Dong Liu dliu at JEEVES.UCSD.EDU
Thu Mar 2 12:50:02 EST 1995

Hi, Dear netters:

Since I asked the question about "How to induce anthocyanin synthesis on
Arabidopsis seedlings which are grown on agar plates", I have got a lot
responses from various people. Here are the massages I would like to share
with you guys. I also want to take this opportunity to thank all the
persons who sent me these valueble information.

Dong Liu

1) From:  bparks at magnus.acs.ohio-state.edu

you need to include sucrose on your plates to get good anthocyanin
production. as per light treatments, continuous white light under a
fluorescent tube works fine. e-mail back to me if you need more details,
but this should give plenty of anthocyanin. the sucrose is the key (2%).
good luck...

2) From:  fac_jmonroe at vax1.acs.jmu.edu

Dong:  I've tried without much luck.  High levels of sucrose and light are
probably required.  I wonder if the plastic prevents the inducing wavelength of
light through since I've seen it under high light intensity in pots but not
plates in the same chamber...

I am interested in hearing what you find out.  I would like to try to screen
for mutants in an undergraduate Plant Physiology course I teach.
If that is what you are looking for also I would be happy to send you any
mutants we would find since that is not my area of research...


Jonathan Monroe
Department of Biology
James madison University
Harrisonburg, VA 22807

3) From:  shirley at vt.edu (internet)

Methods are described in the following papers:

Kubasek, W.L., Shirley, B.W., Mckillop, A., Goodman, H.M., Briggs, W., and
Ausubel, F.M. (1992) Regulation of flavonoid biosynthetic genes in
germinating Arabidopsis seedlings. Plant Cell, 4, 1229-1236.

Feinbaum, R.L., and Ausubel, F.M. (1988) Transcriptional regulation of the
Arabidopsis thaliana chalcone synthase gene. Mol. Cell. Biol., 8,

Feinbaum, R.L., Storz, G., and Ausubel, F.M. (1991) High intensity and blue
light regulated expression of chimeric chalcone synthase genes in
transgenic Arabidopsis thaliana plants. Mol. Gen. Genet., 226, 449-456.

You can use either high intensity light or UV light, and it helps to have
sucrose in the medium if you want high levels of expression (in fact, at
day 3 after germination, the plants do it spontaneously on sucrose).  On
the other hand, it is easy to get induction, even in plants grown in soil,
using high intensity light, as described by Rhonda Feinbaum.

Good luck!   Brenda

* Brenda W. Shirley                                        *
* Assistant Professor           703-231-3013 (phone)       *
* Department of Biology         703-231-9307 (fax)         *
* Virginia Tech                 shirley at vt.edu (internet)  *
* Blacksburg, VA 24061-0406                                *

4) From:  jkreps at biochem.umass.edu

Unintentionally, I have induced anthocyanins (as evidenced by purple coloring)
in seedlings on agar plates in the following ways:  water stress the
seedlings, that is, when you put out the surface-sterilized seeds, leave some
water with the seed and wrap the plate with parafilm; include a small amount
of toxin in the agar (I was working with a alpha-methyl tryptophan resistant
mutant);  grow the seedlings under very high light (>300 uEi).  Basically, any
stress of the seedlings should induce anthocyanin production, so, depending on
what you are studying some stresses may be acceptable and others not.  Best of

Joel A. Kreps
Dept. of Biochem. and Mol. Bio.
U.Mass. at Amherst

5) From: 22676sjg at msu.edu

give them a higher light intensity than they are used to or give them light
plus cold.

Sarah Gilmour

6) From: karin at trna.chem.yale.edu

        You might try stressing the plants by growing them at a
slightly elevated temperature or at high light intensity.  I saw
a lot of anthocyanin produced when I inadvertantly grew my plants
at a temperature 2-3 degrees higher than normal.

Karin Johnson

7) From:  meinke at osuunx.ucc.okstate.edu

Have your tried lowering the temperature?  That certainly works with
plants grown in pots.


David W. Meinke
Department of Botany
Oklahoma State University
Phone: 405-744-6549
FAX:   405-744-7673

Original Email Address Has Been Changed.
Please Note Updated Email Address Below:

8)  From: Greenberg Jean <greenbej at beagle.Colorado.EDU

Dr. Ausubel has published several papers on anthocyanin accumulation in
Arabidopsis.  You should do a search for the references: Feinbaum and
Ausubel in MCB around 1988; Feinbaum, Storz et al in MGG (I'm not sure
when this came out, maybe 1991-1993) and Kubasek et al in the Plant Cell
around 1993.  They used high intensity light-sodium lamps I think.  jean

9) From: abbritt at ucdavis.edu

Anthocyanins aren't normally expressed to a significant degree in Raby
seedlings, but they can be induced by stress.  I've heard that they can be
induced by growing the seedlings in the cold.



Section of Plant Biology
Davis, CA 95616

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