Comparison of growth rates of the filamentous fungus Aspergillus niger L. from contrasting environments
Eighteen strains of filamentous fungus Aspergillus niger were isolated from contrasting environments in Israel: the hypersaline Dead Sea and the temperate «Evolution Canyon» I. A comparison of growth rates under different water activity stress was provided. Clear differences in growth rates were obs...
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Інститут ботаніки ім. М. Г. Холодного НАН України
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| Цитувати: | Comparison of growth rates of the filamentous fungus Aspergillus niger L. from contrasting environments / K. Selezska, L. Brodsky, L. Musatenko, S. Wasser, E. Nevo // Укр. ботан. журн. — 2007. — Т. 64, № 1. — С. 99-106. — Бібліогр.: 22 назв. — англ. |
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irk-123456789-37992009-07-13T12:00:43Z Comparison of growth rates of the filamentous fungus Aspergillus niger L. from contrasting environments Selezska, K. Wasser, S. Nevo, E. Musatenko, L. Brodsky, L. Фізіологія, біохімія та клітинна біологія рослин Eighteen strains of filamentous fungus Aspergillus niger were isolated from contrasting environments in Israel: the hypersaline Dead Sea and the temperate «Evolution Canyon» I. A comparison of growth rates under different water activity stress was provided. Clear differences in growth rates were observed under 5—20 % and 40 % volumes of the Dead Sea water (DSW). No difference was observed when we used 25— 35 % volumes of DSW. Strains from «Evolution Canyon» I grown at lower concentrations of the DSW revealed higher growth rates, while strains from the DSW demonstrated higher activity under higher stress (40 % volumes of DSW). Therefore, the following conclusion can be made: strains of A. niger isolated from the DSW are more adapted to stress associated with low-water activity. Сравнение темпов роста спор изолятов Aspergillus niger L. из контрастных по солености условий существования: гиперсоленых — Мертвого моря и умеренных — Европейского и Африканского склонов "Эволюционного каньона" (Израиль) дало возможность сделать вывод о большой адаптированности спор изолятов A. niger, выделенных из воды Мертвого моря, к стрессу, ассоциированному с низкой активностью воды. Порівняння темпів росту ізолятів Aspergillus niger з контрастних умов існування: гіпер-солоних — Мертвого моря і помірних — "Еволюційного каньйону" (Ізраїль) дало можливість дійти висновку про більшу адаптованість ізолятів A. niger, виділених з води Мертвого моря, до стресу, асоційованого з низькою активністю води. 2007 Article Comparison of growth rates of the filamentous fungus Aspergillus niger L. from contrasting environments / K. Selezska, L. Brodsky, L. Musatenko, S. Wasser, E. Nevo // Укр. ботан. журн. — 2007. — Т. 64, № 1. — С. 99-106. — Бібліогр.: 22 назв. — англ. 0372-4123 http://dspace.nbuv.gov.ua/handle/123456789/3799 uk Інститут ботаніки ім. М. Г. Холодного НАН України |
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Фізіологія, біохімія та клітинна біологія рослин Фізіологія, біохімія та клітинна біологія рослин |
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Фізіологія, біохімія та клітинна біологія рослин Фізіологія, біохімія та клітинна біологія рослин Selezska, K. Wasser, S. Nevo, E. Musatenko, L. Brodsky, L. Comparison of growth rates of the filamentous fungus Aspergillus niger L. from contrasting environments |
| description |
Eighteen strains of filamentous fungus Aspergillus niger were isolated from contrasting environments in Israel: the hypersaline Dead Sea and the temperate «Evolution Canyon» I. A comparison of growth rates under different water activity stress was provided. Clear differences in growth rates were observed under 5—20 % and 40 % volumes of the Dead Sea water (DSW). No difference was observed when we used 25— 35 % volumes of DSW. Strains from «Evolution Canyon» I grown at lower concentrations of the DSW revealed higher growth rates, while strains from the DSW demonstrated higher activity under higher stress (40 % volumes of DSW). Therefore, the following conclusion can be made: strains of A. niger isolated from the DSW are more adapted to stress associated with low-water activity. |
