Journal of Experimental Botany, Vol 49, 817-827, Copyright © 1998 by Oxford University Press
J Newell, R Leigh and J Hall
Oat leaf mesophyll protoplasts were evacuolated and shown to develop acidic
vacuoles when cultured for 3 d. Vacuole development was followed by cell
wall formation. Developing vacuoles, stained with acridine orange, took the
form of a tubular network when viewed by confocal laser scanning
microscopy. The tubules expanded and fused to form a series of
interconnected vacuoles. When thin sectioned material was examined by
transmission electron microscopy, the tubular network appeared as a number
of small, expanding vesicles. The vacuolar
H+-ATPase, H+-PPase and a
membrane integral protein of 23 kDa (VM23) were shown, by Western blotting,
to be removed from protoplasts following evacuolation. After 5 d culture
the H+-ATPase and H+-PPase,
but not VM23, were detectable in microsomal fractions.This study describes,
for the first time, successful vacuole regeneration in a monocotyledenous
plant. This regeneration follows a similar pattern to that seen in
non-cereal protoplasts.Keywords: Evacuolation, oat
leaf mesophyll, protoplasts, vacuole regeneration.
ARTICLES
Vacuole development in cultured evacuolated oat mesophyll protoplasts
School of Biological Sciences, Biomedical Sciences Building, University of Southampton, Southampton SO16 7PX, UK; The Biochemistry and Physiology Department, IACR-Rothamsted, Harpenden, Herts AL5 2JQ, UK; Corresponding author
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