Development of Microbial Biomass and Enzyme Activities in Mine Soils

Transcription

1 Pol. J. Environ. Stu. Vol. 21, No. 3 (2012), Originl Rsrh Dvlopmnt o Miroil Biomss n Enzym Ativitis in Min Soils Mrin Chok 1 *, Mri Niklińsk 2 1 Dprtmnt o Opn-strip Mining, AGH Univrsity o Sin n Thnology, Mikiwiz 30, Krków, Poln 2 Institut o Environmntl Sins, Jgillonin Univrsity, Gronostjow 7, Krków, Poln Riv: 2 April 2011 Apt: 6 Dmr 2011 Astrt This stuy ssss th vlopmnt o miroil iomss, sl rspirtion, n th tivitis o hyrogns, urs, n i phosphomonostrs in sny min soils rlim or orstry n thos vloping unr vgttion rom nturl sussion. Th min soils ontin signiintly lss orgni C (C org ) n totl N (N t ) thn th nturl orst soils. Howvr, in som o thm th miroil iomss n sl rspirtion ttin vlus typil or th nturl orst soils. Th ontnt o N t prov to th most importnt ontrol on th miroil iomss, sl rspirtion, n th tivitis o hyrogns n phosphomonostrs in th min soils. All th miroil proprtis wr positivly rlt lso to C org ontnt. Th tivitis o hyrogns n urs pn strongly on miroil iomss (C mi ). Hn, high tivitis o ths nzyms wr trmin in soils ontining high C mi. Th i phosphomonostrs tivity ws lso positivly rlt to C mi, ut its tivity ws inrs in th soils with low P ontnts. Kywors: min soils, miroil iomss, hyrogns tivity, i phosphomonostrs tivity, urs tivity Introution *-mil: hok@gh.u.pl Aorsttion is ommon wy to rlim post-mining rrns. On o th most importnt gols o rlmtion or orstry is th rstlishmnt o stl n proutiv osystms. This gol n hiv only i soil untionlity is rstor. Soil miroorgnisms r involv in svrl ky soil prosss suh s omposition n ormtion o soil orgni mttr, nutrint yling, n nrgy trnsr [1]. Thror, msurmnt o irnt soil miroil n iohmil proprtis hs n rommn or soil rovry ssssmnt [2]. Soil nzyms r protins tht tlyz spii rtions rquir to pross omplx orgni ompouns into ssimill suunits suh s sugrs, mino is, NH 4+, PO 4 3-, t. [3, 4]. Dtrmining o tivitis o nzyms involv in th yling o nutrints suh s N, P, or S my giv inormtion on nutrint turnovr in soils. Thror, msurmnts o vrious soil nzyms hv n wily us or stimtion o soil rovry tr isturn n or soil qulity ssssmnt [5]. For instn, Mor t l. [6] us th tivitis o hyrogns, rylsults, n β-gluosis to ssss th t o vrious trtmnts on hvy mtl-ontmint soils tr th Aznlollr int in Spin. Blrin t l. [7] stui hronosqun o lyy min soils ovr y spontnously vloping vgttion n rport tivitis o llois, β-xylosis, i phosphts, n rylsults to inrs grully with th sit g. Th ojtiv o this stuy ws to ssss th vlopmnt o miroil iomss, sl rspirtion, n tivitis

2 570 Chok M., Niklińsk M. o soil nzyms involv in N n P yling in sny min soils rlim or orstry n thos vloping unr vgttion rom nturl sussion. Th nturl orst soils wr us s rrn. Mtril n Mthos Stuy Ar Th stuy ws rri out in Uppr Silsi, Poln (19º26' E; 50º16' N), on th grouns o th Szzkow opn-st sn qurry n its surrounings. Th limt is tmprt, with mn nnul pripittion o 700 mm n mn nnul tmprtur o 8ºC. Soils o th stuy r (minly Pozols) vlop rom sns. Th sn posits r luvioglil qutrnry simnts o pr-qutrnry morphologil prssion. Extrtion o sns in Szzkow Qurry rt n opn st ovring ovr 2700 h. Sin th lt 1950s it hs n rlim. Th stnr rlmtion prour inlu orming n lvling o th sur n ing n orgni mnmnt (pprox. 