and 'Phosphate Solubilizing Bacteria' and their Influence on Yicld and ...

Zentralhl. Mikmhiol. 143 (1988). 637-643 VEB Gustnv Flscher Verlap Jena

IDivision of Micrcihiolugy. 1ndla11Agricultural Research InstlNle. New Delhi. Indial

Interaction Between Aiospirillum hrusilense and 'Phosphate Solubilizing Bacteria' and their Influence on Yicld and Nutrient Uptake of Sorghum (Sorghum bicolor L.) A. R. ALAGAWADI and A. C.GAUR

Summary lnteraction between A : o s p ~ r ~ l l ~brusilrnar m and 'Phoaphute soluh~l~z~ng hilcteria' v17, h c , u d o m r ~ ~sfrruru m ~ and Becillur polymjxo, and thew Influence on yleld, nutrient uptake and acetylene reduoiun activity 01' aorghun~were studled insandy.loumalluvvl soil under greenhouse condltlcm, The re.;ulta revealed it sipn~licanltscreaar in the y~eld level ul'sorghum due ai cumhlncd ~noculatlonuver slnglc inoculation lndlcst~npu poaltlvr ~ntcrilclionhc1wcc.n the 2 groups of bacteria. Thc yields were further enhanced markedly hy the uppl~cat~unol 40 ke N as urea and MI kg PIO, in the form of rock phosphate per Iln togcther w ~ t hinoculation treatmentr uver fcnil~ccralonc or inoculation without fenilirers. The possihilitiesof saving40 kg N ilnd replacing the entin: quuntity of super phlnphate u ~ t hn ~ c kphosphate plus innculatinn of A. hrus~lm,seand P . srriuw or B. polymyxu together are discussed.

Zusammenfassung Die Wechselwirkung rwlachen Azo.spirillum hrusilcnse und den phosphatlosenden Btikter~en P.rrudomonur striafu und ~rrnlluspolymyxu. ihr EinfluLi auf den Enrag, die Nnhrstoffaul'nuhn~eund Aktlvitiit der AcelylcnReduktlon vun Sorghum (S[~rghurnbicobr) in einem schluffsand~genalluvialen Boden wurden unter Glashausbedingungcn untersucht. Die Ergebn~ssezeigtcn eine tignifikante Emagserhiihung vun Sorghum be1 dcr komhiniemn Beimpfung mlt he~denOrganlsmon gegenuber jener mil cinzelnen Organismen, was elne positive lnteraktion zw~schenbeiden Bukteriengruppen andeutet. Die Enripe waren auch weiterhin erhoht bei Anwendung von 40 kp Nlha als Hamstoff und 60kg P20riha als Rohphosphat ?usammen niit den Beimpfungsbehandlunpn gegenubcr Dungeminelgaben uhne Beimpfung rder eincr Beimpfunguhne Zusatzdiinpung. Die Moglichkctteneiner Einsparung von 40kg Nlha und einca Austausches von Superphusphat durch Rohphosphat k i glc~ch~xitigerBelmpfung von A, brusilense und P , sfrialo odcr B, polympxu werden diskutien.

Positive role of microorganisms in nutrition and growth of plants is established beyond doubt. The fast depletion of fossil fuel resources, increasing costs of chemical fertilizers and the environmentul pollution caused by them have called for more attention to the use of microorganisms, with the aim of increasing the availability of nutrients to plants. Crop productivity can he improved by qanipulating the rhizosphere microbial population. In this respect, biofertilizdrs prepared from Azofohacter, Rhizohium, ~ z c ~ s ~ i r i ) and l u m'Phosphobacteria' have shown encouragingresults ( G A u Ral.~ ~IYXO,HERNANDEZ and HILL 19x3; SUBBAR ~ o eal. t 1983; PACOVSKY et al. 1985). In recent years, the combined use of chemical fertilizers and biofertiliacrs as a workable technology in p p production has been considered. The present report attempts to study the effect of inore than one biofertilizer in combination with chemical fertilizers on nutrition and growth of sorghum plants.

