Effect of biostimulant sprays on Phaeomoniella chlamydospora and esca proper infected vines under greenhouse and fi eld conditions

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1 Phytopathol. Mediterr. (29) 48, Effect of biotimulant pray on Phaeomoniella chlamydopora and eca proper infected vine under greenhoue and fi eld condition STEFANO DI MARCO and FABIO OSTI Itituto di Biometeorologia, CNR, Via Gobetti 11, 4129 Bologna, Italy Summary. Biotimulant are compound that influence phyiological procee in plant, producing better growth and enhancing tre tolerance. The effect of ome biotimulant on vine wa invetigated over a number of year to ae their effect both on the incidence of eca leaf ymptom in the vineyard and on the growth of Phaeomoniella chlamydopora artificially inoculated into potted vine. Field trial were carried out for 4 7 year in five 15 2-yearold vineyard infected with eca proper. Potted plant were prayed with biotimulant, after which the vine trunk were inoculated with P. chlamydopora, and then the vine were prayed again with biotimulant in the following 2 or 3 growing eaon. On the whole, biotimulant in the field did not reduce foliar ymptom. The percentage of ymptomatic vine that had hown ymptom in previou year wa higher in the biotimulant-prayed plot. In the greenhoue, a certain reduction of internal necroi caued by P. chlamydopora wa een with three of the four biotimulant teted. Propect for biotimulant a a mean control eca are dicued. Key word: grapevine, control, Phaeomoniella chlamydopora. Introduction Although many tudie are currently evaluating different method and trategie to combat eca, control of the eca dieae complex i till lacking. Eca control in mature vineyard, characterized by vine with dark wood treaking (tracheomycoi) and white rot, i till baed on cultural practice, uch a the renewal of the trunk of ymptomatic plant, exciing all the necrotic wood, and removing infected plant and/or pruning debri (Di Marco et al., 2; Surico et al., 26a; 26b). Thee practice reduce the inoculum load and the rik of new infection, but they do not cure already exiting infection (Montert et al., 26). Correponding author: S. Di Marco Fax: ; .dimarco@ibimet.cnr.it The eca complex i correlated with the phyiology of the hot plant. When the vine i phyiologically healthy, eca pathogen live inide the plant without producing ymptom for everal year. When tre condition arie, however, infected vine alo begin to uffer the negative conequence of thee pathogen, coniting in potentially evere foliar ymptom and loe in yield. Eca i therefore a tre-related dieae (Ferreira et al., 1999; Rooney-Latham et al., 25; Surico et al., 26b; Edward et al., 27). Numerou tudie on eca-infected vine have hown that the extent of wood deterioration i not correlated with the expreion of foliar ymptom, but that foliar ymptom are correlated with toxin produced by the tracheomycotic fungi (Mugnai et al., 1999; Surico et al., 2; Abou-Manour et al., 24; Marchi et al., 26; Calzarano and Di Marco, 27; 28). There i, however, a relationhip between foliar ymptom and the quality of both grape clu- 47