300 m 3 h -1 ). Th mnmnt ws mixtur o orst loor (O horizon) n minrl horizons (horizons A h, E, n prtly B), with vrg orgni C ontnt o % ollt rom orst soils in rs to min [8]. Thn th rlim sits wr lim (1.5 Mg olomit h -1 ), n lupin (Lupinus lutus L.) ws ultivt or two yrs. Th lupin ultivtions wr rtiliz with NPK (140 kg N h -1, 300 kg P 2 O 5 h -1, 180 kg K 2 O h -1 ). Atr two yrs, th lupin iomss ws plough into th soil s grn mnur n th sits wr orst with 1-yr-ol Sots pin slings (Pinus sylvstris), ommon irh (Btul pnul) n som othr iuous trs. Ovr th lst 25 yrs, rtin moiitions wr introu to th rlmtion mthos; ths inlu sstion o liming, n rsing th quntity o orgni mnmnt n NPK minrl rtilizrs (inormtion rom th Szzkow Sn Qurry). Chngs in rlmtion prours us th rlim sits smpl in our stuy to irntly trt. Th olst sit ws rlim oring to th stnr prour s sri ov. Th intrmit sit ws not lim ut riv th sm mount o orgni mnmnt n minrl rtilizrs s th olst on. At th youngst rlim sit no liming ws ppli, NPK rtilizrs wr ppli t lowr rts (50 kg N h -1, 140 kg P 2 O 5 h -1, 120 kg K 2 O h -1 ), n th orgni mnmnt (mount rs to pproximtly 30 m 3 h -1 ) ws not spr ovr th ntir rlim r ut ppli irtly unr th slings uring plnting. Th rlmtion trtmnts t ll th smpl min soils r summriz in Tl 1. In th 1970s, 1980s, n 1990s som prts o th opn st wr non n initil soil ormtion gn thr unr th vgttion rom nturl sussion. At th non sits, hrous ommunitis omint y gry hir-grss (Corynphorus nsns L.) wr th irst to stlish. Thy wr ollow y io-groups o trs with Tl 1. Rlmtion msurs t th smpl min soils. Soil Orgni mnmnt Liming ovr 50% omintion y Sots pin (Pinus sylvstris L.) n ommon irh (Btul pnul Roth.), with th osionl trmling poplr (Populus trmul L.) [9]. Soil Smpling Frtiliztion Lupin ultivtion m 3 h m 3 h m 3 h Mg olomit h kg N h -1 2 yrs 140 kg P 2 O 5 h kg K 2 O h kg N h -1 2 yrs 140 kg P 2 O 5 h kg K 2 O h kg N h -1 2 yrs 300 kg P 2 O 5 h kg K 2 O h kg N h -1 2 yrs 300 kg P 2 O 5 h kg K 2 O h -1 Smpls o minrl soil (0-5 m) wr tkn in My 2007 t 10 sits (r = 1000 m 2 ): gr sit prior to rlmtion (), sit tr two yrs o lupin ultivtion (), rlim post-mining sits orst with Sots pin (7, 21, n 29 yrs ol,, n, rsptivly), post-mining sits with spontnously vloping pin orst stns (7, 21, n 28 yrs ol,, n, rsptivly), n two nturl pin orst stns s rrn soils (orst stns unr 31 n 109 yrs ol n, rsptivly). At h sit, ight mix smpls onsisting o iv su-smpls (r o h susmpl = 0.16 m 2 ) wr tkn. At ll th sits (xpt ) th soils wr lssii s sns (sn ontnt 88-99%, silt ontnt 0-9%, ly ontnt 0-2%). At th soil ws lomy sn (87% sn, 9% silt, 4% ly) [10]. Th smpls wr siv (2 mm msh) n ivi into two prts. On prt ws ir-ri n us or hmil nlyss, n th othr ws stor t 4ºC n us or miroil nlyss. Prior to miroil nlyss th smpls wr just to 50% o th mximum wtr holing pity (WHC) n pr-inut t 22ºC or 14 ys.

