Plant Cell Lines in Cell Morphogenesis Research - Plant Cell Morphogenesis: Methods and Protocols - page 208

The advantageous cytological characteristics of these tobacco
cell lines are in contrast to the handicap arising from their “pedi-
gree.” The complicated genome of Nicotiana tabacum L. has not
been fully sequenced yet, and concerning these cell lines them-
selves, detailed gene or even karyological analysis is missing—with
one recent exception documenting pronounced genotype hetero-
geneity of BY-2 cell population, including massive aneuploidy,
chromosomal translocation, and variability in satellite DNA [ 16 ] .
As a logical consequence, there is a continuous effort to derive cell
lines or at least cell suspension cultures of acceptable quality from
the favorite plant experimental model Arabidopsis thaliana Heynh .
According to our knowledge and experience, none of the existing
Arabidopsis cell suspension cultures reach the quality of cytological
characteristics of the tobacco cell lines mentioned above.
Nevertheless, at least some of these cell cultures (e.g., A. thaliana
Heynh ., ecotype Landsberg erecta; ref. [ 17 ]) are applicable for the
experiments aimed for the studies on plant morphogenesis.
Taken together, in populations of in vitro - cultured plant cells,
various morphological parameters, including cell shape, cell size, as
well as orientation of both cell division and elongation, can be
observed directly on a single-cell level. The cell line models allow
also to study dynamics of changes in their intracellular structures,
in parallel with production of material for simultaneous biochemi-
cal analyses. Thus, plant suspension-cultured cell lines represent
advantageous experimental models for studies focused on various
mechanisms affecting cell morphology and the action of relevant
internal and external morphoregulatory factor(s). Their use is
complementary to the use of plant tissues, organs, and intact plants.
Here we demonstrate the procedures for long-term cultivation
of both cell suspensions of tobacco cell lines BY-2 [ 6 ] and VBI-0
[ 5 ] and of A. thaliana , ecotype Landsberg erecta (LE) cell line
[ 17 ], together with detailed protocols for their morphological
characterization and transformation. These are the techniques that
can be used not only in basic research but also in various biotech-
nological applications.

2 Materials

2.1 Plant Material

1. 7-day-old tobacco Nicotiana tabacum L., cv. Bright Yellow 2
(BY-2) cell suspension culture.
2. 14-day-old tobacco Nicotiana tabacum L., cv. Virginia Bright
Italia 0 (VBI-0) cell suspension culture.
3. 7-day-old Arabidopsis thaliana , ecotype Landsberg erecta
(LE) cell suspension culture.
4. 30-day-old BY-2, VBI-0, or LE calli.