Wpływ stymulowanych kwaśnych deszczów na drobnoustroje glebowe

R O C Z N IK I G L E B O Z N A W C Z E T. X L II N R 1/2 W A R S Z A W A 1991: Г7—G8

A L E K SA N D R A PO K O JSK A , M A R IA K A M PER T, H A N N A DAHM , EDM U ND STRZELCZYK, ZBIG N IEW PR U SIN K IEW IC Z

EFFECT OF SIM ULATED ACID R A IN ON SOIL M ICROORGANISM S

of soils scien ce

In stitu te of B iology, N icolau s C opernicus U n iv ersity , Toruń

IN TRO DU CTIO N

It is assum ed that acidic deposition esp ecially at high loading rates,

affects soil organism s and b iological processes in th e soil. T hese effects

m ay be p ositive or n egative, depending upon en viron m en tal condi­

tion s and the organism or process in volved [13]. Som e artificial acidi­

fication exp erim en ts h ave show n reductions in total bacterial popula­

tions in the forest floor [5,

]. O ther w orkers h ave sh ow n no sig n ifi­

cant changes in total num bers, but a significant decrease in m etab oli-

cally active bacteria w ith h igh le v e ls of acidic inp u ts [4].

M ost soil bacteria grow best at pH valu es near neu trality. D ec­

reased pH can affect m em brane p erm eab ility and enzym e activ ity [15]

resu ltin g in low er grow th and activity. N ot all bacteria react n eg a tiv ely

to low pH. A m ong bacteria m any acidophilic sp ecies are know n. A lso

m icroorganism s have th e capability to adapt to altered soil pH [16].

The ob jective of our stu d y w as to learn h ow sim u lated acidic de­

p osition affects m icroorganism s in three soils. The problem of the

effect of acidic deposition on soil m icroorganism s is im portant and

actual because plant grow th, d evelop m en t and h ealth are connected

w ith th e biochem ical a ctiv ity of saprophytic both sym biotic and non

sym biotic soil m icroorganism s and because th e soil receives m ore and

more acidic depositions.

M A TER IA LS A N D METHODS

D E S C R IP T IO N S O F TH E O B JE C T O F S T U D Y

The studies w ere carried out in th e northern part of Tuchola Fo­

rests, Czersk F orest D istrict. This area is situ ated far from greater

58 A. P o k o j s k a i inni

industrial centers and is re la tiv ely free of air p ollution (m ainly sulphur

and nitrogen oxides).

The exp erim en tal plot (42 X 52 m) w as set up on clear cutting

area in a lich en pine forest (C l ad oni o- Pi ne t um) w ith transition to a poor

form of the suboceanic forest (L e u c o b r y o - P i n e t um cladonietosum). The

soil w as classified am ong ru sty soils, w ith hum us of m oder type

(xerom oder). It is form ed from w e ll drainage flu vioglacial sands. The

signs of form er p loughing are v isib le in a profile.

The selected plot w as cleared of stubs and 15 p arallel belts (37 m

long) w ere traced out. Ten ditches (0.5 m deep, 0.5 m w ide) w ere dug

on the belts. F ive of them w ere filled w ith m arly sandy loam and

fiv e —

w ith m ucky soil. The rem aining fiv e b elts w ere lacking of

forest floor and the natural ru sty soil w as left.

Som e p hysical and chem ical properties of the soils used are given in

Tables 1 and

.

One year old pine seed lin gs (Pinus s yl v e s tr i s L.) w ere planted on

th e b elts b etw een 22— 24 of April 1987. D uring 1987— 1989 the soils

w ere w atered m on th ly (from A pril to N ovem ber) w ith solutions of

sulphuric and nitric acids or w ith w ater from the deep -w ater w e ll (con­

trol). A ny contact of the seedlin gs w ith the acids w as avoided. 1 1 of

B ulk d en sity ! Ign ition losses С n i С : N g /cm ;: «!o % 1.48 R usty soil 2.96 1.65 0.052 31.7 M ucky soil 34.26 20.11 0.723 27.8

■Vlarly sa.id y loam (1.5% C a C 0 3)

1.11 0.25 0.014 17.9

T a b l e 2 D yn am ics of pH ch an ges (p Ii-H 20 ) of e x p erim en ta l soils m easured a lw a y s in

N ovem ber (m ean valu es; n = 25)

Soil 1 C om bination j ' 1 1937 ! i 1938 1 1989 control 4,72 4.74 4.60

R usty soil _{acid ified soil 3.70 3.57 3.54}

control 5.20 5.14 5.13

M ucky soil _{acid ified soil 3.89 3.55 3.29}

-control 8.10 8.25 8.20

E ffect of acid rain on m ic r o o r g a n i s m s _{5 9}

0.225 M H

S 0

and 0.5 1 of 0.225 M H N 0

w ere applied each tim e to

1 m

— th is corresponds to 1.5 m m of rainfall.