3 Dvlopmnt o Miroil Biomss n Enzym Anlytil Mthos Mximum wtr holing pity (WHC) ws trmin grvimtrilly [11]. Th ph o th smpls ws msur in 1M KCl solution t 1:10 soil:liqui rtio (w:v) using igitl ph-mtr (CP-401, ELMETRON). Th ontnt o orgni C (C org ) n totl N (N t ) ws trmin y ry omustion using CN nlyzr (Vrio Mx, Elmntr Anlysnsystm GmH). Th ontnt o xtrtl P (P xt ) ws msur tr xtrtion with Mhlih I xtrtnt. Brily, th soil smpls (5 g) wr shkn with 25 ml o Mhlih I xtrtnt (5 N HCl n 25 N H 2 SO 4 ; ph=1.2). Thn, th suspnsions wr iltr through inpor iltr n th inorgni P onntrtion in th iltrts ws trmin olorimtrilly with ontinuous low uto-nlyzr (FIAompt, MLE, Drsn, Grmny). Miroil iomss C (C mi ) ws trmin using sustrt-inu rspirtion mtho [12]. Bsl rspirtion (RESP) ws trmin tr inution o th soil smpls or 24 h t 22ºC in gs-tight jrs. Th volv CO 2 ws trpp in 0.2 M NOH n trmin y titrtion o th xss o hyroxi with 0.1 M HCl in th prsn o phnolphthlin s initor. Dhyrogns tivity ws trmin oring to von Mrsi [13]. Th soil smpls (1 g.w.) wr mix with 1.5 ml Tris ur (ph 7) n 2 ml 0.5% INT (2-pioophnyl-3-p-nitrophnyl-5-phnyl ttrzolium hlori) solution, n inut t 40ºC or 2 h. Th ru ioonitrottrzolium ormzn (INTF) ws xtrt with 10 ml imthyloormmi/thnol (1:1) n msur photomtrilly t 464 nm. Dhyrogns tivity ws xprss s µg INTF g -1 h-1. Ai phosphomonostrs tivity ws msur s sri y Mrgsin [14]. Th soil smpls (1 g.w.) wr mix with 1 ml isoium p-nitrophnyl phospht solution (115 mm) n 4 ml ur solution (ph 6.5) n inut t 37ºC or 1 hour. Th p-nitrophnol rls y phosphomonostrs tivity ws xtrt n olor with NOH n trmin photomtrilly t 400 nm. Ai phosphomonostrs tivity ws xprss s µg p- -1 NP g-1 h n i phosphomonostrs iiny ws lult s i phosphomonostrs tivity normliz to pr µg o C mi sis. Urs tivity ws trmin s sri y Knlr [15]. Th soil smpls (5 g.w.) wr mix with 2.5 ml ur (720 mm) n 20 ml ort ur (ph 10) n inut t 37ºC or 4 hours. Th rls mmonium ws xtrt with iii potssium hlori solution, olour in th moii Brthlot rtion n msur photomtrilly t 690 nm. Urs tivity ws xprss s µg N g -1 h-1 n urs iiny ws lult s urs tivity normliz to pr µg o C mi sis. Clultions n Sttistil Dt Anlyss All nlyss wr prorm in triplit n th mn vlus wr us in urthr lultions. Th t prsnt in th txt r mns vlus o ight rplits pr sit. Dirns in hmil n miroil proprtis twn th orst stns wr tst y on-wy nlysis o vrin (ANOVA). Th Fishr lst signiint irn (LSD) tst or multipl omprisons ws run i signiint irns wr oun (p<5). Rltionships twn th msur hmil n miroil proprtis o th min soils wr stui using Prson orrltions tr xluing th smpls rom nturl orst soils ( n ). Comprison o rgrssion lins ws prorm to hk i th rltionships twn C mi n th RESP n nzym tivitis r onsistnt in th rlim n sussionl soils. Prior to sttistil nlyss th t wr log-trnsorm or squr root-trnsorm to ulill th ssumption o normlity. Th sttistil lultions wr prorm with Sttgrphis Plus vrsion 5.1 (Mnugistis In.). Rsults Chmil Proprtis o Soils Th lowst ontnts o C org n N t wr trmin in th pr-rlmtion min spoil soils () (0.21 mg g -1 n 04 mg g -1, rsptivly) n 2-yr lupin () (0.27 mg g -1 n 11 mg g -1, rsptivly). In orst min soils th ontnts o C org wr signiintly highr n tn to inrs with inrsing sit g (Fig. 1). Howvr, th inrsing trn hs not n osrv or N t. Th ontnts o this lmnt in th rlim soils wr similr ( mg g -1 ) whrs or th min soils unr nturl sussion th lowst N t ws trmin in soil (79 mg g -1 ) n th highst in soil (0.260 mg g -1 ). Th nturl orst soils ontin signiintly mor C org ( mg g -1 ) n N t ( mg g -1 ) thn ny o th min soils (Fig. 1). Th lowst C org -to-n t rtio ( ) ws msur in th youngst orst min soils ( n ) n th highst in th soil (41.5). For th othr min n nturl soils th C org -to-n t rtios wr rltivly similr n vri rom 19.8 to Th highst ontnt o P xt ws msur in th soil (6.31 mg kg -1 ) n th lowst in soil (0.52 mg kg -1 ). Low ontnts o P xt wr msur lso in th sussionl min soils (1.16 mg kg -1 ) n (1.17 mg kg -1 ). In th othr min soils th P xt ontnts wr highr n similr to th P xt onntrtion in th nturl orst soil (Fig. 1). Th highst C org -to-p xt rtios wr msur in th soil (4305) n in th two nturl orst soils (2899 n 3558 or n, rsptivly). Rltivly high C org -to-p xt rtio ws trmin lso in soil (1573). Th highst ph vlus wr msur in th soils n (6.1 n 5.8, rsptivly). Th ph vlus o th orst min soils wr signiintly lowr n vri rom 4.1 to 4.8 (Fig. 1).