Sdl: Unn&dI'i uady-loun dhvid roil & l k fmnn the ladim Apicultunl Research lnuitute. far&, NovDpfhi,~rraad.lhcaoiIhdapHof7~9,~ic~O.39%,7lmgNpnd8.90mgP~gsail.ThesoiI,Sner ~ ~ 2 m m r i c vw c~ m, i ~ m o m u h l y w i ( h R a c l ~ p o c v ~ f ~ n I r d ~ m m a IOtonrlha tUu~of ad l0kg m i l - r a filled in edm pas of30 cm d ~ ~kaa* ' rock phapmre(RP) or super phosphate (SP) at 601uP~ndursl~40udBOknNhwnemied~buuldoselotkDd~aoerthe~eammt~~hed~le. ~~: Smghvm d r (cv.CSH-~) &by d i n g in liquid cuilure fbr 2 h wca sown immediately. Pa onabincd inoculation, Ihc liquid culturea of A, bwiknse .nd P. s& or B. polymyxn were mixed in equal plopabbrusnd the wcdr vne &. I0 skb me sown ar w h mt. Mer n m i u t i o n , thinning was done to ~n4plrmrprpat.lk~pcrimmt w a m n d u n c d u s i n g l l Yi n with 3repli&tions foreach hubmntadstw?cfi.e. at40.80and l u ) d ~ . l h c m c s w c r e w a ~ ~ r m M v t o m a i n ~ ~ i n o M l . Mivily: Actwknc rcduniw lamy of sorghum mafs w k &m at 40 ;Id80 d of plant gmwlh Aoerybnc u dncrlbedby DAYel d.(19151. uslnp Igas c h m t o & p h (AIMIL.Nucon e g c n fluK wnimtion delcnor md W - R r Icolumn temararua of W°C. 'The r t t r l n I.caparsed a\ n mlc, C.H. produd p e r g ~mots l on dry weight basis per hour. ' Dty mstlg and grain yield: At 40 and 80d. the p h t s wcre uprooted gently and the himsphere soil samples were ml!atd for enumeration of Azhrpiril1um and PSB. The plants wea dried at65'C for 72 h and dry weights wea rrcndcd. When (he plants wen complclely manve, hweliting was donc and the grain and smw yields wcre recorded. Nand P upmke: The plant wimples collected at 40 and 80d and the grain and snaw samples after harvest were analyaed for lnal niaogcn using Tcchniwn aumnalyscr (Tcchnicon Monograph 1, 1971) and lotal phosphorus by 1958). Jackson's method (JACKSON The soil ample8 colleaed at the same intervals were snalysed for available phosphorus (OI.SL.N el al. 1954). Rhiwsohere oomlauon of Azos~irillumand PSB: The rhizor~hcresnil mmoles collected frnm all the 3 rcolicate~ were pooled and used for orenunuration at 40 and 80 d of plant growth. EnumerationdAzosprrillum was done by MPN methk usingsemiwlid N-fmmalate medium ( D O ~ E & I N EDAY R ~1974) ~ ~ and that of thePSB was done by dilutron plate method using Pikovskaya's medium (PIKOVSKAYA 1948).

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Results Table 1 shows the grain and s m w yields of sorghum at harvest and the dry matter yield at 40 and 80 d of plant growth. Maximum grain yield wa. recorded in NsoSP, (80 kg N + 60 kg P,Os as super (40 kg N + 60 kg PzO, as rock phosphate) followed by A. hrasilense B, polymyxu + N&P, phosphate) and A. brasilense + P. strraru + N&P, which were statistically at par with each other and superior over other treatments. The combined inoculation treatments with and without NdoRPnu showed a significant increase in the grain yield over single inoculation of A. hru.rilense. P. striaruor B. polymyxu with NaRPm and over P. strialu or 8. polymyxu without N RP respectively. Application of N&PW to single inoculations had no significant effect on grain yield over NmRP, alone, but a significam incredse in the grain yield was noted with combined inoculation plus N a p n a . A similar trend in the straw yield was also obkemed at harvest. The dry matter yield of sorghum plants at 40 and 80 d further suggests the superiority of NnOSPm over all other treatments. Combined inoculation of A. brusilenre and P. striaru or B. polymyxu with showed significant increase in dry matter over single inoculation,of P. srriata or or without N&P, B. polymyxu with or without N&P, at both 40 and 80 d (Table I). Tottil N and P uptakein grain and straw of sorghum as influenced by single and dual inoculation are given in Table 2. The trend in nutrient uptake was the same asobserved in grainand straw yields. The total N and P uptake were highest in NnoSPnafollowed by A. brusilense + B. polymym + N&PW and A. hrasilense P. striaru + N&P, and least in uninoculated control. The increase in N-uptake was 2,4 fold while that of P-uptake was up to 2.9 fold over uninoculated control.

