NUMBER 6 OCTOBER 1992

Effect of Crude Oil on the Growth and AnatomicalFeatures of Chromolaena odorata (L.) K.& R.

L.s. GilLH.Gihmffr"-

Department of BotanyUniversity of Benin

P.M.B.1154Benin City, Nigeria

AbstractInvestigations were carried out on the effect of ForcadosBlend crude oil on seed germination, regeneration ofstumpsandanatomical feahrres of leaves of Chromolaenaodorata (L.) K. & R. Three concentration via. 25.0cm3,50.0cm3 and 75.cm3applied to soil were found to sup-press seed germination, regeneration and caused cellu-lar and stomatal abnormalities. The present study sug-gests that this plant can serve as bioindicator of pollu-tion in oil producing areas.

IntroductionAccording to DeJong (1980) crude oil spillage on

soil makes it unsatisfactory for plant growth. This is dueto insufficient aeration of the soil because of displace-ment of air from the spaces between the soil particles bycrude oil (Rowell,1,977). In Nigeria quite substantialamount of crude oil is spilled annually, For example,Nwankwo (1989) reported 2,000 oil spillages in Nigeriabetween 197 6 andl 988. Drring this period about 2 x 1 05barrels of crude oil were discharged in the environment.

Considering this large quantity of crude oil goinginto the environment, especially farmland, and the factthat the inhabitants of these areas are subsistent farm-ers, there is an urgent need for the various agenciesconnected with oil production in Nigeria to pay moreattention to the problem of oil leakage in future.

(Condnue on Page 2)+Department of Botany, Ondo State University, P.M. B. 5353,Ado-Ekiti, Ondo State, Nigeria

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Fig. la. Mature leafepidermis (Abaxial) ofuntreated plant show-ing normal epidermalcel ls (nec) andanomocytic stoma(ans)..

Fig. lb. Mature epi-dermal peel (abaxial)from the leaf of aplanttreated with 25.0cm3crude oil showing ma-ture anomocytic stoma(mas) and slightly dis-torted epidermal cells(sdec).

Fig.lc. Epidermalpeel(abaxial) from the leafof a plant treated with50.0cm3 crude oi lshowing distorted epi-dermal cells (dec) anddistortedmanue stoma(dms).

Fig. 1d. Epidemalpeel(abaxial) from the leafof a plant rearcd with75.0cm3 crude o i lshowing coalescedepi-dermal cells (ces) andan abnormal stoma withsingle guard cell (s gcs ).

Fig. le. Epidermalpeel(abaxial) from the leafof a plant sprayed withcrudeoil showingbadlydistorted epidermalcel ls (dec) and an

Effect of Crude Oil(Continued from Page 1)

It is known that root stress reduces leafgrowth via stomata conductance (Smith et al.,1989). Furthermore, Epstein (1972) states thatmineral ions absorbed initially by the roots arefinally received by the mesophyll cells of theleaves. According to Udo and Fayemi (1975)

growth of plants growing in oil polluted soilwas generally retarded and chlorosis of leavesresults coupled with dehydration of the plantsindicating water deficiency. The present projecthas been undertaken with three main objectives:(1) to determine the extent of damage caused bycrude oil pollution to the foliar parts ofChromolaena odorata (L.) K. & R., (2) to ascertainthe effect of crude oil on this taxon at the cellularlevel, and (3) to determinehowfarthis plantcanserve as an indicator of oil pollution in Nigeria.

Materials and MethodsPerforated black polythene bags filled with

soil were used for raising plants. Seeds or stumpsof C. odoratawere either sown or transplanted inthese bags. "Forcados Blend" crude oil at 25.0orP50.0cm3 and 75.0cnf with a control of waterwere usd for soil treatment and foliar applica-tion. A foliar treatment with pure crude oil wasalso included. This experiment was laid out ina randomized complete block design with fourreplications.

Growth performances of the plants raisedwere recorded every other day. Anatomicalfeatures of the roots, stem and leaf of the treatedand control plants have been carried out. Theepidermal peels for cuticular studies were ob-tained following the method outlined by Gilland Karatela (1983). Terminology of maturestomata used in this paper is that of Metcalfeand Chalk (1950).

ResultsApart from the seeds in the control which

germinated after three days of sowing, seedssown in varions concentrations of crude oil failedto germinate. Morphological observations car-ried out on plants from sprouted stumps havebeen presented in Table. 1.