4 572 Chok M., Niklińsk M. Miroil n Biohmil Proprtis Th lowst C mi n RESP wr msur in th soil (10.1 µg g -1 n 8.1 µg C-CO 2 g -1 24h -1, rsptivly). Th soil h similr C mi (20.1 µg g -1 ) ut signiintly highr RESP (16.7 µg C-CO 2 g -1 24h -1 ) ompr with th soil (Fig. 2). In th orst min soils, C mi ws similr n vri rom 47.2 µg g -1 to 70.7 µg g -1. Th only xption ws soil, whih ontin signiintly highr C mi (130.6 µg g -1 ). This soil lso xhiit th highst RESP mong ll th stui soils (111.0 µg C-CO 2 g -1 24h -1 ). Rltivly high RESP ws msur lso in soil (81.0 µg C-CO 2 g -1 24h -1 ). Th prntg o C org prsnt s C mi (C mi -to-c org ) ws th highst in th n soils, 8.4% n 5.3%, rsptivly. In th orst min soils th C mi -to-c org ws lowr n rs with sit g. Th lowst prntg o C org prsnt s C mi ws trmin in th nturl orst soils ( %). Th highst hyrogns tivity ws trmin in soil (20.1 µg INTF g -1 h -1 ), ollow y two nturl orst soils (16.7 µg INTF g -1 h µg INTF g -1 h -1 ). In th othr min soils, hyrogns tivity ws lowr n th minimum vlu (1.8 µg INTF g -1 h -1 ) ws msur in soil. A similr pttrn ws osrv or urs tivity: th highst tivity o this nzym ws in soil Corg (%) C org (%) N Nt t (%) C Corg-to-Nt -to-n t ph phkcl Pxt P kg -1 ) Corg-to-Pxt C -to-p (mg kg -1 ) Fig. 1. Mn vlus (n=8) o th orgni C (C org ) n totl N (N t ) ontnts, C org -to-n t rtio, xtrtl P ontnt (P xt ), C org -to-p xt rtio n ph in min soils (,,,,,,, ), n nturl orst soils ( n ). Error rs init stnr rrors. Columns shring th sm lttr o not ir signiintly (p<5).

5 Dvlopmnt o Miroil Biomss n Enzym (2.7 µg N g -1 h -1 ), ollow y th soils (1.9 µg N g -1 h -1 ) n (1.6 µg N g -1 h -1 ), n th lowst ws in soil (0.2 µg N g -1 h -1 ). Howvr, whn urs ws xprss on miroil iomss sis (urs iiny), th pttrn ws quit irnt with th highst urs iiny in soil (Fig. 2). For i phosphomonostrs tivity, th highst vlus wr msur in soils (136.9 µg p-np g -1 h -1 ) n (108.1 µg p-np g -1 h -1 ), n th lowst in soils (5.6 µg p-np g -1 h -1 ) n (8.7 µg p-np g -1 h -1 ). Th youngst orst soils ( n ) xhiit highr i phosphomonostrs tivitis thn th olr ons. Th highst Cmi C kg -1 (mg kg -1 ) ) Cmi-to-Corg C -to-c org (%) APHP APHP (μg (µg p-np g g -1 h h -1-1 ) ) APHP (μg (µg p-np µg μg -1 C Cmi h -1 h -1 ) ) Cmi -1 ) URE (μg N g -1 h ) URE (µg N g -1 h -1 DHG(μg g -1 h (µg INTF g -1 h -1 ) URE (µg N µg (μg N μg h -1 h -1 ) ) (ug g 24h -1 RESP (µg C-CO2 g -1 24h -1 ) Fig. 2. Mn vlus (n=8) o miroil iomss (C mi ), sl rspirtion (RESP), C mi -to-c org rtio, tivitis o hyrogns (DHG), urs (URE), i phosphomonostrs (APHP) n iinis o urs (URE ), i phosphomonostrs (APHP ) in min soils (,,,,,,, ) n nturl orst soils ( n ). Error rs init stnr rrors. Columns shring th sm lttr o not ir signiintly (p<5).

