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Abstract
Malondialdehyde (MDA) concentration is a widely
used method to analyse lipid peroxidation in biological material. In plant tissues, however, certain
compounds (anthocyanins, carbohydrates) may
interfere with measurements which may lead to an
overestimation of the MDA levels. Two methods
were compared for analysing lipid peroxidation,
either uncorrected or corrected for interfering compounds. The comparison was performed in three
separate experiments with respect to cold treatments (snow removal in winter, reacclimation in
summer and cold acclimation in autumn) in bilberry
(Vaccinium myrtillus L.). During winter and autumn
the methods seem to measure different compounds,
but during active growth in the summer the difference
between the methods was less. This is obviously due
to carbohydrates which act as cryoprotectants and
increase in concentration during cold acclimation
as well as due to the anthocyanins. It is thus suggested that the validity of the uncorrected method
to measure MDA and thereby lipid peroxidation is
best in plant tissue which is in an active growth state.
Key words: Lipid peroxidation, stress, Vaccinium myrtillus,
cold acclimation.
Introduction
The major lipid classes of plant tissues are sterols, sterylglucosides, glucocerebrocides, and phospholipids, and
the proportions of free sterols and phospholipids increase
3
2376
Taulavuori et al.
Sampling procedure
In each treatment, the samples were collected and placed
immediately into a styrofoam box filled with ice. They were
weighed and quickly frozen in liquid nitrogen (LN) followed by
storage in a freezer (70 8C).
Lipid peroxidation
2377
Statistical analysis
The results obtained with the methods U and C were compared
with the paired sample T-test. Linear regressions between the
methods were also calculated. The data were arranged in a
hierarchic order for analysis of regression and T-tests, proceeding from general to more categorical levels: first the whole data,
and then all leaves or stems. The procedure was continued until
the split of the data, in order to find out any effects possibly
due to seasonality (Experiment), morphology (stem versus leaf )
and treatment (cold versus warm).
Results
The results are presented in detail in Figs 38. The data
show that the results obtained by the uncorrected (U) and
corrected (C) methods differ significantly (P-0.001) from
each other (Table 1). The findings concern the whole data
and the split categories, although the significance was
lower with bilberry stems in turf patches and excised
stems in a water container in Experiment II (P-0.05 and
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Taulavuori et al.
R2
P-
All
All stems
All leaves (note: only Experiment II)
Stems
Experiment I
Experiment II
Experiment III
Experiment II
All
Bilberries in turf patches: All
Bilberries in turf patches: Stems
Bilberries in turf patches: Leaves
Excised bilberries in water container: All
Excised bilberries in water container: Stems
Excised bilberries in water container: Leaves
All: Cold treatment
Warm treatment
0.037
0.031
0.302
0.001
0.001
0.001
131
100
31
0.001
0.560
0.017
0.001
0.001
0.001
24
28
48
0.398
0.681
0.791
0.649
0.002
0.001
0.000
0.430
0.381
0.001
0.001
0.05
0.001
0.001
0.01
0.001
0.001
0.001
59
30
14
16
29
14
15
30
29
Discussion
In agreement with some other studies (Blokhina et al.,
1999; Hodges et al., 1999), the results suggest that lipid
peroxidation should be corrected against interfering
compounds. The uncorrected and corrected methods
obviously measure different compounds particularly
during snow removal in late winter (Experiment I) and
cold acclimation in autumn (Experiment III). However,
2379
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Taulavuori et al.
Acknowledgement
This work was funded by the Academy of Finland.
References
Blokhina OB, Fagerstedt KV, Chirkova TV. 1999. Relationships
between lipid peroxidation and anoxia tolerance in a range of
species during post-anoxic reaeration. Physiologia Plantarum
105, 625632.
Boo YC, Jung J. 1999. Water deficit-induced oxidative stress
and antioxidative defences in rice plants. Journal of Plant
Physiology 155, 255261.
Chen WP, Li PH, Chen THH. 2000. Glycinebetaine increases
chilling tolerance and reduces chilling-induced lipid peroxidation in Zea mays L. Plant, Cell and Environment 23, 609618.
Dhindsa RS, Plumb-Dhindsa P, Thorpe TA. 1981. Leaf
senescence: correlated with increased levels of membrane
permeability and lipid peroxidation, and decreased levels of
superoxide dismutase and catalase. Journal of Experimental
Botany 32, 93101.
Di Toppi LS, Lambardi M, Pecchioni N, Pazzagli L, Durante M,
Gabbrielli R. 1999. Effects of cadmium stress on hairy roots of
Daucus carota. Journal of Plant Physiology 154, 385391.
Guidi L, Bongi G, Ciompi S, Soldatini GF. 1999. In Vicia faba
leaves photoinhibition from ozone fumigation in light
precedes a decrease in quantum yield of functional PSII
centres. Journal of Plant Physiology 154, 167172.
Heath RL, Packer L. 1968. Photoperoxidation in isolated
chloroplasts. I. Kinetics and stoichiometry of fatty acid
peroxidation. Archives in Biochemistry and Biophysics 125,
189198.
Hodges DM, DeLong JM, Forney CF, Prange RK. 1999.
Improving the thiobarbituric acid-reactive-substances assay
for estimating lipid peroxidation in plant tissues containing
anthocyanin and other interfering compounds. Planta
207, 604611.
Hodges DM, Forney CF. 2000. The effects of ethylene, depressed
oxygen and elevated carbon dioxide on antioxidant profiles
of senescing spinach leaves. Journal of Experimental Botany
51, 645655.
Kingston-Smith AH, Foyer CH. 2000. Bundle sheath proteins are
more sensitive to oxidative damage than those of the