Plant Adaptation to Oil Stress

1067-4136/04/3505- © 2004

MAIK “Nauka

/Interperiodica”0290

Russian Journal of Ecology, Vol. 35, No. 5, 2004, pp. 290–295. Translated from Ekologiya, No. 5, 2004, pp. 330–335.Original Russian Text Copyright © 2004 by Chupakhina, Maslennikov.

To protect the environment from various kinds ofpollution is an urgent problem. As soil resources arelimited, it is especially important to protect the soilfrom pollution with oil and petroleum products spilledin the course of production, transportation, storage,and utilization. Petroleum pollutants impair hydro-logic conditions and physical properties of the soil,drastically reduce the contents of movable nitrogenand phosphorus compounds, and have a toxic effecton plants, causing the destruction of chlorophylls andcarotenoids (Golodyaev and Ivanov, 1988). Fla-vonoids and phenol acids in the plant cell may play animportant role in plant adaptation to various stress fac-tors (Hoch

et al.

, 2001). Thus, the accumulation of fla-vonoids is related to the mechanism protecting thephotosynthetic apparatus from extensive oxidativedamage (Yamasaki

et al.

, 1996; Yamasaki, 1997),which may result from exposure to various pollutants(Lavola

et al.

, 1994). Unlike other phenolic com-pounds of plant origin, anthocyanins have not beenstudied with respect to the influence of petroleum pol-lutants on their synthesis, despite an intensive searchfor prospective biochemical indicators of environ-mental pollution among secondary plant metabolites(Maslennikov, 2001).

Anthocyanins accumulate in the tissues of plantsexposed to stress and change their optical properties,and the use of methods based on this phenomenon maybe most expedient. The contents of riboflavin andascorbic acid are also promising as indicators of thephysiological state of plants. These compounds have avariety of effects on physiological processes in plantcells and, according to recent data, are involved in cellimmunity, which accounts for the resistance of plant

cells to stress factors (Mori and Sakurai, 1995; Chupa-khina, 1997; Powers, 1999; Silva

et al.

, 1999).The purpose of this work was to study the accumu-

lation of anthocyanins, ascorbic acid, and riboflavin inplants growing under conditions of pollution withpetroleum hydrocarbons and crude oil.

MATERIAL AND METHODS

The leaves of

Geum urbanum

L.,

Anthriscussylvestris

L.,

Glecoma hederata

L.,

Taraxacum offici-nalis

L.,

Dactylis glomerata

L., and

Achillea millefo-lium

L. were collected in a zone polluted with petro-leum products (motor and transmission oils, grease,etc.) along railroad tracks. Plants growing 3–5 m awayfrom the railroad were used as the control.

In addition, we studied 25-day-old seedlings of

Hordeum vulgare

L. (cultivar Roland),

D. glomerata

L.(cultivar Asta),

Vicia sativa

L. (cultivar Orlovskaya),

Panicum miliaceum

L. (cultivar Bystroe), and

Zeamays

L. (cultivar Ross 144) grown in soil (light loam)containing the maximum allowable concentrations ofcrude oil (5–10%, depending on the species) from theAleshkino field, Kaliningrad oblast (Dedkov andFominykh, 1999). The seedlings were grown in aTKSh-1 unit at constant illumination (LBU-30fluorescent lamps, 5 J/m

2

s) and room temperature(18

−

22

°

C).The contents of riboflavin in oxidized and reduced

forms (OR and RR, respectively) and anthocyaninswere determined by means of spectrophotometry(Murav’eva

et al.,

1987; Chupakhina, 2000), and thecontents of ascorbic acid (AA), by titration (Chupa-khina, 2000). The results were recalculated per gram

Plant Adaptation to Oil Stress

G. N. Chupakhina and P. V. Maslennikov

Kaliningrad State University, Universitetskaya ul. 2, Kaliningrad, 236040 Russia

Received July 8, 2003

Abstract

—Changes in the contents of anthocyanins, ascorbic acid, and riboflavin were studied in plants grow-ing under pollution with petroleum products along railroad tracks (

Geum urbanum

L.,

Anthriscus sylvestris

L.,

Glecoma hederata

L.,

Taraxacum officinalis

L.,

Dactylis glomerata

L., and

Achillea millefolium

L.) and inseedlings grown in soil containing 5–10% crude oil (

Hordeum vulgare

L.,

D. glomerata, Vicia sativa

L.,

Pani-cum miliaceum

L., and

Zea mays

L.). In the former case, the plants accumulated ascorbic acid and anthocyanins(on average, 2 and 5.2 times those in the norm, respectively), and riboflavin (in both reduced and oxidizedforms). In the latter case oil-induced stress also proved to stimulate the accumulation of all test substances inthe seedlings. The content of anthocyanins is proposed as a test parameter reflecting the degree of environmen-tal pollution, which may be useful for prompt bioindication of pollutants in the ecological monitoring of plantcommunities.