6 574 Chok M., Niklińsk M. Tl 2. Corrltion oiints twn hmil n miroil proprtis in th min soils (n=64, p<5). N t 0.95 C org -to-n t N.D. N.D. C org N t C org -to-n t P xt C org -to-p xt ph C mi RESP DHG APHP URE APHP P xt N.S. N.S. N.S. C org -to-p xt N.S. N.D. ph N.S C mi N.S RESP N.S DHG N.S APHP N.S URE N.S. N.S APHP N.S. N.S N.S N.S. N.S. N.S N.S. URE N.S. N.S N.S. N.S N.S. N.S. N.S N.S. N.S. not signiint N.D. not trmin i phosphomonostrs iiny ws msur or th soil (3.1 µg p-np μg C mi -1 h -1 ). For th othr soils, i phosphomonostrs iiny ws lowr n vri ovr rltivly nrrow rng ( µg p-np μg C mi -1 h -1 ). Thr ws trn or rsing i phosphomonostrs iiny with g in th sussionl n rlim min soils. Rltionships twn Chmil n Miroil Proprtis in th Min Soils Th ontnts o C org n N t wr highly orrlt with h othr (r=0.95, p<001). Howvr, non o ths lmnts orrlt with P xt (Tl 2). Thr ws strong ngtiv rltionship twn C org n N t n soil ph (r=-0.86, p<001 n -0.80, p<001, rsptivly). A wk ut signiint (p=058) ngtiv rltionship lso ws osrv twn soil ph n P xt ontnt (r=-0.26). Th miroil iomss n RESP wr orrlt with h othr (r=0.88, p<001) (Tl 2). Ths two miroil proprtis wr strongly positivly rlt to th ontnts o C org n N t (r= , p<001) n mortly, ngtivly to soil ph (r=-0.58, p<001). Th tivitis o hyrogns, urs, n i phosphomonostrs orrlt positivly with RESP (r= , p<001) n C mi (r= , p<001). Similrly to C mi n RESP, th nzym tivitis pn positivly on th ontnts o C org n N t (r= , p<001), n ngtivly on soil ph (r=-0.57 to -0.64, p<001). Non o th soil nzym tivitis ws rlt to th ontnt o P xt. Howvr, ll proprtis orrlt positivly with C org -to- P xt rtio (r= , p<001). A wk ngtiv rltionship ws osrv twn th i phosphomonostrs iiny n th ontnt o P xt (r=-0.25, p=458). Th urs iiny orrlt positivly with C org -to-n t rtio (r=0.35, p=082). Comprison o th rgrssion lins rvl tht th rgrssion oiints or th rltionships twn C mi n RESP, hyrogns, i phosphomonostrs n urs tivitis i not ir twn th rlim n sussionl soils (Fig. 3). Th p-vlus vri rom 0.54 to 0.93, initing similrity o rgrssion slops or th rlim n sussionl min soils. Disussion Chmil Proprtis o Soils Our rsults onirm inings prviously rport or this r [10]. Th lowst ontnts o C org n N t wr msur in th pur sn prior to rlmtion. Two-yrs o lupin ultivtion i not signiintly inrs th ontnts o ths lmnts. Only in th orst min soils (rlim n sussionl) wr th ontnts o C org n N t signiintly highr ompr with th pur sn, initing th importn o trs or soil vlopmnt in postmining rrns. Among th orst min soils, th highst ontnts o C org n N t wr trmin in soil, whih ontin th highst prntg o ly prtils mong th stui min soils. Soils with highr ly ontnt n stor mor C org u to uiling o orgno-minrl omplxs rsistnt to miroil grtion [16-18]. Our rsults init tht vn smll irn in th ly ontnt my rsult in signiint irns in C org umultion in th rlim min soils. Similrly, Chok n Niklińsk [19] rport tht rlim lomy sns unr

7 Dvlopmnt o Miroil Biomss n Enzym yr-ol orsttions on th xtrnl hp in Błhtów ontin signiintly mor C org n N t thn th sns. Th C org n N t ontnts in ll th min soils wr muh lowr thn in th nturl orst soils, initing tht uilup o orgni mttr n umultion o C org n N t in rlim min soils is long-trm pross [20]. Th orst min soils h signiintly lowr ph ompr with th n soils. This ws proly u to umultion o orgni mttr, whih is otn ompni y rs o soil ph [21]. Furthrmor, Pinus sylvstris, whih ws th ominting tr spis t th xprimntl sits, is known to us soil iiition [22, 23]. A rltivly high onntrtion o P xt ws msur in th soil n th P xt ontnt in soil ws signiintly highr thn in. This inits th t o rlmtion msurs tht inlu P rtiliztion on th hmil proprtis o th min soils. Th P xt ontnts t prtiulr sits i not ntirly orrspon to prviously rport rsults [10]. Howvr, in our rlir stuy th P ontnts wr msur with irnt mtho n thror somwht irnt rsults might hv n otin. Miroil n Biohmil Proprtis Th C mi n RESP wr strongly rlt to h othr oth in th rlim n in th sussionl soils. Th lowst vlus o ths proprtis wr msur in th prrlmtion min spoil. Two yrs o lupin ultivtion h rltivly smll t on ths soil proprtis n inrs only th RESP. In sussionl min soils n, th C mi n RESP rmin onstnt ut pk in th olst sussionl soil,. Th highr C mi n RESP in soil wr proly rlt not only to grul vlopmnt o soil miroil ommunitis, ut lso rsult rom highr ontnt o in prtils (silt n ly) in this soil. Soils with highr ontnt o in prtils usully mintin lrgr miroil ommunitis thn orsly txtur soils, sin thy provi ttr prottion rom unl prtion or miros n mitigt lututions o wtr vilility [16, 17]. For instn, th orst lomy sns t th rlim ovrurn hp in Błhtów ontin signiintly mor C mi n xhiit highr RESP thn th sns [19]. Among th rlim min soils, th highst C mi n RESP wr msur in th youngst soil, monstrting positiv t o rlmtion msurs on soil miroil proprtis. Howvr, this t prov trnsint s in th olr rlim min soils, n, C mi n RESP wr lowr. Dspit th lowr C org n N t ontnts, th miroil iomss n sl rspirtion in soils n rh or vn x th vlus msur in th nturl orst soils. W prsum tht in th nturl orst soils lrg proportion o C org n N t ws in th rlitrnt orm n log(aphp) log(aphp) log(resp) mi 3 log(c mi mi ) 0.8 log(c mi ) Rlmtion y = *x Sussion y = *x p = 0.93 Rlmtion y = *x Sussion y = *x p = log(c mi ) log(dhg) log(ure) Rlmtion y = *x Sussion y = *x p = Rlmtion y = *x Sussion y = *x p = log(c mi )) Fig. 3. Rltionships twn miroil iomss (C mi ) n sl rspirtion (RESP) n tivitis o hyrogns (DHG), urs (URE), i phosphomonostrs (APHP) in th rlim n sussionl soils. Th p-vlus or th omprison o rgrssion oiints r lso prsnt.

