Helena Kubicka*, Tomasz Kubel* THE INFLUENCE OF CHOSEN

OchrOna ŚrOdOwiska i ZasObów naturalnych
nr
43, 2010 r.
Helena Kubicka*, Tomasz Kubel*
THE INFLUENCE OF CHOSEN BIOGENIC ELEMENTS
ON GROWTH OF RYE INBRED LINES
DZIAŁANIE WYBRANYCH PIERWIASTKÓW BIOGENNYCH
NA WZROST LINII WSOBNYCH żYTA (SECALE CEREALE L.)
Słowa kluczowe: cynk, jony, linie wsobne, mangan, żyto.
Key words: inbred lines, ions, manganese, rye, zinc.
Cynk i mangan to biogenne pierwiastki występujące powszechnie w środowisku. Ich niedobór
niekorzystnie wpływa na wzrost i rozwój roślin. W niniejszej pracy badano wrażliwość siewek
pięciu linii wsobnych żyta na zwiększoną zawartość jonów cynku i manganu w podłożu. Uzyskane wyniki wskazują, że stężenie 10-1M obu pierwiastków powoduje znaczne zahamowanie
wzrostu roślin, zarówno korzeni, jak i części nadziemnych. natomiast niższe stężenie 10-2M
jonów cynku lub manganu stymuluje wzrost u niektórych linii. najwyższa tolerancja obu pierwiastków cechowała linię 154. Oceniane linie wsobne żyta zróżnicowanie reagowały na nadmiar jonów cynku lub manganu w podłożu. Bardziej wrażliwe na działanie obu pierwiastków,
aniżeli korzenie, były części nadziemne roślin. Zastosowany w doświadczeniu kwas askorbinowy w kombinacji z cynkiem lub manganem w stężeniu 10-1M powodował zmniejszenie stresu spowodowanego nadmiarem tych pierwiastków w podłożu u niektórych linii.
1. INTRODUCTION
Biogenic elements such as manganese and zinc are parts of many enzymes and are
necessary for development and growth of plants. They are abundant in soil, and their main
natural source is bedrock [CICAD 2004, Broadley et al. 2007].
Levels of anthropogenic zinc and manganese in environment have been rising along
with increasing industrialization. In the nineties emission levels of zinc were close to 2700
* Doc. dr hab. Helena Kubicka, mgr Tomasz Kubel – Ogród Botaniczny – Centrum
Zachowania Różnorodności Biologicznej PAN, ul. Prawdziwka 2, 02-973 Warszawa;
e-mail: [email protected]
65
environment by burning fossil fuels and tyres, using pesticides as well as dumping sewage
sludge and metals coated with anticorrosive
zinc Tomasz
layers [Broadley
et al. 2007].
Helena Kubicka,
Kubel
The main sources of atmospheric manganese contamination are ore mills and burning
fossil
fuels* [Hagelstein
contamination
is mainly
by penetration
kilotons
year-1 while 2009].Manganese
two millennia ago itsoil
was
only 10 kilotons
. The main
areas of
* year-1caused
contamination
are around
mining
sitesand
[Pugh
et al. 2002].
Zinc is also
released
into
ofzinc
manganese
compounds
fromzinc
waste
yards
through
contaminated
rainfalls
[CICAD
environment
by burning
fuels and contamination
tyres, using pesticides
as well
dumping sewage
2004].
A source
of fossil
manganese
is also
theas gasoline
additive
sludge and metals coated with anticorrosive zinc layers [Broadley et al. 2007].
metylocyclopentadienyl manganese tricarbonyl (MMT) that is used in some countries to
The main sources of atmospheric manganese contamination are ore mills and burning
increase
fuel octane
rating2009].
[Kitazawa
et al. 2002].
fossil fuels
[Hagelstein
Manganese
soil contamination is mainly caused by penetra-
tion ofPlant
manganese
from
yards
and through
contaminated
rainfallsspecies
[CItolerancecompounds
to deficiency
of waste
biogenic
compounds
differs
not only between
A source of manganese contamination is also the gasoline additive metylocybutCAD
even2004].
between
lines. One of zinc hyperaccumulators are plants from the Thlapsi genus and
clopentadienyl manganese tricarbonyl (MMT) that is used in some countries to increase fuel
an example of a manganese hyperaccumulator is Phytolacca americana. Inhibition of growth
octane rating [Kitazawa et al. 2002].
and disturbance
in thetomineral
uptake
and distribution
are differs
the main
of excess
Mn
Plant tolerance
deficiency
of biogenic
compounds
notsymptoms
only between
species
butZneven
between
of zincethyperaccumulators
are2007].
plants from the Thlapsi genus
and
in soil
[Lidonlines.
