Journal of Tropical Forest Science 10(2): 212-224 (1997) GROWTH RESPONSE OF WATTLE (ACACIA MEARNSII) SEEDLINGS TO PHOSPHORUS FERTILISATION AND INOCULATIONS WITH GLOMUS DESERTICOLA AND RHIZOBIUM NON-STERILN I . SP E SOIL K. Udaiyan, V. Sugavanam* & S. Manian Microbiology Laboratory, Department of Botany, Bharathiar University, Coimbatore - 641 046, India Received December 1995____________________________________________________ UDAIYAN , SUGAVANAMK. , MANIAN& . ,V . 1997,S . Growth respons f wattleo e (Acacia mearnsii) seedling phosphoruo st s fertilisatio inoculationd nan s with Glomus deserticola d Rhizobiuman n non-sterili . sp e e effecsoil f Th phosphoruo . t ) (P s fertilisatio inoculatiod nan n wit vesicular-arbusculae hth r mycorrhizal (VAM) fungus Glomus deserticola and Rhizobium sp. was studied in containerised seedlings of wattle (Acacia mearnsii). The non-sterile soil used was amended with tricalcium phosphate and superphosphate at the rates of 0, 50, 100, 150, 200 and 250 mg kg-1 soil. The seedlings inoculated with G. deserticola alone but receiving no P fertiliser were comparable in growth to those uninoculated seedlings which received 150 mg kg"1 soil of tricalcium phosphate and 100 mg kg"' soil of superphosphate. Rhizobium inoculated wattle seedlings which receive fertiliseP o n d r were simila n sizi r o t e thos f uninoculateo e d seedlings fertilise mgkg"0 15 d d1 an witsoi 0 tricalciuf lo h 20 m phosphat d superphosphatan e e respectively, wherea e seedlingth s s coinoculated with G. deserticola and Rhizobium sp. without added P fertiliser were more than equivalent in growth to uninoculated control seedlings fertilised with 250 mg kg"' soil of tricalcium phosphate or superphosphate. P fertilisation invariably increased tissue P concentration in the seedlings, reduced mycorrhizal root colonisation but markedly stimulated nodulation e highesTh . t nutrient (N,P,K) concentrationn i s e planth t tissues were recorde e seedlingth n i d s coinoculated wit deserticola. hG and Rhizobium sp. when compared with individual inoculations. Phosphorus utilisation efficiency decreased with increasing P fertiliser doses and inoculations with G. deserticola and Rhizobium sp. Key words : Acacia mearnsii - phosphate fertilisers - Glomus deserticola - Rhizobium sp. - VAM root colonisation - nodulation - phosphorus utilisation efficiency UDAIYAN , SUGAVANAM.VK. , MANIAN.& . 1997,S . Tindak balas pertumbuhan anak benih wattle terhadap pembajaar penginokulatan ida n fosforus dcngan Glomus deserticola dan Rhizobium sp. dalam tanah yang tidak disteril. Kesan pembajaan fosforus (P) dan penginokulatan dengan kulat mikoriza vesikular-arbuskular (VAM), Glomus deserticola Rhizobiumn da . dalasp m anak benih tabling wattle (Acacia mearnsii) dikaji. Tanah yang tidak disteril diubah dengan trikalsium fosfat dan superfosfat pada kadar kg"g m 1 0 tanah , 10025 0 50 ,n ., da 150 Ana0 ,20 k benih yang diinokulat dengan *Present address: Silviculture Division, Institute of Forest Genetics Treeand Breeding, P.O.No. Box 1061, Coimbatore-641 002, India. 212 Journal of Tropical Forest Science 10(2): 212 - 224 (1997) 213 G.deserticola sahaja tetapi tidak menerima baja-P didapati setanding dengan pertumbuhan anak benih yang tidak diinokulat yang menerima 150 mg kg' fosfat 1 trikalsium dan 100 mg kg"1 tanah superfosfat. Anak benih wattle yang dinokulat dengan Rhizobium yang tidak menerima baja-P didapati sama saiznya dengan anak benih yang tidak diinokulat yang dibaja dengan 200 dan 150 mg kg" 1 fosfat trikalsium superfosfan da t sementara anak benih yang dikoinokulat denga deserticola. G n n da Rtiizobium . tanpsp a ditambah baja-P didapati lebih daripada setara dalam pertum- buhan berbanding anak benih yang tidak diinokulat yang clibaja menggunaka0 n25 kg"g m 1 fosfat trikalsium atau superfosfat. Pembajaan-P secara tetap menambahkan kepekatan tis uP dala m anak benih mengurangkan pengkolonian akar mikoriza tetapi dengan jelas merangsang penodulan. Kepekatan nutrien didapati tertinggi (N,P,K) dalam tisu tumbuhan dicatatkan dalam anak benih yang diinokulat dengan G.deserticola dan Rhizobium sp. jika dibandingkan dengan penginokulatan individu. Kecekapan penggunaan fosforus berkurang dengan bertambahny s baja-do an Pda penginokulatan dengan G.deserticola Rhizobiumn da . sp Introduction Microorganism e rhizospherth n si increasy ema r decreaseo e absorptioeth f no inorganic nutrient plany sb t root therd san e appear significane b o st t interactions between s beethemha nt I . clearly shown that vesicular-arbuscular mycorrhiza (VAM improvn ca ) e plant growth through increased uptak f phosphorueo s (P), especially in low-fertile soils (Safir et al. 1972, Mosse et al. 1973). Several studies in recent years have expose e beneficiath d l interaction betwee fungM d an inVA Rhizobium sp. on leguminous plants (Jasper et al. 1988,1989, Reena &: Bagyaraj 1990). Nodulation and subsequent nitrogen fixation by leguminous plants require an optimum level of P in the host tissue (Hayman 1986, Marschner 1986). It is also known that heavy application of P fertiliser can inhibit the percentage infectio fungM f rootVA o ni y (Siqueirb s . 