8 576 Chok M., Niklińsk M. thus ws lss vill or miros. In, th prntg o C org prsnt s C mi in th nturl orst soils ws muh lowr thn in th min soils, initing lowr vilility o C org or soil miroorgnisms. In th min soils th prntg o C org prsnt s C mi rs with g s prviously sri in svrl stuis [24-26], initing th progrssiv umultion o rlitrnt orgni mttr [26]. Dhyrogns tivity orrlt strongly with C mi, n this rltionship ws onsistnt in th rlim n sussionl soils. Consquntly, high hyrogns tivitis wr trmin in th soils with highr miroil iomss. Dhyrognss r intrllulr nzyms tht xist only in th living lls; thror, th msurmnt o hyrogns tivity hs somtims n us s msur o gnrl soil miroil tivity [6, 27, 28]. Th vill ssys my unrstimt hyrogns tivity, sin th sustrts us in th nlyss r lss tiv ltron ptors thn oxygn [3]. Our rsults init, howvr, tht in th min soils th tivity o hyrogns rlts wll thir miroil tivity. Th highst tivitis o urs n i phosphomonostrs wr trmin in sussionl min soils n, rsptivly. Urs n i phosphomonostrs r xtrllulr nzyms involv in th yling o N n P, rsptivly. Urs tlyss th + rls o NH 4 rom ur [4] n i phosphomonostrs tlyss th hyrolysis o orgni P strs to inorgni P [28]. High tivity o soil nzyms my init nutrint limittion or soil miros [29]. Howvr, th rltionships twn xtrllulr nzym tivitis n nutrint ontnts r not lwys lr us nzym tivitis pn oth on nutrint vilility n on th miroil iomss [30]. High nutrint ontnts my supprss synthsis o nzyms ut inrs miroil iomss. Sin th miroil iomss is otn mjor ontrol on miroiologilly mit prosss [31, 32] rs in th nzym synthsis my ompnst y inrsing miroil iomss [30]. In our stuy, th urs n i phosphomonostrs tivitis wr positivly orrlt with miroil iomss n this rltionship ws onsistnt in th rlim n sussionl soils. For urs tivity th rltionship with C mi ws rltivly strong (r=0.70). This my xplin th positiv orrltion o urs tivity with N t us N t ontnt h strong positiv t on th miroil iomss. W think tht high urs tivity in soil rsult minly rom highr miroil iomss us urs iiny in this soil i not ir signiintly rom th othr soils. Th ngtiv t o high ontnts o N t on urs tivity in our stuy ws vint only whn urs tivity ws xprss pr unit o miroil iomss (urs iiny) n mnist s positiv rltionship twn urs iiny n C org -to-n t rtio. Similrly, Allison t l. [30] osrv ngtiv rltionship twn N t ontnt n NAGs tivity in th minrl soils in th orgroun o Frnz Joz glir only whn th tivity o this nzym ws xprss on miroil iomss sis. Surprisingly, th i phosphomonostrs tivity i not orrlt with P xt ontnt, lthough ngtiv orrltions twn tivity o phosphtss n soil P ontnt hv n prviously rport [30, 33]. In our stuy th ngtiv rltionship twn th P xt ontnt n th i phosphomonostrs tivity ws xprss s th positiv rltionship with th C org -to-p xt rtio n th ngtiv orrltion twn i phosphomonostrs iiny n P xt ontnt. High i phosphomonostrs tivity n iiny in soil my xplin y low ontnt o P xt. Lowr P xt ontnt in soil ompr with th two olr rlim min soils ws proly th rson or highr i phosphomonostrs tivity n iiny in this soil. Th msur nzym tivitis wr orrlt with h othr n pn on miroil iomss n th hmil n physio-hmil soil proprtis. Th orrltion nlysis init tht th N t ontnt ws th most limiting tor or th miroil iomss, sl rspirtion n th tivitis o hyrogns n i phosphomonostrs. Only or th urs tivity