| format |
Article |
| author |
Selezska, K. Wasser, S. Nevo, E. Musatenko, L. Brodsky, L. |
| author_facet |
Selezska, K. Wasser, S. Nevo, E. Musatenko, L. Brodsky, L. |
| author_sort |
Selezska, K. |
| title |
Comparison of growth rates of the filamentous fungus Aspergillus niger L. from contrasting environments |
| title_short |
Comparison of growth rates of the filamentous fungus Aspergillus niger L. from contrasting environments |
| title_full |
Comparison of growth rates of the filamentous fungus Aspergillus niger L. from contrasting environments |
| title_fullStr |
Comparison of growth rates of the filamentous fungus Aspergillus niger L. from contrasting environments |
| title_full_unstemmed |
Comparison of growth rates of the filamentous fungus Aspergillus niger L. from contrasting environments |
| title_sort |
comparison of growth rates of the filamentous fungus aspergillus niger l. from contrasting environments |
| publisher |
Інститут ботаніки ім. М. Г. Холодного НАН України |
| publishDate |
2007 |
| topic_facet |
Фізіологія, біохімія та клітинна біологія рослин |
| url |
http://dspace.nbuv.gov.ua/handle/123456789/3799 |
| citation_txt |
Comparison of growth rates of the filamentous fungus Aspergillus niger L. from contrasting environments / K. Selezska, L. Brodsky, L. Musatenko, S. Wasser, E. Nevo // Укр. ботан. журн. — 2007. — Т. 64, № 1. — С. 99-106. — Бібліогр.: 22 назв. — англ. |
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2025-07-02T07:02:19Z |
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| fulltext |
ISSN 0372-4123. Óêð. áîòàí. æóðí., 2007, ò. 64, ¹ 1 99
Ô³ç³îëîã³ÿ, á³îõ³ì³ÿ
òà êë³òèííà á³îëîã³ÿ ðîñëèí
K. SELEZSKA1,2, L. BRODSKY2,
L. MUSATENKO1, S. WASSER1,2
AND E. NEVO2
1 M.G. Kholodny Institute of Botany,
National Academy of Sciences of Ukraine
Tereshchenkivska St., 2, Kyiv, 01601, Ukraine
2Institute of Evolution, University of Haifa
Mt. Carmel, 31905 Haifa, Israel
COMPARISON OF GROWTH RATES
OF THE FILAMENTOUS FUNGUS
ASPERGILLUS NIGER L. FROM
CONTRASTING ENVIRONMENTS
K e y w o r d s: growth rate, Aspergillus niger, stress, water activity
Abstract
Eighteen strains of filamentous fungus Aspergillus niger were
isolated from contrasting environments in Israel: the
hypersaline Dead Sea and the temperate «Evolution Canyon»
I. A comparison of growth rates under different water activity
stress was provided. Clear differences in growth rates were
observed under 5—20 % and 40 % volumes of the Dead Sea
water (DSW). No difference was observed when we used 25—
35 % volumes of DSW. Strains from «Evolution Canyon» I
grown at lower concentrations of the DSW revealed higher
growth rates, while strains from the DSW demonstrated
higher activity under higher stress (40 % volumes of DSW).
Therefore, the following conclusion can be made: strains
of A. niger isolated from the DSW are more adapted to stress
associated with low-water activity.
© K. SELEZSKA,
L. BRODSKY,
L.I. MUSATENKO,
E. NEVO,
S.P. WASSER, 2007
ÓÊÐÀ¯ÍÑÜÊÈÉ
ÁÎÒÀͲ×ÍÈÉ
ÆÓÐÍÀË
ISSN 0372-4123. Ukr. Botan. Journ., 2007, vol. 64, ¹ 1100
Introduction
Aspergillus niger van Tieghem is one of the most common filamentous fungus reported
from a great variety of natural habitats. It was found in different soils, food, and indoor
and outdoor environments. A. niger is the third commonest species associated with
invasive pulmonary aspergillosis and aspergilloma [9, 13]. It is also cultured for the
industrial production of some chemical compounds. Various strains of A. niger are used
in the industrial preparation of citric acid (E330), gluconic acid (E574), and the enzymes
glucoamylase and α-galactosidase (Beano®, Bean-zyme™). In Israel, Aspergillus niger
was documented from all soils that were subjected to mycological investigations [4—
7, 22]. Thus, A. niger can be easily isolated from a wide range of habitats.
