46th Croatian & 6th International Symposium on Agriculture - hr

Toxic influence ORIGINAL of nickel (Ni) SCIENTIFIC on germination PAPER
of two cultivars of wheat (Triticum aestivum L.)
Toxic influence of nickel (Ni) on germination of
two cultivars of wheat (Triticum aestivum L.)
Snežana BRANKOVIĆ 1 , Milan STANKOVIĆ 1 , Vera ĐEKIĆ 2 , Marina TOPUZOVIĆ 1
1 University of Kragujevac, Faculty of Science, Radoja Domanovića 12, 34000 Kragujevac, Serbia,
(e-mail: pavsnez@yahoo.co.uk)
2 Agriculture Research Institute Serbia, Small Grains Research Center, Save Kovačevića b.b., 34000 Kragujevac, Serbia
Abstract
The paper presents results of researches of the effect of several doses of NiCl2 (0.00195, 0.00391,
0.0078, 0.0156, 0.0313, 0.0625, 0.125, 0.25, 0.5 and 1M) on% of germination, the length of root
and shoot of two wheat Triticum aestivum L. cultivars - Kruna and Toplica. The results
indicated that% of germination, root and shoot length of investigated cultivars were remarkable
reduced with the rise of NiCl2 concentration after 3. and 5. days of incubation. The average
values of germinated seeds illustrated higher% of germination of cultivar Kruna than Toplica in
both measurements by about 20%. The analysis of the average length of root and shoot showed
remarkable differences in their length between cultivars and also between measurements after 3.
and 5. incubation days of each cultivar. Results obtained by analysis of variance present high
statistic differences (p≤0.01) among concentrations of% of germination after 3. and 5. days, as
both root and shoot length after 3. days of incubation. Our results also illustrate statistical
significant differences (p≤0.05) of% germination between cultivars after 3. days and of the shoot
length after 5. days of incubation.
Key words: wheat cultivars, germination, nickel
Introduction
Some toxic metals at low doses are essential micronutrients for higher plants, among them is nickel (Ni). In
higher doses heavy metals may causes metabolic disorder and growth inhibition for most of the plant species
(Fernandes and Henriques, 1991; Claire et al., 1991). High levels of Ni in plants growth media are phytotoxic.
Slight increases in the growth of a number of plants species were attributed to low levels of Ni, but its
function is not known. Possible benefitic effects of Ni on plant growth are poorly defined and not
understood. Some studies have confirmed that the nickel is essential for activation of urease, an enzyme
involved with nitrogen metabolisam that is required to process urea. Possibly urease and, therefore, Ni might
be required for the mobilization of stored seed-nitrogen through ureides or arginin during early stage of
seedlig growth. Without Ni, toxic levels of urea accumulate, leadind to the formation of necrotic lesions
(Welch, 1981; Barker and Pilbeam, 2007). The present manuscript reports results of the percentage of
germinated seeds, the root and shoot length of two cultivars Kruna and Toplica of wheat (Triticum aestivum
L.) which were treated with different concentration of NiCl2.
Materials and methods
In order to reasearch effects of nickel to germination of wheat seeds NiCl2 (6H2O) which was obtained from
Merck, Germany was used. The effect of several doses of NiCl2 (0.00195, 0.00391, 0.0078, 0.0156, 0.0313,
0.0625, 0.125, 0.25, 0.5 and 1M) on% of germination, the length of root and shoot of two cultivars Kruna and
Toplica of wheat (Triticum aestivum L.) was studed. For the nickel treatment of seed, the initial solution of
NiCl2 in distilled water at the concentration of 1M was prepared. From the initial solution, 10 dilutions with
descending gradients of concentration were prepared up to the concentration of 0.00195M NiCl2. The seeds
were germinated in Petri dishes with double layer of filter paper soaked in distilled water (control) and 1M -
Proceedings. 46 th <strong>Croatian</strong> and 6 th <strong>International</strong> <strong>Symposium</strong> on Agriculture. Opatija. Croatia (413-416)
Section 3 . Genetics, Plant Breeding and Seed Production 413

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Snežana BRANKOVIĆ, Milan STANKOVIĆ, Vera ĐEKIĆ, Marina TOPUZOVIĆ
0.00195M NiCl2 solutions. Petri dishes were kept under standard laboratory coditions (light/darkness - 16/8h,
temperature 22/18ºC and humidity 55/65%). Data are the results from three separate analysis with 100 seeds
in each Petri dishes. Statistical analysis was performed based by STATISTICA. The data were analyzed by
analysis of variance (ANOVA) to determine the effect of treatments. The percentage of germinated seed and
the length of root and shoot were recorded for each sample three and then five days after seeding in the same
experiment.