D etailed data on th e exp erim en tal area are given by P rusinkiew icz,

K w iatk ow sk a and Pokojska [17].

M I C R O B I O L O G I C A L S T U D I E S

The sam ples of three soils for m icrobiological stu d ies — from con­

trol and acidified area — w ere collected on spring, sum m er, autum n

during 1987— 1988 and on spring 1989. The surface layer of the soil

w as rem oved and sam ples w ere taken to a depth of 5 cm.

The follow in g an alyses w ere perform ed:

1. Total num ber of bacteria on soil extract agar [12]. N y sta tin and

actidion in am ount of 50 mg/1 w ere added to th e autoclaved m edium .

2

. N um ber of fu n gi on M artin’s m edium w ith rose bengal (33.3 mg/1)

and aureom ycin (30 mg/1).

3. Id en tification of fu n gi w as p erform ed according to G ilm an [11]

and B arnett [

] m anuals.

4. For enum eration of num bers of m icroorganism s belonging to

d ifferen t p hysiological groups:

am m onifiers,

denitryfires, m icroorga­

n ism s reducting of su lp h ates and decom posing of cellu lose, th e m ost

probable num ber m ethod has been applied [18]. The m edia used for

grow in g the above m icroorganism s w ere the sam e as recom m ended by

A llen [3].

R ESU L TS

R esu lts on total num ber of bacteria and fu n gi in control and acidi­

fied soils are p resented in Table 3. D epending on the season of the

year, soil typ e and the degree of acidification big d ifferen ces in the

q u an titative occurrences of these m icroorganism s w ere noted. In g e­

neral th e m arly sandy loam and m ucky soil contained more bacteria

than the ru sty one, but the num ber of fungi w as the sm alest in the

m arly sandy loam (Table 3).

A cidification of th e soil affected stronger the bacteria than fungi.

The n um erosity of bacteria in control and acid treated soils d iffered

sta tistically (S tu d en t’s test) w hen exam in ed at d ifferen t seasons of

th e year (Tabela 3). In the rusty and m ucky soils acidification caused

a significant decrease in th e num ber of bacteria but an increase in the

m arly sandy loam. D ifferen ces in num ber of fu n gi in control and aci­

d ified soils in su ccessive seasons w ere often sta tistica lly not significant.

In th e ru sty soil acidification caused a decrease o f th e num ber of

T a b i c 3 N um bers of bacteria and fu n gi in d iffeven t soils (in th ou san ds per 1.0 g of d iy boil)

a — n u m b e r s o f b a c te r ia , b — n u m b e r s o f fu n g i. * s ig n if i c a n t d if fe r e n c e s a t P < 0.05.

— n o s ig n ifi c a n t d if f e r e n c e s a t P < 0.05. Date

of sam pling

Rusty soil 1 M ucky so il M arly sandy loam

control 1 acid ified soil | control 1 a cid ified soil control j acid ified so il 1987 3 VI a b 356* 271 " 110* 251 8 562* 219-3 190* 144 4 029" 100“ 4 894“ 75“ 5 V III a 1 686~ 2 006" 10 734* 3 713* 68 639- 85 727-b 346- 334- 177- 162“ 34* 104* 7 X a 2 244* 1 367* 7 410" 6 840- 7 405* 12 159* Ъ 271* 144* 69- 96- 45“ 45-1988 19 IV a 633* 283* 10 236* 909* 3 789“ 5 179-b 301- 235- 72“ 165- 44- 38-1 V II a 456“ 257“ 9 210* 3 451* 20 866* 37 668* b 176* 70* 194* 350* 5 0 - 6 0 -4 X a 1 4Ö5* 157* 11 686* 1 070* 5 762* 10 797* b 201" 195" 257“ 278* 76* 184* D V a 1 361* 44* 2 441* 602* 3 710* 9 052* b 195* 61* 115* 215* 40* 84*

T a b l e 4 T w o-factor an a ly sis of varian ce (ANOVA) com paring th e e ffects of seasons and a cid ifica tio n on n u m bers of bacteria

and fu n g i in d ifferen t soils

E ffects of: D egrees of freedom

R u sty soil M ucky soil M arly san d y loam

F 1 P F 1 P F

₁

6

0.00000

0.00002

0.00000

1

0.00000

0.00000

6

0.00000

6

0.00002

0.00002

0.00001

1

0.00111

6

62 A. P okojskci i in n i

fungi. On the other hand in th e m ucky soil and m arly sandy loam

increasing acidification affected

th e n u m erosity of fungi.

