The effect of detergent as polluting agent on the photosynthetic activity and chlorophyll content in bean leaves

doi:10.4236/health.2010.25059

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Vol.2, No.5, 395-399 (201

doi:10.4236/health.2010.25059

0) Health

Openly accessible at/HEAL H

The effect of detergent as polluting agent on the

photosynthetic activity and chlorophyll content

in bean leaves

Branislav R. Jovanić1*, Srdjan Bojović2, Bratimir Panić1, Božidar Radenković3,

Marijana Despotović3

1Institute of Physics, Belgrade University, Zemun, Serbia; *Corresponding Author: branislav.jovanic@ipb.ac.rs

2Institute for Biological Research “Siniša Stanković”, Belgrade, Serbia

3Faculty of Organization Science, Belgrade, Serbia

Received 28 October 2009; revised 10 January 2010; accepted 12 January 2010.

ABSTRACT

The paper investigates effects of detergent for

domestic use on the photosynthetic activity and

chlorophyll content in intact bean leaves. The

plants were watered for 21 days with a solution

of domestic washing powder of 0.60 g r/l. It was

established that the activity of photosynthetic

apparatus in the plant leaf PhACNorm [%] decrea-

ses exponentially with the length of plant treat-

ment/watering. At the end of the treatment (21st

day) the activity of photosynthetic apparatus in

the dosed plant leaf was no more than 45% of

that in control plant (those which were not wa-

tered with detergent solution). With increased

plant treatment duration the changed chloro-

phyll concentration ΔChlNorm [%] rose non-linearly

in plant leaves. The highest change ΔChlNorm [%]

was observed on the 21st day and amounted to

12%.

Keywords: Chlorophyll; Detergent; Plant;

Photosynthesis; Pollution; Water

1. INTRODUCTION

High technologies and technological processes are al-

ways accompanied with products which pollute the en-

vironment to varying extents. Very few of these products

are not pollutants. This is the reason why environmental

study is becoming increasingly important for the sur-

vival of plant and animal world and ultimately of hu-

mankind itself. It should be noted that a culprit for envi-

ronmental pollution should not be sought only in out-

dated or new technologies. Sources of pollution can be,

which is often ignored, some domestic processes in ur-

ban environment, such as food preparation or personal

hygiene. The subject of this study is an investigation into

water pollution resulting from everyday domestic hygi-

enic procedures. There are hardly any households with-

out a washing machine connected with pipes to sewage

for discharge of used water with detergent. Used water is

discharged into the nearest river or a lake and together

with it detergent. Undoubtedly, with time detergent con-

centration in the river/lake goes up and the direct conse-

quence of this is a dramatic change in the biosphere.

Significant pollution in ground water was observed in

Tehran [1]. There are other numerous examples of pol-

luted rivers and lakes with industrial detergents. For

example it was found out that the Caspian Sea waters

and Volga Terek, and Sulak rivers were extremely pol-

luted with a high detergent concentration [2]. The Аsa

River in Nigeria is dramatically polluted with industrial

detergents [3]. Likewise the Coastal Zone of the Sea of

Okhotsk and Avacha Bay are polluted with detergents

[4]. Regardless whether it is river or lake water, it is used

in gardening for watering vegetables used for human

consumption. Doubtless, this water will cause changes in

vegetables which can have an adverse effect on people

eating them.

2. MATERIAL AND METHODS

2.1. Methods

Bean (Phaseolus vulgaris L.) seeds were grown for 3

weeks. They were placed in a growth chamber adjusted

to the identical growing conditions (humidity, lighting,

temperature, nutrition of soil). The seedlings were wa-

tered daily during all investigated periods with tap water.

After this period the plants were divided in two groups:

control and stress. Growing conditions were also identi-

cal for control and stressed samples and the only differ-

ence was the presence or absence of detergent in soil.

Concentration of domestic use detergent in the water

B. R. Jovanić et al. / HEALTH 2 (2010) 395-399

396

Openly accessible at/HEALH

used for watering was 0.60 g r/l. The control samples

were watered with water without detergent. Therefore,

any differences between fluorescent spectra and fluores-

cence induced curve for stressed versus control plants

could only be the result of the presence of detergent.

In all experiments the plant’s leaves remain intact

(cutting is an additional stress) and we could make sev-

eral measurements on the same plant at any time. In fur-

ther text, the subscripts (S) and (C) will denote the

stressed or control (nonstressed) conditions, respectively.