+

+

tm#xun mots, st 40 d, w u sigmficantlymore in plann inoculated with A. braJh#s'withorw&mtPSB md N&PW(Table 3). Combiaed inoculation aad NMRpMhad iPlbvoce m nitmactivity at 40 d over single inoculation of A. bmsilense. &&W, af Wdzhe anyme activity wan &m in A. bmdl.nre 8. polymyxa N&PM -by A. b d h e P. snhto N & P a . A. brasilense B polymyxaand.4, braslknse P. w m e48Ibtidy 8 i g n i r i t over rest of the trcahncols The a d v ~ t ywas lowest

+

+

+

+

+

+

h , N & F d a t 8 0 d . lYMP 3. NIaDgrmmaclivhy in wqbum mots an ~nfluenadby a~ngkand dud ~noculetlonw ~ t hA brnsilcnse and phOqb.lsrohbiluin(l~fPSB)

Nilmgamc activity (n moles CzH, f d g

Ihy-) 40dayn

80days

cml A, brarlkrue (A.b)

P. miam p.r) B, poryrnym ( a . ~ )

+

A.b P.a ~ . +ba . p

N&Pm (40 @ N + 60 kg P20, as mck phospbale/ha) A.b + N&Pm P.s + N & P m 0 . p N&Pm A.b + P.s NmRPm A.b + 0 . p N&P, NnSPm (80 kg N 60 kg P20, as super phosphama)

+

+ +

+

The population counts of Arospirillum and PSB in the rhizosphere indicated that the population of both thegroups of bacteria were generally more at 80 d than st 40 d (Table 4). Seed intxulation by Azospirillum brasilense resulted in increased population of ArospiriNum while that of P,srriara or B. polymym showed more population of PSB in the rhiwsphere. Fertilizer application also enhanced the rhizosphere population of both the groups. The available P content in soil increased following inoculation with P. srriara or B. polymyxo (Table 5 ) . The increase was more prominent in combined inoculations than in single inoculations. further augmented the available P content in soil. However. the soil The addition of N&P, available P content in NgoSPsn'treatmentshowed a gradual decrease i.e. from 12.49 mg Plkg soil at 40 d to 12.3 1 mg at 80 d and 9.96 mg after harveSt sugg'esting a continuous removal of available P by the plants in the absence of any replenishment.

Discussion Tlie results of this investigation clearly indicate the positive interaction of the 2 groups of P. striara or A. polymyxa) resulting in simultaneously inoculated bacteria (A. brasilense enhanced sorghum yields due to their better establishment in the rhizosphere, increased C2H2 reduction activity and available P content in soil and due to increased N and P uptake compared to RAOet aI. 1979; SAHAR SINCHet 81. 1980; single inoculation of the 2 groups. It is reported (SUBRA

+

Interaction

641

Table 4. Populationof Azospirillum and PSB in sorghum rhizospherc as influenced hy sinslc and dud Incxruhtionr Tm~nmt

Awrpirillum

PSR

( x 10'1~soil) 40days

( x IO7lBsoil) 40dayr ROdaya

80dayr

Table 5. Available P amtenr In \o!l (mglkgl under .;orghum or dlficrenl \age* of Ihr rxpL.nmrnl 811dry, allcr plsnl!sp Control A, hr,~srlenrr(A h) P,srrrr1ru (P $1 8 . polymvxu (H.pl

A b + P.r A.b + H.p N~~+R%I (40 kg N + M kg I',O, aa mck phosph;aleiha) A.b + NJI&PMI P s + N4,,RPn, B.p + NmKl'hl, A.b+P.\+N,,,RP,,, A b B . p + N,RP, NxcBPu, (80 kg N ti) kg P?Oqa* super phorphalcihal