2001,One
Broadley
al. 2007, Mina et al.
and an example of a manganese hyperaccumulator is Phytolacca americana. Inhibition of
The objective of this research was to evaluate the influence of zinc and manganese in
growth and disturbance in the mineral uptake and distribution are the main symptoms of ex-
the growth medium on growth of rye seedlings .
cess Mn and Zn in soil [Lidon 2001, Broadley et al. 2007, Mina et al. 2007].
The objective of this research was to evaluate the influence of zinc and manganese in
the growth medium on growth
of rye seedlings.
MATERIALS
AND METHODS
Five, genetically different inbred lines (L176, 154, L29, CH7/99, L230, M353) of rye
MATERIALS
METHODS
were used in the experiment2. [Kubicka
et AND
al. 2006].
Four-day-old rye seedlings were
transferred
the Hoagland’s
with(L176,
addition
zincCH7/99,
or manganese:
the medium
Five, to
genetically
different medium
inbred lines
154,ofL29,
L230, M353)
of rye
were used
in the
experiment
[Kubicka
al. as
2006].
Four-day-old
transwithout
zinc or
manganese
addition
was et
used
the control:
10-1Mrye
andseedlings
10-2M Znwere
or Mn.
Two
ferred to the Hoagland’s medium with addition of zinc or manganese: the medium without
combinations were prepared with addition of ascorbic acid
(10-2M) -2and zinc or manganese at
-1
zinc or manganese addition was used as the control: 10 M and 10 M Zn or Mn. Two com-
-1
M. Thewith
length
of roots
and shoots
was(10
measured
seven
and fourteen
a concentration
-2
M) and after
zinc or
manganese
at
binations were10prepared
addition
of ascorbic
acid
-1
days.
The tolerance
waslength
calculated
according
to the
equation:
M. The
of roots
and shoots
was
measured after seven and foura concentration
10index
teen days. The tolerance index was calculated according to the equation:
Tolerance index 
value before metal treatment
 100%
value after metal treatment
RESULTS
3. RESULTSAND
ANDDISCUSSION
DISCUSSION
Although Zn and Mn are biogenic elements, exposition of rye inbred lines seedlings to
Although Zn and Mn are biogenic elements, exposition of rye inbred lines seedlings to
excess
ZnZn
or or
MnMn
resulted
mainly
– 4).
excess
resulted
mainlyinininhibition
inhibitionof
of seedlings
seedlingsgrowth
growth (Fig.
(Fig. 11–4).
Theshoots
shootsofoftested
testedrye
ryeinbred
inbredlines
lineswere
weremore
moresusceptible
susceptibletotoZnZnororMn
Mn
than
roots.
The
than
thethe
roots.
Distinguishedgrowth
growthinhibition
inhibition was
was observed
especially on
Distinguished
observed especially
on the
the growth
growthmediums
mediumswith
withhigher
higher
metal concentration (10-1M). The plants exposed to lower metal concentration (10-2M) devel-
metal concentration (10-1M).-2 The plants exposed to lower metal concentration (10-2M)
oped better. Manganese (10 M) stimulated growth of 7 and 14 days old seedlings of inbred
66
-2-
The influence of chosen biogenic elements on growth of rye inbred lines
developed better. Manganese (10-2M) stimulated growth of 7 and 14 days old seedlings of
line 154. Lines L29 and M353 showed high tolerance to manganese at a 10-2M -2concentrainbred line 154. Lines L29 and M353 showed high tolerance to manganese at a 10 M
tion – 7 days old seedlings were almost of the same length as the control seedlings. The bigconcentration – 7 days old seedlings were almost of the same length as the control seedlings.
