Advances in Environmental Biology

Advances in Environmental Biology, 8(7) May 2014, Pages: 2366-2371
AENSI Journals
Advances in Environmental Biology
ISSN-1995-0756
EISSN-1998-1066
Journal home page: http://www.aensiweb.com/aeb.html
Joint effect of salinization and copper ions on anatomical organization of leaves
and roots of wheat’s variety (Triticum aestivum L.)
Saule Jumagalievna Atabayeva, Akmaral Nurmahanova, Saule Sagidullaevna Aidosova, Aigul
Akhmetova, Saule Sagyndikovna Kenzhebayeva, Saltanat Shyntayevna Asrandina, Ravilya Alybayeva,
Sabina Alimzhanovana Shoinbekova, Ayzhan Zhumagazievna Beisenova
Al-Farabi Kazakh National University 050040 Almaty, Kazakhstan, Al-Farabi, 71
ARTICLE INFO
Article history:
Received 25 March 2014
Received in revised form 22 April
2014
Accepted 15 May 2014
Available online 10 June 2014
Key words:
ABSTRACT
The objective of this work was to explore anatomical features of 5 wheat varieties
(Triticum aestivum L.), resistant and non-resistant to joint effect of copper ions and
salinization. When exposed to salinization (NaCl) and copper ions (Cu) under their
separate and joint effect, anatomical showings of wheat varieties were undergoing
changes depending on plant’s variety. It was stated that the following anatomical
features may be used for checking plants’ resistance to salinization and heavy metal
actions: diameter of conducting bundles, ratio between thickness of exoderm and
thickness of endoderm. Key words: wheat, salinization, cuprum, anatomical features.
© 2014 AENSI Publisher All rights reserved.
To Cite This Article: Saule Jumagalievna Atabayeva, Akmaral Nurmahanova, Saule Sagidullaevna Aidosova, Aigul Akhmetova, Saule
Sagyndikovna Kenzhebayeva, Saltanat Shyntayevna Asrandina, Ravilya Alybayeva, Sabina Alimzhanovana Shoinbekova, Ayzhan
Zhumagazievna Beisenova., Joint effect of salinization and copper ions on anatomical organization of leaves and roots of wheat’s variety
(Triticum aestivum L.). Adv. Environ. Biol., 8(7), 2366-2371, 2014
INTRODUCTION
Substantive areas of soils and waters, particularly near big cities and large industrial complexes of
Kazakhstan, are polluted with heavy metals [1,2]. Another topical ecological problem for Kazakhstan is
salinization. Irrigated farming constitutes a main threat for water resources on the south and south-east of the
country [3].
Earlier the authors had stated that in conditions of acute pollution and medium salinization plants
accumulate a large amount of heavy metals and their translocation into above-ground organs increases [4]. By
contrast to this, content of Zn and Cd in roots of Tamarix maritima, A. tripolium increased threefold in the area
of acute salinization [5]. Other plants, while being exposed to both stressors, didn’t show the damage symptoms,
though their biomass decreased by 40-70% [6].
Salinization leads to anatomical changes in the structure of cell wall. Cotton plants demonstrated
suberinization of Casparian strips [7-9], decrease in central cylinder to cortical parenchyma ratio, which points
to decrease in diameter of central cylinder, thickening of exoderm and endoderm, expansion of intercellular
space in cortex and significant lignification of exoderm cells [10-13].
For successful development of agriculture in problem areas it is reasonable to cultivate croppers and
varieties of crops, which would be tolerant to exposure to these factors. The objective of this work was to
explore anatomical features of wheat varieties, so that it is possible to discover anatomical features that would
become sample features for resistant varieties.
MATERIALS AND METHODS
Experimental design, plant material and treatments. The objects of research were five wheat varieties:
Kazakhstanskaya-3, Kazakhstanskaya rannaya, Melturn, Kaiyr and Shagala. Seeds of plants were germinated in
a growing chamber at 25ºC. Plants were grown in hydroponic conditions during 7 days in solutions containing
various concentrations of copper. Four treatments were defined as: Control, 50 µm NaCl; 0,25 µm СuSO4;
NaCl (50 µm)+ СuSO4 (0,25 µm).