The present study is focused on Aspergillus niger isolated from the hypersaline
and temperate environments (the Dead Sea water and «Evolution Canyon» (EC)
I). We comparatively investigated natural populations of A. niger under different
stress, associated with low-water activity (aw) on local (opposite slopes of «Evolution
Canyon» I) and regional («EC» I vs DSW) scales.
Evolutionary biology focuses a lot on how genetic diversity is maintained in
nature. Stressful conditions can bring higher rates of mutation and recombination,
which are very important in the evolution process [11, 19]. Stress-induced genetic
variation may serve as an important source of material for natural selection to
produce novel adaptations, and differential selection regimes in natural populations
may cause complex spatial variation in population structure, and in the level and
spectrum of genetic variation [8, 11]. Micro- and macrosite ecological contrasts
are excellent critical tests for evaluating biodiversity patterns and dynamics and for
assessing the relative importance of different forces in evolution [16].
Materials and methods
Study sites
(I) «Evolution Canyon» I is located at lower Nahal Oren, Mt Carmel National
Park, Israel (32° 43’N, 34° 58’E). «EC» I has two slopes: the south-facing (tropical,
xeric, «African», AS) and north-facing (temperate, mesic, «European», ES) slopes;
the distance between slopes is 100 m at the bottom and 400 m at the top. The
opposite slopes of lower Nahal Oren, designated «Evolution Canyon», display
dramatic biotic contrasts. Higher solar radiation on the south-facing slope (up to
600 %) makes it warmer, drier, and spatiotemporally more heterogeneous than the
north-facing slope. Consequently, local biodiversity differentiation across several
hundred meters displays global patterns of divergence [14, 15].
(II) The Dead Sea is located in the Syrian-African rift valley between Israel
and Jordan. The climate is very arid, with a mean annual rainfall near 60 mm and
mean temperatures of the hottest and coldest months from 32—35 °C and 14—
16 °C, respectively [1]. The DSW contains 340 g/l total dissolved salts. It has very
aw of < 0.669 with a pH that is relatively low (5.5—6.0) [12]. The increased drying
of the DS can be observed nowadays. Divalent cations (Mg+2 and Ca+2) now
ISSN 0372-4123. Óêð. áîòàí. æóðí., 2007, ò. 64, ¹ 1 101
dominate over monovalent cations (Na+ and K+). Cl- makes up 99 % of the anion
sum. Mycological studies of the water samples from the DS revealed soil mycobiota
comprising 77 species from 26 genera [17].
Sampling
(I) «Evolution Canyon» I
Fourteen soil samples from «EC» I were taken from each of the 6 stations (3 stations
on each slope from the AS and ES). Samples were taken from shady niches under
trees (ES) and in sunny open niches (AS), from the upper soil layer (on 1—5 cm
depth). Altogether, 84 samples were collected in spring (April) 2005.
(II) The DS water — from its surface and depth (50 m and 250 m)
Samples were collected in autumn (October) 2005, four times at seven different
sites along the western shore of the Dead Sea. They were withdrawn from near-
shore aquatic localities and from deep and surface water from the center of the
sea, about 8 km northeast of En Gedi («En Gedi 320»). Samples were collected by
means of Go-Flo sampling bottles. Afterwards, water for the experiments was
collected the same way four times per year.
Fungal isolation
Aspergillus niger strains were isolated using the soil dilution plate method [3].
The dilution for the samples was 1:10 by weight. Ten grams of each soil sample
taken were suspended in sterile water. For isolation of fungi from the DSW, 2 ml
of the DSW (from the depth and surface) were poured into Petri dishes and mixed
by rotation with molten agar media (MEA).