gest inhibition of growth, on the medium with excess Mn and Zn, was observed in line L230
The biggest inhibition of growth, on the medium with excess Mn and Zn, was observed in line
(Fig. 1–2). Shoots of tested lines were more sensitive to Zn: they grew much shorter than
L230 (Fig. 1-2). Shoots of tested lines were more sensitive to Zn: they grew much shorter
in the lines with Mn added to the medium. Zinc concentration of 10-2M -2had less influence
than in the lines with Mn added to the medium. Zinc concentration of 10 M had less
on rye seedling growth. Lines 154, L230 and L29 had the highest tolerance index, ranging
influence on rye seedling growth. Lines 154, L230 and L29 had the highest tolerance index,
from 60% (line L29) to 80% (line 154). Some lines growing on the medium with ascorbic acid
ranging from 60% (line L29) to 80% (line 154). Some lines growing on the medium with
showed higher tolerance to Zn and Mn. Shoot increment was noted in lines: CH7/99 and
ascorbic acid showed higher tolerance to Zn and Mn. Shoot increment was noted in lines:
L230 (7 days, Zn), L154 and L176 (7 and 14 days, Mn).
CH7/99 and L230 (7 days, Zn), L154 and L176 (7 and 14 days, Mn).
120%
Tolerance index
100%
80%
60%
40%
L176
L154
L29
L230
10-2M
10-1+witC
10-1M
10-2M
10-1M
CH7/99
10-1+witC
10-2M
10-1+witC
10-1M
10-2M
10-1+witC
10-1M
10-2M
10-1+witC
10-1M
10-2M
10-1M
0%
10-1+witC
20%
M353
Inbred lines
Zn 7dni
Zn 14 dni
Fig. 1.
The influence
of zinc
the length
of shoots
7 and
dayseedlings
seedlings ofofrye
Fig. 1. The
influence
of zinc ions
on ions
the on
length
of shoots
of 7ofand
1414day
rye inbred
inbred lines in comparison to control [%]
Wpływ jonów cynku na długość kiełków 7 i 14 dniowych siewek linii wsobnych
Rys. 1. Wpływ jonów
na długość
kiełków
żyta cynku
w porównaniu
do kontroli
[%] 7 i 14 dniowych siewek linii wsobnych żyta w po-
lines in comparison to control [%]
Rys. 1.
równaniu do kontroli [%]
Tolerance index
120%
100%
80%
60%
40%
20%
L176
L154
L29
CH7/99
L230
10-2M
101+witC
10-1M
10-2M
101+witC
10-1M
10-2M
101+witC
10-1M
10-2M
101+witC
10-1M
10-2M
101+witC
10-1M
10-2M
101+witC
10-1M
0%
M353
Inbred lines
Mn 7 dni
Mn 14 dni
-3Theofinfluence
of manganese
on the of
length
of shoots
of 714and
day
Fig. 2. Fig.
The2.influence
manganese
ions on ions
the length
shoots
of 7 and
day14seedlings
of
seedlings of rye inbred lines in comparison to control [%]
rye inbred lines in comparison to control [%]
Rys. 2
Wpływ jonów manganu na długość kiełków 7 i 14 dniowych siewek linii
Rys. 2. Wpływ jonów
manganu
długość kiełków
7 i [%]
14 dniowych siewek linii wsobnych żyta
wsobnych
żyta na
w porównaniu
do kontroli
w porównaniu do kontroli [%]
The excess of both biogenic molecules had less influence on growth of roots than
shoots (Fig. 3 – 4). The biggest growth inhibition for Zn and Mn was observed at a
concentration of 10-1M. The rye inbred lines differed in their reaction to tested metals. Lines
L176 and L230 appeared to be most susceptible and line 154 most tolerant. The root tolerance
67
Helena Kubicka, Tomasz Kubel
The excess of both biogenic molecules had less influence on growth of roots than
shoots (Fig. 3–4). The biggest growth inhibition for Zn and Mn was observed at a concentration of 10-1M. The rye inbred lines differed in their reaction to tested metals. Lines L176
and L230 appeared to be most susceptible and line 154 most tolerant. The root tolerance
index of inbred line 154 exceeded 100%: at a concentration of 10-2M Zn or Mn, after 7 days
at a concentration 10-1M (Zn or Mn) and after 14 days at a concentration 10-1M with addition
of ascorbic acid. After 7 days of observation ascorbic acid lowered the stress caused by ex-
L176
154
L29Inbred
linesCH7
Inbred lines
Zn 7dni
Zn 7dni
L230
M353
10-2M
10-2M
10- 101+wit.C
1+wit.C
10-1M
10-1M
L230
10-2M
10-2M
10- 101+wit.C
1+wit.C
10-1M
10-1M
CH7
10-2M
10-2M
10- 101+wit.C
1+wit.C
10-1M
10-1M
L29
10-2M
10-2M
10- 101+wit.C
1+wit.C
10-1M
10-1M
154
10-2M
10-2M
10- 101+wit.C
1+wit.C
10-1M
10-1M
L176
10-2M
10-2M
0%
10- 101+wit.C
1+wit.C
140%
120%
140%
100%
120%
80%
100%
60%
80%
40%
60%
20%
40%
0%
20%
10-1M
10-1M
Tolerance
index
Tolerance
index
cess Zn or Mn in lines M353, L176 and 29.
