floral dip surfactants--Responses

[Steve Clough]

<x-rich>Hello,


I posted a question asking the Arabidopsis community for alternative
surfactants that one could use in the floral dip transfromation
protocol. I asked not because I was looking for something better, but
because several researchers have asked for other options that might be
easier to find or cheaper.  I received replies stating that Tween 20
and  Extravon have both been used as effective substitutes for Silwet
L-77.


Many thanks to those that responded. Parts of their letters are copied
below.


Sincerely,

Steve Clough


Summary:

--------------

In my laboratory, we have been using Tween20 (the final concentration
is

0.02%), and it works very well as same as Silwet L-77.

Good luck!


Naoto Kawakami

kawakami at isc.meiji.ac.jp


--------------

When I heard the first rumors about your "floral dip" transformation

a couple of years ago, I could not find "Silwet" in Switzerland.......

I tested a commercial wetting agent used as an additive in crop

protection, "Extravon" (formerly from Ciba-Geigy, then Novartis Crop

Protection, I don't know who sells it now).


After some fine tuning, I used Extravon for "floral dip" traformation
of

Columbia at 0.05% in the infiltration medium (10 mM MgCl2, 5%

sucrose), basically following your procedure. I established a T-DNA

activation-tagged collection of about 20.000 independent Arabidopsis

Columbia transformants, so I can say it definitely worked. I never

did a side-by side comparison with Silwet, but was happy with my

transformation frequency (0.3 %).


According to the product information, "Extravon" is a 25 % solution

of a isooctylphenyl-polyethylenglycol-ether, a non-ionic detergent.

Chemically, it should therefore be quite similar to Triton X-100 -

did anyone try that?


I would imagine that other wetting agents for crop protection would

also work, unless they are toxic for Agro.


Regards, Jan Lucht

luchtjan at uni-freiburg.de


-------------

The history of floral dip and vacuum infiltration seems to be tied into
the old practices of vacuum infiltrating viruses into plants and the use of
surfactants to enhance the penetration of various xenobiotics.  In 
that regard, a
targeted query to individuals in pesticide companies who test various
surfactants in conjunction with herbicides and insecticides may give 
some insights.

Several years ago, I had the same question.
I finally followed up on my curiosity, searched through the web and 
found several investigations on the efficacy of surfactants.

I have not tested these compunds in the floral dip.  The listed compunds are
just ones I found and are not necessarily the best candidates.

Silwet L-77
silicone polyalkyleneoxide copolymer
OSi Company,
Tarrytown, NY 10591


Silwet 408
silicone polyalkyleneoxide copolymer
OSi Company,
Tarrytown, NY 10591


Sylgard 309
silicone adjuvant mixture of
2-(3-hydroxypropyl)-heptamethyltrisiloxane,
ethyloxylated, acetate EO glycol, -allyl, acetate
Dow Corning
Corporation, Midland,
MI 48684


Kinetic blend of polyalkyleneoxide modified polydimethysiloxane and
nonionic surfactant
Helena Chemical
Company, Memphis,
TN 38137


Dyne-Amic
blend of polyalkyleneoxide modified polydimethysiloxane, nonionic
surfactant and methylated vegetable oil
Helena Chemical
Company, Memphis,
TN 38137


X-77
a mixture of alkylarylpolyoxyethylene glycols, free fatty acids, and
isopropanol
Loveland Industries,
Greeley, CO 80632


MSO
methylated soybean oil concentrate
Terra Riverside,
Sioux City, IA 51101


Rigo Oil Concentrate (ROC)
83% paraffin base petroleum oil and 17% nonionic surfactant/emulsifier
Wilbur-Ellis
Company, San
Francisco, CA 94104


Agri-Dex
83% paraffinic mineral oil and 17% polyoxyethylene sorbitan fatty acid
esters
Helena Chemical
Company, Memphis,
TN 38137


Jim Tokuhisa, Ph.D.

tokuhisa at ice.mpg.de


_________________



**************************************************

Steven J. Clough

Dept Crop Sciences, U of Illinois

1201 W. Gregory Dr., Lab 356 ERML

Urbana, IL 61801


Phone: 	217-244-3929 (Functional Genomics' Lab)

          or	217-244-6150 (Dr. Lila Vodkin's lab)

FAX:   	217-333-4582

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