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ANALYSIS OF VIABILITY THROUGH TZ TEST OF SEED OF TWO ALYSSUM SPECIES OF IMPORTANCE FOR PHYTOMINING FROM SERPENTINES IN ALBANIA. View project
Erasmus + Programme, KA1, 2015/2016 ; Faculty of Agriculture and Environment at Agricultural University of Tirana and Faculty of Biologjy at Sofia University View
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20°00′09″E (700 m a.s.l.) and from Kukës (K), [20] and Ranal et al. [21]. All treatements were per-
42°00'35"N, 20°17'44"E (1300 m a.s.l.) in 2014. The formed with three replications.
soil chemical composition of these sites was previ-
ously analyzed [1, 18] and reflected their ultramafic Data analysis. The data were analysed with
origin, i.e. between 1100-2150 mg kg-1 of Cr, 1080- one-way analysis of variance (ANOVA) to deter-
3100 mg kg-1 of Ni and 104 -280 mg kg-1 of Co. Seed mine which treatments differed from each other
material was collected from 50 individuals per pop- when germinability, root and hypocotyl elongation
ulation. The seeds were extracted manually from were used as variables. The data for both species and
fruits and those of bad quality (damaged, empty and their population were analysed separately. In cases
immature) were rejected. The received seeds from A. when the ANOVA showed significance of differ-
markgrafii were low in comparison to A. murale, ences (P < 0.05), pairwise mean comparisons of var-
probably due to lower seed production. The material iables, a least significant difference (LSD) test was
was collected and stored according to international performed. The statistical analysis was performed
guidelines [19]. Vouchers were deposited in the Ag- using Microsoft Excel 2010.
ricultural University of Tirana.
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© by PSP Volume 27 – No. 3/2018 pages 1345-1352 Fresenius Environmental Bulletin
markgrafii was more spread out through time, with a Root and hypocotyl elongation. The root and
peak of germination at ~12th and 14th days respec- hypocotyl elongation of seeds treated with Ni was
tively for both populations. The germination was also affected by elevated concentrations in both spe-
less spread out in time in A. murale and a strong ger- cies (Fig. 1). In general, the length of root treated
mination peak occurred at the 4th day for seeds from with different Ni concentrations decreased compared
the first population and at the 6th day after beginning to the control. The length of the root after Ni appli-
of the experiment for the second population (Table cation was significantly different from the controls
1). in A. murale (LSD = 2.88, P < 0.05 and LSD = 2.10,
The mean germination rate (MR) calculated as P < 0.05, respectively) and A. markgrafii (LSD =
the reciprocal of the mean germination time (MT) 4.39, P < 0.05 and LSD = 7.72, P < 0.05, respec-
was the highest for A. murale (Prrenjas) and lowest tively) populations (Figs. 1A and C). The effect of
for A. markgrafii (Kukës). The seeds of A. murale inhibition on the root growth was higher after treat-
presented greater homogeneity (lower value of CVt) ments with ≥ 2 mM Ni in both species. The concen-
and higher synchrony of germination (lower value of tration of 1 mM Ni reduced root length about 50%
U and higher value of Z) compared to A. markgrafii. for A. murale and A. markgrafii from Gjegjan. For
The differences of the germination process were the seeds of A. markgrafii (Kukës) this concentration
found not only between the species but also between was 0.5 mM Ni. The root length was more or less
their populations, higher between A. murale popula- equal when the seeds were treated with 6 and 8 mM
tions. Ni and no significant differences were found.
TABLE 1
The calculated values for germinability (G%), mean germination time MT (days), coefficient of variation
of the germination time (CVt%), mean germination rate (MR), uncertainty (U) and synchronization of
germination (Z) for A. murale and A. markgrafii populations. Values are the means of three replicates per
population ± SD (n=6). Different letters show significant differences based on one-way ANOVA and LSD
test (P < 0.05) performed separately for each population.