+

+

X 10 X 34 4 91 Ill I8 10 119 I0 4X I1 ? I Y

13

1 1 .?7 Ill 14

I ? 411 13 14 I? i l

TII.AKet 81. 1982; PACOVSKY et al. 1085) t h d innculation o f wrghum with A

hrrtsrlrn.,r ha.; increased the yields. Sirnilarly, inoculation, with PSI1 have increased the yields of rice and whci11 (SHARMA and SINGH 197 1 :GAURet al. 1980). The increased yields o f sorghum due to A. h r o . \ i l ~ ~ r t . ~ ~ ~ and that of rice and wheat due l o PSB were ataihuted respeclively to the nitrogen fixalion (BI.KKIIM and BOHLOOL. 1980) and enhanced uptake o f NO,-.K' and HzW40Ji n thc roots of s(1rghurn inoculated with Arospirillum (LIN el al. 1983) and increascd phosphate avo~lahilityin PSI1 inoculated soils ( G A ~1972). R Apart from making available N and Ptothe planlb, both A . hro.sil[,nsi,

kemium lrorrtB . m, d b6:&ma&bd¶o!& hmaed yields.

paaetiasd m t a m p w r d. 1979; HARWNN et d OAWR1987) sod ad@AYA ct d, 1981) which m n h t Fainintundona of ArdrilIwn wrth Azo&cr tgu-a J. t978: mu d. 1982). 19833 and VAM f u (SUSSA ~ I*o & 1.115) plan! m also Imown. Howevcr, there are fewer spans on the of^kuilauc mid PSB and tbek influeacr on plant growth A & ¶ ? m e in the grab awl dry maUu y e l d and mlarent uptake m sorghum due t o rmsbjPdda#mc&m N & P m over canlnncdtnoculattw w ~ t h wN&Pw t seems qurte obvrous rc tks plmb are MU applied with both N and P rcsultmg m better growth and yreld of sorghum plant#.DawE red KONDE(1984) reported that peed bactermtion wrth Azosprrrllum along wrth 66 & Nlhapmduculh t similar y e i d as that of 100kg Nlha in sorghum tndtcahng a net savtng of 34 kg N h due to Azospirillum laoeulatlon. Similarly, G Aand~OSTWAL (1972) reported that the response of w h a t to B, polymyxrr + rock phosphate was comparable wrlh that of superphosphrtc application.OAURa el. (1980) also reported r n m a s e rn the gram yreld of wheat when rock phwphmwas spplied to so11 and seeds w m rnoeulated wlth P. stnafa, where the response of the cmptobrctcnal rnoculatlon was q u ~ v a l c n to t 50 kg PZOJha as superphosphate In lrne wlth these reports, in the p r e e n 1 investlgatlon, applrcatlon of 40 kg N + 60 kg P 2 0 , as rock phosphate (I e N&Pw) with stmultaneous moculatlon of A brasrlense + P srrtara or B polymyxa has glven almost similar yteld as that wlth 80 kg N + 60 kg PzOS as superphosphate Hence, rt appears possible to save m u n d 40 kg N and 60 kg P 2 0 ~as superphosphate by combtned rnoculatron of A brasllenrc and P strtata o r B polymym wtth N&Pm appltcatron Crop response to combrned use of A. brostlense and P srrrata or B polymyxa wrth and wtthout N4~RPb0studled under field con&trons wtll be presented rn a forthcoming separate repon SATT-