T w o-factor an alysis of variance (ANOVA) has sh ow n a significant

effect of the season of th e year and acidification on num bers of fu n gi

and esp ecially on bacteria in particular soils (Table 4). U su ally the

effect of th e season w as sta tistic a lly m ore significant than acidification.

No sign ifican t d ifferen ces w ere noted in kind com position of fu n gi

am ong th e control and acidified soils (Table 5). Such d ifferen ces w e re

connected rather w ith the soil typ e than w ith acidification.

T ables

show num bers of am m onifires,

den itryfires, su lp h ate

reducers and cellu lose decom posers. N um bers of d en itryfiers and am m o-

n ifiers d iffered considerable in d ifferen t seasons. In the soils under

stu d y cellu lose decom posers and sulphate reducers w ere not num erous.

S tatistical evalu ation s (M ann-W hitney U test) have show n that th e

effect of sim ulated acid rain on d ifferen t p hysiological groups of m i­

croorganism s w as m ost o ften unsignificant.

T a b l e 5 P red om in an t kin d s of fu n g i in the soils under stu d y

Soil Control A cid ified soil

R usty soil P é n ic illiu m sp. P é n ic i ll iu m sp. M ortier ella sp. T r ic h o d e r m a sp.

M ucky soil P énic illiu m sp. M o r tier ella sp. T r ic h o d e r m a sp. T r ic h o d e r m a sp.

Mucor sp. P é n ic illiu m sp.

P u llula ria sp. Mucor sp.

C la d o s p o r iu m sp.

M arly sandy loam P é n icilliu m sp. P é n ic illiu m sp. M ortierella sp. T r ic h o d e r m a sp. T r ic h o d e r m a sp. l l y a l o p u s sp.

Mucor sp. P ullula ria sp.

Mucor sp.

D ISC U SSIO N

R eview s on th e effects of acidic deposition on soil organism s or

n utrien t cyclin g have been p ublished by A lexan d er [1, 2], Colem an

[10], Olson [14]. An ex cellen t rev iew on th e effects of acidic deposition

on soil organism s has been p ublished by M yrold [13]. M any of the

stu d ies on the effect of atm ospheric deposition on bacteria have also

m easured fu n gal populations. In general m oderate to high acidic inputs

T a b l e б N um bers of m icroorganism s b elon gin g to d ifferen t p h ysiological groups in th e ru sty soil (per 1 .0 g of dry soil)

Date of sam pling

D enitryfiers _{A m m on ifiers} M icroorganism s red u cin g of su lp h ates

M icroorganism s d ecom p osin g of cellu lo se

control acidified soil control acid ified so li control a cid ified soil control 1acid ified soil 1987 3 VI 5 VIII 7 X 7 900 108 400 1 000 26 000 106 700 490 10 000 5 100 4 800 98 000 168 000 500 0 8 0 0 3 0 0 5 10 4 0 0 1988 19 IV 11 800 46 000 260 4 600 4 4 4 4 1 V II 260 000 21 000 260 40 0 26 0 4 4 X _{800 50 260 25} 0 9 46 0 1989 5 V 455 000 150 4 500 40 9 0 4 0 M ann-W hitney U test (significance ^ ~ level) = 0.28 P == 0.66 _{P = 0.66 P = 0.14}

T a b l e 7 N um bers of m icroorganism s belonging to d ifferen t p h ysiological group in the m ucky soil (per. l.ü g of dry soil)

D ate of sam pling

D en itry fiers A m m o n ifiers M icroorganism s reducing _{of su lp h ates} M icroorganism s decom p osin g o f cellu lose

con trol acid ified soil control acid ified soil con trol acid ified soil con trol a cid ified soil 1987 3 VI 150 000 6 700 70 500 44 800 0 10 150 220 5 V III 4 570 000 350 000 820 000 260 000 7 12 820 12 7 X 40 000 2 400 18 400 24 000 0 0 150 10 1988 19 IV 790 000 41 300 3 900 24 800 89 10 390 120 1 VII 1 8G0 000 147 000 15 400 4 000 32 62 650 310 4 X 4 500 1 500 3 700 3 100 67 17 1 100 46 1989 5 V 99 500 2 600 12 600 5 800 33 19 12 0 M ann-W hitney U test U test P = 0.06 P = 0.91 P =0.66 P = 0.04 (significance level)

