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1 Biologil Control 52 (21) Contents lists ville t SieneDiret Biologil Control journl homepge: Suppressive effet of non-erte ompost tes on folir fungl pthogens of tomto Souleymne B. Koné, Antoine Dionne, Russell J. Tweell, Hni Antoun,, Tyler J. Avis,, * Centre e reherhe en hortiulture, Université Lvl, Quée, Qué., Cn G1V A6 Déprtement es sols et e génie grolimentire, Université Lvl, Quée, Qué., Cn G1V A6 Deprtment of Chemistry, Crleton University, Ottw, Ont., Cn K1S 5B6 rtile info strt Artile history: Reeive 7 July 29 Aepte 27 Otoer 29 Aville online 3 Otoer 29 Keywors: Botrytis inere Compost te Disese suppression Miroil ntgonists Oiium neolyopersii Compost tes re fermente wtery extrts of omposte mterils tht re use for their ility to erese plnt isese. Non-erte ompost tes (NCT) prepre from five types of ompost were teste for their ility to inhiit the growth of Alternri solni, Botrytis inere, n Phytophthor infestns in vitro. Weekly pplitions of NCT were lso use in greenhouse trils to ssess their suppressive effet on powery milew (Oiium neolyopersii) n gry mol (B. inere) on tomto plnts. All NCT signifintly inhiite the myelil growth of A. solni (37 66%), B. inere (57 75%), n P. infestns (1%), wheres sterilize tes i not inhiit growth of the teste pthogens. Although NCT file to effiiently ontrol powery milew, they were le to ontrol tomto gry mol for up to 9 weeks in greenhouse experiments. Among the teste ompost tes, NCT prepre from sheep mnure ompost onsistently provie the highest inhiition of myelil growth n the highest isese suppression, in prtiulr of gry mol (>95% isese reution). The overll reltive effiy of the vrious NCT i not orrelte well with miroil ommunities or physio-hemil omposition of the prepre NCT. Results lso suggest tht the presene of the miroorgnisms in the NCT is prerequisite for inhiition. Ó 29 Elsevier In. All rights reserve. 1. Introution Compost tes re fermente wtery extrts of omposte mterils tht re use for their enefiil effet on plnts (Litterik et l., 24). The most wiely esrie enefit of ompost tes is their ility to erese plnt isese when use s soil renhes or folir sprys (Sheuerell n Mhffee, 22, 24). In this respet, ompost tes re viewe s potentil lterntives to the use of syntheti hemil fungiies s they provie mens of ontrolling plnt pthogens tht re eeme sfer for helth n the environment (Siiqui et l., 29). Although isese suppressive effets of ompost tes hve een reporte in numerous griulturl systems, their effiy remins vrile (Sheuerell n Mhffee, 26). The potentil prmeters tht ffet the effiy of ompost tes re twofol: the trget pthosystem (pthogen n host plnt) n the preprtion methoologies of the tes (ertion, ompost type, nutrient itives, urtion of fermenttion, et.) (Sheuerell n Mhffee, 26). Among ompost te preprtion ftors, the ompost type hs een stuie to etermine its effet on the effiy of the prepre te. In previous reports, the type of ompost is eeme ruil ftor in the effiy of the proue te. Inee, * Corresponing uthor. Aress: Deprtment of Chemistry, Crleton University, 1125 Colonel By Drive, Ottw, Ont., Cn K1S 5B6. Fx: E-mil ress: tyler_vis@rleton. (T.J. Avis). mny uthors reporte tht tes prepre from mnure-se omposts re generlly more effetive ginst plnt pthogens thn tes from plnt-se omposts (Weltzien, 1991; Al-Dhmni et l., 23; Hggg n Ser, 27). Conversely, others hve onlue tht the type of the ompost oes not iretly ffet the effiy of the te n tht oth mnure-se omposts n other ompost types n provie n effiient te to ontrol plnt pthogens (El n Shtienerg, 1994; Sheuerell n Mhffee, 26). Compost tes re reporte to ontrol plnt pthogens through ifferent mehnisms. The most reporte ftor influening the