The plants were conserved according to Strasburger-Fleµming method [14]. Preservative liquid was the
mixture “alcohol-glycerin-water” pro rata 1:1:1. Fixation was performed in 96% ethanol. Aboveground and
underground vegetative organs of test cultures were fixed.
Corresponding Author: Saule Jumagalievna Atabayeva, Al-Farabi Kazakh National University 050040 Almaty,
Kazakhstan, Al-Farabi, 71
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Advances in Environmental Biology, 8(7) May 2014, Pages: 2366-2371
Anatomical products were made with the help of microtome with freezing device ТОС-2. Sections were
placed onto glycerin and balm in accordance with standard practices [14-16]. Thickness of anatomical sections
was 10-15 μm. For quantitative analysis morphometric research was performed with the use of eyepiece
micrometer МОВ-1-15 (object lens was х 9 and magnification was х 10,7).
Statistical analysis. Study data was analysed statistically with the use of two-way ANOVA with varieties
and treatment as main effects on anatomical parameters. All the values were expressed as the mean value of
three measurements for each treatment. The data was analysed statistically using two-way ANOVA with species
and treatments as main effects on shoot and root anatomical parameters. All the values were expressed as the
mean value of three measurements for each treatment. Values represent mean ± standard error (SE).
RESULTS AND DISCUSSION
Preliminary study of wheat varieties’ resistance to joint effect of salinization and copper ions on growth
parameters displayed that Kazakhstanskaya rannaya and Kazakhstanskaya-3 varieties have the best resistance to
these stressors, and Shagala variety is the most perceptible to these stressors. Other varieties appeared to have
medium resistance. Determination of anatomical structure of wheat lamina, when exposed to joint effect of
salinization and copper ions, showed that thickness of lower epidermis of Kaiyr variety increased by 23%,
thickness of Kazakhstanskaya rannaya variety kept being at control level, and other varieties demonstrated
thickness decrease in comparison with control: Melturn variety showed the decrease by 8%, Kazakhstanskaya-3
– by 11% and Shagala – by 13%, though, the difference between the varieties in terms of this index were not
verified (р > 0.05). Upper epidermis thickness of Kaiyr, Kazakhstanskaya rannaya and Melturn with respect to
control increased by 24, 21, и 4%, respectively. Shagala’s upper epidermis thickness kept being close to control
level (98%), and Kazakhstanskaya-3 showed decrease of 4% (Pictures 1,2) (р < 0,05).
Совместное действие двух стрессоров у сорта Казахстанская ранняя не вызывало изменения
толщины нижнего эпидермиса, равно как и их раздельное действие (102, 100 и 100% при раздельном
действии NaCl (50 мМ), ионов меди и совместном действии, соответственно). У сортов Казахстанская-3
и Шагала совместное действие усугубляло негативный эффект раздельного действия двух стрессоров.
У сорта Кайыр совместное действие снимало ингибирующий эффект раздельного действия стрессоров.
Joint effect of two stressors on Kazakhstanskaya rannaya variety didn’t lead to changes in thickness of
lower epidermis, equally as their separate effect (102, 100 and 100% when exposed to separate effect of NaCl
(50 µm), copper ions and joint effect, respectively). Kazakhstanskaya-3 and Shagala, when exposed to joint
effect, were more perceptive to negative impact of separate stressors. When Kaiyr variety was exposed to joint
effect, the latter annihilated inhibitive effect of separate actions of stressors.
When exposed to stressors’ joint effect, almost all the varieties had the upper epidermis thickness higher
than control: Kazakhstanskaya rannaya (121%), Kaiyr (124%), Melturn (104%) and insignificantly declined for
Kazakhstanskaya-3 (96%) and Shagala (98%). Inhibitory effect of stressors’ separate actions was annihilated by
joint effect for almost all the varieties.