1984al t e a, Thomso . 1986)al t e n . However, when the initial concentration of P is extremely low, small additions may favour infection (Sander Tinkers& s 1973, Schubert &Hayman 1986). Restoration and maintenance of soil fertility is a basic and critical problem, especiall e tropicth n i ys where nutrient deficient soil frequente ar s . Such soils reclaimee ar d through afforestation programmes with multipurpose forest tree species which require large quantitie f gooo s d quality seedlings f commerciaI . l application f mycorrhizao s lo becom t fung e ar irealitya e must i , demonse tb - trated that mycorrhizal fung economicalln ca i y substitut fertilisereP . Hencee th , objectives of the present study were to (a) examine the growth benefits of phosphorus fertilisatio inoculationd an n s wit e mycorrhizahth l fungus Glomus deserticola and Rhizobium sp. on wattle (Acacia mearnsii De Wild.) seedlings, (b) equat e inoculatioth e n benefit e seedlingth f o s s wit e fertilisatioth h n dosef o s tricalcium phosphate and superphosphate, and (c) relate the tissue nutrients concentrations with VAM root colonisation, sporulation and nodulation. 4 21 Journal of Tropical Forest Science 4 (1997 22 10(2) - 2 ) 21 : Materials and methods Soil Sandy loam collected fro experimentae mth l Bharathiae fieldth f o s r University, Coimbatore (1:10 8. ,f soil:water)o H ,p Indiaa d ha , electric conductivitS m 2 y0. cm'1, nitrogen 10.9 mg kg"1 , phosphorus 0.5 mg kg"1 and potassium 24.2 mg kg"1. d availabltotae an Th lN wereP e determined respectivel micro-Kjeldahl'e th y yb s method and the molybdenum blue method of Jackson (1973). Exchangeable K was extracted from the soil in an ammonium acetate solution (pH 7.0) and measured wit digitaha l flame photometer (Jackson 1973)naturaa soie d ha lTh . l mycorrhizaA V l population (Acaulospora scrobiculata, Glomus fasciculatum, geosporum. G and G. mosseae) of 72 spores 100 g"1 dry soil. Phosphorus fertilisation Phosphoru applies e for wa sth f commerciam o n di l acid soluble tricalcium phosphate [Ca3 (PO4).J and water soluble superphosphate [Ca (H^PO^ H2O] at the rate of 0, 50,100,150, 200 and 250 mg kg"' soil and uniformly mixed. One milligra f mtricalciuo m phosphat superphosphatd ean e contains 0.6 0.8d g 11an m of phosphate respectively. Plant material Wattle (Acacia mearnsii) seeds were procured fro e Institutmth f Foreso e t Genetics and Tree Breeding, Coimbatore, India. Healthy, uniform seeds were surface sterilise H.,0.,% 5 n d,i treated with boiling wate r 30sfo r , then soaken di water for about 24 h and sown in 10 X 15 cm polythene bags, each containing 3 kg unsterilised soil. Initially , three seeds were planted per bag but the seedlings were thinned to one on the tenth day. Inoculum The VAM fungus Glomus deserticola Trappe, Bloss & Menge, isolated and re- corde dominana s da t specie rhizosphere th n si e soil sample f Acaciaso mearnsii found growin e plantatioth n gi n fores f Kodaikanalo t , Tamil Nadu, Indias wa , multiplied in pot culture in the roots of Sorghum vulgare and the soil containing the extramatrical hyphae, chlamydospores and infected root segments served as mycorrhizal inoculum. Fifty gram inoculum containin infectiv0 50 2 1 . ec g pro - pagules, based on a most probable number estimate (Porter 1979) was placed in each polythene bag about 2 cm below seed level. Fresh nodules of wattle (Acacia mearnsii) were collected from the plantation forest, Kodaikanal, Tamil Nadu, India. Rhizobium isolates wa maintaine. d dsp an d in yeast extract mannitol broth. Ten millilitre broth was added to each polybag. 5 21 Journal of Tropical Forest Science (19974 22 10(2) - 2 ) 21 : Experimental design The experimental design was 4x2x6 factorial consisting of 4 levels of endo- phyte inoculations, 2 phosphate sources and 6 levels of P fertiliser doses. Thus in all there were 48 treatments and each treatment was replicated 5 times. The treatments were arrange completela n di y randomised block design. Harvesting and measurement Plants were harvested after 120 days and their dry weights were recorded. Root subsample segment0 lengtm (1 sc 1 hf o seedling"s ) were processe r microdfo - 1 scopic observation following the procedure of Phillips and Hayman (1970) and the percentage mycorrhizal infectio determines nwa rooe th ty dslidb e techniquf eo Read et al. (1976). The dried plant material (shoot and root) was ground and used for the analysis of N, P and K. Total Kjeldahl nitrogen was detected on a Kjeltec Auto Analyser (1030); phosphorus determination was done by the vanadomolybdate phosphoric yellow colour method and potassium content was determined by flame photometer (Jackson 1973). Phosphorus utilisation efficiency (PUE) was calculated usin formule th g f Siddiquao d Glas plan= an i y ( 1981E s dr t PU : ) weight/P content.