effiy of ompost tes in inhiiting the evelopment of plnt isese is their miroil ontent. The miroorgnisms present in the te my t s pthogen ntgonists through their ility to ompete for spe n nutrients (Al-Mughri et l., 28), to estroy pthogens y prsitism (El-Msry et l., 22), to proue ntimiroil ompouns, or to inue systemi resistne in plnts (Zhng et l., 1998). Other work hypothesize tht the physiohemil properties of the ompost tes, nmely nutrients n orgni moleules suh s humi or phenoli ompouns (Hoitink et l., 1997; Siiqui et l., 28), my protet the plnt ginst isese through improve nutritionl sttus, iret toxiity towr the pthogen or inue systemi resistne. In this stuy, non-erte ompost tes (NCT) prepre from vrious ompost types were teste for their effet on the growth of ifferent tomto (Solnum lyopersium L.) folir pthogens n for their effet on the evelopment of two tomto folir /$ - see front mtter Ó 29 Elsevier In. All rights reserve. oi:1.116/j.ioontrol

2 168 S.B. Koné et l. / Biologil Control 52 (21) iseses nmely powery milew (Oiium neolyopersii) n gry mol (Botrytis inere) in n effort to gin insight into their suppressive nture. NCT ws fvore over erte ompost tes (ACT) sine they re regre s the simplest tehnology for the preprtion of ompost tes (no speilize ertion equipment neessry) while generlly onsiere to e s effetive s ACT in ontrolling isese (Welke, 25; Sheuerell n Mhffee, 26). 2. Mterils n methos 2.1. Miroil mteril Four tomto pthogens were use to ssess the suppressiveness of the prepre NCT. Alternri solni Soruer n B. inere Pers. were mintine on potto extrose gr (PDA; Beton Dikinson, Sprks, MD) t 24 C. Phytophthor infestns (Mont.) e Bry ws mintine on rye gr (Klimzk n Prell, 1984) ontining 2 g/l oile (9 min) rye grin filtrte, 5 g/l extrose, n 12 g/l gr t 24 C. These three isoltes were otine from the Lortoire e ignosti en phytoprotetion (MAPAQ, Quée, Cn). The iotroph O. neolyopersii L. Kiss ws mintine on tomto plnts uner greenhouse onitions Preprtion of ompost tes Commerilly-ville omposts proue from hiken mnure, sheep mnure (four soures; SM1 SM4), ovine mnure, shrimp power, or sewee were use for the proution of NCT. Eh NCT ws prepre y mixing 1 L of homogenize ompost with 5 L of wter (from n griulturl well) in 1-L ontiner oring to moifie metho of El n Shtienerg (1994). The mixture ws loosely overe n store in the rk t 18 C for 14 ys. The NCT were stirre on the 7th n 14th y of inution. Following the inution perio, eh mixture ws filtere through eight lyers of heeseloth n onserve t 4 C until use Physio-hemil n miroil nlysis of NCT The minerl ontents of the proue NCT were nlyze using stnr methos. NH 4 n PO 4 were etermine y olorimetry oring to the methos of Nkonge n Bllne (1982) n Trn n Simr (1993), respetively. NO 3,SO 4, n Cl were quntifie y ioni hromtogrphy (Dionex, Sunnyvle, CA). K n N were etermine y tomi emission n C, Mg, Fe, Cu, Mn, n Zn were quntifie y tomi sorption spetrosopy (Perkin-Elmer Atomi Asorption Spetrometer 33, Ueerlingen, Germny). The eletril onutivity (EC) n ph were lso etermine for eh NCT. The presene of oliforms n Esherihi oli in the prepre NCT were nlyze in uplite on E. oli/coliform Petrifilm ount pltes (3M, Lonon, Cn) oring to the mnufturer s iretions. The popultions of fungi, tinomyetes, n teri (totl, Grm negtive, n fluoresent pseuomons) were nlyze in eh NCT on ifferent seletive ulture mei. Fungi were grown on Mlt gr (Beton Dikinson) mene with.3 g/l streptomyin (Sigm Alrih, St. Louis, MO). Atinomyetes were ulture on Atinomyete Isoltion Agr (Beton Dikinson) mene with glyerol (1 g/l; Sigm Alrih). Totl teri were grown on Trypti Soy Agr (Sigm Alrih) n Grm negtive teri were ulture on Trypti Soy Agr mene with rystl violet (.2 g/l; Sigm