The varieties are sorted by decrease of these parameters when exposed to combined action of copper ions
(0,25 µm) and salinization (50 µm NaCl) in the following way (% to control): lower epidermis – Kaiyr (123) >
Kazakhstanskaya rannaya (100) > Melturn (92) > Kazakhstanskaya-3 (89) > Shagala (87%) (р > 0,05); upper
epidermis – Kaiyr (124) > Kazakhstanskaya rannaya (121) > Melturn (104) > Shagala (98%) >
Kazakhstanskaya-3 (96%) (р < 0,05) (Pictures 1, 2).
Combined action of copper ions and salinization on diameter of conducting bundles didn’t demonstrate
significant negative impact (р > 0,05). When exposed to combined action of salinization (NaCl) and copper,
diameter of lamina conducting bundles increased for Kaiyr (by 11%), Kazakhstanskaya rannaya (by 3%), and
Kazakhstanskaya-3 and Shagala varieties this ration was almost at the control level (98% to control), except
Melturn variety, which showed decrease of diameter of conducting bundles by 15%.
Picture. 1: Effect of joint action of salinization (NaCl) and copper on lower epidermis thickness of wheat
leafage
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Picture 2: Effect of joint action of salinization (NaCl) and copper on upper epidermis thickness of wheat
leafage
Sizes of conducting bundles directly depend on diameters of xylary vessels. Thus, decrease in diameter of
conducting bundles is directly connected with decrease in xylary vessels’ areas, which, in their turn, being
conducting elements, distinctly respond to conduction of various substances by changing its diameter. In this
case, changes in conducting bundles’ diameters positively correlate with varieties’ resistance according to
growth parameters.
When exposed to combined action of salinization (50 µm NaCl) and copper ions, the diameter of
conducting bundles decreased in the following sequence: Kaiyr(111%) > Kazakhstanskaya rannaya (103) >
Kazakhstanskaya-3 (98%) = Shagala (98%) > Melturn (85%) (р < 0,05) (Picture 3).
Picture. 3: Effect of joint action of salinization (NaCl) and copper on conducting bundles’ diameter of wheat
Joint effect of copper and salinization on this index annihilated negative impact of stressors’ separate
actions.
Resistant (in terms of growth parameters) varieties Kaiyr, Kazakhstanskaya rannaya and Kazakhstanskaya3 demonstrated increase or stability of this index. Shagala variety had low index of biomass of above-ground
organs, though the diameter of conducting bundle stayed at control level. Thus, when exposed to joint effect of
copper and salinization, not all the varieties demonstrated changes on tissue level reflected at organismic level.
While studying morphometric indices of wheat roots, all the varieties demonstrated thickness of exoderm
decreased. The least decrease was seen on Kazakhstanskaya-3 (by 8%) and the largest decrease was seen on
Melturn variety (by 49%). The varieties are sorted by changes in thickness of root exoderm in the following
way: Kazakhstanskaya-3 (92%) > Kaiyr (84%) > Shagala (80%) > Kazakhstanskaya rannaya (67%) > Melturn
(51%) (р < 0,05) (Picture 4).
Melturn demonstrated the most significant decrease in thickness of exoderm, and it corresponded to the
most significant decrease in biomass of above-ground organs and roots of this variety in preliminary studies.
Combined action of stressors at Kazakhstanskaya rannaya (67%), Kaiyr (84%) and Melturn (51%) increased
negative effect of their separate influence on exoderm thickness. Combined action of stressors at
Kazakhstanskaya-3 and Shagala mildly lowered negative effect of separate action of stressors on exoderm
thickness.
Thickness of endoderm increased with Kazakhstanskaya rannaya (121%), Shagala (106%) and Melturn
(103%) varieties. Kaiyr and Kazakhstanskaya-3 varieties showed decrease of endoderm thickness by 44 and
52%? respectively (Picture 5).