Results and discussion
The analysis data of the percentage of germination showed that the number of germinated seeds depended
on the concentration of NiCl2 in the medium. In comparison with control, (sample without NiCl2),% of
germination, root and shoot length of investigated cultivars were considerably reduced with the rise of NiCl2
concentration in both measuremets (Tab. 1 and 2). The average values of germinated seeds illustrated
higher% of germination of cultivar Kruna than Toplica in both investigated days by about 20%. Cultivar
Kruna showed the biggest average value of germinated seeds after 3. days (63.4%). The germination wasn’t
found in the samples of 1 M solution of NiCl2 (except at cultivar Toplica after 5. days of incubation). The
average value of percentage of germinated seeds of both cultivars didn’t remarkably change between first and
second measurements. Seed germination and seedling growth inhibition by heavy metals has also been
reported by (Eskew et al., 1984; Prasad, 1995; Lee et al., 1999; Reeves and Baker, 2000; Liu et al., 2004; Jun et
al., 2009; Stanković et al., 2010).
Table 1. Percentage of germination, root and shoot length after three days of incubation
% of germination1 root length1 (mm) shoot length1 Concentration
(mm)
of NiCl2(M) Kruna Toplica Kruna Toplica Kruna Toplica
1 0 0 0 0 0 0
0.5 3.33±2.31 1.33±2.31 0 0 0.01±0 0.003±0.006
0.25 10±3.46 5.33±3.01 0 0 0.01±0 0.01±0
0.125 9.33±1.15 0 0.007±0.011 0 0.015±0.003 0
0.0625 81.33±7.6 9.33±9.01 0.01±0.01 0.003±0.005 0.010±0.001 0.008±0.007
0.0313 96±3.46 13.33±2.3 0.022±0.025 0.007±0.011 0.011±0.001 0.012±0.004
0.0156 97.33±1.1 41.3±18 0.059±0.026 0.017±0.02 0.012±0.002 0.011±0.002
0.0078 100±0 92.67±2.3 0.034±0.005 0.018±0.017 0.017±0.005 0.011±0.002
0.00391 100±0 98±0 0.039±0.009 0.045±0.007 0.037±0.003 0.033±0.009
0.00195 100±0 99.33±1.2 0.038±0.009 0.125±0.091 0.037±0.036 0.272±0.20
0 100±0 96±1 0.096±0.01 0.135±0.01 0.22±0.01 0.278±0.10
¯¯х 63.4±44.7 41.15±43 0.028±0.031 0.032±0.055 0.04±0.06 0.058±0.11
1 Values expressed as mean ± standard error (n=3)
The analysis of the average length of root and shoot showed remarkable differences in their length between
cultivars and also between measurements after 3. and 5. incubation days of each cultivar. Considering, root
and shoot length of cultivar Toplica showed higher average values in both of investigated days than of
cultivar Kruna. After the first measurement, the average values of root length of cultivar Toplica was 0.032
mm, and second mesurement showed remarkable difference in value and was 2.26 mm. Significant reduction
in root and shoot growth of both cultivars with the increase in concentration of Ni treatment were also
observed as compared to control. Only in the sample with 0.00195 M of NiCl2 the length of root and shoot
was higher than control sample which is in accordance with earlier researches (Welch, 1981). The root didn’t
develop at both cultivars in the samples with 1 M till 0.25 M solution of NiCl2 in both studies days.
On the other hand, the shoot didn’t develop with both cultivars in the sample with 1 M solution of NiCl2 in
both study days (except at cultivar Toplica after 5. days of incubation). The average shoot length of cultivar
Toplica was 0.58 mm after 3. days, and 5.28 mm, nearly 10 times bigger, after 5. days of incubation. Nickel in
high concentration affects many plant metabolic processes. As a consequence of the Ni 2+ toxicity, reduction
in root and shoot size were evident. This is due to reduction in both new cell formation and cell elongation in
the extension region of the root and shoot (Prasad, 1995; Liu et al., 2004).
46 th <strong>Croatian</strong> and 6 th <strong>International</strong> <strong>Symposium</strong> on Agriculture

Toxic influence of nickel (Ni) on germination of two cultivars of wheat (Triticum aestivum L.)