Alrih). Finlly, fluoresent pseuomons were grown on Pseuomons Agr F (Beton Dikinson) mene with glyerol (1 g/l; Sigm Alrih), mpiillin (.5 g/l; Sigm Alrih), hlormpheniol (.125 g/l; Sigm Alrih), n hlorothlonil (1.1 g/l; Syngent, Guelph, Cn). Eh NCT ws 1-fol seril ilute (1 1 7 ) n 5 ll of eh ilution were plte on the seletive mei. After n inution perio of 2 4 ys t 25 C, the most prole numer tehnique (MPN) (Woomer et l., 1988; Min, 1999) ws use to estimte the ifferent miroil popultions on eh seletive meium. Four replites for eh NCT were performe to etermine MPN Effet of NCT on myelil growth To etermine their effet on myelil growth of A. solni, B. inere, n P. infestns, the proue NCT or griulturl well wter (ontrol) were inorporte into PDA (15% vol:vol) (Diánez et l., 26). To prepre the NCT ontining mei, PDA ws sterilize n oole to 45 C. The NCT were then mixe with the oole PDA n immeitely poure into the Petri ishes. In ition, the effet of sterilize NCT on myelil growth ws evlute y inorporting utolve (121 C for 2 min) or mirofiltere (.2 lm) NCT into PDA (15% vol:vol), prepre s esrie ove. Agr plugs (.5 m imeter) overe with tively growing myeli of the three pthogens were iniviully inoulte on the mei n inute 6 7 ys in the rk t 25 C. After the inution perio, myelil growth of eh fungus ws mesure s the verge of two perpeniulr imeters of the olony. All experiments were onute oring to ompletely rnomize esign with four repetitions Effet of NCT on tomto isese severity Tomto sees (ultivr Bush Beefstek) were sown in rok wool. After 2 weeks, the tomto seelings were iniviully trnsplnte into 3-m pots ontining Promix sustrte (Premier Teh, Rivière-u-Loup, Cn). Seelings were grown uner greenhouse onitions t 19 C (night) n 23 C (y) with 16-h photoperio n 85% reltive humiity. Plnts were fertilize ily with nutrient solution ontining lg/ml of N P K. When tomto plnts rehe the four lef stge, leves were rtifiilly inoulte with either B. inere or O. neolyopersii spores 3 ys fter (preventive) or efore (urtive) the initil tretment with NCT. Botrytis inere inoulum ws prepre s esrie in Sheuerell n Mhffee (26), juste to onii/ml suspension, n uniformly sprye onto eh lef of eh plnt (2 ml per plnt). Tomto leves hevily infete with O. neolyopersii were shken 1 y prior to inoultion to ensure tht freshly forme onii were ville. Lef segments ering onii were hrveste n use to inoulte the experimentl plnts y mnully shking the infete lef segments over the plnts to isloge the onii. Non-inoulte plnts were lso inlue to evlute potentil phytotoxiity of the teste NCT. Eh of the five teste NCT (ovine mnure, sheep mnure SM1, hiken mnure, shrimp, n sewee ompost NCT) or griulturl well wter (ontrol) ws pplie weekly throughout the experiment y sprying of the entire folir surfe until runoff. Disese severity ws evlute weekly s the perentge (%) of visile symptom re of gry mol (B. inere) or powery milew (O. neolyopersii) on the fourth true lef of eh plnt. Symptoms of phytotoxiity (sorhing, urning, or spotting of tomto plnt leves) were note n plnt height ws mesure. The experiments were onute oring to ompletely rnomize lok esigns with five repetitions. The experiments were repete twie Sttistil nlysis Anlysis of vrine (ANOVA) ws rrie out with the GLM proeure of SAS (SAS Institute, 1999) n, when signifint (P <.5), tretment mens were ompre using Fisher s protete lest signifint ifferene (LSD) test. In the tomto isese

