Pollen Mutagenesis

Koji Goto gotok at TANSEI.CC.U-TOKYO.AC.JP
Fri Jul 15 10:00:48 EST 1994


Dear Arabido-netters;
        Following is a summary of answers to my query of pollen
mutagenesis.  These would help others.  I thank all who send me
infomations.

1)
Here is the irradiation conditions I have experience with.
Inflorescences were harvested and placed in closed plastic tubes to avoid
desiccation during the irradiation period.  Inflorescences were irradiated
at room temperature shortly after harvest with up to 150 krad gamma rays
from a 137 cs source (Gammacell-1000 Elite, Nordion International inc.,
Kanata, Ontario, Canada) which provided 0.923 krad/min.
Mature flowers (stage 13-14) were selected from inflorescences and used
immediately to pollinate multimarker lines.  Where necessary (plants not
carrying the ms1 mutation), multimarker lines were emasculated and
pollinated under a stereo microscope.
Pollen treated with between 30 and 60 krad resulted in siliques
containing 50% aborted seeds, while complete seed abortion was observed
only at doses above 100 krad.  The optimal dose is 60 krad (20% of Fl seed
was viable). More details are published in the paper ,Isolation of
Deficiencies in the Arabidopsis Genome by gamma-irradiation of Pollen
Genetics, 8, 1994 (in press).
Sincerely,
Igor Vizier
plxiv at uk.ac.nottingham.ccc.vax
molecular Biology Group
Department of Life Science
University of Nottingham
NG7 2RD
England

2)
Pollen Mutagenesis: To mutagenize Arabidopsis pollen, whole flowers from
axr2-1/axr2-1 (ecotype Columbia) were picked and placed in a small petri
dish.  The entire dish was placed in the irradiator (J.L. Shepherd.
Glendale, CA) containing 137Cs for the appropriate time period.  Flower
from +/+ (ecotype" Niederzenz) were emasculated and the mutagtenized pollen
was placed on the stigma surface.  In vitro germination tests indicated
that pollen was viable for at least 24 h when stored at 4 C @ irradiation
(Turner and Estelle, unpublished).

Mark Estelle 
Dept. Biology
Indians University
Bloomington, IN
=================================================
Koji GOTO
Molecular Genetics Research Laboratory (MGRL)
Science Bldg. #7, Univ. of TOKYO, Hongo, 113, Japan
e-mail:  gotok at tansei.cc.u-tokyo.ac.jp
FAX: +81-3-5684-0785,  Phone: +81-3-3814-4937




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