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PertanikaJ. Trop. Agric. ScLI8(2): 95-101(1995) ISSN: 0126-6128© Universiti Penanian Malaysia Press

Vesicular-Arbuscular Mycorrhizal Colonization andGrowth of Tomato (Lycopersicon esculentum) in Autoclaved Soil

S. MANIAN, THOMSON T. EDATHIL and K. UDAlYAJ'\l

MicrobioloIDI Unit, Department of Botany, Bharathiar University,

Coimbatore - 641 046, Tamil Nadu, India

Keywords: VAM, autoclaving, soil microbes, tomato, biomass

ABSTRAK

Kesan 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.

ABSTRACT

The 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.

INTRODUCTION

Partial 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 develop­ment 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. UDAlYAN

MATERIALS AND METHODS

Preparation of substrates and raising of seedlings

Moderately 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 con­secutive 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 seedlings

Two 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 seed­lings in the autoclaved potted soil were trans­planted 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 (Treat­ment 3), and non-autoclaved soil (Treatment 4).111ere were six replicates for each treatment.

All the transplanted seedlings under dif­ferent treatments were allowed to grow for an·other 60 days (90 days in total) without addingany fertilizer, when they were harvested for esti­mation of VAM colonization, sporulation andgrowth parameters.

Laboratory analysis

Soil 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 coloni­zation index (VAMl) was estimated after stain­ing the root samples following the method ofPhillips and Hayman (1970) and using the scor­ing method of Edathil et al. (1994). The numberof VAM fungal spores of the rrozosphere soil ofdifferent treatment plants was estimated by wet­sieving 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 re­corded. ' 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 1995

VAM COLONIZATION IN AUTOCLAVED SOIL

RESULTS

The 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 colo­nization. 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 grown

in autoclaved soil while lowest growth wasrecorded in YAM-free plants grown inautoclaved soil. There was no marked differ­ence 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 plants

Treatment Per ceot colonization index(PDt)

Spore countsg"l soil

YAM-free plants in autoc1aved soilVAM plants in amoc1aved soilVAM plants in autoc1aved soil amended

with YAM-free filtrate of non-amoc1aved soilVAM plants in non-autoc1aved soil

o55.11b

56.94b

87.78'

P >0.01CD 16.03

o11.54"9.51 b

12.39'

P >0.05CD 1.83

Values with same alphabet in the same column are not significantly different.

TABLE 2VAM fungal species recorded fl"Om the rhizosphere soil of

VA1\1 tomato plants after 90 days of growth

1. 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 & Schenck

PERTANIKAJ. TROP. AGRlC. SCI. VOL. 18 NO.2, 1995 97

S. MANIAN, THOMSON T EDATHIL AND K UDAIYAN

P deficient soil. The suppressed growth ofVAMplants with the addition of soil microbes (non­autoc1aved 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.

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