Eighteen strains were taken for this experiment. Six are from the middle station
of the AS (from sunny, opened niches); six from the middle station of the ES (from
shady niches, under trees and bushes), and six from the DSW: four from a depth
(50 m and 250 m) and 2 froma surface distant from the shore.
Experimental design
The linear growth rate on solid substrate, e.g., agar medium, is a good
approximation of biomass increase in liquid culture. In the present experiment,
strains were grown on Petri dishes with 10—12 ml of glucose yeast extract agar (GY)
(10 g/L glucose, 1 g/L yeast extract, 20 g/L agar; pH 6.0—6.5). Agar disks (4—
5 mm in diam) were cut out from 4-day-old cultures on agar plates and inoculated
at the center of Petri dishes with GY (35 g/L agar), prepared with different
concentrations of the (DSW); each strain in two repetitions. Volumes of the DSW
differed from 0 % till 80 %, with the step of 5 %. Water activity was modified with
calculated amounts of the DSW and distilled water. Water activity of the DSW is
0.669 and aw of the distilled water is 0.99. Calculations were made according to
the following formula:
aw = ((100 - X) • 0.99 + 0.669 X) / 100
aw — final water activity; X — used volume of the Dead Sea water
ISSN 0372-4123. Ukr. Botan. Journ., 2007, vol. 64, ¹ 1102
In the case of the medium prepared without DSW, MEA (Malt Extract Agar)
(30 g/L Malt extract, 3 g/L peptone, 15 g/L agar; pH 6.0—6.5) was used. For the
media prepared with 10 % volumes of the DSW, both GY and MEA were used.
Measurements of the mycelium growth were recorded in two orthogonal
directions every 24 hours until the Petri plates were completely colonized or if the
log-phase was observed.
Data analysis
Under the same salinity treatment the divergence between species specific
cumulative distributions was checked by the non-parametric Kolmogorov-Smirnov
test. The pvalue of the test could be detected with the following statistic formula:
max( ( ) ( ))
i i
i
N S x F x⋅ −
that has Kolmogorov distribution. N — number of thresholds in the series;
i th
x i−
threshold of growth rates; ( )
i
S x — fraction of rates of the first distribution that are
less than threshold
i
x ; ( )
i
F x — fraction of rates of the second distribution that are
less than threshold
i
x .
Results
The present experiment is the first of its kind in which ecological comparisons have
been made between strains of fungi isolated from the Dead Sea and «Evolution
Canyon» I.
Data analysis showed that the minimum water-activity limited growth was 0.779.
After two months of incubation at 30 °C, none of the strains performed growth in
the range of 0.779—0.728 aw (65 %—80 % of volumes of DSW). At the range aw
0.845—0.796 (45 %—60 % of volumes of DSW) colonies grew very slowly and no
measurements could be obtained.
As the control we took aw — 0.99 and fungus isolated from the DSW demonstrated
higher growth rate compared with the «EC» I strains. Due to some methodological
problems, A. niger strains at the control were grown on the MEA instead of GY. In
order to be sure that the difference in the strains’ behavior was not influenced by the
type of media used in the experiment, the experiment with 10 % volumes of the DSW
(aw 0.958) was repeated with MEA. Results were the same from those with GY.
Optimal growth for all strains was observed at 0.974 water activity (aw) — 5 %
of volume of DSW (Fig. 1). Strains from different environments (AS, ES, and DSW)
demonstrated a significant difference between their growth rates on the medium
with that of water activity. The same difference was observed at 0.958, 0.942, and
0.926 aw — 10 %, 15 % and 20 % of DSW volume correspondingly. On the media
with these salinities, fungus from «Evolution Canyon» I obviously grew faster than
strains from the Dead Sea.
A different pattern was observed on the media with higher volumes of DSW.