Photosynthetic activities were determined using well

known Kautczhy method. Photosynthesis measurements

of light-adapted plants, non-destructive measurements of

potential quantum yield (Fv/Fm), were taken using a

photodiode connected with 14bit AD card for collecting

modulated fluorescence. In front of the photodiode was

placed interference filter 690 nm  5 nm. Excitation

source for fluorescence induction curve was high inten-

sity LED 470 nm/12 mW. The beans were transferred to

a darkened laboratory for 5 minutes for adaptation be-

fore measuring fluorescence kinetics at 690 nm [5]. Each

point in Figure 1 and Figure 2 represents mean value of

15 measurements on different leaves which completely

satisfies the demand for value measurement and calcula-

tion precision [ΔChl(a,b) and PhAcNorm] to be higher

than 1% [6].

For excitation the leaves and obtained fluorescence

spectra the leaf was irradiated by high power LED (470

nm/12 mW). The fibre inlet was placed 15 mm from the

leaf surface. The LED beam diameter on the leaves was

~10 mm. The LED light beam was always directed onto

the upper surface of the leaves at a 90 angle of the leaf

axis, and the optical fibre was set at a 90 angle to the

leaves’ surface on the same side. Fluorescence emitted

radiation from intact leaf was collected and directed

through an optical fibre (N.A. of 0.22 and 1000 µm di-

ameter) that was coupled to a portable 2048-element

CCD spectrometer (AVANTES 1000 PC). Data collec-

tion and spectrum processing were conducted in real

time with microcomputer and commercial software OOI

Base (AVANTES Inc.). The results for each groups of

bean represent an average of the measurement of ten

leaves. Fluorescence measurements took 1.5 min for

each measured leaf.

Chlorophyll content Chl(a,b) in bean leaf was deter-

mined from experimentally obtained fluorescence spec-

tra and well known relation between chlorophyll content

and fluorescence intensity ration. It is well known that

the ratio of the two chlorophyll fluorescence peaks

(F730/F690) in leaves correlates well with amount of chlo-

rophyll content in the bean plant leaves [7]. Therefore

chlorophyll content was determined using: a) Fluores-

cence bean spectra and b) relation between chlorophyll

content and the fluorescence intensity ratio FIR defined

as the ratio of the fluorescence intensity measured at

730 nm (F730) and 690 nm (F690) FIR = FIR690/FIR730.

For the bean linear correlation (r2 = 0.954) between

chlorophyll(a,b) content and FIR is Chl(a,b) = 42.93 –

12 FIR and was obtained from literature data [5]. In or-

der to eliminate errors which can appear due to differ-

ences in individual chlorophyll contents in different bean

samples we introduced a relative change of the chloro-

phyll(a,b):





UVCUV C

Chl(a, b)Chl(a, b)Chl(a, b)12FIRFIR 

FIRC and FIRUV are the fluorescence intensity ratio for

control plant which were not exposed to the UV radia-

tion and the plant which were exposed to the UV radia-

tion. This method was successful in the experiment with

a pumpkin exposed to the γ-nuclear radiation [8].

Figure 1. Change in chlorophyll content ΔChl(a,b) in the bean

leaves during irrigation with water contain detergent.

Figure 2. Change of the normalized photosynthesis activity

PhAcNorm in the bean leaves as function of time exposition to

radiation with detergent.

B. R. Jovanić et al. / HEALTH 2 (2010) 395-399

397

3. RESULTS

To avoid effects of plant age on chlorophyll concentra-

tion the control and dosed plants were of the same age,

To avoid determining real chlorophyll concentration,

relative chlorophyll concentration reduction was deter-

mined: chlorophyll concentration in the dosed plants was

marked ΔChlNorm as opposed to the control samples of

the same age which were not treated with detergent. The

reduction in chlorophyll concentration in the marked

values ΔChlNorm over detergent treatment time is shown

in Figure 1. Figure 1 shows that with time relative

change of chlorophyll concentration increases quickly to

reach its peak on the seventeenth day at about 11.5%.

Obviously in this period the detergent has adverse effect

on chlorophyll by destroying it constantly. After this the

relative change of chlorophyll concentration remains

steady at the same value. This pattern can be explained

with a hypothesis that the plant adapted to the given un-

favourable conditions and developed a mechanism to

maintain the reduced chlorophyll concentration.

When determining photosynthetic activity PhAc of

photosynthetic apparatus in a plant leaf, equally when

determining reduction in chlorophyll concentration in

control and dosed plants, the plants were of the same age

to avoid the effect of plant age. To avoid determining

absolute photosynthetic activity, relative reduction of

PhAc was determined. The PhAc in detergent treated

plant was marked PhAcNorm to distinguish them from

control samples of the same age which were not treated

with detergent. The obtained changes in marked values

PhAcNorm over the detergent treatment time are shown in

Figure 2. Unlike chlorophyll concentration, photosyn-

thetic activity PhAc constantly, exponentially decreases

to reach only 45% of initial activity on the 21st day. This

pattern indicates that this water has adverse effects on

plants.