ws C org ontnt mor importnt. Th N-limittion o miroil growth in soils is ommon phnomnon n hs n rport or irnt soils [1]. All th msur miroil proprtis orrlt ngtivly with soil ph. Soil ph is n importnt tor ting soil miros, ut usully positiv rltionships twn ph n miroil proprtis hv n rport [34-36]. In our stuy th ngtiv rltionship o soil miroil proprtis to soil ph rsult pprntly rom th t tht th umultion o orgni mttr ws oupl with rsing ph. Th C org n N t ontnts r mor importnt ontrols on soil miroil proprtis thn soil ph [1]. Conlusions Th min soils ontin signiintly lss C org n N t thn th nturl orst soils. Howvr, in som o th min soils th miroil iomss n sl rspirtion ttin th vlus typil or th nturl orst soils. Th ontnt o N t ws th most importnt ontrol on th miroil iomss, sl rspirtion, n th tivitis o hyrogns n phosphomonostrs in th min soils. All th miroil proprtis wr positivly rlt lso to C org ontnt. Th tivitis o hyrogns n urs pn strongly on C mi. Thror, high tivitis o ths nzyms wr trmin in soils ontining high C mi. Th i phosphomonostrs tivity ws lso positivly rlt to C mi, ut its tivity ws nhn in th soils with low P ontnts. Aknowlgmnts Th stuy ws inn y th Polish Ministry o Sin n Highr Eution, grnt Nos. 2P04G02230 n

9 Dvlopmnt o Miroil Biomss n Enzym Rrns 1. BAUHUS J., KHANNA P.K. Th signiin o miroil iomss in orst soils. In: N. Rstin, J. Buhus (Es.), Going Unrgroun Eologil Stuis in Forst Soils, Rsrh Signpost, Trivnrum, Ini, pp , HARRIS J.A. Msurmnts o th soil miroil ommunity or stimting th suss o rstortion. Eur. J. Soil Si. 54, 801, NANNIPIERI P., KANDELER E., RUGGIERO P. Enzym tivitis n miroiologil n iohmil prosss in soil. In: Burns RG n Dik RP (E.) Enzyms in th nvironmnt. Ativity, ology n pplitions. Mrl Dkkr, Nw York, pp 1-33, CALDWELL B.A. Enzym tivitis s omponnt o soil ioivrsity: rviw. Poiologi 49, 637, GIL-SOTRES F., TRASAR-CEPEDA C., LEIRÓS M.C., SEOANE S. Dirnt pprohs to vluting soil qulity using iohmil proprtis. Soil Biol. Biohm. 37, 877, DE MORA A.P., ORTEGA-CALVO J.J., CABRERA F., MADEJÓN E. Chngs in nzym tivitis n miroil iomss tr in situ rmition o hvy mtl-ontmint soil. Appl. Soil Eol. 28, 125, BALDRIAN P., TRÖGL J; FROUZ J., ŠNAJDR J., VALÁŠKOVÁ V., MERAHUTOVÁ V., CAJTHAML T., HERINKOVÁ J. Enzym tivitis n miroil iomss in topsoil lyr uring spontnous sussion in spoil hps tr rown ol mining. Soil Biol. Biohm. 40, 2107, STRZYSZCZ Z. Th soil lss rlmtion mtho o th tr-inustril rs in Silsin provin hivmnts n thrts. Soil Sin Annul. 25, 405, 2004 [In Polish]. 9. PIETRZYKOWSKI M. Chrtristis o slt turs o rorsnt vgttion in rlim rs n in rs lt or sussion s xmplii y xprimntl plots in th Szzkow sn min xvtion. At Agr. Silv. sr. Silv., 63, 1, 2005 [In Polish]. 10. CHODAK M., PIETRZYKOWSKI M., NIKLIŃSKA M. Dvlopmnt o miroil proprtis in hronosqun o sny min soils. Appli Soil. Eol. 41, 259, SCHLICHTING E., BLUME H.O. Bonkunlihs Prktikum. Pul Pry, Hmurg, ANDERSON J.P.E, DOCH K.H. A physiologil mtho or th quntittiv msurmnt o miroil iomss in soils. Soil Biol. Biohm. 10, 215, VON MERSI W. Dhyrogns tivity with th sustrt INT. In: Mthos in soil iology. Springr Vrlg: Brlin, Hilrg, pp , MARGESIN R. Ai n lklin phosphomonostrs tivity with sustrt p-nitrophnyl phospht. In: Mthos in soil iology. Springr Vrlg: Brlin, Hilrg, pp , KANDELER E. Urs tivity y olorimtri thniqu. In: Mthos in soil iology. Springr Vrlg: Brlin, Hilrg, pp , FRANZLUEBBERS A.J., HANEY F., HONS F.M., ZUBERER D.A., Ativ rtions o orgni mttr in soils with irnt txtur. Soil Biol. Biohm. 