Rice Transformations?

[Chris Ashcraft <AshcraftC@EDENBIO.COM>]

Hello Everyone -

I am experiencing a rather different problem with Rice transformations. I am
using a Kanamycin-based Agro protocol, and typically end up with massive
amounts of callus, and later shoots that have no chlorophyll.

I am transfecting scutellum derived callus, but suspect the source tissue is
able to bloom under selection, and interfere with transformed callus
development. I have recently switched to transfecting only compact callus
that typically remains attached to the scutellum, and am avoiding all
friable material from future work.

I will be switching to Hygromycin as soon as the construct is ready, but
other than that suggestion, are there any other insights from the group??

Christopher W. Ashcraft
Plant Transformation Specialist
EDEN Bioscience
11816 North Creek Parkway N.
Bothell, WA 98011-8205
Phone 425-806-7300
FAX 425-806-7400
Cell 206-465-1635


-----Original Message-----
From: Boosani Chandra shekhar [mailto:boosani2001@YAHOO.COM]
Sent: Thursday, February 08, 2001 12:43 AM
To: plant-tc@tc.umn.edu
Subject: Problem in selection, please suggest.

Dear all,
I am working on trasformation of indica rice using
Agrobacterium.
I am transfering the cocultivated callus to selection
medium containing Cefotaxim at 500mg/L and Hygromycin
at 50mg/L along with 3mg/L concentration of 2,4-D.
All the time i could find the callus turning into pale
brown and are not proliferation even after 6 weeks on
selection.
please suggest with some clues that may help me in
proliferation the transformed callus on selection
medium.

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