3 S.B. Koné et l. / Biologil Control 52 (21) severity tests, ANOVA of the experimentl t showe no signifint ifferene etween the two experiments. The t were therefore omine n nlyze s single experiment. 3. Results 3.1. Physio-hemil n miroil nlysis of NCT Physio-hemil properties of the prepre NCT re shown in Tle 1. ph of NCT rnge from 5.92 (sheep mnure SM1) to 6.57 (hiken mnure). The griulturl well wter use to proue the NCT h ph of The highest EC ws foun in NCT from hiken mnure ompost (1.5 ms) followe y NCT from sewee ompost (.48 ms). NCT from ovine mnure, sheep mnure (SM1), n shrimp omposts showe lower EC (.8.1 ms). Wter h n EC of.2 ms. Overll, NCT proue from hiken mnure ompost h the highest quntities of mro- n miroelements (Tle 1). NCT from sewee ompost h the highest onentrtions of Fe, Mn n the seon most elevte quntities of NH 4,PO 4, Cl, SO 4, K, C, Mg, N, Fe, Cu, n Zn. NCT from ovine mnure, sheep mnure (SM1), n shrimp omposts ll h omprle mro- n mironutrient onentrtions, whih were generlly equivlent to wht ws foun in the griulturl well wter ontrol. Esherihi oli ws not present in ny of the NCT or in the griulturl well wter ontrol (t not shown). NCT from hiken mnure n sewee omposts showe n verge of 28 n 5 oliforms/ml, respetively, wheres no oliforms were etete in the other prepre NCT or in the griulturl well wter. Miroil popultions in the proue NCT re shown in Tle 2. NCT from sheep mnure (SM1) ompost provie the highest onentrtion of totl teri, Grm negtive teri, tinomyetes, n fluoresent pseuomons. NCT from shrimp n sewee ompost h the lowest onentrtion of totl teri. Chiken mnure ompost NCT h the lowest Grm negtive teri. Finlly, the NCT from ovine mnure ompost lso h the highest onentrtion of Grm negtive teri s well s the lowest fluoresent pseuomons onentrtion. When NCT from ifferent soures of sheep mnure ompost were evlute, there ws no signifint ifferene in miroil popultions (totl teri, Grm negtive teri, tinomyetes, fluoresent pseuomons, n fungi) etween ny of the four teste NCT (SM1 SM4) (t not shown) Effet of NCT on myelil growth All NCT inhiite the in vitro growth of the three teste pthogens when ompre to the wter ontrol. NCT from sheep mnure (SM1), ovine mnure, n shrimp ompost provie the highest inhiitory effet on A. solni n B. inere, reuing myelil growth y pproximtely 66% n 75%, respetively (Fig. 1). NCT from hiken mnure ompost h lower inhiitory effet thn those from sheep mnure (SM1) n shrimp ompost. NCT from sewee ompost showe the lowest inhiitory effet mong the teste NCT. All teste NCT emonstrte 1% inhiition of P. infestns (t not shown). When sterilize y utolving or y mirofiltrtion, there ws no signifint inhiition of ny of the sterilize NCT on the myelil growth of the three teste pthogens (t not shown) Effet of NCT on tomto isese severity When NCT were sprye on non-inoulte tomto plnts, there ws no pperne of phytotoxi symptoms (sorhing, urning, or spotting of tomto plnt leves) in ny of the tretments (t not shown). Moreover, the pplition of NCT to the non-inoulte tomto plnts showe no signifint effet on plnt height for ny of the tretments (t not shown). A preventive pplition of the five NCT (3 ys efore rtifiil inoultion with O. neolyopersii) provie signifint erese in powery milew severity over the first 4 weeks of the trils (Fig. 2). NCT from sheep (SM1) n hiken mnure omposts emonstrte the lowest isese severity fter 3 n 4 weeks, reuing powery milew y 45 62% when ompre to the wter ontrol. NCT from ovine mnure, shrimp, n sewee omposts lso signifintly reue powery milew severity fter 3 n 4 weeks ut to lesser extent (23 35% isese reution) thn sheep (SM1) n hiken mnure ompost NCT. A urtive pplition of the NCT (3 ys fter rtifiil inoultion) lso emonstrte reution in powery milew severity (Fig. 3), leit to lesser extent thn the preventive tretments. Throughout the urtive tretment tril, NCT from sheep mnure (SM1) ompost provie the lowest powery milew severity mong the NCT (48% n 21% isese reution for weeks 2 n 3, respetively). There ws no signifint ifferene in powery