T a b l e 8 N um bers of m icroorganism s b elon gin g to d ifferen t p h y sio lo g ic a l groups in the m arly sandy loam (per 1 . 0 g of dry soil)

D ate

D en itryfiers A m m on ifiers M icroorganism s reducing of su lp h ates

M icroorganism s decom p osin g of cellulose

of sam p lin g

control acid ified soil control acid ified soil control acid ified soil control acid ified soil

1987 3 VI 25 800 9 700 46 500 30 600 9 0 0 15 5 V III 10 600 8 400 28 000 16 700 0 0 280 280 7 X 2 000 78 000 11 900 4 700 0 0 2 800 4 700 1988 19 IV 460 000 1 000 000 46 000 21 000 3 0 97 16 1 V II 483 000 451 000 2 700 250 4 0 8 95 4 X 26 000 46 000 2 600 1 500 4 0 26 97 5 V 30 000 45 000 750 4 000 4 0 0 11 M ann-W hitney U test (sign ifican ce P = 0.57 p = 0.57 P = 0.02 P =: 0.49 level)

6 6 A. P o k o j s k a i in ni

h a v e not been found to effect fu n gal populations in organic or m ineral

soil orizons [7, 9]. The sam e phenom enon w as stated in our studies-

Soil acidification affected stronger bacteria than fungi. In rusty and

m u ck y soils acidification caused a decrease in num ber of bacteria but

an increase in m arly sandy loam soil.

Acidic deposition m ay affect soil organism s directly or indirectly.

The greatest attention received acidity. The acidity is h ow ever com ­

p licated by association of H + w ith various anions, esp ecially N O “ and

S O “ . Indirest effects of atm ospheric inputs on soil organism s m ay be

concerned w ith alternation of the soil chem ical en viron m en t and altered

organic m atter inputs [13]. Thus it seem s that both th e direct and

indirect im pact of acidic deposition on soil m icroorganism s requires

fu th er studies.

It is su ggested by M yrold [13] that fu th er research should be done

on the effects of realistic chronic inputs, because m uch work on the

effect of acidic deposition on soil organism s has in volved u n realistica lly

h igh inputs over short tim e periods.

R EFERENCES

[1] A l e x a n d e r M. E ffects of acid ity on m icroorgan ism s and m icrob ial pro­ cesses in soil. (In.) E ffects of A cid P recip ita tio n on T errestia l E cosystem s. (H utchinson Т. C. and H avas M. eds). P len u m P ress, N e w Y ork 1980. [2] A 1 с X a n d e r M. E ffects of acid p recip ita tio n on b io ch em ica l a c tiv itie s in

scil. (In:) E cological Im pact of A cid P recip itation . Proc. Intern. Conf. (Dra- bloes D. and T ollan A. eds), SN SF, Oslo 1980, N orw ay.

[3] A 11 e n O. N. E x p erim en ts in soil b acteriology. B urgess Publ. Co., M inneapolis 1951.

[4] B e a c h E., L u n d g r e n B., S ö d e r s t r ö m B. E ffects of a rtific ia l acid rain on m icrobial a c tiv ity and biom ass. B ull. Env. C ontam . T oxicol. 1979, 23: 737— 740.

[5] B a a t h E., B e r g U., L o h m B., L u n d k v i s t T., R o s w a 11 B., S ö d e r ­ s t r ö m B.. W i r e n A. S oil organism s and litter decom p osition in a Scots pine forest. E ffects of e x p erim en ta l a cid ifica tio n . (In:) E ffects of A cid P r e ­ cipitation on Terrestial- E cosystem s. (H utchinson T. C. and H avas M. eds). P len u m Press. N ew Y ork 1980.

[6] B a a t h E., B e r g B., L o h m U., L u n d g r e n B., L u n d k v i s t H., R o s - w a l l B., S ö d e r s t r ö m B., W i r e n A. E ffects of ex p erim en ta l a c id ifi­ cation and lim in g on soil organ ism s and decom p osition in Scots p in e forest. Pedobiol. 198U, 20: 85— 100.

[7] B a a t h E., L u n d g r e n B. S ö d e r s t r ö m B. F ungal pop u lation in podzo- lic so il ex p e r im e n ta lly a cid ified to sim u late acid rain. M icrob. Ecol. 1984, 10: 197—203.

