The effect of autoclaving soil an uesicular-arbuscular mycorrhizal (VAAl) Jungal colonization and biomass

production of tomato (Lycopersicon esculentum Mill) was assessed in pot experiments under greenhouse

conditions. Four treatments were given viz., (1) VAM-free plants in autoclaued soil, (2) VAM plants in

autoclaued soil, (3) VAM plants in autoclaved soil amended with VAM-free filtrate of non-autoclaved soil, and

(4) VAM plants in non-auloclaved soil. The VAM plants grown in non-autoclaued soil showed the highest root

colonization of 87.78% while those under (2) and (3) showed only 55.11 % and 56.94% respectiuely. On the

other hand, significantly higher shoot length (105.4 cm2/plant), wtal kaJ area (740.3 cm2/plant) and biomass

(8.43 g/plant) were obtained in VAM plants grown in autoclaved soiL VAM-free plants in autaclaved sail had

reduced growth. In treatments 3 and 4 plant growth was intermediate. The results indicate that VAM

colonization and sporulation were favoured under natural conditions, but VAM plants under autoclaved soil

conditions produced maximum growth

K., Udaiyan

S., Manian

Thomson T., Edathil

Universiti Putra Malaysia Institutional Repository

PertanikaJ. Trop. Agric. ScLI8(2): 95-101(1995) ISSN: 0126-6128© Universiti Penanian Malaysia PressVesicular-Arbuscular Mycorrhizal Colonization andGrowth of Tomato (Lycopersicon esculentum) in Autoclaved SoilS. MANIAN, THOMSON T. EDATHIL and K. UDAlYAJ'\lMicrobioloIDI Unit, Department of Botany, Bharathiar University,Coimbatore - 641 046, Tamil Nadu, IndiaKeywords: VAM, autoclaving, soil microbes, tomato, biomassABSTRAKKesan tanah auwklaua terhadap pmgkownian kulat uesikal-arbuskular mikoma (VAM) dan pmgeluarantomato biojisim tetah dinilai dalam bekas yang diuji di rwnah hijau. Empat rawatan telah diberi iaitu (1)Tumbuhan bebas-VAM dalam tanah autoklaua (2) Tumbuhan VAM dalam tanah autoklaua (3) TumbuhanVAM dalam tanah autoklaua yang diubah suai dmgan tanah tanpa autoklaua filtrat bebas-VAM (4)Tumbuhan VA.A1 dalam tanah tanpa autoklava. Tumbuhan VANf yang membesar dalam tanah tanpaautoklava menunjukkan pengkolonian akar paling tinggi iaitu 87.78%, manakala (2) dan (3) musing-masinghanya 55.11 % dan 56.94%. Di samping itu, panjang tunas (105.4 em/tanaman), jumlah ruang daun(740.3 cm'/tanaman) dan biojisim (8.43 gltanaman) diperolehi dalam tumbuh-tu,nbuhan VAM yang membesardalam tanah autoklava. Tumbuhan bebas VANf dalam tanah autoklava kurang rnembesar. Dalam TQwatan (3)dan (4) pembesaran tumhuhan adalah seder-hana. Keputusan menunjukkan bahawa pengkolcnian VAM danpengsporaan sesuai dalam keadaan semulajadi tetapi tumbuhan VAM dalam keadaan tanah autoklavamenghasilRan pertwnbuhan yang maksimum.ABSTRACTThe effect oj autoclauing soil an uesicular-arbuscular mycorrhizal (VAAl) Jungal colonization and biomassproduction of tomato (Lycopersicon esculentum Mill) was assessed in pot experiments under greenhouseconditions. Fou,· treatments were gium viz., (1) VAM-jree plants in autoclaued soil, (2) VAM plants inautoclaued soil, (3) VAM plants in autoclaued soil amended with VAM-jree filtrate oj non-autoclaued soil, and(4) VAM plants in non-auloclaued soil. The VAM plants grown in non-autoclaued soil showed the highest rootcolonization oj 87.78% while those under (2) and (3) showed only 55.11 % and 56.94% respectiuely. On theother hand, significantly higher shoot length (105.4 em/plant), wtal kaJ area (740.3 em'/plant) and biomass(8.43 glplant) were obtained in VAM plants grown in autoclaued soiL VAM-jree plants in autaclaued sail hadreduced growth. In treatments 3 and 4 plant growth was intermediate. The results