milew severity etween the wter ontrol n shrimp n sewee ompost NCT fter 3 weeks n etween the ontrol n NCT from hiken mnure ompost fter 4 weeks. A preventive pplition of the five NCT (3 ys efore rtifiil inoultion with B. inere) provie signifint erese in gry mol severity over 9-week perio when ompre to the wter ontrol (Fig. 4). NCT proue from sheep mnure (SM1) gve the lowest gry mol severity, reuing isese y 99% n 97% fter 8 n 9 weeks, respetively. NCT from hiken mnure ompost gve the seon lowest isese severity (88 92% reution) fter 8 9 weeks. NCT from ovine mnure n shrimp omposts signifintly reue gry mol severity etween 72% n 85% fter 8 9 weeks ompre to the ontrol. Sewee ompost NCT generlly showe the highest isese severity mong teste NCT. A urtive pplition NCT from hiken mnure, sheep mnure (SM1), n ovine mnure (3 ys fter rtifiil inoultion with B. inere) provie signifint erese in gry mol severity Tle 1 Physio-hemil properties of non-erte ompost tes (NCT) from hiken mnure, ovine mnure, sheep mnure, sewee, n shrimp omposts. NCT ph EC Mro- n mironutrient onentrtion (mg/l) NO 3 NH 4 PO 4 Cl SO 4 K C Mg N Fe Cu Mn Zn Wter Chiken mnure Bovine mnure Sheep mnure (SM1) Sewee Shrimp Within olumns, mro- n mironutrient mens with the sme letter re not signifintly ifferent oring to Fisher s protete LSD test (P =.5). EC, eletril onutivity in millisimens (ms)., elow etetion limits.

4 17 S.B. Koné et l. / Biologil Control 52 (21) Tle 2 Miroil popultions of non-erte ompost tes (NCT) from hiken mnure, ovine mnure, sheep mnure, sewee, n shrimp omposts. NCT Totl teri (1 6 ) Grm negtive teri (1 4 ) Atinomyetes (1 4 ) Fluoresent pseuomons (1 1 ) Fungi (1 3 ) Chiken mnure 3.2 ( ) 1. (.3 3.8) 1.8 (.5 6.8) 7. ( ) 3.2 ( ) Bovine mnure 3.2 ( ) 28 ( ) 7. ( ) 6. ( ) 2.8 (.7 1.6) Sheep mnure (SM1) 12. ( ) 28 ( ) 23. ( ) 1 ( ) 1.4 (.4 5.3) Shrimp 1.2 (.3 4.6) 7. ( ) 2.2 (.6 8.4) 7. ( ) 2.8 (.7 1.6) Sewee 1. (.3 3.8) 5. ( ) 3.2 ( ) 9. ( ) 3.2 ( ) Mens were lulte y the most prole numer metho in MPN/ml. Vlues in rkets represent the low n high onfiene limits (95%). Myelil growth (m) A. solni B. inere Fig. 1. Effet of non-erte ompost tes on myelil growth of Alternri solni n Botrytis inere in vitro. Compost tes were inorporte into PDA (15% vol:vol). For eh fungl pthogen, mens with the sme letter re not signifintly ifferent oring to Fisher s protete LSD test (P =.5). Chiken mnure ompost h; sheep mnure (SM1) ompost ; ovine mnure ompost ; shrimp ompost ; sewee ompost ; wter ontrol j. over 5- to 9-week perio when ompre to the wter ontrol (Fig. 5). Throughout the urtive tretment tril, NCT from sheep mnure (SM1) ompost provie the lowest isese severity followe y either hiken mnure ompost NCT or ovine mnure ompost NCT. While initilly le to reue gry mol severity, sewee ompost NCT n shrimp ompost NCT i not signifintly erese severity s ompre to the ontrol fter 8 n 9 weeks of ulture, respetively (Fig. 5) Effet of ifferent sheep ompost NCT on myelil growth In ition to NCT prepre from sheep mnure ompost SM1, three other NCT were prepre from ifferent soures of ommerilly-ville sheep mnure omposts (SM2, SM3, SM4) n teste ginst A. solni, B. inere, n P. infestns in vitro. NCT from SM1, SM2, SM3, n SM4 inhiite the myelil growth of A. solni n P. infestns to lmost equivlent levels (Fig. 6). NCT from SM3 n SM4 provie signifint reution in B. inere myelil growth, lthough these were signifintly less inhiitory thn SM1 n SM2. When sterilize y utolving or y mirofiltrtion, there ws no signifint inhiition of ny of the sterilize sheep mnure NCT on the myelil growth of the three teste pthogens (t not shown). 