Species &
Ni
popula- G (%) MT (day) CVt (%) MR (1/day) U Z
(mM)
tions
0 87.33±1.15a 3.00±1.06a 33.95±15.47a 0.35±0.10a 1.04±0.13ab 0.63±0.05a
0,5 80.67±5.03ab 2.78±0.85ab 24.53±3.23a 0.37±0.09a 1.13±0.33a 0.52±0.15b
1 86±4a 2.97±0.92bc 24.47±1.18a 0.34±0.09a 1.36±0.50bc 0.45±0.18b
A. murale
2 82±2ab 3.28±0.82c 28.05±4.51a 0.31±0.07a 1.67±0.17cd 0.31±0.04bc
(P)
4 84±7.21ab 3.26±1.00c 31.74±11.28a 0.32±0.08a 1.64±0.23c 0.35±0.07bc
6 83±5.03ab 3.57±0.80c 31.55±4.67a 0.28±0.05a 1.95±0.18d 0.26±0.04c
8 78±4b 3.58±0.99c 30.27±6.12a 0.29±0.07a 1.85±0.25d 0.30±0.07bc
0 68±4a 4.15±0.14a 22.13±1.35a 0.23±0.05a 1.77±0.13ab 0.31±0.06a
0,5 64±11.13ab 4.52±0.29a 24.19±0.43a 0.21±0.01b 2±0.08ab 0.24±0.01bc
1 55±17.00abc 4.94±0.27a 20.96±4.14a 0.19±0.01bc 1.87±0.24ab 0.29±0.03ab
A. murale
2 51±10.06bc 5.15±0.42a 20.14±2.37a 0.18±0.01c 1.96±0.14ab 0.25±0.02bc
(Po)
4 55±1.15abc 5.07±0.25a 22.81±3.85a 0.19±0.01bc 2.00±0.13a 0.24±0.05bc
6 43±7.02c 5.27±0.48a 22.26±0.75a 0.18±0.02c 1.99±0.08b 0.23±0.02bc
8 57±9.86abc 5.3±0.1a 22.20±1.48a 0.18±0.05c 2.13±0.03b 0.21±0.01c
0 51±5.13a 10.52±0.75abc 27.65±1.05a 0.09±0.01ab 2.63±0.19ab 0.08±0a
0,5 48±5.03ab 9.59±0.82ab 41.69± 0.87ab 0.10±0.01a 2.86±0.51ab 0.09±0.06a
A. 1 49±5.54ac 9.77±0.77a 44.67±9.42b 0.09±0.01ab 2.75±0.27ab 0.08±0.02a
markgrafii 2 30±3d 11.61±0.40bc 38.43±4.93ab 0.08±0.00ab 2.61±0.20ab 0.04±0.02a
(G) 4 26.33±11.54d 10.75±2.18abc 29.19±4.54aa 0.09±0.02ab 2.19±0.61a 0.09±0.06a
6 34±4.04bcd 12.41±0.92c 32.32±5.41ab 0.07±0.01b 2.68±0.35b 0.08±0.04a
8 38.00±15.58bad 12.63±2.65c 31.24±18.29ab 0.07±0.02ab 2.63±0.48b 0.07±0.02a
0 50±5.77a 8.87±1.13a 38.87±9.76a 0.11±0.01a 2.62±0.50abc 0.12±0.07a
0,5 38.67±5.13ab 10±1a 32.05±6.29a 0.10±0.01ab 2.71±0.13abc 0.08±0.01ab
A. 1 45.67±14.01ab 9.90±1.26a 38.47±6.23a 0.09±0.01ab 2.99±0.31a 0.06±0.02ab
markgrafii 2 33±10b 11.27±0.45a 29.60±5.29a 0.08±0.01b 2.62±0.25abc 0.06±0.04ab
(K) 4 31±7.21b 11.64±2.12a 31.89±13.89a 0.08±0.01b 2.49±0.39b 0.01±0.04ab
6 21±13.32b 11±1.80a 32±8.07a 0.09±0.01b 2.15±1.01c 0.03±0.02b
8 21.00±8.54b 11.76±0.48a 34.11±13.26a 0.08±0.00b 2.00±0.10c 0.12±0.07ab
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FIGURE 1
Root and hypocotyl length in Alyssum murale (A and B) and Alyssum markgrafii (C and D) populations.
Values are the means of three replicates per population ± SD (n=6). Abbreviations are used to indicate
different populations: Prrenjas (P), Pogradec (Po), Gjegjan (G) and Kukës (K). Different letters show
significant differences based on one-way ANOVA and LSD test (P < 0.05) performed separately for
each population.