indicate that VAMcolonization a.nd spornlat-ion were favoured under natural conditions, but VAM plants under autoclaved soilconditions produced maximum growth.INTRODUCTIONPartial or complete sterilization of soil oftenchanges its nutrient status and structure (Lopesand Wollurn, 1976; Mulder, 1979;]akobsen andAndersen, 1982). Sterilization also removes someor all of the microorganisms (Bowen and Rovira,1969). However, enhanced fertility due toautoclaving is available to plants only duringtheir initial stage of growth.Mosse et aL (1969) obseIVed that soilsterilization had a positive effect on the develop-ment of vesicular-arbuscular mycorrhizal (VAM)infection and plant growth, but Wilson (1984)reported that autoclaving of soil inhibited VAMspore germination. Little is known about theeffect of autoclaving of soil on VAM fungalcolonization and biomass production in tomato(Lycopersicon esculentum Mill.) which this studyattempted to elucidate.S. MANIAN. THOMSON T. EDATHIL AND K. UDAlYANMATERIALS AND METHODSPreparation of substrates and raising of seedlingsModerately fenile sandy loam soil collected fromthe experimental fields of Bharathiar University,Coimbatore (pH 8.1, EC 0.1 m Scm'" N 10.5 mgkg', P 1.7 mg kg', K 38 mg kg'), mixed with sandat 1: 1 proportion was used as the substrate forplant growth. The substrate was sterilized incloth bundles in an autoclave at 1.5 kg sq em-Ipressure (121 "C) for 1 hour each on three con-secutive days and left in the laboratory for sevendays 1O facilitate release of any toxic substancesproduced during heating. Ten presterilized potsof 18 cm diam. were filled with 6 kg par' ofautoc1aved soil and another 30 pots were filledwith non-autoclaved soil-sand mixture.The field soil contained VAM fungalspores predominantly of Acaulospora bireticulataRothwell & Trappe; A. sporocarpiaBerch; Glomusdesertieola Trappe, Bloss & Menge; G.Jasciculatum (Thaxter & Gerdemann)Gerdemann & Trappe; G. geosporum (Nicolson& Gerdemann) Walker; G. tenue (Greenall)Hall and G. sinosum (Gerdemann & BakshiAlmeida & Schenck (= ScieTOeystis pakistanicaIqbal & Bushra), having a total spore count of20.43 (± 0.73) spores g' dry soil. The VAMfungal species were identified using synoptickeys (Hall, 1984; Morton 1988; Schenck andPerez, 1987) for spores and sporocarps.All pots were sown with unifonn sizedseeds of tomato cv. Co. 1 at a density of 5 perpot and w"atered regularly. One week aftergermination, they were thinned to maintain onehealthy seedling per pot and allowed to grow for30 days under greenhouse conditions. VAMfungal infection was detected by the method ofPhilips and Hayman (1970) in the seedlingsraised on non-autoclaved soil, while those fromautoclaved soil were free from any infection.Transplanting of seedlingsTwo sets of potted soil, i.e. 30 pots of autoclavedand 10 non·autodaved, were prepared as in theprevious experiment. Of these, 10 pots ofautoclaved soil were drenched with soil filtrate ofnon-autoclaved soil at the rate of 100 ml pori tofacilitate VAM and mycophagous animal freemicrobial action in the soil. The filtrate wasprepared by adding 500 ml of sterilized waterto 350 g of non-autoclaved soil and thoroughlymixing. The liquid portion of mixture was de-canted and fIltered through a 38 mm mesh thatretained VAM fungal spores and mycophagousanimals but not other microbes (Azcon·Aguilarand Barea, 1985).The 3Q-day-old VAM-free tomato seed-lings in the autoclaved potted soil were trans-planted into 10 pots of autoc1aved soil with oneseedling per pot as Treatment 1, and out of the30 VAM seedlings in the non-autoclaved pottedsoil, 10 pots each with one seedling were trans·planted