28, 1367, MÜLLER T., HÖPER H. Soil orgni mttr turnovr s untion o th soil ly ontnt: onsquns or mol pplitions. Soil Biol. Biohm. 36, 877, SOLLINS P., HOMANN P., CALDWELL B.A. Stiliztion n stiliztion o soil orgni mttr: Mhnisms n ontrols. Gorm 74, 65, CHODAK M., NIKLIŃSKA M. Et o txtur n tr spis on miroil proprtis o min soils. Appl. Soil Eol. 46, 268, ŠOURKOVÁ M., FROUZ J., FETTWEIS U., BENS O., HÜTTL R.F., ŠANTRŮČKOVÁ H. Soil vlopmnt n proprtis o miroil iomss sussion in rlim post mining sits nr Sokolov (Czh Rpuli) n nr Cottus (Grmny). Gorm 129, 73, BOLAN N.S., HEDLEY M.J., WHITE R.E. Prosss o soil iiition uring nitrogn yling with mphsis on lgum s psturs. Plnt Soil 134, 53, ARNOLD G. Soil iiition s us y th nitrogn uptk pttrn o Sots pin (Pinus sylvstris) Plnt Soil 142, 41, EMMER I.M., SEVINK J. Tmporl n vrtil hngs in th humus orm proil uring primry sussion o Pinus sylvstris. Plnt Soil 167, 281, INSAM H., DOCH K.H. Rltionships twn soil orgni ron n miroil iomss on hronosqun o rlim sits. Miro. Eol. 15, 177, ŠANTRŮČKOVÁ H. Miroil iomss, tivity n soil rspirtion in rltion to sonry sussion. Poiologi 36, 314, GRAHAM M.H., HAYNES R.J. Orgni mttr sttus n th siz, tivity n mtoli ivrsity o th soil mirolor s initors o th suss o rhilittion o sn uns. Biol. Frtil. Soils 39, 429, PASCUAL J.A., GARCIA C., HERNANDEZ T., MORENO J.L., ROS M. Soil miroil tivity s iomrkr o grtion n rmition prosss. Soil Biol. Biohm. 32, 1877, TAN X., CHANG S.X., KABZE R. Soil omption n orst loor rmovl ru miroil iomss n nzym tivitis in orl spn orst soil. Biol. Frtil. Soils 44, 471, SINSABAUGH R.L., ANTIBUS R.K., LINKINS A.E., MCCLAUGHERTY C.A., RAYBURN L., REPERT D., WEILAND T. Woo omposition: nitrogn n phosphorus ynmis in rltion to xtrllulr nzym tivity. Eology 74, 1586, ALLISON V.J., CONDRON L.M., PELTZER D.A., RICHARDSON S.J., TURNER B.L. Chngs in nzym tivitis n soil miroil ommunity omposition long ron n nutrint grints t th Frnz Jos hronosqun, Nw Zln. Soil Biol. Biohm. 39, 1770, ŠANTRŮČKOVÁ H., VRBA J., PICEK, T., KOPACEK. Soil iohmil tivity n phosphorus trnsormtions n losss rom iii orst soils. Soil Biol. Biohm. 36, 1569, SELTMANS P.C., HART S.C., BOYLE S.I., STARK J.M. R lr (Alnus rur) ltrs ommunity-lvl miroil untion in onir orsts o th Pii Northwst, USA. Soil Biol. Biohm. 37, 1860, ALLISON S.D., VITOUSEK P.M. Rsponss o xtrllulr nzyms to simpl n omplx nutrint inputs. Soil Biol. Biohm. 37, 937, PRIHA O., SMOLANDER A. Nitrogn trnsormtions in soil unr Pinus sylvstris, Pi is n Btul pnul t two orst sits. Soil Biol. Biohm. 31, 965, ANDERSSON S., NILSSON S.I. Inlun o ph n tmprtur on miroil tivity, sustrt vilility o soil solution tri n lhing o issolv orgni ron in mor humus. Soil Biol. Biohm. 33, 1181, QUILCHAO C., MARAŃÓN T. Dhyrogns tivity in Mitrrnn orst soils. Biol. Frtil. Soils 35, 102, 2002.

10 Copyright o Polish Journl o Environmntl Stuis is th proprty o Sintii Invstigtion Committ n its ontnt my not opi or mil to multipl sits or post to listsrv without th opyright holr's xprss writtn prmission. Howvr, usrs my print, ownlo, or mil rtils or iniviul us.