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Preserving Arisaema Pollen

Hi fellow Aroiders and Arisaema nuts,
I just had a Kaichen1999 A-05 rebloom for me. This time it had 2 shoots and
one bore a male inflorescense. A-05 has been variously described as
Arisaema odoratum (I can't smell anything) or saxatile. It looks more like
saxatile to me. After nearly 2 weeks the anthers decided to shed their
pollen and as it collected at the bottom of the spadix tube and the bloom
started to fade I cut it off yesterday.

Having no female Arisaema flowers available yet I decided to try
cryopreservation of the pollen. I cut off the long pendant spadix and all
but the spathe base and tapped out all loose pollen into 2 sterile
polypropylene 1.5 ml conical microcentrifuge tubes, about a grape pit sized
yield each. I did not weigh the yield. To one tube was added 0.5 ml sterile
30% sucrose solution in distilled water and to the second 0.5 ml sterile
50% glycerol. Tubes were capped. Some vortexing (rapid circular shaking)
and quick spin down in a lab benchtop microcentrifuge of the sucrose tube
was necessary to get the pollen off the inside walls but the glycerol
solution suspended the pollen nearly homogeneously after a few minutes.
The tubes were refridgerated (4C) until now, approx. 24 hours. The
remaining spadix was folded in a small piece of black construction paper
and placed in a 50ml plastic conical centifuge tube with cap which had a
small amount of Drierite (anhydrous calcium sulfate), a dessicating agent,
at the bottom. This tube will be kept refridgerated and hopefully more
pollen will shed and may be used in a month or so when my other species

Cryoprotectant tube pollen was observed to have settled to the bottom of
the tubes slightly today and was resuspended by gentle mixing. A few large
anther remnants were still observed. Both tubes were simultaneously dipped
to their 0.5 ml liquid level in -70C methylbutane for 30 seconds (snap
frozen) and immediately stored in a -70C freezer. The sucrose solution
froze solid but the glycerol solution remained liquid.

The sucrose method protects animal tissue for histology analyses from
freezing damage, and bacterial liquid cultures are frozen with the glycerol
method (though at a lower concentration of glycerol, and frozen
immediately). This is not as long term a solution as liquid nitrogen
cryopreservation, but is used to store biochemistry and microbiology
specimens for over a year. Tubes will be quick thawed in room temperature
water and pollen solution applied as is to receptive stigmas on the
spadixes (spadices?, spadae?; or in Dubbya's terminology, spadixices? (not
very subliminibable!)) of other species when they are ready.

I hope this works.

Bonaventure W. Magrys
Research Assistant,
Neuroscience Institute
Seton Hall University
Department of Graduate Medical Education
South Orange, NJ

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