In general, the hypocotyl length in the control in both species and populations were more sensitive
was lower compared to the length of hypocotyl for to Ni compared to hypocotyl. Although the length of
seeds treated with lower concentrations of Ni (0.5 the roots in the controls was higher compared to the
and 1 mM) (Figs. 1B and D). After treatment with length of the hypocotyl, it decreased quickly and
such concentrations the length of the hypocotyl de- reached lower values in samples treated with 6 and 8
creased and reached 50% inhibition for seeds treated mM Ni.
with 8 mM Ni compared to the control in both pop-
ulations of A. markgrafii. For A. murale 50% inhibi- Nickel impact on seedlings. Seeds of both spe-
tion was not reached in both populations. The statis- cies and populations treated with different Ni con-
tical analysis and pairwise mean comparisons of var- centrations showed abnormalities in their germina-
iables demonstrated significantly differences be- tion, including chlorosis, necrosis, coiling of the root
tween the control and seeds treated with lower Ni and the hypocotyl, lack of hairs in the zone of root
concentrations only in A. murale from Prrenjas (LSD hairs, a short zone of root elongation and the appear-
= 1.94, P < 0.05). In the population of A. murale from ance of root deviation very near to the root tip, brown
Kukës the hypocotyl length in the control was sig- cotyledons and hypocotyl of the seedlings or three
nificantly different from seeds treated with higher cotyledons instead of a pair (Fig. 2). These abnor-
concentrations (LSD = 0.85, P < 0.05). For both pop- malities varied considerably for each Ni treatment.
ulations of A. markgrafii the hypocotyl length in the Soon after germination in 0.5 mM Ni, the root and
control was significantly different from seeds treated hypocotyl started coiling (Figs. 2D and E) and brown
with higher concentrations (LSD = 2.08, P < 0.05 color of the hypocotyl and the cotyledons appeared
and LSD = 2.14, P < 0.05, respectively). (Figs. 2G and H). The root coiling was more than
The effect of Ni on root and hypocotyl length 70% of the seedlings of A. murale treated with 0.5
was different in both species and their populations. mM Ni while for A. markgrafii it was more than
Lower concentrations of Ni stimulated hypocotyl 50%. The percentage of the root coiling increased
growth but inhibited root elongation. Root seedlings and varied between 90% and 100% when the seeds
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were treated with 8 mM Ni in both species and pop- Most often roots without hairs appeared in both spe-
ulations. The percentage of the hypocotyl coiling (in cies when treated with Ni concentrations above 1
the area near to the cotyledons) was lower compared mM. The seed germination stopped soon after the
to the root coiling in both species and all Ni treat- appearance of radical in higher Ni concentrations
ments. Frequently, cotyledons with curved margins (Fig. 2I). Also, a short zone of root elongation (Fig.
were found (Fig. 2F). The cotyledons demonstrated 2J) and the appearance of root deviation very near to
also chlorosis and necrosis when Ni concentrations the root tip (Fig. 2K) were found in seeds treated
were higher (Figs. 2B and C). The percentage of the with 6 and 8 mM Ni. Rarely, three cotyledons in-
cotyledons with chlorosis was higher in A. murale. stead of a pair were observed (Fig. 2L).
FIGURE 2
Different patterns of abnormally germinated seed of Alyssum species after Ni application. A: normally
germinated seedling; B-C: chlorosis and necrosis on the leaves; D-E: coiling root and hypocotyl; F: cot-
yledons with curved margins; G-H: brown color of hypocotyl and cotyledons; I: seed germination
stopped after the appearance of radical in higher Ni concentrations; J: short zone of root elongation;
K: root deviation very near to the root tip; L: appearance of three cotyledons instead of a pair. Scale in
millimeters (mm).
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© by PSP Volume 27 – No. 3/2018 pages 1345-1352 Fresenius Environmental Bulletin
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(2015) Seed weight and germination behavior of
Dolja Pavlova
the submerged plant Potamogeton pectinatus in
Department of Botany, Faculty of Biology,
the arid zone of northwest China. Ecol. Evol. 5,
University of Sofia blvd. Dragan Tzankov 8,
1504–1512.
1164 Sofia – Bulgaria
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