and Application.
Kluwer Academic Publishers. $61.50.
Lab design, info on labs worldwide, in depth discussions of
problems. Not for the beginner.
Donnelly, D.J., and W.E.Vidaver, 1988. Glossary of Plant Tissue
Culture, Portland,
OR. Timber Press, $22.95.
Good definitions of tissue culture terms.
Kyte, Lydiane and J. Kleyn, 1996. Plants from Test Tubes: An
Introduction to Micropropagation, 3rd ed., Timber Press, 1996
$29.95.
Good basics for the beginning amateur or grower.
Smith, Roberta H., 1992. Plant Tissue Culture-Techniques and
Experiments. Academic Press. $35.00.
Good introduction and broad base for college course.
Trigiano, Robert N, and Dennis J. Gray, eds.1996,Plant Tissue
Culture Concepts and Laboratory Exercises. CRC Press. $65.00.
For the advanced student.
Sources of supplies:
Carolina Biological
Edmond Scientific
PhytoTechnology Laboratories
This company specializes in plant tissue culture supplies. Downloadable documents (choose MS Word or PDF format) on Media Preparation, Setting Up a Tissue Culture Lab, Basic Laboratory Procedures, and more are available in the "Technical" section of the web site.
Sigma, 1996. "Plant Tissue Culture Catalog".
In addition to media ingredients,
premixes, equipment and supplies, this catalog contains a media comparison chart, procedures for media preparations.references and other valuable data.
preparation | laboratory protocol
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