into autodaved soil (Treatment 2),autoclaved soil amended vvith filtrate (Treat-ment 3), and non-autoclaved soil (Treatment 4).111ere were six replicates for each treatment.All the transplanted seedlings under dif-ferent treatments were allowed to grow for an·other 60 days (90 days in total) without addingany fertilizer, when they were harvested for esti-mation of VAM colonization, sporulation andgrowth parameters.Laboratory analysisSoil pH, EC and nutrient starns were analysedusing standard procedures (Misra, 1968;Jackson,1973). For estimation ofVAM colonization, atharvest, 0.20 g (wet weight) was excised fromeach root system. The rest of the root systemwas kept for observing dry weight. VAM coloni-zation index (VAMl) was estimated after stain-ing the root samples following the method ofPhillips and Hayman (1970) and using the scor-ing method of Edathil et al. (1994). The numberof VAM fungal spores of the rrozosphere soil ofdifferent treatment plants was estimated by wet-sieving and decanting method (Gerdemann andNicolson. 1963). Spore counting was done with100 g dry soil and spore density was expressed asnumber of spores per gram of soil.The total leaf area was measured using aleaf area meter and the mean of six replicates ofeach treatment calculated.Root length, shoot length, root dry weight,shoot dry weight, root/shoot (R/S) ratio for dryweight and mycorrhizal dependency were re-corded. ' Dry weights were obtained by dryingfor 24 h at 45°C in a hot air oven. Mycorrhizaldependency (M.D.) was calculated using theformula of Plenchette et al. (1983):(DM o[YAM plant - DM of YAM-free plant)M.D.= x 100(DM of YAM-free plant)where DM = dry mass.96 PERTANlKAJ. TROP. AGRIC. SCI. VOL. 18 NO. 2 1995VAM COLONIZATION IN AUTOCLAVED SOILRESULTSThe highest percentage ofVAM colonization andspore density was observed in the VAM plantsgrown in non-autoclaved soil, while those inamoc1aved soil (amended or non-amended withsoil filtrate) showed significantly lower root colo-nization. However, the number of spores wassignificantly reduced only in the filtrate amendedtreatment (Table I). Altogether 17 species ofVAM fungi belonging (03 genera were recordedfrom the rhizQsphere soils of the YAM plantsirrespective of treatment conditions (Table 2).For plant growth, significantly higher(P < 0.01) shoot length. total leaf area andbiomass were obtained in VAM plants grownin autoclaved soil while lowest growth wasrecorded in YAM-free plants grown inautoclaved soil. There was no marked differ-ence in root length or R/S ratio among VAMplants and in autoclaved and non-autoclavedsoils. However RjS ratios were higher inYAM-free plants and in VAM plants grown inthe substrate amended with soil microbes(Table 3; Fig. 1).The VA.1\1-free plants in amoc1aved soilshowed good growth initially; they becamestunted in due course, but retained a lushgreen colour unlike plants in non-autoclavedsoil which had stunted growth and were pale bythe end of the experiment.TABLE 1Effect of amoclaving of soil on the VAM fungal infectivity on tomato plantsTreatment Per ceot colonization index(PDt)Spore countsg"l soilYAM-free plants in autoc1aved soilVAM plants in amoc1aved soilVAM plants in autoc1aved soil amendedwith YAM-free filtrate of non-amoc1aved soilVAM plants in non-autoc1aved soilo55.11b56.94b87.78'P >0.01CD 16.03o11.54"9.51 b12.39'P >0.05CD 1.83Values with same alphabet in the same column are not significantly different.TABLE 2VAM fungal species recorded fl"Om the rhizosphere soil ofVA1\1 tomato plants after 90 days of growth1. Acaulospura bireticulata Rothwell & Trappe2. A. trappe-; Ames & Linderman3. Gigaspora alhidn