Key words

: petroleum pollutants, anthocyanins, ascorbic acid, riboflavin, bioindication.

RUSSIAN JOURNAL OF ECOLOGY

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No. 5

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PLANT ADAPTATION TO OIL STRESS 291

dry tissue weight. The values shown in the figures(means and mean square deviations) were obtained infour to six independent experiments, each performed inthree biological replications.

RESULTS AND DISCUSSION

Pollution along the railroad proved to stimulate theaccumulation of anthocyanins in all plant species(Fig. 1). The contents of these pigments were mark-edly higher than in the control plants (on average, bya factor of 5.2). The greatest and smallest differencesfrom the control were observed in the cases of

G. urbanum

and

G. hederata

: by factors of 16.9 and1.4, respectively.

The contents of AA in the leaves of plants from thepolluted zone also proved to exceed the control level by

factors of 2.3 (

G. urbanum

), approximately 2 (

A. mille-folium, A. sylvestris, T. officinalis

, and

G. hederata

),and 1.6 (

D. glomerata

) (Fig. 2).

A similar result was obtained when analyzing thecontents of riboflavin: the accumulation of both RR andOR proved to be activated in the polluted zone. Thecontents of OR in the leaves of

G. urbanum, A. millefo-lium, A. sylvestris, T. officinalis, G. hederata

, and

D. glomerata

exceeded the control level by a factor of1.2–1.5 (Fig. 3a). The contents of RR in the leaves of

G. urbanum, G. hederata

, and

D. glomerata

exceededthe control level by a factor of 1.3; and in the leaves of

A. millefolium, A. sylvestris

, and

T. officinalis

, by a fac-tor of 1.5–2 (Fig. 3b).

A positive correlation was observed between thecontents of anthocyanins and AC. This correlation was

Dactylis glomerata

Glecoma hederata




Geum urbanum

0 1 32 4 5 6Anthocyanins, mg/g

Clean zone

Polluted zone

Fig. 1.

Effect of pollution with petroleum products along the railroad on the anthocyanin content in different plant species.

Dactylis glomerata

Glecoma hederata




Geum urbanum

0 50 100 150 200Ascorbic acid,

µ

g/g

Clean zone

Polluted zone

Fig. 2.

Effect of pollution with petroleum products along the railroad on the ascorbic acid content in different plant species.

292


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CHUPAKHINA, MASLENNIKOV

of medium strength in the plants from the control(clean) zone (

r

= 0.54–0.68) and strong (

r

= 0.87–0.98)in the plants from the polluted zone. Likewise, the cor-relation between the contents of riboflavin and anthocy-anins was stronger in the latter (

r

= 0.69–0.81) than inthe former (

r

= 0.12–0.34).To study the effects of petroleum products on plant

growth and the synthesis of test substances, seedlingsof

H. vulgare, D. glomerata, P. miliaceum

, and

Z. mays

were grown in soil containing the maximumallowable concentrations of crude oil: 10% for

H. vul-gare, Z. mays, P. miliaceum

, and

V. sativa

and 5% forthe less tolerant

D. glomerata

(Dedkov and Fom-inykh, 1999). At this concentration of oil in the soil,the emergence of

D. glomerata

seedlings was retardedby two to seven days, the first true leaves appeared oneto three days later, and the height of two-week-oldplants grown in the soil contained 5% crude oil was29% smaller than in the control. The growth anddevelopment of

V. sativa, H. vulgare, Z. mays

, and

P. miliaceum

seedlings in the soil containing 10% oilwere retarded. In

H. vulgare

, seedlings appeared one

or two days later; in

V. sativa, P. miliaceum

, and

Z. mays

, six to seven days later than in the control.The emergence of the first and second true leaves in

V. sativa

was retarded by seven and four days; in

P. miliaceum

, by six and seven days; and in

H. vulgare

, by four and six days; plant height on day14 was 56, 83, and 45% smaller than in the controlplants of the same species, respectively. Higher concen-trations of oil in the soil prevented seed germination.