At water activities of 0.91—0.878 (25 %—35 % volumes of the DSW), no significant
difference between the strains’ behavior was observed. Still, when aw reached 0.862
ISSN 0372-4123. Óêð. áîòàí. æóðí., 2007, ò. 64, ¹ 1 103
Figure 2. Growth rate of A. niger strains from African and European slopes of «Evolution Canyon»
I and the Dead Sea on the GY medium with 40 % volume of DSW (aw 0.862)
Figure 1. Growth rate of A. niger strains from African and European slopes of «Evolution Canyon»
I and the Dead Sea on the GY medium with 5 % volume of DSW (aw 0.974)
5 % (AS,ES vs. DSW) KS pvalue < 0.0000001
0
0.2
0 .4
0 .6
0 .8
1
1.2
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G r o w t h r a t e p e r d a y
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(40 % volume of the DSW), salt-tolerant strains from the Dead Sea started to show
higher growing rates (Fig. 2). These factors are pretty good indicators for the
appropriateness of media.
ISSN 0372-4123. Ukr. Botan. Journ., 2007, vol. 64, ¹ 1104
Discussion
Experimental studies on the combined influence of water activity and temperature
on growth and spore production of Aspergillus niger strains [2, 18] demonstrated
that the studied strains grew optimally at aw 0.97—0.98.
Comparative investigation of the survival rate of conidia of the A. niger strains
from the northern «EC» II (Upper Galilee) and the DS shore in different
concentrations of the DS water showed 1.4—2.5-fold higher spore viability of the
DS shore strains. Therefore, the potential for the introduction of foreign fungal
species into the Dead Sea is very low. Examining and comparing species isolated
from the Dead Sea to the same species from other environments can answer the
question as to what extent have the species isolated from the Dead Sea adapted to
its habitat, hypersaline brine [10].
This is the first study where such a complicated mixture with large amounts of
different chemical elements as DSW was used for adapting aw. Previously, in all
similar experiments, the water activity of the medium was modified with calculated
amounts of the non-ionic solute glycerol and the ionic solute NaCl. The use of
glycerol to modify media water availability produced a higher growth rate than with
NaCl, probably because glycerol can be utilized as a carbon source and can act
directly as a compatible solute. In contrast, high concentrations of NaCl can be
toxic, and this may explain the differential growth patterns observed [18]. These
studies showed that the wild-type strains grew optimally at 0.95 aw on glycerol-
modified media, but optimally at 0.99 aw when NaCl was used [18]. All the strains
grew in CYE (Czapek Yeast Extract) medium from 0.86 to 0.99 aw (when glycerol
was used). The germination of A. niger strains has been reported at 0.77 aw [20] and
0.80 aw. The slow growth of A. niger isolates was reported at 0.76 aw on malt extract
agar (MEA) [22], whereas in another study the minimum water-activity limit for
growth was 0.82 using the same culture medium [18]. The water of the Dead Sea
has a very complicate structure. Therefore, it was impossible to predict fungal growth
rates on the medium prepared with different volumes of the DSW.
Nevertheless, our data correlate good with the abouve said. Optimal growth
for this fungus was not 0.99 aw, but lesser water activity — 0.974 aw (5 % volumes
of DSW). Limiting growth aw was 0.779 (65 % volumes of DSW). Thus, in spite of
extremely high concentration of NaCl in the DSW brine, fungal growth is influenced
not only by this salt. Therefore, media adjusted with DSW allow A. niger to grow
with the rate that is in between, NaCl and glycerol adjusted media.
Results of the comparison between different populations («<EC» I vs DSW)
seemed confusing. Low concentrations of DSW (5—20 % volumes of DSW; 0.974—
0.926 aw) gave advanatge to «EC» isolates, growth in media with moderate stress
(25—35 % volumes of DSW; 0.91—0.878 aw) didn’t show any differences in strains’
behavior. Nevertheless, when aw reached 0.862 (40 % volumes of DSW), population
isolated from DSW began to show higher growth rate.
ISSN 0372-4123. Óêð. áîòàí. æóðí., 2007, ò. 64, ¹ 1 105
Two hypotheses could be used to explain such interesting behavior. The first is
that low concentrations of the Dead Sea water do not activate salt-resistant pathways
in the salt-tolerant strains, while high volumes do. A. niger is considered to be a
xerotolerant species. Therefore, high-water activity is a bit stressful for A. niger. Due
to the constant salt stress of the habitat, to which strains from the Dead Sea are
subjected, A. niger are more resistant to the stress associated with low-water activity.