4. Disussion Compost tes re pereive s potentil lterntives to syntheti hemil fungiies (Siiqui et l., 29). Although most ounts esrie ompost tes proue from vrious ompost types emonstrting n inhiitory effet on plnt pthogens in numerous plnt pthosystems, these effets remin vrile (Sheuerell n Mhffee, 26) n therefore iffiult to use relily. In reent work, the mehnisms of tion unerlying the effiy of ompost tes to ontrol plnt pthogens hve een reporte s single or multiple mehnisms involving miroil ntgonism (through ntiiosis, prsitism, ompetition for nutrients/spe or inue plnt resistne; Zhng et l., 1998; El-Msry et l., 22; Al-Mughri et l., 28) or suppressive physiohemil properties (improve nutrient sttus of the plnt, toxi Week fter tretment Fig. 2. Effet of preventive pplition of non-erte ompost tes on the severity of tomto powery milew (Oiium neolyopersii) in greenhouse ssys. Compost tes were pplie until runoff on rtifiilly-inoulte tomto plnts. For eh week, mens with the sme letter re not signifintly ifferent oring to Fisher s protete LSD test (P =.5). Chiken mnure ompost h; sheep mnure (SM1) ompost ; ovine mnure ompost ; shrimp ompost ; sewee ompost ; wter ontrol j.

5 S.B. Koné et l. / Biologil Control 52 (21) Week fter tretment Fig. 3. Effet of urtive pplition of non-erte ompost tes on the severity of tomto powery milew (Oiium neolyopersii) in greenhouse ssys. Compost tes were pplie until runoff on rtifiilly-inoulte tomto plnts. For eh week, mens with the sme letter re not signifintly ifferent oring to Fisher s protete LSD test (P =.5). Chiken mnure ompost h; sheep mnure (SM1) ompost ; ovine mnure ompost ; shrimp ompost ; sewee ompost ; wter ontrol j e e Week fter tretment Fig. 4. Effet of preventive pplition of non-erte ompost tes on the severity of gry mol of tomto (Botrytis inere) in greenhouse ssys. Compost tes were pplie until runoff on rtifiilly-inoulte tomto plnts. For eh week, mens with the sme letter re not signifintly ifferent oring to Fisher s protete LSD test (P =.5). Chiken mnure ompost h; sheep mnure (SM1) ompost ; ovine mnure ompost ; shrimp ompost ; sewee ompost ; wter ontrol j. ompouns, or inue resistne; Hoitink et l., 1997; Siiqui et l., 28). In this stuy, NCT prepre from vrious ompost types were evlute for their ility to ontrol tomto pthogens to gin insight into properties involve in their suppressive effets. In vitro ssys showe tht the five teste NCT provie signifint inhiition of myelil growth of A. solni, B. inere, n P. infestns. Among the teste NCT, those prepre from sheep mnure (SM1), ovine mnure, n shrimp omposts gve the highest inhiition wheres NCT prepre from hiken mnure n, in prtiulr, sewee ompost lwys h lesser effet. This suggests tht the NCT (n their intrinsi omponents) hve iret inhiitory effet on the teste pthogens. When NCT were sterilize y mirofiltrtion or utolving, they h ompletely lost their iret inhiitory effet on the myelil growth of the teste pthogens. This inites tht toxi ompouns (extrte from the orgni n inorgni ompost mterils or proue y the miroil popultions within the NCT) were not initilly present in the NCT or t lest not t levels tht impee the growth of the teste pthogens. This lso implies tht the presene of miroil ntgonists in the NCT is neessry for iret inhiition of myelil growth. These ntgonists oul potentilly use ompetition or prsitism to exert their iret effet on the pthogens n my lso proue ntimiroil ompouns (ntiiosis) when in iret onfronttion with the pthogens. Along these lines, previous reports hve shown tht iosynthesis of toxi metolites in teri n fungi oul e enhne or newly inue in the presene of ompeting miroorgnism (Sonnenihler et l., 1994; Mrtinez et l., 26). In in vivo trils, ll teste NCT emonstrte inhiition of the evelopment of oth iseses, in prtiulr gry mol (B. inere). Although powery milew (O. neolyopersii) severity ws signifintly lower thn the ontrol, pplition of the teste NCT file to effiiently ontrol this isese. The preventive pplition of NCT 3 ys efore inoultion y the pthogens ws more effetive in ontrolling evelopment of the iseses thn urtive pplition (3 ys fter inoultion with the pthogens). This woul inite tht the NCT suppressive effet is prtly euse of inhiition of fungl spore germintion

