EFFECT OF ZINC AND COPPER SOIL CONTAMINATION ON THE TRANSPIRATION INTENSITY AND STOMAL INDEX OF WINTER CROP WHEAT SEEDLINGS

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1 J. Elementol. 2007, 12(1): EFFECT OF ZINC AND COPPER SOIL CONTAMINATION ON THE TRANSPIRATION INTENSITY AND STOMAL INDEX OF WINTER CROP WHEAT SEEDLINGS Ann Stolrsk, Jcek Wróbel, Ann WoŸnik, Brbr Mrsk 1 Chir of Plnt Physiology Agriculturl University in Szczecin Abstrct In the study, the physiologicl response of whet plnts to the effect of metl slts ws exmined by mesuring gseous exchnge trnspirtion prmeters, such s trnspirtion intensity, stomtl conductivity nd stomt count per 1mm 2 of the lef under- nd upperside. The study ws crried out using the gley soil [blck erth] of grnulometric composition of lom collected from the plough-humus horizon of 0-10 cm depth. The soil ws contminted with different pollutnt doses, i.e. 0 (control), 0.05, 0.5, 5 nd 50 mmol kg -1. Ech combintion ws seeded with 10 whet grins. Two-fctor nlysis of vrince ws used, with the type of pplied slt (ZnSO 4 nd CuSO 4 ) being the first fctor nd its dose the second one. The significnce of fctors ws exmined with Tukey s test (p<0.05). A delyed germintion of seeds ws observed in reltion to the control in soil smples contminted with 5 mmol kg -1 (both slts). The seedlings exmined showed lrge differentition in their response to Zn nd Cu; in the cse of CuSO 4, trnspirtion ws much lower thn under the effect of ZnSO 4. The trnspirtion intensity ws the lowest fter ppliction of copper slts in concentrtion of 5 nd 50 mmol kg 1, wheres the highest one ws observed in zinc concentrtion of 5 mmol kg -1. The soil contmintion with dose of 0.05 nd 50 mmol kg -1 resulted in significnt decrese of stomtl conductivity in reltion to the control. The exmined hevy metls significntly ffected the stomtl count only on the lef underside, where it incresed t the highest concentrtion of hevy metls, i.e. 50 mmol kg -1. Key words: cooper, trnspirtion, stomtl conductivity, stomt count. dr Ann Stolrsk, Ktedr Fizjologii Roœlin AR w Szczecinie, ul. S³owckiego 17, Szczecin e-mil: stolrsk@gro.r.szczecin.pl, tel.: (091)

