Demidchik, V. V.

Przhevalskaya, D. A.

Zvanarou, S. A.

BSU Digital Library

Тезисы докладов Стендовой сессии / Poster Session Abstracts                                   June 25-26,2015 
 
 
46 
After 15 min the optical density of the sample was measured at 524 nm. Next, 
we set the curves for Trolox and studied compounds. 
2. ABTS radical scavenging activity. 
ABTS was dissolved in distilled water to a 7 mM concentration. ABTS 
radical cation (ABTS•+) was produced by reacting ABTS stock solution with 
2.45 mM potassium persulfate (final concentration) and allowing the mixture 
to stand in the dark at room temperature for 16 h before use. Then a solution 
of 0.180 ml of dilute ABTS • + (A734nm = 0.700 ± 0.020) was added 0.2 ml test 
sample. Then the sample was incubated 15 min. The optical density of the 
sample was measured at 734 nm. Next, we set the curves for Trolox and stud-
ied compounds. 
3. Ferric reducing-antioxidant power (FRAP) assay. 
To 0.180 ml of the working compound (0.3 M acetate buffer pH 3.6,  
10 mM 2,4,6-Tris (2-pyridyl) -s-triazine TPTZ 40 mmol/l HCl; 20 mM FeCl3, 
in ratio of 10:1:1) was added 0.02 ml test sample. After 15 min the optical 
density of the samples were measured at 593 nm and set the curves. 
The obtained results indicate different activities tested compounds. The 
problem is more complicated if there are many mixed compounds. 
SALINITY INDUCES PRODUCTION OF SUPEROXIDE ANION  
RADICALS IN PHYSCOMITRELLA PATENS  
Zvanarou S.A., Przhevalskaya D.A., Demidchik V.V. 
Department of Plant Cell Biology and Bioengineering, Biological Faculty,  
Belarusian State University, Minsk, Belarus  
The moss Physcomitrella patens is a model plant, which is widely used 
to investigate physiological reactions of plant cells at the molecular level and 
to produce pharmacologically active compounds. This moss has a dominant 
haploid phase allowing direct forward genetic analysis and manipulations with 
bioengineering tools. Physcomitrella patens can be easily cultured and main-
tained in the non-differentiated juvenile growth form (protonema form), which 
is convenient for study of developmental programmes, role of mutations and 
DNA damage response. Adult growth form (gametophores) contains leaf- and 
stem-like structures, and rhizoids (root-like organs) without vascular tissues. 
Leaves, rhizoids, and protonemal filaments consist of one layer of cells that 
facilitates microscopic observations of stress-induced and developmental mod-
ifications. Apart from annotated genome, the genomic resources for this plant 
include ESTs and full-length cDNA collections and microarrays. Growing in 
wet environment, this moss is normally not exposed to high salinity or 
Тезисы докладов Стендовой сессии / Poster Session Abstracts                                   June 25-26,2015 
 
 
47 
drought. Nevertheless, global warming leading to increased drought periods 
and soil salinisation can also affect mosses. At the moment, very few data can 
be found on how salt stress affects nonvascular plants, such as mosses and 
ferns. The aim of this study was to explore changes in growth and primary salt 
stress responses in Physcomitrella patens. In higher plants, NaCl (>40 mM) 
induces complex osmotic and ionic perturbations leading to oxidative stress. 
The mechanism of the salt-induced radical imbalance is related to the damage 
of electron transport chains and biosynthesis of oxygen-derived radicals de 
novo by HADPH oxidases, class III peroxidases and apoplastic oxidases. 
Here, superoxide anion radical production in response to NaCl was tested us-
ing fluorescent probe dihydroethidium, which is believed to be specific to su-
peroxide, and fluorescent microscopy (Nikon Eclipse TS100F). Growth meas-
urements showed that Physcomitrella patens protonema extension and game-
tophore expansion are significantly inhibited by NaCl starting from 100 mM. 
Stop of growth was found at 400 mM NaCl. This  indicative of relatively high 
salt tolerance of Physcomitrella patens. Fluorescent microscopy measure-
ments using dihydroethidium demonstrated that superoxide is produced after 
treatment of moss with NaCl concentrations over 200 mM. Surprisingly, 100 
mM did not cause superoxide generation. The pharmacological analysis of this 
effect demonstrated that it is sensitive to thiourea, reduced gluthathione, poly-
amines, gadolinium ions and superoxide dismutase. Here, the model of molec-
ular and cellular mechanisms of NaCl induced superoxide generation is pro-
posed. Overall, this study showed that high [NaCl] is capable of inducing ini-
tial reaction of the oxidative stress (superoxide production) in the moss 
Physcomitrella patens which correlates with inhibition of growth in the same 
species. 
БИОРАЗНООБРАЗИЕ ЛЕСНЫХ ЭКОСИСТЕМ ЦЕНТРАЛЬНО-
КАЗАХСКОГО МЕЛКОСОПОЧНИКА – ПЕРСПЕКТИВНЫЕ  
ИСТОЧНИКИ ПРИРОДНЫХ АНТИОКСИДАНТОВ 
Айдарханова Г.С., Кожина Ж.М. 
Евразийский национальный университет им. Л.Н.Гумилева, Астана, Казахстан 
Территория Центрально-Казахского мелкосопочника (ЦКМ) вклю-
чает целый ряд глобально значимых экосистем. Лесные экосистемы ост-
ровками, ленточной полосой расположены во всех природных зонах Ка-
захстана. Северные пустыни, реликтовые еловые леса и уникальные гор-
ные экосистемы, расположенные на территории ЦКМ  вошли в Глобаль-
ный Перечень Всемирного Фонда дикой природы. Международные, ре-


Salinity induces production of superoxide anion radicals in physcomitrella patens

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Salinity induces production of superoxide anion radicals in physcomitrella patens

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