M353
Zn 14 dni
Zn 14 dni
Inbred linesCH7
Inbred lines
Mn 7 dni
Mn 7 dni
M353
M353
10-2M
10-2M
10-101+wit.C
1+wit.C
10-1M
10-1M
L230
L230
10-2M
10-2M
10-101+wit.C
1+wit.C
10-1M
10-1M
CH7
10-2M
10-2M
10-101+wit.C
1+wit.C
10-1M
10-1M
L29
L29
10-2M
10-2M
10-101+wit.C
1+wit.C
10-1M
10-1M
154
154
10-2M
10-2M
10-101+wit.C
1+wit.C
10-1M
10-1M
L176
L176
10-2M
10-2M
10-101+wit.C
1+wit.C
140%
120%
140%
100%
120%
80%
100%
60%
80%
40%
60%
20%
40%
0%
20%
0%
10-1M
10-1M
Tolerance
index
Tolerance
index
Fig. 3.
The
of zinc of
ions
theon
length
of roots
of 7 of
and
14 14
dayday
seedlings
Fig.
3. influence
The influence
zinconions
the length
of roots
7 and
seedlingsofofrye
ryeinbred
Fig.
3. in comparison
The influence
zinc ions
oncontrol
the length
inbred
lines intoof
comparison
[%]of roots of 7 and 14 day seedlings of rye
lines
control
[%]to
inbred lines
in comparison
to control [%] 7 i 14 dniowych siewek linii wsobnych
3.
Wpływ
jonów
na długość
Rys. Rys.
3. Wpływ
jonów
cynku
nacynku
długość
korzenikorzeni
7 i 14 dniowych siewek linii wsobnych żyta w poRys. 3.
Wpływ
jonów
cynku
na
długość
żyta w porównaniu do kontroli [%]korzeni 7 i 14 dniowych siewek linii wsobnych
równaniużyta
do w
kontroli
[%]
porównaniu do kontroli [%]
Mn 14 dni
Mn 14 dni
Fig. 4.
The influence of manganese ions on the length of roots of 7 and 14 day seedlings
4. influence
The
of manganese
ionsthe
oncontrol
the length
of roots
7 and
dayseedlings
seedlings of rye
Fig. 4.Fig.
The
of manganese
ions on
length
of roots
of 7ofand
1414day
of
ryeinfluence
inbred
lines
in comparison
to
[%]
of
rye
inbred
lines
in
comparison
to
control
[%]
inbred
in comparison
to control
[%]
Rys.
4. lines
Wpływ
jonów manganu
na długość
korzeni 7 i 14 dniowych siewek linii
Rys. 4.