2 S. Di Marco and F. Oti ter and the wine preed from them. Symptomatic vine have a lower quality, wherea aymptomatic vine, even if they are in fact dieaed, produce grape with quality-characteritic imilar to the grape of healthy vine (Calzarano et al., 21; Calzarano et al., 24b). Therefore, trategie aimed at reducing the incidence and/or everity of foliar ymptom would alo limit lo in quality. Biotimulant are non-nutritional compound that promote favourable plant repone (Zhang, 1997; Miller and Gange, 23; Schmidt et al., 23), booting phyiological procee, producing fater growing and healthier plant, improving crop yield and quality, and enhancing tre tolerance (Blake, 22; Fregoni and Fregoni, 25). A recent Italian legilative decree publihed in the Gazzetta Ufficiale (26) defined biotimulant a product that are added to other fertilizer and/or to the oil and/or to plant, that promote or regulate the aborption of nutrient by plant, and that improve phyiological anomalie. Phytohormone, or active ingredient with a pecific phytoanitary function, are not biotimulant (Ciavatta and Cavani, 26). In ome preliminary tudie, biotimulant were prayed on eca-infected vineyard in Emila Romagna and Abruzzo. The treatment were applied for only a hort period (two year), and they did not lower eca everity (Di Marco and Oti, 25a; Calzarano et al., 27). Over a coniderable number of year, biotimulant were experimentally prayed on vineyard affected with eca proper, in order to ae their effect on eca incidence in the field. Thee biotimulant were alo teted in greenhoue trial on potted vine inoculated with Phaeomoniella chlamydopora, conidered a repreentative tracheomycotic agent cauing eca. Thi paper report on the finding obtained in thoe experiment. Material and method Greenhoue trial Two experiment were carried out over a fiveyear period, the firt in 23-25, and the econd in Twelve replicate per treatment, each coniting of one potted vine, were et up. Plant material One-year-old vine, cv. Trebbiano, grafted on SO4, (firt experiment) and cv. Cabernet Sauvignon Omega, grafted on 113P, (econd experiment), were obtained from a commercial nurery and planted in 2 cm diameter platic pot containing 7% of a commercial mixture of peat with nutrient and element at low concentration (Floragard TKS2, Oldenburg, Germany) and 3% perlite. Potted plant were grown outdoor, watered normally, treated with copper or ulphur-baed compound, and protected from hail with creen. Biotimulant praying and fungal inoculation Vine were prayed on the leave with a portable atomizer prayer (capacity 5 l). They were prayed four time every year at 1- to 14-day interval, and following manufacturer doage intruction, for 3 year, except for Brotomax, for which the vineyard praying chedule recommended by the manufacturer wa followed (Table 1). In the firt year, even day after the lat praying, potted vine were inoculated with P. chlamydopora train 56 (CBS ). A hole 5 mm in diameter wa drilled into the trunk and a plug from a 2-week-old PDA fungal culture wa inerted (a terile plug wa inerted for the control). The hole were covered with Amojell lubricant (Sigma-Aldrich) and protected with Parafilm M (American National Can, Chicago, IL, USA) to prevent drying. The econd experiment followed the ame biotimulant praying chedule but ued Marvita rather than Fitotim a the biotimulant. Evaluation of praying, fungal re-iolation, and data analyi The effect of the biotimulant pray wa determined by aeing the vegetative characteritic of the prayed vine during each growing eaon, the internal longitudinal pread of the necroi through the vine, and the re-iolation of the pathogen from the dicoloured wood. Necrotic leion were meaured during each growing eaon, and a final aement (baed on at leat 7 vine per type of pray given) wa made 3 month after inoculation in Experiment 1, and 18 month after inoculation in Experiment 2. For the final aement, the trunk were cut lengthwie from the inoculation ite and the length of the necrotic leion wa meaured. The data were expreed a the average length of necroi and tatitically analyzed uing Duncan Multiple Tet, P=.5. Soon after the leion were meaured, 28 frag- 48 Phytopathologia Mediterranea

3 Effect of biotimulant pray on eca dieaed vine ment of necrotic wood (4 per plant) from each pray treatment for the firt experiment, and 21 fragment (3 per plant) for the econd experiment, were cut aeptically and placed on PDA amended with treptomycin ulfate (Merck) at 8 mg l -1 in order to ae the viability of P. chlamydopora from the inoculated plant. Plate were maintained at 25±2 C in the dark. The percentage of fertile fragment of P. chlamydopora wa calculated. Vineyard trial The trial wa conducted in in 5 vineyard. Biotimulant were prayed with a preure atomizer. Half of the inpected vine in each vineyard wa prayed. Detail on field praying are ummarized in Table 1. In each vineyard, the trunk of ome vine were cut and the wood inpected to determine wood dicoloration and decay caued by eca. Every growing eaon, between the end of Augut and the third week of September, at the time of the maximum expreion of eca ymptom, the incidence of eca wa calculated in each vineyard by dividing the number of vine with viible ymptom by the total number of inpected vine (excepted dead vine). Data were expreed a annual and a cumulated incidence (Calzarano and Di Marco, 27). The percent frequency of vine with foliar ymptom wa alo calculated, and expreed a the ratio in a given year of the number of ymptomatic vine that had hown ymptom in previou year over Table 1. Biotimulant ued: compoition, application rate and chedule, plant phenological tage and vineyard detail. Trade name Compoition Plant phenological tage a (application chedule) Application Rate b No. c Year Cultivar Vine inpected (No.) Brotomax Aluminium lignin ulphate, gluconic acid, microelement I: hoot 25 cm long II: 2 day after I III: berrie beginning to touch IV: end of harvet 1 l hl Pignoletto 445 Fitotim Aminoacid, peptide, peptone From pre-flowering to majority of berrie touching (14 day-interval) 15 ml hl Pignoletto 29 Kendal Glutatione, oligoaccharine From pre-flowering to majority of berrie touching (1- to 14-day interval) 3 ml hl Albana Sangiovee Marvita Achophyllum nodoum extract From pre-flowering to majority of berrie touching (1- to 14-day interval) 3 g ha Pignoletto 476 a The phenological tage are decribed according to BBCH cheme. b The application rate refer to the commercial formulation. c No. of praying per year. Vol. 48, No. 1 April, 29 49

4 S. Di Marco and F. Oti the total number of ymptomatic vine aeed in that year, and multiplying by 1. Thi percentage wa firt calculated the year after the biotimulant pray were initiated. Vineyard were alo inpected for eca-related mortality (occurrence of dicoloration and decay in dead vine wood). Vine that died in the firt year of the tudy were excluded becaue it wa not certain whether thee plant died in that year or in a previou year. Data were expreed a annual and a cumulated incidence of mortality. For each year of aement, the annual cumulated incidence of mortality, the incidence of foliar ymptom, and vine mortality were ubjected to Chi-Square tatitical analyi (P=.5) uing SAS verion 8.1 (Anonymou, 199). Reult Greenhoue trial Reult of greenhoue trial are ummarized in Table 2. At the end of the firt experiment (3 month after inoculation) biotimulant-prayed vine howed a certain reduction in the necrotic treak caued by P. chlamydopora. Thi reduction wa ignificant (P<.5) only with vine prayed with Fitotim (7.4 cm), a compared with unprayed inoculated control plant (11. cm). At the end of the econd experiment (18 month after inoculation) vine prayed with Brotomax and Marvita howed a ignificant (P<.5) reduction in necrotic leion from P. chlamydopora (4.9 cm and 4.1 cm repectively) in comparion with the unprayed inoculated control vine (6.7 cm). Phaeomoniella chlamydopora wa iolated from the necrotic leion of both prayed and unprayed vine. However, vine prayed with biotimulant had lower percentage of fragment with P. chlamydopora: ranging from 6.7 % (with Kendal and Brotomax, firt experiment) to 76.2% (econd experiment) compared with the control (75.% %). Vineyard trial Both vineyard prayed with Kendal had a higher annual incidence of eca than the unprayed vineyard (P>.5), but the difference Table 2. Effect of pray of biotimulant on Phaeomoniella chlamydopora (Pch) inoculated in the trunk of potted grafted grapevine. Treatment Average necroi length (cm) Experiment 1 (3 m.a.i. a ) Experiment 2 (18 m.a.i. a ) Fertile Pch fragment (%) Average necroi length (cm) Fertile Pch fragment (%) Brotomax 9.8 a b b b 71.4 Fitotim 7.4 b 64.3 nt nt Kendal 1. a a 76.2 Marvita nt nt 4.1 b 76.2 Untreated control 11. a a 85.7 a Month after inoculation. b Value in column followed by the ame letter do not differ ignificantly according to Duncan Multiple Range tet (P=.5). nt, Not teted. 5 Phytopathologia Mediterranea

5 Effect of biotimulant pray on eca dieaed vine wa not ignificant (Fig. 1A, B). The cumulated incidence gave contradictory reult: the percentage of ymptomatic Albana vine in the Kendalprayed plot increaed, though not ignificantly (P>.5), wherea the percentage of Sangiovee vine decreaed, a compared with the unprayed plot. In the firt four year of Brotomax praying (21 24), the percentage of ymptomatic vine 45 4 A 35 Symptomatic vine (%) B 25 Symptomatic vine (%) Annual incidence unprayed Cumulated incidence unprayed Annual incidence prayed Cumulated incidence prayed Fig. 1. Effect of application of Kendal on annual incidence (bar) and cumulated incidence (line) aeed in vineyard cv. Albana (A) and Sangiovee (B). For each treatment and year of aement, value of annual incidence or cumulated incidence are not ignificantly different according to Chi Square tet (P>.5). Vol. 48, No. 1 April, 29 51

6 S. Di Marco and F. Oti in the prayed plot marginally increaed and the cumulated eca incidence wa tatitically higher than in the unprayed plot (Fig. 2A). Starting from 25, the annual incidence in the prayed plot lightly decreaed, and thi decreae became ignificant (P<.5) in 27. The cumulated incidence A Symptomatic vine (%) B Symptomatic vine (%) C Symptomatic vine (%) Annual incidence untreated Cumulated incidence untreated Annual incidence treated Cumulated incidence treated Fig. 2. Effect of application of Brotomax (A), Fitotim (B) and Marvita (C) on annual incidence (bar) and cumulated incidence (line) aeed in three different vineyard cv. Pignoletto. For each treatment and year of aement, value of annual incidence or cumulated incidence marked with aterik or S repectively, differ ignificantly according to Chi Square tet (P<.5). 52 Phytopathologia Mediterranea

7 Effect of biotimulant pray on eca dieaed vine increaed le trongly in the prayed plot than in the unprayed plot. The percentage of annual and cumulated eca incidence calculated for the period of wa ignificantly greater (P<.5) in the plot prayed with Fitotim than in the unprayed plot (Fig. 2B). Both the annual and the cumulated incidence of eca in the Marvita-prayed plot wa lower than that in the unprayed plot (Fig. 2C). In 27 the percent cumulated incidence wa ignificantly different (P<.5) between the prayed and the unprayed plot. In each year of the tudy, ymptomatic vine that had hown ymptom in previou year were more frequent in prayed plot than in unprayed plot. Thi wa found with all biotimulant prayed, except Kendal in 25 ( Albana vineyard trial), and Brotomax in 24 and 27 (Fig. 3 and 4). The percentage of vine that died from eca after 7 6 A 5 Vine (%) B 3 Vine (%) Unprayed Sprayed Fig. 3. Effect of application of Kendal on the frequency of ymptomatic vine that howed ymptom in previou year aeed in vineyard cv. Albana (A) and Sangiovee (B). For each treatment and year of aement, value of frequency are not ignificantly different according to Chi Square tet (P>.5). Vol. 48, No. 1 April, 29 53

8 S. Di Marco and F. Oti 8 7 A 6 Vine (%) B 4 Vine (%) C Symptomatic vine (%) Unprayed Sprayed Fig. 4. Effect of pray of Brotomax (A), Fitotim (B) and Marvita (C) on the incidence of ymptomatic vine that had hown ymptom in previou year calculated in 3 different vineyard cv. Pignoletto. For each biotimulant prayed and each year, bar marked with an S differ ignificantly according to the Chi-Square tet (P=.5). 54 Phytopathologia Mediterranea

9 Effect of biotimulant pray on eca dieaed vine Table 3. Effect of biotimulant (Brotomax, Kendal and Marvita) pray on the percentage of eca-related mortality on different grapevine cultivar. Top: vine that died in a given year (annual mortality). Bottom: vine that died in one of the year of the tudy (cumulated mortality). Data on the vineyard treated with Fitotim are not given becaue the number of vine that died from eca in thi vineyard wa too low. Year of tudy Annual mortality Brotomax Kendal Marvita Cv. Pignoletto Cv. Albana Cv. Sangiovee Cv. Pignoletto Unprayed Sprayed Unprayed Sprayed Unprayed Sprayed Unprayed Sprayed nt a nt nt nt nt nt nt nt nt nt nt nt nt nt.6.2 nt nt nt nt nt nt.5.5 Year of urvey Cumulated mortality Brotomax Kendal Marvita Cv. Pignoletto Cv. Albana Cv. Sangiovee Cv. Pignoletto Unprayed Sprayed Unprayed Sprayed Unprayed Sprayed Unprayed Sprayed nt nt nt nt nt nt nt nt nt nt nt nt nt nt.6.2 nt nt nt nt b nt nt a For each treatment and cultivar, value in row followed by an aterik differ ignificantly according to the Chi-Square tet (P=.5). nt, Not teted. Vol. 48, No. 1 April, 29 55

10 S. Di Marco and F. Oti the firt year of biotimulant praying i hown in Table 3. The incidence of annual mortality in Brotomaxprayed vine wa lightly lower than in unprayed vine. Spraying with Kendal gave contradictory reult: in the Sangiovee vineyard it reduced the percentage of annual and cumulated mortality, though the decreae wa ignificant (P<.5) only in 26, but in the Albana vineyard, it increaed vine mortality, though not ignificantly, a compared with the unprayed vine. An increae in percent mortality wa alo found with Marvita-prayed vine. Thi increae wa tatitically ignificant in 27 (P<.5), but only for cumulated mortality. Dicuion The primary objective of thi tudy wa to determine whether biotimulant pray over a number of year would limit foliar ymptom in grapevine uffering from eca proper. On the whole, the biotimulant teted did not limit foliar ymptom. Similar unatifactory reult were alo recently obtained in Autralia, where nutrient fertilizer and Brotomax prayed on eca-infected grapevine did not limit dieae ymptom or progre (Edward and Pacoe, 25). In our trial, prayed vine often howed a certain increae in the annual and cumulated incidence of eca. Thi finding i conitent with Calzarano et al. (27), who found that frequent praying of foliar fertilizer and bioactivator (eleven per year) on eca-proper infected vine during the growing eaon increaed both the incidence and the everity of the dieae. In our tudy biotimulant were prayed four or five time a year (except for Brotomax, which wa prayed following the manufacturer pray chedule) on vine with mot of their leave aymptomatic and till photoynthetically efficient (Petit et al., 26). The phenological growth tage at which biotimulant were prayed, and the mall number of praying, could not have caued any imbalance or exce of nutrient in the leave, which generally make plant more uceptible to tre (Scheir and De Bruyn, 24). Calzarano et al. (27) potulated that nutritional imbalance increaed foliar ymptom expreion. Mot likely, the increae in foliar ymptom from biotimulant found in thi tudy wa mainly due to an increae in plant phyiological proce: a a reult of which fungal toxin moved to the leave more readily and/ or in greater amount, thu raing the incidence of foliar ymptom (Surico et al.). Further evidence for thi increae in phyiological procee wa the fact that prayed vine had a greater incidence of ymptomatic vine that had hown ymptom in previou year. Thi higher incidence led to a clear reduction in the fluctuation of foliar ymptom from year to year. The way in which Brotomax act on eca and aociated pathogen wa invetigated by Del Rio et al. (21; 24). In our tudy, after five year of praying Brotomax in a vineyard without achieving any poitive effect, the percent annual and cumulate eca incidence wa reduced for three year. Another tudy likewie found that when Brotomax wa prayed on grapevine affected with Eutypa dieback the yield of the vine improved only after the biotimulant had been prayed for three year (Sonowki et al., 25). It eemed that Brotomax in the field wa only effective if it wa prayed for everal year, and thi limit it potential a a biotimulant to be ued in infected vineyard. Moreover, the reduction in foliar ymptom found in our tudy may alo have been favoured by the low percentage of ymptomatic vine and by the proper management of the vineyard. Such condition proved to be important alo for the activity of foetyl Al on foliar ymptom expreion of eca (Di Marco et al., 1999; Di Marco and Oti, 25b). Biotimulant baed on extract from the eaweed Acophyllum nodoum (uch a Marvita) were reported to have a poitive effect when prayed on cultivar of vegetable and flowering bedding plant, conifer and broadleaf tree, and turfgra (Poincelot, 1993; Thompon, 24; Erwin et al., 24). In the preent tudy, Marvita reduced eca ymptom in the vineyard. However, thi i only a preliminary finding that will need ome more year of invetigation before it effect on eca in the vineyard can be known for certain. The data obtained in thi tudy made it impoible to acertain the effect of biotimulant on eca-related mortality becaue of the low number of dead vine found in the vineyard tudied. Mot biotimulant prayed on the potted plant ignificantly reduced the necrotic leion caued by the fungal pathogen. Leion in artificially inoculated vine were alo reduced by foliar, oil or trunk injection with the triazolic fungicide, foetyl Al (Di 56 Phytopathologia Mediterranea

11 Effect of biotimulant pray on eca dieaed vine Marco et al., 1999; 2) and biological control agent uch a Trichoderma pp. (Di Marco et al., 24; Di Marco and Oti, 27). All thee reduction were achieved only in imple and/or localized form of infection occurring in young potted vine inoculated with P. chlamydopora, which are obviouly different from eca proper a it occur in the vineyard. Although the mode and mechanim of action of each biotimulant are different, and will be the object of further invetigation, the reduction of necroi in prayed plant probably reult from an interaction between the ingredient of the biotimulant and the phyiological procee of the hot plant. Thi interaction ha a beneficial effect on the vine, in the ame way a Trichoderma pray have a beneficial effect on vine in the nurery (Fourie et al., 21; Di Marco et al., 24; Fourie and Halleen, 24, 26; Di Marco and Oti, 27). In concluion, biotimulant were not effective againt eca proper in infected vineyard. However, further tudie are needed to ae the practical feaibility of biotimulant to control eca in plant material with no or low level of infection. Acknowledgement Reearch tudy commiioned from ARSIA- Tocana (Regional Agency for Development and Innovation in Agriculture and Foret) on behalf of fourteen adminitrative Region and one autonomou province, and financed with fund provided by the Minitero per le Politiche Agricole e Foretali (Minitry for Agriculture and Foretry Policy) to implement the inter-regional Project Grapevine eca: reearch and experiment in the nurery and in the field for prevention and cure. The author alo thank Stefano Bongiovanni (CAA G. Nicoli ) Stefano Forbicini (Conorzio Agrario di Bologna e Modena) and Guido Ghermandi (AgriTe), for help given during the trial. Literature cited Anonymou, 199. SAS/STAT Uer Guide, Categorical Data Analyi. SAS Intitute Inc., Cary, NC, USA. Abou-Manour E., E. Couché and R. Tabacchi, 24. Do fungal naphthalenone have a role in the development of eca ymptom? Phytopathologia Mediterranea 43, Blake TJ., 22. Antioxidant and natural biotimulant enhancement of eedling growth and tre tolerance in conifer eedling. Journal of the Minitry of Foret Britih Columbia 14(1-2), Calzarano F., C. Amalfitano, L. Seghetti and V. D Agotino, 27. Foliar treatment of eca-proper affected vine with nutrient and bioactivator. Phytopathologia Mediterranea 44, Calzarano F., A. Cichelli and M. Odoardi, 21. Preliminary evaluation of variation in compoition induced by eca on cv. Trebbiano D Abruzzo grape and wine. Phytopathologia Mediterranea 4, S443 S448. Calzarano F. and S. Di Marco, 27. Wood dicoloration and decay in grapevine with eca proper and their relationhip with foliar ymptom. Phytopathologia Mediterranea 46, Calzarano F. and S. Di Marco, 28. Approfondimenti ulla dinamica di colonizzazione e ulla manifetazione dei intomi fogliari nell'eca propria. Micologia Italiana 37(2), Calzarano F., Di Marco S. and A. Ceari, 24a. Benefit of fungicide treatment after trunk renewal of vine with different type of eca necroi. Phytopathologia Mediterranea 43, Calzarano F., L. Seghetti, M. Del Carlo and A. Cichelli, 24b. Effect of eca on the quality of berrie, mut and wine. Phytopathologia Mediterranea 43, Ciavatta C and L. Cavani, 26. Problematiche per l inerimento dei biotimolanti nella legilazione dei fertilizzanti. Atti Convegno Biotimolanti in agricoltura: apetti agronomici, analitici, normativi 7-8 July 26, Montegridolfo (RN), Italy, Fertilita Agrorum 1(1), Del Río J.A., A. González, M.D. Futer, J.M. Botía, P. Gómez, V. Fría and A. Ortuño, 21. Tyloe formation and change in phenolic compound of grape root infected with Phaeomoniella chlamydopora and Phaeoacremonium pecie. Phytopathologia Mediterranea 4, S394 S399. Del Río J.A., P. Gómez, A. Baidez, M.D. Futer, A. Ortuño, and V. Fría, 24. Phenolic compound have a role in the defence mechanim protecting grapevine againt the fungi involved in Petri dieae. Phytopathologia Mediterranea 43, Di Marco S., A. Mazzullo, F. Calzarano and A. Ceari, In vitro tudie on the phophorou acid-viti tilbene interaction and in vivo phoetyl-al activity toward Phaeoacremonium pp. grapevine wood decay agent. In: Modern Fungicide and Antifungal Compound II (H. Lyr, P.E. Ruel, H.-W. Dehene, H.D. Siler ed.), Intercept Ltd, Andover, UK, Di Marco S., A. Mazzullo, F. Calzarano and A. Ceari, 2. The control of eca: tatu and perpective. Phytopathologia Mediterranea 39(1), Di Marco S. and F. Oti, 25a. Eperienze di lotta al mal dell eca. Il Divulgatore 28(5), Di Marco S. and F. Oti, 25b. Effect of foetyl Al foliar application toward eca fungi in grapevine. Phytopathologia Mediterranea 44(1), (abtract). Di Marco S. and F. Oti, 27. Application of Trichoderma to prevent Phaeomoniella chlamydopora infection in organic nurerie. Phytopathologia Mediterranea 46, Di Marco S., F. Oti and A. Ceari, 24. Experiment on Vol. 48, No. 1 April, 29 57

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