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THE PLANT CELL, Vol 9, Issue 9 1527-1545, Copyright © 1997 by American Society of Plant Biologists
A Novel Cell Ablation Strategy Blocks Tobacco Anther Dehiscence
T. P. Beals and R. B. Goldberg
Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, California 90095-1606
We utilized a new cell ablation strategy to ablate specific anther cell
types involved in the dehiscence process. The tobacco TA56 gene promoter is
active within the circular cell cluster, stomium, and connective regions of
the anther at different developmental stages. We introduced a cytotoxic
TA56/barnase gene into tobacco plants together with three different
anticytotoxic barstar genes. The anticytotoxic barstar genes were used to
protect subsets of anther cell types from the cytotoxic effects of the
TA56/barnase gene. The chimeric barstar genes were fused with (1) the
tobacco TP12 gene promoter that is active at high levels in most anther
cell types; (2) the soybean lectin gene promoter that is active earlier in
the connective, and at lower levels in the circular cell cluster and
stomium, than is the TA56 promoter; and (3) the tobacco TA20 gene promoter
that is active at high levels in most anther cell types but has a different
developmental profile than does the TP12 promoter. Normal anther
development and dehiscence occurred in plants containing the TA56/barnase
and TP12/barstar genes, indicating that barstar protects diverse anther
cell types from the cytotoxic effects of barnase. Anthers containing the
TA56/barnase and lectin/barstar genes also developed normally but failed to
dehisce because of extensive ablation of the circular cell cluster,
stomium, and contiguous connective regions. Anthers containing the
TA56/barnase and TA20/barstar genes failed to dehisce as well. However,
only the stomium region was ablated in these anthers. The connective,
circular cell cluster, and adjacent wall regions were protected from
ablation by the formation of barnase/barstar complexes. We conclude that
anther dehiscence at flower opening depends on the presence of a functional
stomium region and that chimeric barnase and barstar genes containing
promoters that are active in several overlapping cell types can be used for
targeted cell ablation experiments.
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