Schenck & Smith4. Glomus aggregatum (Schenck & Smith) Koske5. G. deserlicola Trappe, Bloss & Menge6. G. fascUulatum (Thaxter & Gerdemann) Gerdemann & Trappe7. G. Julvurn (Berk.) Pat.8. G. geosporum (Nicolson & Gendermann) Walker9. G. intraradices Schenck & Smith10. G. maculosum Miller & Walker11. G. macrocarpum (Tul. & Tul) Nicolson & Gerdemann12. G. manihotis Howler, Sieverding & Schenck13. G. m£/ollosparttm Gerdemann & Trappe14. G. microaggregatum Koske, Gemma & Olexia15. G. mosseae (Nicolson & Gerdemann) Gerdemann & Trappe16. G. tenue (Greenall) Hall17. G. sinosum (Gerdemann & Bakshi) Almeida & SchenckPERTANIKAJ. TROP. AGRlC. SCI. VOL. 18 NO.2, 1995 97S. MANIAN, THOMSON T EDATHIL AND K UDAIYANP deficient soil. The suppressed growth ofVAMplants with the addition of soil microbes (non-autoc1aved soil filtrate) ''VaS also evident. VAMefficiency was mainly the result of the presenceof extramatrical hyphae and the efficient VAMsymbiosis. which reduced RjS ratio, and not theextent of per cent root colonization.REFERENCESNEK, D., JA. MENGE and E.L.V. JONHSON. 1990.Effect of Pythium ultimum and metalaxyl treat-ments on root length and mycorrhizal coloni-zation of cotton, onion and pepper. PlantDisease 74: 117-120.ATIlANO, RA, JR. RICH, H. FERRIS and JA. MENGE.1986. Effect of Meloidogyne arenaria onendomycorrhizal grape (Vilis vinijera) rootings.Journal of Nematowgy 8: 278.AzON-AGUILAR, C. and JM. BAREA. 1985. Effect ofsoil microorganisms on formation of vesicular-arbuscular mycorrhizas. Transactions of BritishMycowgical SOcUty 84: 535-537.BE~'TIVENGA. S. 1988. Effect of Inorganic NutrientTreatment, Soil Microorganisms, and Vesicular-arbuscular Mycorrhizae on the Growth of BigBluestem (Andropogon gerardii). Master's The-sis, Illinois State University.BOWEN, C.D. and AD. RoVIRA. 1969. The influenceof microoganisms on root growth and metabo-lism. In Wittington WJ. (ed): Root Grawth.London: Butterworths.CLARKE, C. and B. MaSSE. 1981. Plant growthresponse to vesicular-arbuscular mycorrhiza. Xli.Field inoculation responses of barley at two soilP levels. New Phytowgist 87: 695-703.DA~IELS, B.A. and JA. MENGE. 1980.Hyperparasitization of vesicular-arbuscularmycorrhizal fungi. Phytapathowgy 70: 585.D"'~lElS, BA and JM, TRAPPE. 1980, Factorsaffecting germination of vesicular-arbuscularmycorrhizal fungus Glomus epigaeum. Mycologia72: 457-471.DOUDS, D.D. and W.R. CHANEY. 1982. Correlationof fungal morphology and development tohost growth in a green ash mycorrhiza. NewPhytowgist 92: 519-526.EDATHIL, T.T., S. MANIAN, and K. DDAlYAN. 1994.The effect of vesicular-arbuscular mycorrhizalexposure period on their colonization of andspore production in tomato seedlings(Lycopersicon esculentum MilL), and on hostbiomass. Agriculture, Ecosystems and Environ-rrumt 51: 287-292.FITTER, A.H. 1977. Influence of mycorrhizal infec·tion on competition for phosphorus and po-tassium by the grasses. New Phytologist 79: 119-125.GERDEMA.!'l"N,j.W. and T.H. NICOLSON. 1963. Sporesof mycorrhizal endogone species extractedfrom soil by wet sieving. Transactions ofBritishMycowgical Society 46: 234-235.GROTH, D.E. and C.A. MARTINSON. 1983. Increasedendomycorrhizal infection in maize andsoybeans after soil treatment and metalaxy.Plant Disease 67: 1377-1378.HALL, I.R. 1984. Taxonomy of VA mycorrhizalfungi. In Powell 1.,1., Bagyaraj DJ. (eds) VAM)'corrhiza. CRC Press Inc., Boca Raton, Florida.pp. 57-94.HALL, I.R., P.D. JOHNSTONE and R. DOLLY, 1984.Interactions between endomycorrhizas and soilnitrogen and phosphorus on the growth ofrye-grass. New Phytowgist 97: 447-453.HERlGK, BAD., D.G. KIn and G,T. WIlSON. 