Stomal distortions observed havebeen pre-sented in figs. 1a - e. Analysis of variance of data

collected on growth rate and stomatal frequency,length, breadth and pore size were given inTable 2. Crude oil treatment of C. odorata re-sulted indistortion of cells of the epidermalandcortical regions of root, stem and petiole. In thecase of the leaf lamina the distortion of cells wasmore pronounced in the spongy mesophyll andabaxial epidermal region. The concentration of75.0cm3 of crude oil treatment was most effec-tive.

DiscussionEnvironmental pollution has been shown

to have adverse effects on plant growth andthese may range from morphological aberra-tions, reduction in biomass to stomatal abnor-malities (Sharma et al., 1980). Similar reportshave been made by Gill and Sandota (1976),

Ghouse et al., (1980), Wallace (1951)'andVostokova et al., (1961). Atuanya 0987) re-ported that waste oil to cause a breakdown ofsoil texture followed by soil dispersion. How-ever, during the present study fresh crude oilrather showed a coagulatory effect on the soil,binding the soil particles into a water impreg-nable soil block which seriously impair waterdrainage and oxygen diffusion. Atuanya (1987)furtherstates that seeds sown in such soil failedto germinate. The results of the present studywhere seeds failed to germinate even after 30days of sowing confirm the report of Atuanya(1,987)

Udo and Fayemi (1975) reported that oilhas adverse effect on plant growth. the presentstudy gave similar results where there was asignificant difference in growth between theuntreated and treated plants. A positive rela-tionship has been observed between the extentof retardation in growth and concentration ofcrude oil applied to the soil. these findings areinaccordance with previous reports of Atuanya(1987),Ghouse et al., (1980) and Gill & Sandota(197O.

Results of the present shrdy shows that leafstomata were grossly affected by crude oil. Theeffect was manifested either in the distortion orreduction in the number of stomata per unit areaof the leaf. A strong relationship between de-

gree of disruption and concentration of crude oilin the soil was noticed. This further confirmsearlier report of Sharma et al. (1980).

Since uptake of water and salts (ions) iscarried out by the roots, the untreated plantswith roots undisturbed grew norrrully whilethe treated plants suffered morphological andanatomical aberrations. Cell disruption in rootsand other organs were eminent. furthennore,anatomical studies of the treated plants revealedthe presence of oil films in the epidermal andcortical regions of the root, stem and leaves.Similarfindings have been reportedby Anoliefo(1991). However, the absence of oil films in thevascular region suggest that movement of oilfilms from root to leaves could be via theapopplastic pathway.

From the foregoing, it is not clear whetherthe damage done to the cells was by toxic hydro-carbons and natural gases contained in crude oilor by smouldering and osmotic effect of thecrude oil liquid. It seems to the present authorsthat the adverse effect noticed on the treatedplants may be due to unfavorable conditionscreated in the soil resulting in drought condi-tions as well as nonavailability of nutrientsratherthan cellular destruction by toxic substancescontained in the crude oil. the present authorsare of the opinion that enlargement of cells in thevarious tissues is due to oxygen starvation ofthose tissues whereby the cells coalesce andthus forming large cells in these tissues.

AcknowledgementWe are grateful to the kind gesture of Prof. F.I.Opute of the Department of Botany,Universityof Benin who kindly procured for us the crudeoil used in this study from the Shell PetroleumCompany, Nigeria (Ltd.).

ReferencesAnoliefo. G.O. 1991. Forcados blend crude oil effect

in respiration, metablism, elemental compositionand growth of Citrullus vulgaris (Schrad). Ph.D.Thesis, Benin. pp. 6-30.

Atuanya, E.I. 1987. Effects of waste engine oilpollution on physical and chemical properties ofsoil: A case study of waste oil contaminated Deltasoil in Bendel State. Nigerian. J. Appl. Sci. 5:155-116.

3De Jong, E. 1980. The effect of a crude oil spill on

cereals. Environ. Poll. 22:187 -196.

Epstein, E. 1972. Mineral nutrition of plants, prin-ciples and penpectives, John Wiley, New York.pp. 15-21.

Ghouse, A.K.M., S.H. Zaidi & A. Atique. 1980.Effect of pollution on the foliar organs ofC alistemon citrinus Stap-f. J. Sci. Res . 2:207 -209.

Gill, L.S. & Y.Y. Karatela. 1983. Stomatogenesis inthe leaf of Cc lotropis procera Ait. (Asclepidaceae).Feddes Repert. 94:519-525.

Gill,L.S. &R.M.A. Sandota 1976. Effectof foliarlyapplied ccc on the growth of Phaseolus aureusRoxb. (mung or green gram). Bangladesh Joum.Biol. Sci. 15:35-40.