et

m

+

Refcrrnecs B A L T K N ~ ~ A. ~ RA.. G SCHANK. ~R. S. C.. SMITH. R. L.. LITTELL. R. C., BOUTON, I. H.. DUDECK. A E.: Effect of inoculUion with Azospirillwn and Azorobacnr on turf-lyp Bermuda gcnolypes. Cmp. SCI 18 (1978). 1043-1045. BARE*. I. M.. N~vAauo,E.. M O N ~ Y E.: A , Muction of plant grnwth regulators by rh~zospherephosphale solubilizing bacteria. I. Appl. Baclcriol. 40 (1976). 129- 134. BAYA.A. M.. BOETHLING. R. S., RAMOS-CORMENZANA. A,: Vitamtn pmduction In relrlion to phnspharc wlubilizslion by soil bacteria. Soil Biol. Biochem. 13(1981), 527-531 BeruuM. P. VAN. BOHLOOL.B. 0.:Evduarion of nitrogen firation hy bacteria in a..m,ialion with rmLI uf tmpical gnsses. Mimbiologicd Rev. 44 (1980). 491 -517. DAY,J. M., NEVES. M. C. P., WBEREINER, I.: Nitmpnaseactivity on the nmts of Uopical tomgc grasses. Soll Biol. Biockm. 7 (1975). 107-1 12. D ~ S A LA. E ,G.. KONDE, B. K.: Response of sorghum to seed bacterisalion with nitrogen levels. J MaharashIra Apri. Univ. 9 (1984). 169-170. WesRetNen. I., DAY,J M.: Associative symbimis in tropical grasses. Chwrcterizatlon of microorpan~smaand dinimgen fixlng sitcs. Inlcrnatl. Symp. N,-Fixation-lnrerdisclplinary discussion. 3-7 June. 1974 Washington State University, Pullman. Washtnglon. USA GAUR.A. C.: Rok of phosphate solubiliztng mirroorgan~smsand organlc maner In soil pmducl~v~ty. Symp. Soil pmductivity, Nad. Acad. Sci. lndia (1972). 259-268. - OSWAL.K. P.: lnflucncc of Phosphate dissolving bacilli on yield and phosphate uptake of whcat crop Indian J. Exp. Biol. 11 (1972). 427-429. - - M ~ r n mR.. S.: Save superphosphateby using phosphate solubilizing culIure*and m k phosphate. Kheti 32 (1980). 23-25. HAITMANN, A,. SINOH. M.. KLINGMILLER. W.: Isolation and charaetenzation of indole acetic acid ovm pmducing m~runrsofAzospirillum.2nd lnternatl. Symp. on Nz fixation with non.legumes. 5-10 Sep.. I982 (Abstracrcls). BanR. Canada. HERNANDEZ. L G.. HILL,G. D.: EflCcl of plant population and inoculat~anon yield and yield components of chickpea (Cicer arierinwn L.) Roc. Agmn. Soc. NZ 13 (1983). 75-79. JACKSON, M. L.: Soil chemical Analysis. h n t i c e Hall Inc. Englewood Cliffs N.J. (1958) pp 151-153 and 331-334.

UN,W.. OKON.Y.. HARDY.R. W. F.: 6 n M mined uplake by Zcomoys aadSor~hvnbtcolur mots inoeulatcd with AzospiriUwn brarilcnsr. Appl. Environ. M i d i o l . 45 (19831, 1775-1779. OLSON.S. R.. COLE.C. V.. WAT~NME.F S.. DEAN.L . A,: Btimation ofsva~lahlcphos phosphnms tn mil hy extmctlon with sodium hicutmue. Unitcd Sues Dep.of Agriculture Circular No. 939 (1945). P ~ c o w r v .R. S., PAUL. E. A,. B~THLENFALVAY. G. I.: Nuhitlon d mghum plmtb fenhlir.4 with n*n>gcnn imruhtcd with Azospirillm hmsilmrr. P h t and Sod 85 (I9851. 145- 148. PIKOWUYA. R. 1.: Mobiliatiion of phosphaus in soil in connection with vital activlry of xnnc n~~nubial rpccier. Micmbiologiya 17 (19481, M2-370. RAI. R.: E f f ~ a c yof asmimivc Nz-fixalron by rmpomycln resarunl mutnnls of Aa,rptrillum bmriirnre wlth p o l y p s of chickpea Rhirvbium atrains. I . Agric. Sci. Camb. I W (1983). 75-00 SAHABSINGH. GANGULY, T. K.. NEBLKANTHAN. KAMAL.A SINGH. AMARIEET SINOH. TOMBR.I' S : Note on the rcsponsc of sorghumand cowpcaloA:osptrillm broriienrc and nimgen. lnd J. Agnc Scl 50 ( IYUOI. 721-7?4 SATTAR.M. A.. GAUR.A. C.. Production of Auxins and Gihbercllins by Phosphate d~s%olv~ng mw'norganlsnis Zcnurlbl. Miluobiol. 142 (19871, 393-395. SHAM*. J . P.. SINGH. M.: Respon~of rtcc to phmphatlc and nmugenour feniltnca wtth and wtlhoul phosphabanerinculturc, lndzan I . A w n . 16 (19711. IS- I8 Sunn~ RAO.N. S., TILAK. K. V 11. R., LAKsHMIKoMkRl. M.. SINCH. C. S.: Respnsc of tcw c~(mon~ic ~pccic 01~ graminaceous plants to tnoculal~onwith Ar,,spiriNum brrrrilrnrr Cum. Scl. 48 (19791, 133- 124. SINGH. C. S.: Synergiliticcffcclof Vcricular Arhuscularmyc~,rrh~za* andA:,,.sp,riilum hmribnsr on the grnwth of barley in pols. Soil Bnal. Brwhem. 17 (IYU5). II9- 121 - - LINGEOOWDA, B. K . F l ~ l rrsponsz d of finger millet (Elrurmc rnracano) 10 ~mxulata,nwllh A:orpirillsm hrailmse. Cum Sci. 52 (1087). 439-440. Techniwn Monogmph 1 (19711 Technicon Private Ltd Tanytnnir, Ncw Ywk. TIEN, T. M., GASKINS.M. H.. HUBBELL.. D.H.. Plant growth suhstancer pnxluced by A:~uptrtllum hrorrlt.nrr und thar cffen on the grnwth of par1 millct (P.-nnnrltm omerrronum I. ) Appl Envlmn Mlrrohtol 37 (19701, 1016-1024. TILAK, K. V. B. R.. SINCH. C S., ROY.N K SUBBARAU. N. S : A:,r.vp~rillum hrusilcnsr and Azofohrrrrrr chnw,coccum ~noeulum: Effecton yleld of matm (Zeoma.yrJand sarghum lSer,qhumhtlolor) Stril Blol. Hn,ohcn~. 14 (1982). 147- 148.