The varieties are sorted by thickness of endoderm in the following way: Kazakhstanskaya rannaya (121%)
> Shagala (106%) > Melturn (103%) > Kaiyr (56%) > Kazakhstanskaya-3 (48%) (р < 0,05).
Joint action of stressors towards Kazakhstanskaya rannaya, Melturn and Shagala varieties annihilated
negative impact of separate copper action towards endoderm thickness, and exceeded control level by 21.3 and
9%, respectively.
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Picture. 4: Effect of joint action of salinization (NaCl) and copper on thickness of wheat exoderm
Ratio of exoderm thickness to endoderm thickness is an important index of plants’ resistance. According to
authors’ opinion, the higher this index is, the higher is plants’ resistance [17]. Thickening of exoderm indicates
adaptive reactions against stressors [18]. This index decreases when the plant is exposed to joint effect of
salination and copper in the following sequence (% to control level): Kazakhstanskaya-3 (191) > Kaiyr (146%)
> Shagala (71) > Kazakhstanskaya rannaya (55%) > Melturn (49) ( р < 0,05).
Kazakhstanskaya-3 and Kaiyr had their exoderm-endoderm ratio higher than control level by 91 and 46%,
respectively. These varieties demonstrated the least decrease in roots’ biomass – by 25%/
This suggests that in conditions of chemical stressors’ actions exoderm thickness increases with respect to
endoderm, which may reduce absorption of toxicants by the root cells. Under stress exoderm provides peripheral
barrier on the way of ingress of dissolved matters to apoplast [19].
Picture. 5: Effect of joint action of salinization (NaCl) and copper on thickness of wheat endoderm
The least exoderm-endoderm ratio compared to control was detected with Melburn variety (by 58%), which
correlated with low biomass indices of this variety.
Most of the varieties didn’t have their diameter of central cylinder changed significantly (р > 0,05). Indices
of Kazakhstanskaya rannaya and Shagala were a bit higher than control value (104%). When these varieties
were exposed to joint action of the stressors mentioned, negative impact of separate actions of stressors is
annihilated. The largest decrease in diameter of central cylinder was seen with Melturn variety – by 37%
(Picture 6).
The varieties are sorted by diameter of central cylinder in the following way: Melturn (103) >
Kazakhstanskaya rannaya (98%) = Kaiyr (98) > Shagala (94) > Kazakhstanskaya-3 (93) (р > 0,05).
Melturn’s index also positively correlates with low growth indices. Combined action of stressors at
Kazakhstanskaya-3, Kaiyr and Melturn increased negative effect of their separate influence. Combined action of
stressors at other varieties mildly lowered negative effect of separate action of stressors.
Thus, resistant variety Kazakhstanskaya rannaya showed good results in endoderm thickness (121% in
relation to control values) and diameter of central cylinder (98% of control values). Other resistant variety
Kazakhstanskaya-3 showed good results in thickness of root exoderm (92% in of control values) and ratio of
exoderm thickness to endoderm thickness (191% of control values).
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Picture. 6: Effect of joint action of salinization (NaCl) and copper on diameter of central cylinder of wheat
roots
Shagala as a fickle variety had low indices as for diameter of central cylinder (94% of control values).
Melturn showed perceptibility to stressors and had indices as for thickness of exoderm (51% of control values).
Summary:
Exploration of anatomical features of wheat leafage and roots, exposed to combined effect of copper ions
and salinization (NaCl), showed that in conditions of this stress structure of leafage and roots undergo
substantial changes. Changes in anatomical structure correlated with plants’ resistance in growth indices.
Conclusions: Anatomical features, which correlated with resistance to a greater or lesser extent – diameter
of conducting bundles, ratio of exoderm thickness to endoderm thickness, diameter of central cylinder – may be
used as a test, revealing tolerance to salinization and heavy metals.
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