Metcalfe, C.R. & A. Chalk. 1950. Anatomy of thedicotyledons vols I & II (lst edn.) 1500 p.Clarendon Press, Oxford.

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Rowell, MJ.1977. The effect of crude oil spills onsoils. A review of literature. In: Toogood, J.A.(ed.). "The reclamation of agricultural soils afteroil spills" Part 1, Edmonton, Canada pp. 1-33.

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Vostokova, Y.A., D.O. Vyshivkiri, M.S. Kasynova,N.G. Nesvelaglova & A.M. Shvyryayera.1961.Geobotanical indicators of bitumen. Int. geologi-cal Revision, 3 :598-608.

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Table 1. Morphological changes observed in Chromolaena odorata due to crude oil treatment

Treatment Observable effects Percentage survival

Control NilTwelve plants survived 100.0

25.0 cm3 (i) Stunted growth(ii) Leaf buming with

resultant brown coloration(ii) Death of one plant out of twelve 91.3

50.0 cm3 (i) Drastically reduced growth rate(ii) Leaf burning leading to brown

coloration(iii) Death of four plants out of twelve 66.7

75.0 cm3 (i) Terminated growth(ii) Leaf burning leading to brown

coloration(iiD Death of four plants out of twelve 66.7

Foliage (i) Terminated growthsprayed with (ii) Leaf buming, color of leavespure crude tumed blackoil (iiD Stem turned pale

(iv) All 12 plants died 0.0

Table 2. Analysis of variances of effect of crude oil on Chromolaena odorata

Character 'F' Value

Growth rateStomotal frequencyStomotal lengthStomatal breadthStomatal pore size

20.40 **45.90 **45.69 **26.50 **

106.39 **

** p=0.01

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rishikeshensis Grover (Diptera) in India. Joumal of Biological Control 2:89-91.Akinluyi, T.O. & O. Odeyemi. 1989. The effect of Chromolaena odorata on the biomethanation of five

animal manures. Microbios Letters 4 I : 103- 105.Akinyemiju,O.A. &T. Alimi. 1989. Economicsofmaize (Zeamays)prroductionunderdifferentweedcontrol

methods. Nigerian Joumal of Weed Science 2:51-55Ahmed, M. 1989. Feeding diversity of Myllocerus viridanus Fab. (Coleoptera; Curculionidae) from South

India. Indian Forester 1 15:832-838.Amakiri, M.A. & E.A. Ajioye. 1989. Evaluation of three soil-applied herbicides for weed control inGmelina

arborea. Nigerian Joumal of Weed Science 2:4549Ambika,S.R.&Jayachandra. l989.InfluenceofstorageonseedgermipationinChromolaenaodorara. Seed

Research 17:143-152.Baggayan, R.L. Jr. 190. Fertilizationof Leucoenn diversifulia (ScNecht.) Benth. in Carranglan grassland.

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Nigeria. Cafe Cacao The 3l: 303-310.Erasmus, D.J. & D.J.R. Noel. 1989. Chemical control of Chromolaena odorata g-.) K. & R. efficacy of

herbicides applied to stumps. Applied Plant Science. 3:18-21.Eze, J.M. O. & L.S. Gill. 1992. Chromolaeana odorata - A problematic weed. Compositae Newsletter 20l

2 I : 17 -21 .Escarlos, J.A. 1990. Survey of leaf stomates and trihchomes of some problem weeds of Bukidnon,

Philippines. B IOTROP Special Publications 3 8 : 8 1 -84.

Gauntier, L. & R. Spichiger. 1990. Contact foret-savane en Cote D'Ivoire Centrale: Role de Chromolaenaodorata(L.) King &Robinsondansladynamiquedelavegetation. Cahiers,Fac. Sci. Univ. Genive 2O:75-89.

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43:241-245.Hulugalle, N.R., R. Lal & M. Gichuru. 1990. Effect of five yean of no-tillage and mulch on soil properties

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Kluge, R.L. 1989. Insects released against triffed weed in Natal. Plant Protection News 15:5.Lal,L. 1987. Studies on natural repellents against potato tuber moth, (Phthorimaea operculella7*l7er) in

country stores. Potato Research 3O:329-334.Lucas, E.D. 1989. Siam weed (Chromolaena odorata ) and crop production in Nigeria. Outlook on

Agriculture 18: 133- 138.Lyla, K.R., P.J. Joy & K.C. Abraham. 1987. Insectpests of Chromolaenaodorata (Eupatoriumodoratum).

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6Marks,M.K. &A.M. Seddon. 1988. Interactionbetween Chromolaenaodorata(Compositae) andTnnocerus

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