Wpływ jonów
na długość
7 i 14 dniowych siewek linii
wsobnych
żyta wmanganu
porównaniu
kontrolikorzeni
Rys. 4. Wpływ jonów
manganu
na długośćdo
korzeni
7 [%]
i 14 dniowych siewek linii wsobnych żyta
wsobnych żyta w porównaniu do kontroli [%]
w porównaniu do kontroli [%]
The results show that line 154 is the most tolerant to the excess Zn and Mn. However,
The results show that line 154 is the most tolerant to the excess Zn and Mn. However,
it has a higher tolerance to Mn than Zn. Depending on plant organs, the difference in
it has a higher tolerance to Mn than Zn. Depending on plant organs, the difference in
68 tolerance
was observed. Shoots were more susceptible to excess Zn or Mn than roots, though
tolerance was observed. Shoots were more susceptible to excess Zn or Mn than roots, though
Zn had a greater inhibiting effect. The roots of line 154 expressed especially high tolerance to
Zn had a greater inhibiting effect. The roots of line 154 expressed especially high tolerance to
The influence of chosen biogenic elements on growth of rye inbred lines
The results show that line 154 is the most tolerant to the excess Zn and Mn. However,
it has a higher tolerance to Mn than Zn. Depending on plant organs, the difference in tolerance was observed. Shoots were more susceptible to excess Zn or Mn than roots, though
Zn had a greater inhibiting effect. The roots of line 154 expressed especially high tolerance
to Zn ions. The tolerance index for the lower concentration of Zn reached 120% and was
greater than 100% for Mn. Growth of line 154 roots was stimulated by the lower concentration (10-2M) of manganese or zinc and shoot growth was stimulated by the lower concentration of manganese. This can be caused by an important role of Mn ions in the PS II
complex, an increased efficiency of photosynthesis and better distribution of minerals. According to Monnet et al. [2001] an elevated concentration of manganese caused a disorder
in the uptake of important minerals – including iron, which is necessary for plant growth and
development. It is important to notice that the tolerance of roots is higher than shoots. This
is in accordance with suggestions of Lidon [2001] about storing manganese in different plant
organs. Lower Zn tolerance is probably caused by oxidative stress induction and mineral
uptake disorders of Mn, Mg, or Fe, what is proved by Broadley et al. [2007].
To test whether Zn and Mn do induce oxidative stress, an experiment was conducted
where the growth medium was enriched with ascorbic acid and Zn or Mn at a concentration
of 10-1M. It is well known that ascorbic acid is an important dietary ingredient. Plants are rich
in this compound and it is especially abundant in chloroplasts. It mainly works as an antioxidant [Smirnoff 2000]. Because heavy metals induce oxidative stress, the higher tolerance
index was expected when plants grew on the medium enriched with ascorbic acid. The results show slightly better growth of some lines on the medium with ascorbic acid. It is true
for both 7 and 14 day-old seedlings.
Tested rye inbred lines differed in their reaction to excess Zn or Mn in the growth medium. Line 154 showed to have the highest tolerance among all tested lines. The most susceptible was line L230.
4. CONCLUSIONS
1.
Inbred lines of rye responded differently to excess of Zn or Mn in the medium.
2.
Ascorbic acid in combination with Zn or Mn at a concentration 10-1M in the medium reduced the stress caused by the excess of analized heavy metal ions.
REFERENCES
BROADLEY M.R., WHITE P.J., HAMMOND J.P., ZELKO I., LUX A. 2007. Zinc in plants.
New Phytol. 173: 677–702.
World Health Organization, Manganese and its Compounds: Environmental Aspects.
2004. CICAD.
69
Helena Kubicka, Tomasz Kubel
HAGELSTEIN K. 2009. Globally sustainable manganese metal production and use. Environ. Manage. 90: 3736–3740.
KITAZAWA M., WAGNER J.R., KIRBY M.L., ANANTHARAM V., KANTHASAMY A.G. 2002.
Oxidative Stress and Mitochondrial-Mediated Apoptosis in Dopaminergic Cells Exposed to Methylcyclopentadienyl Manganese Tricarbonyl. JPET. 302: 26–35.
LIDON F.C. 2001. Tolerance of rice to excess manganese in the early stages of vegetativegrowth. Characterisation of manganese accumulation. J. Plant Physiol. 158: 1341–
1348.
MINA Y., BOQINGB T., MEIZHENC T., AOYAMA I. 2007. Accumulation and uptake of manganese in a hyperaccumulator Phytolacca americana. Minerals Engineering 20: 188–
190.
MONNET F., VAILLANT N., VERNAY P., COUDRET A., SALLANON H., HITMI A. 2001. Relationship between PSII activity, CO2 fixation, and Zn, Mn and Mgcontents of Lolium
perenneunder zinc stress. J. Plant Physiol. 158: 1137–1144.
PUGH R.E., DICK D.G., FREDEEN A.L. 2002. Heavy Metal (Pb, Zn, Cd, Fe, and Cu) Contents of Plant Foliagenear the Anvil Range Lead/Zinc Mine, Faro, Yukon Territory. Ecotoxicol. Environ. Saf. 52: 273–279.
SMIRNOFF N. 2000. Ascorbate biosynthesis and function in photoprotection. Phil. Trans. R.
Soc. Lond. B. 355:1455–1464.
70