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Aulhors' address: Dr. A. C. GAUR.Prnfessor and Head. lkpt. of Mtcmbiology. Indian Agricultural Rewarch In\itute. New l k l h l I1001?.India: Dr A R. AI.A~AWADI.Avstxiale h f e r u l r . Dept of Agriv Macrnh~olngy.l l n w of Agnc Sclmuer. Dharwad-S8OW5. Kamalaka, Indla.

lJI&r8chutz und Landnutzung EruMkt von 5 Fachwlwenschaftlfm. Hem-

von Prof. Dr. sc. nat.

WQOWNFTscmcE, Halie (Saaie) 1W.MSdB#r, 100 Abblldungen, 48 Tabelen, 17 cmx24 an,Leinen, DDR 95,- M; Au8hd 48,-DM Wellnummer: 5342637

I I

Wle kann, wle mu8 die Menschheit ihre Entwickluno ienken. will sie nicht ihrer n a ~ l l i d s nLebenabedingungen verlustig gehen?-~ragen'des Umwek- und Natumh~~tzes wMren heute m l zu den wicfllten Roblemen, denen sich alle biker gegenrlberstehen. Durchdie fur seine~xlstenznotwendige Nutzung des Landes verlndert der Mensch stlndig seine Urnwelt. Eine tragflhige Lbung ist daher nur in der schbpferischen Zusammenarbeit zwischen Landnutruna und Natursohutz in der Bereitschaft zu aemeinsamem Handeln zu tinden.bin*n Anllegen 1st vorliegendes Buch geGidmet. ais Beitrag ru einer Annniherungteilweise noch diver~ierenderStandpunkte. ErOrtert werden drei umfanoreiche Komolexe: ~aturr6sourcenin der Umweit der menschlichen ~eselbhaftaus ikologischer Sicht (also Boden, Wasser, Luft, Vegetation, Tierwelt); sodann als zentrales Thema Gestaltung, Nutzung und Schutz der nattiriicl;en Umwelt (Wasserwirtschaft, Bergbau ind lnduscie, Siedlung und Verkehr, Erholungswesen, Landschattsplanung); schiieBiich der spezielle Naturschutz aisTeii der soziaiistischenLandeskultur. Rechtsfragenin der DDR, ein internationaler Ausblick sowie umfassende LiteraturangabenbeschlieBen diese lange erwartete, informativeGemeinschaftswerk. Bestellungen nur an den Buchhandel erbeten

VEB GUSTAV FISCHER VERLAG JENA