[8] B a r n e t t H. L. Illu strated genera of im p erfect fu n gi. B urgess Publ. Co., M in n eap olis 1985.

[9] В a w 1 e y R. J., P a r k i n s o n D. E ffects of sulphur d ioxid e p ollu tion on fo rest soil m icroorgan ism s. Can. J. M icrobiol. 1984, 30: 1— 10.

[10] C o l e m a n D. C. T he im p acts of acid d ep osition on so il biota and С c y c ­ ling. Env. Exp. Bot. 1983: 23, 225— 233.

E ffe c t of acid rain on m ic r o o r g a n i s m s _{6 7}

[11] G i l m a n J. C. A m an u al of soil fu n gi. The Iow a S tate C ollege Press, U SA , 1957.

[12] L o c h h e a d A. G., C h a s e F. E. Q u alitative stu d ies o f soil m icroorganism s. V. N u trition al req u irem en ts of the p red om in an t b acterial flora. S o il Sei. 1943, 84: 395.

[13] M y r o l d D. D. E ffects of acidic d ep osition on soil organism s. N C A SI, Tech. B ull. 1987, 527: 1— 29.

[14] O l s o n R. A. The im pacts of acid d ep osition cn N and S cycling. Env. Exp. Bot. 1983, 23: 211— 223.

[151 P a d i n E. A daptation of b acteria to e x te r n a l pH. (In:) Current P ersp ectiv es in M icrobial E cology. (K lug M. J. and R eddy C. A. eos). A m er. Soc. for Ivlicrobiol., W ashington 1981.

[16] P a r k i n T. B., S e x t o n e A. J., T i e d j e J. M. A d ap tation of d en itri­ fy in g p opulations to lo w soil pH. A ppl. Env. M icrobiol. 1985, 49: 1053— 1056. [17] P r u s i n k i e w i c z Z., K w i a t k o w s k a A., P о к o j s к a U. D yn am ik a

zm ian odczynu gleb pod w p ły w em k w a śn y ch d eszczów (w druku).

[18] R o d i n a A. M ikrobiologiczne m etod y badania w ód. PW RiL, W arszaw a 1968.

А. П окойска, М. К ам перт, Г. Дам, Э. С трж ельчы к, 3. П руси нкеви ч В Л И Я Н И Е И С К У С С ТВ Е Н Н Ы Х К И С Л Ы Х Д О Ж Д Е Й Н А П О Ч ВЕН Н У Ю М И К РО Ф Л О РУ З а в ед ен и е М икробиологии и Зав ед ен и е П оч вов еден ия И нститута Биологии У н иверситет Н иколая К оперни ка, Т орунь

Р е з ю м е

6 8 A. P o k o j s k a i in ni

A . P O K O J S K A , М. К AM P E R T . H. D A HM, E. ST R Z E L C Z Y K , Z. P R U S IN K IE W IC Z W PŁYW SY M U LO W A N Y C H K W A ŚN Y C H DESZCZÓW

N A D R O BN O U STRO JE GLEBOW E

Zakład M ikrobiologii i Z akład G leb ozn aw stw a In stytu tu B iologii U n iw ersy tetu M ikołaja K opernika w Toruniu

S t r e s z c z e n i e

Przeprow adzono a n a lizy m ik rob iologiczn e trzech gleb (piaszczysta, m u rszow a i glin iasta) k on troln ych i zak w aszon ych . Stw ierd zon o, że za k w a szen ie gleb w p ły ­ w ało siln iej na liczeb n ość b ak terii niż grzybów . W glebach p iaszczystej i m ur- szow ej n otow an o isto tn y spadek liczeb n ości b ak terii pod w p ły w em zak w aszan ia, a w g leb ie g lin ia stej — w zrost ich liczebności.

W pływ pór roku na liczeb n ość b ak terii i grzyb ów b ył n ajczęściej bardziej isto tn y sta ty sty czn ie niż w p ły w zak w aszan ia.

W pływ sy m u lo w a n eg o k w a śn eg o deszczu na różne grupy fizjo lo g iczn e d rob ­ n ou strojów był n ieisto tn y sta ty sty czn ie.

D r A. P o k o j s k a P r a c a w p ł y n ą ł a d o r e d a k c j i w l i p c u 1990 r. Z a k ł a d M i k r o b i o l o g i i

U n i w e r s y t e t M i k o ł a j a K o p e r n i k a w T o r u n i u 87-100 T o r u ń , G a g a r i n a 9