6 172 S.B. Koné et l. / Biologil Control 52 (21) Week fter tretment Fig. 5. Effet of urtive pplition of non-erte ompost tes on the severity of gry mol of tomto (Botrytis inere) in greenhouse ssys. Compost tes were pplie until runoff on rtifiilly-inoulte tomto plnts. For eh week, mens with the sme letter re not signifintly ifferent oring to Fisher s protete LSD test (P =.5). Chiken mnure ompost h; sheep mnure (SM1) ompost ; ovine mnure ompost ; shrimp ompost ; sewee ompost ; wter ontrol j. Myelil growth (m) n/or penetrtion insie the plnt ells s hve een shown previously (Diánez et l., 27; Siiqui et l., 29). NCT from sheep mnure (SM1) ompost lwys provie the highest inhiition of isese evelopment in ll trils. In prtiulr, preventive pplition of NCT from sheep mnure (SM1) ompost ws le to reue gry mol severity y more thn 95% over 9-week perio. NCT from hiken mnure ompost onsistently provie the seon highest isese inhiition wheres NCT from sewee ompost ws lwys the lest inhiitory. NCT from ovine mnure, hiken:ovine (1:9 vol:vol) mnure, n grpe mr omposts using similr proution methoology (1:5 ompost/wter rtio; 14-y inution) h een previously shown to reue gry mol on tomto y out 74 8% in growth hmers n NCT from ovine mnure reue gry mol in greenhouse tomtoes y 3% n 37% fter 6 n 8 weeks, respetively (El n Shtienerg, 1994). This sme work showe signifint inhiition (4 52%) of the evelopment of nother powery milew fungus, Leveillul turi, on tomto plnts with weekly pplitions of ovine mnure omposts. A. solni B. inere P. infestns Fig. 6. Effet of non-erte sheep ompost tes prepre from ifferent ommeril sheep mnure omposts on myelil growth of Alternri solni, Botrytis inere, n Phytophthor infestns in vitro. Compost tes were inorporte into PDA (15% vol:vol). For eh fungl pthogen, mens with the sme letter re not signifintly ifferent oring to Fisher s protete LSD test (P =.5). Sheep mnure omposts: SM1 h; SM2 ; SM3 ; SM4 ; wter ontrol j. When tken together, in vitro n in vivo results ppere to initilly orrelte well with totl teri onentrtions in iniviul NCT. The most effiient NCT from sheep mnure (SM1) ompost showe the highest totl teri onentrtion, whih ws pproximtely fourfol higher thn the next most effiient NCT (hiken n ovine mnure omposts). NCT from sheep mnure (SM1) ompost lso h 1- n 12-fol higher totl teri onentrtion thn NCT from shrimp n sewee omposts, respetively, whih were the lest effiient NCT in this stuy. Although, NCT from sheep mnure (SM1) ompost lso h the highest onentrtion of Grm negtive teri, tinomyetes n fluoresent pseuomons, these prtiulr ftors i not orrelte well with the reltive effiy of the other NCT. Previous stuies h lso postulte tht high totl teri ontent ws linke to the effiy of vrious NCT (Weltzien, 1991; Diánez et l., 27; Al-Mughri et l., 28). However, results from ssys with NCT prepre from four soures of sheep mnure ompost h vrile effiy ginst the teste pthogens in vitro lthough they h similr miroil popultion ounts inluing totl teri. Contrry to the previous ssys, this inite tht totl teril ounts in the NCT i not orrelte with their reltive effiy. Other work esrie tht prtiulr teril isoltes from NCT oul effetively ontrol gry mol when pplie lone (El n Shtienerg, 1994) n tht inresing the teril ounts in NCT through the use of fermenttion itives i not neessrily orrelte