The second explanation could be in the different pathways that the DSW strains
and «EC»s strains use to resist osmotic stress. DSW strains are more resistant to
stress associated with water availability. Probably, the pathway that is used by DSW
strains gives A. niger advantages in surviving under salt stress. Any of these hypotheses
support the suggestion that Aspergillus niger from the Dead Sea water is a salt-tolerant
species which has some genetic advantages over the A. niger from «Evolution
Canyon» I.
In any case, future investigations will be provided to confirm or disprove each
of these hypotheses.
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Submitted 05.09.2006
Ê. Ñåëåçñüêà1,2, Ë. Áðîäñüêèé2, Ë. Ìóñàòåíêî1, Ñ. Âàññåð1,2 ,Å. Íåâî2
1 ²íñòèòóò áîòàí³êè ³ì. Ì.Ã. Õîëîäíîãî ÍÀÍ Óêðà¿íè, ì. Êè¿â
2 ²íñòèòóò åâîëþö³¿, Óí³âåðñèòåò Õàéôà, ²çðà¿ëü
ÒÅÌÏÈ ÐÎÑÒÓ ²ÇÎËßҲ ASPERGILLUS NIGER
Ç ÊÎÍÒÐÀÑÒÍÈÕ ÓÌΠ²ÑÍÓÂÀÍÍß
Ïîð³âíÿííÿ òåìï³â ðîñòó ³çîëÿò³â Aspergillus niger ç êîíòðàñòíèõ óìîâ ³ñíóâàííÿ: ã³ïåð-
ñîëîíèõ — Ìåðòâîãî ìîðÿ ³ ïîì³ðíèõ — «Åâîëþö³éíîãî êàíüéîíó» (²çðà¿ëü) äàëî ìîæ-
ëèâ³ñòü ä³éòè âèñíîâêó ïðî á³ëüøó àäàïòîâàí³ñòü ³çîëÿò³â A. niger, âèä³ëåíèõ ç âîäè Ìåð-
òâîãî ìîðÿ, äî ñòðåñó, àñîö³éîâàíîãî ç íèçüêîþ àêòèâí³ñòþ âîäè.
Ê ë þ ÷ î â ³ ñ ë î â à: ð³ñò, Aspergillus niger, ñòðåñ, âîäíà àêòèâí³ñòü
K. Ñåëåçñêàÿ1,2, Ë. Áðîäñêèé2, Ë. Ìóñàòåíêî1, Ñ. Âàññåð1,2,Ý. Íåâî2
1 Èíñòèòóò áîòàíèêè èì. Í.Ã. Õîëîäíîãî ÍÀÍ Óêðàèíû, ã. Êèåâ
2 Èíñòèòóò ýâîëþöèè, Óíèâåðñèòåò Õàéôà, Èçðàèëü
ÒÅÌÏÛ ÐÎÑÒÀ ÈÇÎËßÒÎÂ ASPERGILLUS NIGER
ÈÇ ÊÎÍÒÐÀÑÒÍÛÕ ÓÑËÎÂÈÉ ÑÓÙÅÑÒÂÎÂÀÍÈß
Ñðàâíåíèå òåìïîâ ðîñòà èçîëÿòîâ Aspergillus niger èç êîíòðàñòíûõ óñëîâèé ñóùåñòâîâà-
íèÿ: ãèïåðñîëåíûõ — Ìåðòâîãî ìîðÿ è óìåðåííûõ — «Ýâîëþöèîííîãî êàíüîíà» (Èçðà-
èëü) äàëî âîçìîæíîñòü ñäåëàòü âûâîä î áîëüøåé àäàïòèðîâàííîñòè èçîëÿòîâ A. niger,
âûäåëåííûõ èç âîäû Ìåðòâîãî ìîðÿ, ê ñòðåññó, àññîöèèðîâàííîìó ñ íèçêîé àêòèâíîñ-
òüþ âîäû.
Ê ë þ ÷ å â û å ñ ë î â à: ðîñò, Aspergillus niger, ñòðåññ, âîäíàÿ àêòèâíîñòü
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