2 80 WP YW ZANIECZYSZCZENIA GLEBY CYNKIEM I MIEDZI NA INTENSYWNOŒÆ TRANSPIRACJI ORAZ INDEKS SZPARKOWY SIEWEK PSZENICY OZIMEJ Ann Stolrsk, Jcek Wróbel, Ann WoŸnik, Brbr Mrsk Ktedr Fizjologii Roœlin Akdemi Rolnicz w Szczecinie Abstrkt Rekcjê fizjologiczn¹ roœlin pszenicy n dzi³nie soli metli bdno mierz¹c prmetry wyminy gzowej, tkie jk: intensywnoœæ trnspircji, przewodnoœæ szprkow orz liczb prtów szprkowych n 1 mm 2 dolnej i górnej epidermy blszki liœciowej. Bdni przeprowdzono z u yciem czrnej gleby o sk³dzie grnulometrycznym gliny, pobrnej z poziomu orno-próchniczego z g³êbokoœci 0-10 cm. Glebê znieczyszczono ró nymi dwkmi polutntów: 0 kontrol; 0,05; 0,5; 5; 50 mmol kg -1. Do wszystkich wrintów wysino po 10 nsion pszenicy. Zstosowno dwuczynnikow¹ nlizê wrincji: pierwszym czynnikiem by³ rodzj zstosownej soli (ZnSO 4, CuSO 4 ), drugim dwk. Istotnoœæ czynników testowno testem Tukey n poziomie 0,05. Zobserwowno opóÿnione kie³kownie nsion w stosunku do kontroli w glebie znieczyszczonej dwom rodzjmi soli w iloœci 5 mmol kg -1. Stwierdzono, e bdne roœliny ró nie regow³y n zstosowny metl. W przypdku soli miedzi trnspircj roœlin by³ zncznie mniejsz ni w przypdku soli cynku. Intensywnoœæ trnspircji by³ njmniejsz po zstosowniu soli miedzi w stê eniu 5 i 50 mmol kg 1, ntomist njwiêksze ntê enie trnspircji wykzno po zstosowniu soli cynku w stê eniu 5 mmol kg -1. Znieczyszczenie gleby dwk¹ 0,05 i 50 mmol kg -1 spowodow³o znczne zmniejszenie przewodnoœci szprkowej w stosunku do kontroli. Bdne metle ciê kie wp³ynê³y istotnie n liczbê prtów szprkowych jedynie n dolnej epidermie blszki liœciowej, gdzie ich liczb zwiêkszy- ³ siê po zstosowniu dwki 50 mmol kg -1 metli ciê kich. S³ow kluczowe: miedz, cynk, trnspircj, przewodnoœæ szprkow, indeks szprkowy. NTRODUCTION Industry nd intensive griculture cuse incresed environmentl pollution with hevy metls. High level of contmintion occurs most frequently in the vicinity of emitters (GORLACH 1991, FABER, WARTA 1981). According to the Stte Inspectorte for Environmentl Protection (1997), soil contmintion with copper nd zinc results first of ll from the impct of metllurgicl, electrochemicl nd chemicl industries, wheres zinc pollution is dditionlly cused by col, wste nd refuse combustion s well s hevy rod trffic. Hevy metls vilble to plnts cn ffect their growth nd physiology in different wy. While in threshold quntities, they frequently stimulte physiologicl processes, they will become toxic fter exceeding permissible mounts, first of ll disturbing the stbility of cytoplsmtic membrnes nd cell structures (GORLACH 1991, HARBORNE 1997, STROIÑSKI 2002).

3 The study imed t determining the effect of different zinc nd copper concentrtions in soil on the trnspirtion intensity, stomtl conductivity nd stomtl index of whet seedlings. 81 MATERIAL AND METHODS In , n experiment ws crried out under lbortory conditions with cv. Toncj winter crop whet involving 10-hour-long invrible light intensity PAR t 400 µmol m -2 s -2 nd men ir temperture of o C. Soil mss per one pot ws 0.5 kg. The substrte consisted of typicl blck erth, lomy in grnulometric composition, collected from the plough-humus horizon of 0 10 cm depth in the Experimentl Sttion of Agriculturl University in Szczecin in Do³uje. Soil rection (ph KCl ) ws 6.4 nd the hydrolytic cidity ws 1.2 me 100 g -1. The content of soil zinc equlled mg kg -1, wheres tht of copper to 9.60 mg kg -1. The soil ws contminted with hevy metls in the form of CuSO 4 nd ZnSO 4 slts in the mounts of 0.05, 0.5, 5 nd 50 mmol kg -1 of soil. The control ws soil free of ddition of zinc nd copper slts. Ech combintion ws seeded with 10 whet grins of cv. Toncj. The experiment ws crried out in four replictions. After germintion, mesurements were mde of the intensity of trnspirtion process (H 2 O m -2 s -1 ) nd of the stomtl conductivity (mmol m -2 s -1 ) using gs nlyser LCA-4. Stomt were counted with n opticl microscope Olympus CX-41on the upper nd the lower lef epidermis, converting them into n re of 1 mm 2. The results underwent sttisticl elbortion by mens of two-fctor nlysis of vrince, with type of the pplied slt type (ZnSO 4 nd CuSO 4 ) being the first fctor nd Zn nd Cu dose the second one. The significnce ws exmined with Tukey s test t significnce level of RESULTS AND DISCUSSION The soil contmintion with copper nd zinc slts clerly modified the vlues of physiologicl prmeters exmined, i.e. of trnspirtion intensity, stomtl conductivity nd stomtl index. Hevy metls dded to substrte in lrger doses resulted in cler osmotic stress, thus impeding the wter intke from substrte by germinting seeds nd cusing weker sprouting of seedlings. According to STOLARSKA et l. (2006), soil contmintion with copper nd zinc slts results in delying of whet grin germintion, which is brought bout by the suppression of wter ccess from substrte to germinting seeds nd then to seedlings. The