with the reution of gry mol (Sheuerell n Mhffee, 26). This my inite tht speifi miroorgnisms (potentil iologil ontrol gents) woul e more importnt in the suppressive effet thn the high totl ounts of teri. In some ses, it ws shown tht NCT were more effiient in reuing the severity of gry mol on tomto plnts thn it ws in inhiiting the myelil growth of B. inere in vitro. The itionl iniret effet of these NCT ginst B. inere in vivo might hve een result of their higher onentrtion in nutrients (Tle 1). Although improve nutritionl sttus my protet the plnt ginst isese (Siiqui et l., 28), there ws no inrese in plnt height in plnts NCT tretment n solute onentrtions of mro- n mironutrients woul mke the prepre NCT wek fertilizers t est. It is more prole tht this iniret effet ws provie y prtiulr miroorgnisms or ompouns within the NCT tht, while not iretly inhiitory to B. inere myelil growth

7 S.B. Koné et l. / Biologil Control 52 (21) in vitro, h possile effet on the inution of intrinsi plnt efenses tht help stve off gry mol s hs een reporte in other systems (Zhng et l., 1998; Hggg n Ser, 27; Siiqui et l., 29). The higher nutrient onentrtion in some NCT my hve lso supporte the growth of the ntgonisti miroorgnisms on the lef surfe. With respet to potentil miroil sfety issues of NCT, lthough E. oli ws not present in ny of the NCT, it is noteworthy tht NCT prepre from hiken mnure n sewee omposts onsistently showe the presene of oliforms. Both of these NCT (hiken mnure NCT in prtiulr) showe oliform ounts tht re ove mny USA stte or Cnin guielines for irrigtion wter. Although oliforms in NCT hve een reporte to fll elow etetle levels when pplie to plnts (Welke, 25), re shoul e tken to limit or eliminte potentil humn pthogens from ompost tes, in prtiulr if preprtion inlues itives estine to enhne miroil growth (Ingrm n Millner, 27). Overll, results from this stuy showe tht NCT from vrious ompost types provie signifint inhiition of the myelil growth of folir pthogens of tomto n signifintly reue the evelopment of powery milew (O. neolyopersii) n, in prtiulr, gry mol (B. inere) on tomto plnts. NCT prepre from sheep mnure ompost (SM1) onsistently provie the highest inhiition throughout the stuy. The reltive effiy of ll the prepre NCT ppere to initilly orrelte well with the onentrtion of totl teri in the respetive NCT, ut i not orrelte well with other miroil ommunities (Grm negtive teri, fluoresent pseuomons, tinomyetes, fungi) or with mron mironutrient onentrtions. However, results from the vrious sheep mnure ompost NCT woul inite tht speifi miroorgnisms (potentil iologil ontrol gents) woul e more importnt in the suppressive effet thn the high totl ount of unspeifie teri. Although the loss of in vitro effiy upon steriliztion of the NCT inites tht miroil ommunities must e present to exert their ntgonisti effet (ompetition, prsitism, ntiiosis) ginst the pthogens n suggests tht miroil ommunities or prtiulr miroorgnisms re involve iretly in isese suppression, the inution of plnt efense retion y prtiulr miroorgnisms or y orgni n inorgni ompouns in the NCT nnot e rule out. Future work will ttempt to ientify miroil ntgonists in highly suppressive NCT s well s ientify ompost te preprtion prmeters tht enhne popultions of these ntgonists. Aknowlegments The uthors thnk Krine Guette for tehnil ssistne. S.B.K. ws supporte y sholrship from the Islmi Development Bnk. This work ws supporte y the Ministère e l Agriulture, es Pêheries et e l Alimenttion u Quée (MAPAQ). Referenes Al-Dhmni, J.H., Assi, P.A., Miller, S.A., Hoitink, H.A.J., 23. Suppression of teril spot of tomto with folir sprys of ompost extrts uner greenhouse n fiel onitions. 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