Under conditions of oil pollution, the contents ofanthocyanins in the seedlings of all species increasedby a factor of 2.4, on average (Fig. 4). The contents ofAA increased by a factor of 2–2.4 in

P. miliaceum,D. glomerata

, and

H. vulgare

and by a factor of 1.7–1.8in

Z. mays

and

V. sativa

(Fig. 5). The riboflavin poolproved to increase in the seedlings of all species, withOR exceeding the control level by a factor of 1.8–2.5(Fig. 6a), and RR, by a factor of 1.6–2.2 (Fig. 6b). Inboth experimental and control plants, a direct correla-tion of medium strength between the contents of antho-cyanins and total riboflavin (

r

= 0.29–0.64) and a strong

Dactylis glomerata

Glecoma hederata




Geum urbanum

0 0.5 1.0 1.5 2.0 2.5

(‡)

Dactylis glomerata

Glecoma hederata




Geum urbanum

0 1 2 3 4 5

(b)

Riboflavin, mg/g

Clean zone

Polluted zone

Fig. 3.

Effect of pollution with petroleum products along the railroad on the contents of (a) oxidized and (b) reduced forms of ribo-flavin in different plant species.


Vol. 35

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2004


correlation between the contents of anthocyanins andAA (

r

= 0.89–0.99) were revealed.Thus, soil pollution with petroleum products and

crude oil promoted the accumulation of anthocyanins,AA, and riboflavin (both OR and RR) in

G. urbanum,A. sylvestris, G. hederata, T. officinalis, D. glomerata,A. millefolium, H. vulgare, V. sativa, P. miliaceum

, and

Z. mays

plants.The accumulation of anthocyanins and riboflavin

under the effect of soil pollution with crude oil (in

V. sativa, D. glomerata, H. vulgare, Z. mays

, and

P. miliaceum

) and petroleum products along the rail-road (in

G. urbanum, A. sylvestris, G. hederata, T. offi-cinalis, D. glomerata

, and

A. millefolium

) may be dueto numerous disturbances of soil conditions: impairedaeration resulting from air displacement by oil; activa-tion of anaerobic microorganisms; a disturbed water

balance in the soil–plant system; poisoning with sul-fides and excess manganese released upon decompo-sition of some hydrocarbons; impaired hydrologicconditions; and changes in the physical, morphologi-cal, and agrochemical soil properties entailing a sharpdecrease in the contents and accessibility of movablenitrogen, phosphorus, and calcium compounds(Maslennikov and Chernova, 2001). In addition, oilpollution leads to sodium chloride salinization of thesoil, and its absorption capacity decreases (Golodyaevand Ivanov, 1988).

The parallel accumulation of anthocyanins andascorbic acid—compounds differing in functional com-partmentalization within the cell—in the plants grow-ing under conditions of oil pollution may contribute tothe efficiency of the antioxidant system in neutralizing

Hordeum vulgare

Panicum miliaceum

Zea mays

Dactylis glomerata

Vicia sativa

0 1 2 3 4Anthocyanins, mg/g

Experiment

Control

5

Fig. 4.

Effect of oil pollution on the accumulation of anthocyanins in the leaves of test plants.

Hordeum vulgare¸

Panicum miliaceum

Zea mays

Dactylis glomerata

Vicia sativa

0 50 100 150 200Ascorbic acid,

µ

g/g

Experiment

Control

250

Fig. 5.

Effect of oil pollution on the accumulation of ascorbic acid in the leaves of test plants.

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CHUPAKHINA, MASLENNIKOV

the products of oxidative stress and, hence, to plantresistance against the effect of pollutants.

The content of endogenous anthocyanins may beused as an indicator of the physiological state of plantcells, tissues, and ecosystems. It is a promising testparameter characterizing the degree of environmentalimpact of various pollutants, which may be useful fortheir prompt bioindication in the ecological monitor-ing of plant communities. Changes in the optical prop-erties of plant tissues caused by the accumulation ofanthocyanins under stress can be monitored byremote-sensing methods. Their use will allow special-ists to reveal changes in individual components of anecosystem, and on this basis predict the course of itsevolution.

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Maslennikov, P.V., Bioindication of Oil Pollution on theBasis of the Pigment Apparatus, Materialy I Mezhdunarod-noi nauchnoi konferentsii “Sovremennye problemy orga-nicheskoi khimii, ekologii i biotekhnologii.” T. 2. Ch. 1.Ekologiya i ratsional’noe prirodopol’zovanie (Proc. 1st Int.Sci. Conf. “Current Problems in Organic Chemistry, Ecol-

Hordeum vulgare

Panicum miliaceum

Zea mays

Dactylis glomerata

Vicia sativa

0 0.2 0.4 0.6 0.8

Experiment

Control

1.0

(‡)

Hordeum vulgare

Panicum miliaceum

Zea mays

Dactylis glomerata

Vicia sativa

0 0.5 1.0 1.5 2.0

Experiment

Control

2.5

(b)

Riboflavin, µg/g

Fig. 6. Effect of oil pollution on the accumulation of (a) oxidized and (b) reduced forms of riboflavin in the leaves of test plants.

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Plant Adaptation to Oil Stress