4 82 ccepted limit vlue of copper content in soils under griculturl use is 150 mg kg -1, wheres tht of zinc is 300 mg kg 1 (Ordinnce of the Minister of Environmentl Protection of 9 September 2002). In contminted soils, these doses cn be severl-fold exceeded (TRZASKOŒ, DZIDA 1995). In the own reserch, delyed germintion of whet grins ws observed with the soil contmintion with zinc nd copper slts in the mounts of 5 nd 50 mmol kg -1 in reltion to the control. This is confirmed by studies of PRZYBULEWSKA nd STOLARSKA (2004), who stte tht mong the symptoms of excessive zinc concentrtions in plnts re suppression of seed germintion nd slower seedling growth. This experiment demonstrted tht reduction of trnspirtion intensity in whet leves ws nerly proportionl to the soil contmintion with copper slts (Fig. 1). Fig. 1. Influence of vried zinc nd copper rtes in substrte on trnspirtion t winter crops whet sedling seedling Rys. 1. Wp³yw ró nych stê eñ cynku i miedzi n intensywnoœæ trnspircji siewek pszenicy ozimej According to PRZYBULEWSKA nd STOLARSKA (2004), cerel plnts re unusully susceptible to incresed concentrtion of tht chemicl element is soil. The effect of ZnSO 4 in dose of 0.05 mmol kg -1 ws not significnt. The rise of zinc sulphte dose resulted in considerble increse of the intensity of trnspirtion process, not until lrge concentrtion of tht zinc slt ws used (50 mmol kg -1 ), which clerly inhibited this process (Figure 1). Similr results were obtined by BRUNE et l. (1994). According to PRZYBULEWSKA nd STOLARSKA (2004), cerels re one of the plnts tht re most susceptible to the concentrtion of tht metl in soil. PANDEY nd SHARAMA (2002) report tht lrge content of hevy metls in soil considerbly decreses the intensity of trnspirtion process in plnts. The whet seedlings clerly showed greter differentition of trnspirtion intensity depending on metl type thn on its doses, since it ws demonstrted tht fter ppliction of 5 nd 50 mmol kg -1 copper slt the trnspirtion intensity ws significntly lower thn fter pplying zinc slt in the mount of 5 mmol kg -1 (Figure 1). The stomtl conductivity of whet seedlings resulting from the soil contmintion with copper nd zinc slts ws shped like (Figure 2). However, the ex-

5 83 Tble 1 Tbel 1 Influence of zinc nd copper slts (mmol kg - 1 of soil ) on trnspirtion of winter crops whet seedling Wp³yw soli cynku i miedzi (mmol kg -1 gleby) n intensywnoœæ trnspircji siewek pszenicy ozimej Dose Dwk A verge Œredni Metl Cu Zn A verge Œredni b Fig. 2. Influence of vried zinc dn cooper rtes in substrte stomtl conductnce t winter crops whet seedling Rys. 2. Wp³yw ró nych stê eñ cynku i miedzi n przewodnoœæ szprkow¹ siewek pszenicy ozimej mined plnts clerly responded to dose of hevy metl rther thn to its type. A decrese in stomtl conductivity ws found fter ppliction of 0.05 nd 50 mmol kg -1 doses in reltion to the control (Tble 2). Similr results were obtined by BURZYÑSKI nd K OBUS (2004), who found tht t dose of copper slt of 0.02 mmol kg -1 the stomtl conductivity ws similr to tht in control plnts, wheres rise of metl dose to 0.05 mmol kg -1 considerbly reduced this index. According to MONNET et l. (2001), very low concentrtion of copper slts resulted in strong reduction of stomtl conductivity versus the control. Slight elevtion of slt concentrtion increses the stomtl conductivity, while very strong concentrtion of tht chemicl element in soil suppresses it (MONNET et l. 2001).