4. DISCUSSION

Scientific papers offer data indicating different effects of

different detergents on plants. In most cases detergents

have adverse effects on plant pigments and morphology

and inhibit metabolic processes. It was found the toxic

effect of sodium dodecyl sulfate (SDS) and the house-

hold synthetic detergents (HSDs) Kristall and Tix (0.1, 1,

and 10 mg/l) on the diatom Alga Thalassiosira pseu-

donana [9]. By the presence in water of detergents for

wool domestic washings the native enzyme lost 50% of

activity after 20 min of incubation [10]. The effect of

detergent on plants varies depending on how the plant is

exposed to it. For example, when bean was watered with

0.01% (w/v) solution the nondenaturing, zwitterionic

detergent [3-{3-cholamidopropyl}-dimethylammonio]-

1-propane-sulfonate) 0.01% (w/v) it induced root hair

deformation [11]. Also, in mungbean (Vigna radiata)

seeds synthetic detergent induced reduction in dehydro-

genase activity [12]. The leaves completely lost their

turgor pressure and displayed chlorosis when they are

treated with detergents [13]. The biophysical character-

istics of the membrane were changed after detergent

(Brij 58) treatment [14]. Detergent inhibited growth,

metabolic activity took place only for 1 to 5 days, after

which metabolic activity also ceased [15]. Also, high

concentration of detergent might cause loss of the native

configuration of β-carotene [16]. Cell growth and fission

inhibition, as well as morphological changes and block-

ing of chlorophyll a synthesis, were recorded at 10 mg/L

concentration of detergent of household synthetic ab-

stergent (HSA) [17]. When β-carotene is treated with a

high concentration of detergent [18] this might cause

loss of the native configuration [18]. Higher plant thyla-

koid membranes can be fractionated with various deter-

gents [19].

As can be seen from our data chlorophyll is sensitive

to detergent which tallies with other research results. The

plant treated with water content detergents showed high

inhibitory effect on chlorophyll content in sunflower

leaves [20]. The aggregation of chlorophyll is partly

inhibited by detergent Triton X-100 [21]. In addition to

reducing its concentration, detergents have other effects

on chlorophyll. Studies on light harvesting complexes

LHCs show that detergent-induced dissociation of LHCs

and caused decline in bonding Chl b and Chl a [22]. The

results on pigment-protein complexes of Pisum sativum

thylakoids treated with detergent Triton X-100 and

n-octyl β-D-glucopyranoside show that reversible disso-

ciation of pigment-protein complexes occur [23]. Cell

growth and fission inhibition on cryptophytic alga

Chroomonas salina (Wils.) Butch. (Cryptophyta), as well

as morphological changes and blocking of chlorophyll a

synthesis, were recorded at 10 mg/L concentration of

household synthetic abstergent (HSA) «Tix» [17] .

In addition to the above discussed effect of detergent

on chlorophyll, it was to be expected that similar possi-

bly even identical effect will be observed on photosyn-

thesis. Detergent-induced reversible denaturation of the

photosystem reaction [24]. Detergents have strong effect

on the fluorescence properties of the light-harvesting

complexes of photosystem II [25]. Detergent (Triton

X-100) has effect on relaxation dynamics of photosys-

tem II [26]. Some results have shown that low concen-

tration of nonionic detergent Triton X is sufficient for

saturation of photosynthesis in terrestrial higher plants

[27]. Even a short exposure to detergent effects causes

extensive changes in photosynthesis. For example ex-

posing for 10 min in water containing a few drops of

liquid detergent induce the increase of photosyntesis

[28]. It was concluded that detergent (Triton X-100)

causes damage of the donor part of photosystem 2 in

B. R. Jovanić et al. / HEALTH 2 (2010) 395-399

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isolated chloroplasts [29]. Detergent treatment of the

membranes resulted in loss of PS I activities [30].

Non-ionic detergent n-dodecyl-α, D-maltoside cause

disintegration of the potosystem II (PS II) into separated

PS II in stacked and unstacked thylakoid membranes

from spinach [31]. The addition of the detergent Triton

X-100 to the ‘chromatophore’ facilitated the photooxida-

tive destruction of the antenna BChl [32]. In addition to

inhibition of activities PHII detergent can cause mor-

phological changes of these centres. Detergent treatment

of stacked thylakoid or BBY membranes usually gives to

PS II–LHC II varying size [31].

5. CONCLUSIONS

Detergents in the water for watering plants have adverse

effect. In the bean plants which were watered with de-

tergent water, significant changes were observed. Chlo-

rophyll concentration dropped by 12%. The activity of

photosynthesis apparatus in leaves decreased by around

45%.

6. ACKNOWLEDGEMENTS

This work was supported by Grants No. 141007 of MNRS. Authors

want to thank Mr. P. Hiddinga on kindness in supporting equipments.

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