1986.The influence of phosphorus fertilizer,drought, fungal species and non-sterile soil onthe mycorrhizal growth response in tall grassprairie. Canadian Journal of Botany 64: 1199-1203,HERTRICK, BAD., GT. WIlSON, D.G. Krrr and A. P.SCHWAB. 1988. Effect of soil microorganismson mycorrhiza contribution to growth of bigbluestem grass in non-sterile soil. Soil BioloF!Jand Biochemistry 20: 501-507.JACKSON, N.L. 1973. Soil Chemcal Analysis. PrenticeHall of India Pvt. Ltd. New Delhi.JAKOBSEN, Land AJ. ANDERSEN. 1982. Vesicular-arbuscular mycorrhiza and growth in barley.Effects of irradiation and heating of soil. SoilBiowgy and Biochemistry 14: 171.100 PERTANlKA]. TROP, AGRlC. SCI. VOL. 18 NO.2 1995VAM COLONIZATION [ AUTOCLWED SOILKHAuL, S., T.E. Lon'ACHEN, and H.S. Me NABB JR.1992. Colonization of soybean by mycorrhizalfungi and spore population in Iowa soils.Agrollomy Journal 84: 832-836.LOPES, A.S. and A.G. WOLLU~f. 1976. Comparativeeffecti\'eness of methylbromide. propylene ox-ide and autoclave sterilization on specific soilcharacteristics. Tunialha 26: 351.MEREDITH, J.A. and R.C. ANDERSON. 1992. Theinfluence of varied microbial substrate condi-tions on the growth and mycorrhizal coloniza-tion of liule bluestem. New Phylologist 121:235-242.MISRA, R. 1968. EcoliJgy Work Book. Oxford, IBHPublishing Co. Pvt. LTD., New Delhi.MORTO~, J.B. 1988. Taxonomy of VA mycorrhizalfungi: Classification, nomenclature and identi-fication. iWycotaxon 32: 267-324.MaSSE, B., D.S. !iAY>w; and GJ. IDE. 1969. Growthresponse of plants in unsterilized soil to inocu-lation ,vith vesicular-arbuscular mycorrhiza.Natnre (London) 224: 1031-1032.MULDER, D. (Ed.), 1979. Soil Disinfestation: SomeSeieTltiftc and Practical Contributions in the Field ofSoil Disi1ifestalion. Elsevier: Amsterdam.PHlU1PS. J.M. and D.S. H.mlA.'I. 1970. Improvedprocedures for clearing roolS and staining para-sitic and vesicular·arbuscular mycorrhizal fungifor rapid assessment of infection. Tra1lSactionsof British Mycological Society 55: 158-161.PUNCHETTf:, C., J.A. FORTIN and V. FURL"'. 1983.Growth response of several plant species tomycorrhizae in a soil of moderate P~fertility %mycorrhizal dependency under field condi-tions. Plallls and Soi170: 199-209.Sc:HLl\fCK, '.C., R.A. KIl'LOCK and D.W. DICKSON.1975. Interaction of endomycorrhizal fungiand root·knol nematode of soybean. InEndc1n)'corrhiUlS, Sanders, Masse, B. and Tinker,P.B. (Eds). Academic Press: London.SCHENCK, N.C., and Y. PEREZ. 1987. A Mllnual ojIdentification of Vesicular-arbuscular MJcorrhhalFungi. I VAM, University of Florida,Gainesville, Florida.SEQ, H.-A., R.C. M"DERSON and A.E. LIBERTA. 1988.Influence of varied soil microbial and inor·ganic nutrient conditions on the growth andVA colonization of little bluestem (Schizachyriumscoparium). Biology and Fertility of Soils 6: 1-5.WAR.'IOCK, AJ., A.H. FITTER and M.B. USHER. 1982.The influence of a spring tail Fol.somia candida(Insect, Collembola) on the mycorrhizal asso-ciation of leek Allium porrum and the vesicular-arbuscular mycorrhizal endophyte Glomusfasciculatum. New PhytoliJgist 90: 285.WIL50K, J.M. 1984. Inhibition of germination ofspores of a Gigaspora species in sterilized soils.Soil Biology and Biochemistry 16: 433-435.(Received 19 July 1994; accepted 30 August 1995)PERTANIKAJ TROP. AGRle. SCI. VOL 18 NO.2. 1995 101

Vesicular-Arbuscular Mycorrhizal colonization and growth of tomato (Lycopersicon esculentum) in autoclaved soil

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Vesicular-Arbuscular Mycorrhizal colonization and growth of tomato (Lycopersicon esculentum) in autoclaved soil

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