1021-4437/02/4903- $27.00 © 2002
Russian Journal of Plant Physiology, Vol. 49, No. 3, 2002, pp. 406–412. Translated from Fiziologiya Rastenii, Vol. 49, No. 3, 2002, pp. 451–458.
Original Russian Text Copyright © 2002 by Stepanova, Polyakova, Dolgikh, Vartapetian.
Oxygen deﬁciency in the environment, i.e., hypoxia
and anoxia, considerably affects higher-plant metabo-
lism, because these plants, being obligate aerobes,
require a constant supply of exogenous molecular oxy-
gen. The plants on overmoistened and ﬂooded soils
generally experience oxygen deﬁciency because of the
low solubility and diffusion rate of oxygen in water.
Such soils occupy extensive territories in many coun-
tries of the world . Anaerobic stress frequently
results in the injury and even large-scale destruction of
agricultural crops and wild plants and, therefore, in
substantial economical and ecological damage. Never-
theless, many higher-plant species permanently inhabit
overmoistened and even ﬂooded soils. Therefore, the
elucidation of the mechanism of damage to some plant
species and the strategy of adaptation of other ones to
these extreme conditions, as well as the use of both
classic and modern approaches to produce anaerobio-
sis-resistant plants, are the most urgent problems in
both plant physiology and biotechnology.
Physiological and biochemical aspects of the effects
of hypoxia and anoxia in higher plants, i.e., the mecha-
nisms of damage and adaptation of plants under condi-
tions of anaerobic stress, have been actively discussed
within the past 10–15 years in many reviews and mono-
graphs [2–8]. There are at least two main strategies of
higher-plant adaptation to the conditions of anaerobic
tolerance, i.e., the adaptation at the
level of whole organism, when the anatomical and mor-
phological characteristics of an organism promote the
transfer of molecular oxygen from the aerated organs
into the organs localized in an anaerobic medium; and
tolerance, i.e., metabolic adaptation realized at
the molecular level and in the absence of oxygen in the
medium [2, 5, 6, 9]. This concept, put forward about
20 years ago,  is now universally accepted [4, 6, 8, 10].
There are many plant species tolerant to anaerobic
stress. Therefore, it is basically possible and promising
to look for the means of creating more resistant forms,
in which the mechanisms of either true or apparent tol-
erance (avoidance of anaerobiosis) are realized. Within
the past 20 years, much interest has been awakened in
plant anaerobiosis, and this problem is being actively
studied from various viewpoints. The experimental
investigations are mainly performed using either whole
plants or detached plant organs. Biotechnological tech-
niques, in particular cells cultured
, were rarely
used for this purpose up to now [11–13]. In this connec-
tion, it is interesting to note a recent attempt to activate
The Response of Sugarcane (
Cells to Anoxia and the Selection of a Tolerant Cell Line
A. Yu. Stepanova, L. I. Polyakova, Yu. I. Dolgikh, and B. B. Vartapetian
Timiryazev Institute of Plant Physiology, Russian Academy of Sciences, Botanicheskaya ul. 35, Moscow, 127276 Russia;
fax: 7 (095) 977-8018; e-mail: email@example.com
Received March 15, 2001
—The effect of anoxia on the sugarcane (
L.) cultured cells was studied in
order to elaborate a technique for
selection of cell lines, which would be tolerant to anaerobic stress.
Inhibitory and lethal doses of anaerobic incubation were established from the state of the mitochondrial ultra-
structure during the anaerobic incubation of cells either with or without exogenous glucose, as well as from the
pattern of the post-anaerobic callus growth. An intact state of the mitochondrial ultrastructure and the viability
of some cells in the presence of 3% glucose were shown to be maintained for at least 14 days of anaerobic incu-
bation, while the index of post-anaerobic growth decreased by almost 50% even after 72-hour-long anaerobio-
sis. In the absence of exogenous glucose, a marked destruction of mitochondria and a twofold decrease in the
callus growth index were observed as early as after six-hour-long anaerobic stress. A 48-hour-long incubation
under these conditions resulted in the maintenance of the intact ultrastructure only in 7–10% of cells, while a
96-hour-long anaerobiosis brought about the complete degradation of the subcellular structure and cell death.
A 48-hour-long anaerobiosis without exogenous glucose was chosen for selecting the anoxia-tolerant cell lines.
After three cycles of selection, the anoxia tolerance of the selected cell line exceeded the respective index of
the initial callus several-fold. In selected line, about 50% of cells retained viability and could resume growth
even after 96-hour-long anaerobic incubation. The experimental results obtained were used to determine the
possible causes of the heterogeneity of callus cells as regards their anoxia resistance.
Key words: Saccharum ofﬁcinarum - tissue culture - anoxia - cell selection
: MS—Murashige and Skoog nutrient medium.