Table 2. Percentage of germination, root and shoot length after five days of incubation
% of germination1 root length1 (mm) shoot length1 concentration
(mm)
of NiCl2 (M) Kruna Toplica Kruna Toplica Kruna Toplica
1 0 1.33±0 0 0 0 0.033±0.03
0.5 4.67±4.61 2.67±1.15 0 0 0.047±0.046 0.027±0.011
0.25 10±4 6.67±1.15 0 0 0.10±0.04 0.067±0.011
0.125 7.33±3.1 1.33±1.15 0 0 0.073±0.031 0.013±0.011
0.0625 86.67±7.6 9.33±9.01 0.027±0.046 0 0.90±0.11 0.093±0.09
0.0313 85.33±5 31.33±8.1 0.087±0.11 0 0.90±0.12 0.313±0.08
0.0156 96±0 60±22.71 0.14±0.11 0.02±0.034 1.15±0.197 0.68±0.09
0.0078 98.67±2.3 95.33±2.3 0.58±0.158 0.36±0.06 2.18±0.39 1.5±0.35
0.00391 100±0 96.67±1.1 2.57±0.29 1.81±0.53 4.95±0.83 8.07±1.83
0.00195 100±0 97±1 2.35±1.89 15.83±0.03 5.22±5.92 28.35±0.03
0 94±1 99±1 4.68±0.02 6.81±0.099 19.04±0.02 18.88±0.02
¯х 62.1±43.8 45.7±43.2 0.95±1.59 2.26±4.79 3.14±5.62 5.28±9.31
1Values expressed as mean ± standard error (n=3)
Table 3. ANOVA for% of germination, root and shoot length after three and five days of incubation
Cultivars Concentration of NiCl2
F - calculated value after 3. days after 5. days after 3. days after 5. days
% germination 5.226* 4.407 7. 261** 10.261**
The lenght of root 0.145 1.1217 5.060** 2.1972
The lenght of shoot 1.154 1.012 9.566** 3.978*
F1,10;0.005 = 4.96, F1,10;0.001 = 10.04, F10,10;0.05 = 2.94, F10,10;0.001 = 4.85
F-test of significance level: * p≤0.05; ** p≤0.01
Results obtained by analysis of variance (ANOVA) present high statistic differences (p≤0.01) among
concentrations of% of germination after 3. and 5. days, like both root and shoot length after 3.days of
incubation (Tab. 3). Our results also illustrate statistical significant differences (p≤0.05) of% germination
between cultivars after 3. days and of shoot length after 5. days of incubation.
Conclusion
The results show that increasing concentration of NiCl2 significantly inhibited seed germination and early
growth of wheat root and shoots. Cultivar Kruna showed higher average value of% of germination after 3.
and 5. days of incubation than cultivar Toplica. On the other hand cultivar Tolica had higher average value
of root and shoot length after 3. and 5. days of incubation than cultivar Kruna. Results obtained by analysis
of variance (ANOVA) show high statistic differences among concentrations of% germination after 3. and 5.
days, like both root and shoot length after 3. days of incubation. Our results also illustrate statistical
significant differences of germination between cultivars after 3. days and of shoot length after 5. days of
incubation.
References
Barker A.V., Pilbeam D.J. (2007). Handbook of Plant Nutrition. C.R.C. Taylor and Francis.
Claire L.C., Adriano D.C., Sajwan S.L., Abel D.P., Thoma D.P., Driver J.K. (1991). Effect of
selected trace metal on germination seeds of six plant species. Water, Air and Soil Pollution
59: 231-240.
Eskew D., Welch R. M., Norvell W. (1984). Nickel in Higher Plant. Plant Physio. 76: 691-693.
Fernandes J.C., Henriques F.S. (1991). Biochemical, physiological and structural effects of
excess copper in plants. The Botanical Rewiew 57: 246-273.
Jun R., Ling T., Guanghua Z. (2009). Effect of chomium on seed germination, root elongation
and coleoptile growth in six pulses. Int. J. Environ. Sci. Tech. 6 (4): 571-578.
Lee Y.Z., Suzuki S., Kawada T., Wang J., Koyama H., Rivai I.F., Herawati N. (1999).: Content of
cadmium in carrots compared with rice in Japan. Bull. Environ. Contam. Toxicol. 63: 711-
719.
Section 3 . Genetics, Plant Breeding and Seed Production 415

416
Snežana BRANKOVIĆ, Milan STANKOVIĆ, Vera ĐEKIĆ, Marina TOPUZOVIĆ
Liu D., Jiang W., Gao X. (2004). Effects of cadmium on root growth, cell division and nucleoli
in tips of garlic. Physiologia Plantarum 47: 79-83.
Prasad M.N.V. (1995). Cadmium toxicity and tolerance in vascular plant. Environmental and
Experimental Botany 35: 525-545.
Reeves R.D., Baker A.J.M. (2000). Metalaccumultig plants. In: Raskin and B.D. Ensly (Ed.).
Phytoremediation of toxic metals: using plants to clean up the environment. John Wiley and
Sons, Inc, Toronto, Canada, p.303.
Stanković M., Topuzović M., Marković A., Pavlović D. (2010). Influence of Zn on germination
of wheat (Triticum aestivum L.). Biotech. and Biotech. Equipments 4: 236-239.
Welch R.M (1981). The biological significance of Ni. Journal of Plant Nutrition. 3 (1-4): 345-
356.
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46 th <strong>Croatian</strong> and 6 th <strong>International</strong> <strong>Symposium</strong> on Agriculture