6 84 Tble 2 Tbel 2 Influence of zinc nd copper slts (mmol kg - 1 of soil ) on stomtl conductnce of winter crops whet seedling Wp³yw soli cynku i miedzi (mmol kg- 1 gleby) n przewodnoœ æ szprkow¹ siewek pszenicy ozimej Dose Dwk A verge Œredni b b b Metl Cu Zn A verge Œredni Influence Wp³yw soli Tble 3 Tbel 3 of zinc nd copper slts (mmol kg -1 of soil) on stomtl pprtus in le f underside nd lef upperside of winter crops whet seedling cynku i miedzi (mmol kg -1 gleby) n liczbê prtów szprkowych dolne j i górnej strony liœci siewek pszenicy ozimej Lef upperside Stron górn Dose Dwk verge œredni L ef underside Stron doln d ose dwk verge œredni b b L ef upperside Stron górn M etl verge œredni metl Cu 442 Cu Zn 443 Zn L ef underside Stron doln verge œredni

7 The size of hevy metl dose hd significnt effect on the stomt count only on the lef underside. After ppliction of hevy metls in dose of 50 mmol kg -1, the number of stomt ws significntly lrger thn when exposing plnts to lower doses of tht metl, i.e. 0.05, 0.5 nd 5 mmol kg -1 (Tble 3). However, no significnt differences were found in reltion to the control. It is difficult to explin such response of whet seedlings to the incresed concentrtion of zinc nd copper slts, s vilble reference dt re short of findings tht refer to their effect on the number of stomt Fig. 3. Influence of vried zinc nd copper rtes in substrte on stomtl pprtus in lef underside nd lef upperside of whet seedling winter crops whet seedling Rys. 3. Wp³yw ró nych stê eñ cynku i miedzi n liczbê prtów szprkowych dolnej i górnej strony liœci siewek pszenicy ozimej CONCLUSIONS 1. The intensity of trnspirtion ws more evidently slowed down fter ppliction of copper slts in the mounts of 5 nd 50 mmol kg -1 thn fter ppliction of zinc slts in dose of 5 mmol kg -1. Most probbly, the whet seedlings considerbly suppressed trnspirtion fter ppliction of copper slts in order to use wter in the photosynthesis process rtionlly. 2. The soil contmintion with zinc nd copper slts in the mounts of 0.05 nd 50 mmol kg -1 resulted in significnt decrese of stomtl conductivity in reltion to the control object.

8 86 3. After soil contmintion with hevy metls in dose of 50 mmol kg -1, the men stomt count on the lef underside ws over twice s high s tht in control plnts. REEFERNCES BURZYÑSKI M., K OBUS G Chnges of photosynthetic prmeters cucumber leves under Cu, Cd nd Pb stress. Photosynthetic, 42 (4): BRUNE A., URBACH W., DIETZ K.-J Comprtmenttion nd trnsport of zinc in brley primry leves s bsic mechnismus involved in zinc tolernce. Plnt, Cell Environment, 17: FABER A, WARTA Z Wp³yw znieczyszczeñ przemys³owych zwierj¹cych cynk, o³ów, kdm n przyrodnicze wrunki rolnictw. Post. Nuk Rol., 3: , GORLACH E Metle ciê kie w glebie i roœlinch. Aur, 11: HARBORNE J.B Dictionry of plnt toxins. John Wiley & Sons, pp MONNET F., VAILLANT N., HITMI A., COUDRET A., SALLANON H Endophytic Neotyphodium lolii induced tolernce to Zn stress in Lolium perenne., Physol., Plnt., 113: Ocen sk eni œrodowisk Polski metlmi ciê kimi przy u yciu mchów jko biowskÿników PIOŒ. Wrszw. PANDEY N., SHARMA CH Effect of hevy metls Co 2+, Ni 2+ nd Cd 2+ on growth nd metbolism of cbbge. Plnt Sc., 163: PRZYBULEWSKA K., STOLARSKA A Wp³yw stê eni metli ciê kich (Hg, Pd, Cu) w glebie n wzrost i rozwój siewek jêczmieni. J. Elementol., 9 (3): Rozporz¹dzenie Ministr Œrodowisk z dni 9 wrzeœni 2002 r. w sprwie jkoœci gleby orz stndrdów jkoœci ziemi. Dz.U. Nr 165 poz STOLARSKA A,. WRÓBEL J., WO NIAK A., MARSKA B Influence of copper nd zinc in soil on Physiologicl Rection of Whet seedling. Ecol. Chem. Eng. (in press). STROIÑSKI A Odpornoœæ roœlin n stres wywierny przez metle ciê kie. Biotechnology, 3 (58): TRZASKOŒ M., DZIDA M Metle ciê kie w glebie n ³¹kowo-pstwiskowych terench przyleg³ych do utostrdy. Zesz. Prob. Post. Nuk Rol., 421: