48 Katarzyna Gleń show that foliar fertilizers may either inhibit or stimulate plant pathogen development under in vitro conditions [711]. Fungi which

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1 ECOLOGICAL CHEMISTRY AND ENGINEERING A Vol. 15, No Katarzyna GLEŃ* COMPARISON OF FOSTAR AND WAPNOVIT FOLIAR FERTILIZERS EFFECT ON PHYTOPATHOGENIC FUNGI OF GENUS PORÓWNANIE ODDZIAŁYWANIA NAWOZÓW DOLISTNYCH FOSTAR I WAPNOVIT NA GRZYBY FITOPATOGENNE NALEŻĄCE DO RODZAJU Summary: : The investigations aimed at comparing the effect of various concentrations of foliar fertilizers, Fostar and Wapnovit, on linear growth, biomass increment and sporulation of phytopathogenic fungi: Fusa- rium solani (Mart.) Sacc., coeruleum (Sacc.) Booth, culmorum (W.G. Smith) Sacc. and graminearum Schwab under in vitro conditions. The conducted research revealed a lack of uniform reaction of tested fungi species to the analyzed foliar fertilizers and their concentrations used in the experiment. Fostar foliar fertilizer revealed stronger fungistatic properties apparent as considerable limiting of surface growth in all species of tested fungi and total inhibition of sporulation process in coeruleum, culmorum and graminearum.. On the other hand, Wapnovit in 0.1 and 0.01 mm 3 /cm 3 concentrations slightly inhibited linear growth and conidia production in coeruleum,, whereas when applied in 1 mm 3 /cm 3 it stimulated both linear growth and biomass increment in coeruleum, culmorum and graminearum.. Among the tested fungal organisms F. graminearum proved the species most sensitive to the applied foliar preparations. Both foliar fertilizers, irrespective of applied concentrations, blocked the conidia production process and in most combinations inhibited biomass increments. Keywords: fungi, foliar fertilizers, linear growth, biomass, sporulation The main task of foliar fertilizers introduced to agrocenoses is increasing the amount of obtained yields [14]. Plants receiving foliar nutrition accumulate lesser quantities of nitrates(v) and nitrates(iii)(nitrites) harmful for homoiothermic organisms [5]. According to Nowosielski [6] foliar fertilizers may cause non-specific changes in plants result- ing in their altered susceptibility to diseases. Moreover, the protective activity of foliar fertilizers is their direct influence on the pathogen. Few studies conducted in this field * Departament of Agricultural Environment Protection, Agricultural University of Krakow, al. A. Mickie- wicza 21, Kraków, rrglen@cyf-kr.edu.pl

2 48 Katarzyna Gleń show that foliar fertilizers may either inhibit or stimulate plant pathogen development under in vitro conditions [711]. Fungi which belong to genus are common in the natural environment and infect many plant species, they cause fusarium wilt, seedling and foot blight, rot of various plant organs and pollute plant products with toxins [1214]. The studies aimed at comparing the influence of two different concentrations of foliar fertilizers, Fostar and Wapnovit, on linear growth, biomass increment and sporulation of phytopathogenic fungi: solani (Mart.) Sacc., coeruleum (Sacc.) Booth, culmorum (W.G. Smith) Sacc. and graminearum Schwab under in vitro conditions. Material and methods Foliar fertilizers Fostar and Wapnovit produced by Intermag Enterprise from Olkusz, PL were selected for the laboratory experiment. Fostar contained: 14.8 % P, 34.1 % P 2 O 5 (500.3 g P 2 O 5 /dm 3 of fertilizer), ph 1.4, whereas Wapnovit: % CaO, (256 g CaO/dm 3 ), 10 % nitrogen (N-NO 3 ), 0.48 % MgO), 0.05 % B, 0.02 % Zn, ph 2.7. solani (Mart.) Sacc., coeruleum (Sacc.) Booth, culmorum (W.G. Smith) Sacc. and graminearum Schwab pathogenic fungi from the collection of Agricultural Environment Protection Department Agricultural Univer- sity of Krakow were used for the experiment. The phytopathogenic fungi were cultured in vitro on PDA (glucose-potato) medium at 23 C in five replications. PDA medium with added solutions of Fostar and Wapnovit was prepared in the way allowing to ob- tain the following concentrations in 1 cm 3 : 0.01, 0.1 and 1.0 mm 3. The media reaction was measured prior to their inoculation. The ready media were inoculated with an agar circle, 5 mm in diameter, overgrown with the two-week old mycelium of the tested or- ganism. Petri dishes with the medium without fertilizers served as the control. The influence of individual foliar fertilizers on the linear growth of the tested fungal organisms was presented as a difference between the diameter of the fungus colony on the control dishes and the diameter of mycelium colony on dishes with individual concentrations of foliar fertilizers. Subsequently the result was converted into the index of biomass growth inhibition/stimulation in percentage according to the formula by Ab- bot [15]: where: I index of linear mycelium growth inhibition; K diameter of fungus colony on control dish; A diameter of fungus colony on dish with determined concentration of foliar fertilizer. After three weeks of fungi culturing on PDA media with foliar fertilizers and on the control, spore numbers were assessed in Thom hemocytometer.

3 Comparison of Fostar and Wapnovit Foliar Fertilizers Effect on Phytopathogenic Fungi Growth of the fungi biomass was maintained for 21 days in 300 cm 3 Erlenmayer flasks on 100 cm 3 of the modified PDA medium (without agar) with added foliar fertilizers in the same concentrations as in the presented experiment. Then the post-cul- ture liquid together with mycelium was filtered through filter paper. The mycelium was dried on sterile glass at 80 C until constant weight was obtained and then weighed. The effect of individual foliar fertilizers on the biomass of fungal organisms was presented as a difference between fungus biomass in the control flasks and mycelium biomass in flasks with individual concentrations of foliar fertilizers. Subsequently the result was converted into a of biomass inhibition/stimulation index according to the formula presented in the description of linear growth. The results were elaborated statistically using analysis of variance for two-factor ex- periments (1 factor foliar fertilizer, 2 factor fertilizer concentration) in independent design. Significance of differences between combinations was assessed on the basis of the t-student test. Results and discussion The conducted research revealed that foliar fertilizers: Fostar and Wapnovit influenced differently linear growth, biomass and sporulation of the tested fungi. Other authors [711, 16] confirm that the in vitro effect of foliar fertilizers is primarily depend- ent on the fungus species and the kind and concentration of fertilizer preparation. Among the tested foliar fertilizers Fostar revealed strong fungistatic properties, particularly towards solani,, irrespective of its concentrations used. It was re- flected by between % and % inhibition of its colony surface growth, by between 3.19 and % limitation of its biomass growth and reduction of sporulation (Fig. 1, Table 1, 2). Fig. 1. Effect of foliar fertilizers on linear growth of solani (concentrations in [mm 3 /cm 3 ]). Negative values denote growth stimulation.

4 50 Katarzyna Gleń Table 1 Fertilizer Influence of foliar fertilizers on tested fungi biomass [g] Concentration [mm 3 /cm 3 ] solani Fostar a* 26.66** bc b Wapnovit e Control d b 8.92 Fungi biomass [g] coeruleum d b c e c bc culmorum d b c e bc +93, d graminearum e a bc f b cd c a a de * values in columns marked with the same letters do not differ significantly; ** coefficients of biomass increment inhibition/stimulation; + denotes biomass increment stimulation Fertilizer Influence of foliar fertilizers on tested fungi sporulation Concentration [mm 3 /cm 3 ] Fostar Wapnovit 1.0 * no sporulation solani Number of macroconidia in 1 cm coeruleum * culmorum Table 2 graminearum Control Fungistatic activity, particularly of higher concentrations (1.0 and 0.1 mm 3 /cm 3 ) of Fostar was visible also for surface growth of the other tested fungi species and as complete inhibition of their sporulation process (Fig. 24, Table 2). However, biomass increments in coeruleum and culmorum on liquid media with Fostar sup-

5 Comparison of Fostar and Wapnovit Foliar Fertilizers Effect on Phytopathogenic Fungi Fig. 2. Influence of foliar fertilizers on linear growth of coeruleum (concentrations in [mm 3 /cm 3] ). Negative values denote growth stimulation Fig. 3. Influence of foliar fertilizers on linear growth of culmorum (concentrations in [mm 3 /cm 3 ]). Negative values denote growth stimulation plements in various concentrations were significantly greater than in the control (Table 1). Especially intensive biomass increments of 89.5 % for F. coeruleum and % for F. culmorum were noted also for concentrations where linear growth was the most limited. Diversification between biomass increment and linear growth may be explained by the fungus colony growth retardation observed in the first days of the experiment. It is worth mentioning that the time of fungi culturing on the solid medium was 58 days and on the liquid one 21 days. In the opinion of Kwaśna et al. [13] the optimum ph for fungi ranges between 4 and 6.5. In the Authors own investigations the ph

6 52 Katarzyna Gleń Fig. 4. Influence of foliar fertilizers on linear growth of graminearum (concentrations in [mm 3 /cm 3 ]). Negative values denote growth stimulation value of the media with 0.01 mm 3 /cm 3 concentrations was 2.53, 2.75 and 3.32, respectively. A strongly acid reaction of the medium did not favour the growth of the tested phytopathohenic fungi species, especially in the short experiment conducted on Petri dishes. It was confirmed by our former research [10]. Foliar fertilizers characterized by low ph and applied in 1.0 mm 3 /cm 3 concentrations caused a complete inhibition of F. avenaceum, F. coeruleum and F. graminearum growth. There is also a well known opinion that fungal organisms are highly tolerant to the environment reaction [13, 1719]. Therefore, stimulation of F. coeruleum and F. culmorum biomass increment might have been the effect of these species adaptation to the environment. Wapnovit, unlike Fostar, is a multicomponent fertilizer and beside calcium it also contains nitrogen, magnesium, boron, copper and zinc. Moreover, the media ph com- bined with Wapnovit ranged between 4.51 and 5.75, which is the range optimal for fungi. The tested fungi species revealed high variability in their response to various concentrations of this fertilizer preparation. Lower concentrations of Wap- novit in the medium (0.1 and 0.01 mm 3 /cm 3 ) limited surface growth of F. coeruleum, by and % respectively and its sporulation but stimulated biomass increment (Fig. 2, Table 1). Irrespective of the applied concentration the fertilizer affected the intensity of sporulation in F. solani.. On the other hand the highest tested Wapnovit concentration slightly (9.76 %) inhibited linear growth and simultaneously stimulated biomass in this species. A reverse reaction was observed for F. graminearum,, since the 0.01 mm 3 /cm 3 Wapnovit supplement in the medium slightly stimulated linear growth (6.27 %) but limited biomass increment by %. Juffery and Burgers [20] think that the amount of energy substances and nutrients in the medium might be one of the caus- es of weak density of F. graminearum aerial mycelium. Therefore, despite stimulation of linear growth, no increment of aerial mycelium occurs and biomass in effect. In the Author s own investigations solani revealed a similar relationship between

7 Comparison of Fostar and Wapnovit Foliar Fertilizers Effect on Phytopathogenic Fungi linear growth and biomass at the lowest concentration of Wapnovit (Fig. 1, Table 1). Also in [9] it observed stimulation of F. solani growth after application of 0.03 mm 3 /cm 3 of multicomponent foliar fertilizers such as Insol-7 and Yeald to the medium. Linear growth and biomass growth of F. coeruleum, F. culmorum and F. graminearum on media with 1.0 mm 3 /cm 3 of Wapnovit (field dose) were strongly stimulated. Other authors investigations point rather to inhibition of fungi development at high concentrations of multicomponent fertilizers [8, 10, 21]. Conclusions 1. Among the tested fungal organisms F. graminearum proved the species most sensitive to applied fertilizer preparations. Fostar and Wapnowit fertilizers blocked the process of conidia formation, irrespective of their concentrations and in most combinations also inhibited biomass increments. 2. Fungistatic properties of Fostar were apparent as strong inhibition of surface growth in all tested fungi species and total inhibition of sporulation in coeruleum, culmorum and graminearum. 3. Wapnovit foliar fertilizer applied in 0.01 and 0.01 mm 3 /cm 3 concentrations slightly reduced linear growth and conidia formation in coeruleum.. Its concentration of 1.0 mm 3 /cm 3 stimulated both linear growth and biomass increment in all tested fungi species. References [1] Jabłoński K. and Bernat E.: Wpływ dolistnego nawożenia Mikrosolem Zn na kształtowanie się plonu ziemniaka i jego jakość oraz możliwość ograniczenia stosowania fungicydów do zwalczania zarazy ziemniaka.. Progr. Plant Protect., 2001, 41(1), [2] Boligłowa E.: Wpływ dolistnego dokarmiania ziemniaka na plon jego strukturę, zdrowotność i trwałość przechowalniczą bulw.. Acta Agrophys., 2003, 85,, [3] Tobiasz-Salach R. and Borecka-Jamro D.:Wpływ wieloskładnikowych nawozów dolistnych na plono- wanie i skład chemiczny owsa.. Acta Agrophys., 2003; 85,, [4] Chwil S. and Szewczuk C.: Wpływ dolistnego dokarmiania buraka cukrowego na jego plon i niektóre cechy jakościowe.. Acta Agrophys., 2003; 85,, [5] Sady W.: Nawożenie warzyw polowych. Kraków 2000, [6] Nowosielski O.: FloroGama (FGO) Pierwszy nawóz nalistny ochroniarski. Ochr. Rośl, 1986, 7, [7] Przeździecki Z., Wojciechowska-Kot H., Mikołajska I. and Murawa D.: Wpływ mikroelementów na roz- wój grzybni, zarodnikowanie i zawartość kwasów tłuszczowych i avenaceum.. Acta Acad. Agricult. Tech., Olst., 1991, 53,, [8] Weber Z. and Wyrwa P.: Wpływ nawozu dolistnego Bonga na wzrost czterech gatunków grzybów in vitro.. Rocz. Akad. Roln. Poznań, Roln., 1993, 42(247), [9] Boligłowa E. and Gleń K.: Wpływ nawozów dolistnych na wzrost i zarodnikowanie grzybów patogenicz- nych z rodzaju w warunkach in vitro.. Acta Agrophys., 2003, 85,, [10] Gleń K. and Boligłowa E.: Effect of Mikrovit Fe and Mikrovit Mn on development of culmorum (W.G. Smith) Sacc.. Ecol. Chem. Eng., 2006, 8(13), [11] Gleń K. and Boligłowa E.: : Assessment of Tytanit in vitro effect on selected phytopathogenic fungi. Ecol. Chem. Eng., 2006, 7(13), [12] Rataj-Guranowska M. and Wiatroszak J.: Niekonwencjonalne metody identyfikacji grzybów z rodzaju.. Rocz. Nauk. Rol., Seria ET. 1988, 18(1), 5160.

8 54 Katarzyna Gleń [13] Kwaśna H., Chełkowski J. and Zajkowski P.: Grzyby (Mycota), t. XXII, Sierpik (), War- szawakraków [14] Cook R. J.: : Diseases, biology and taxonomy.. Pensylv. St. Univ. Press., Park and London, [15] Kowalik R. and Krechniak E.: Szczegółowa metodyka biologicznych i laboratoryjnych badań środków grzybobójczych.. [in:] Materiały do metodyki badań biologicznej oceny środków ochrony roślin, Inst. Ochr. Rośl., Poznań [16] Kotlińska T.: Wpływ Florogamy ochronnej na wzrost i rozwój kolonii grzybów-patogenów roślin.. Progr. Plant Protect., 1988, 28(2), [17] Kirlay Z., Klement Z. and Solymosy F.: Fitopatologia. Wybór metod badawczych. PWRiL. Warszawa [18] Kochman J. and Węgorek W.: Ochrona roślin. Plantpress, Kraków [19] Borecki Z.: Nauka o chorobach roślin. PWRiL, Warszawa [20] Jeffery S. and Burges L. W.: Growth of graminearum Schwabe group 1 on media amended with atrazine, chlorosulfuron or glyphosate in relation to temperature and osmotic potential.. Soil Biol. Biochem., 1990, 22(5), [21] Boligłowa E. and Gleń K.: Response of some fungi to Indol-7 and Ekosol-K itrafoliar fertilizers added into the medium.. Ecol. Chem. Eng., 2000, 9(4), PORÓWNANIE ODDZIAŁYWANIA NAWOZÓW DOLISTNYCH FOSTAR I WAPNOVIT NA GRZYBY FITOPATOGENNE NALEŻĄCE DO RODZAJU Streszczenie z e e Celem podjętych badań było porównanie wpływu różnych stężeń nawozów dolistnych, tj. Fostar i Wapno- vit na wzrost liniowy, przyrost biomasy i zarodnikowanie grzybów fitopatogennych: solani (Mart.) Sacc, coeruleum (Sacc.) Booth, culmorum (W.G. Smith) Sacc., graminearum Schwab. w warunkach in vitro. W przeprowadzonych badaniach stwierdzono brak jednoznacznej reakcji testowanych gatunków grzybów na analizowane nawozy dolistne i użyte ich stężenia. Nawóz dolistny Fostar odznaczał się silniejszymi właś- ciwościami fungistatycznymi. Wyrażało się to silnym ograniczeniem rozrostu powierzchniowego wszystkich gatunków grzybów testowych i całkowitym zahamowaniem procesu sporulacji: coeruleum, Fusa- rium culmorum i graminearum. Natomiast Wapnovit w stężeniu 0,1 i 0,01 mm 3 /cm 3 nieznacznie hamował wzrost liniowy i wytwarzanie konidiów coeruleum. Z kolei zastosowany w koncentracji 1 mm 3 /cm 3 stymulował zarówno rozrost liniowy, jak i przyrost biomasy coeruleum, culmorum i graminearum.. Spośród testowanych organizmów grzybowych, F. graminearum okazał się gatunkiem najbardziej wrażliwym na stosowane preparaty nawozowe. Oba nawozy dolistne niezależnie od zastosowanego stężenia blokowały proces wytwarzania konidiów oraz w większości kombinacji hamowały przyrosty biomasy. Słowa kluczowe: fungi, foliar fertilizers, linear growth, biomass, sporulation