21. THE EFFECT OF BIOCHIKOL 020 PC ON MICROORGANISM COMMUNITIES IN THE RHIZOSPHERE OF FABACEAE PLANTS

Transcription

1 21. THE EFFECT OF BIOCHIKOL 020 PC ON MICROORGANISM COMMUNITIES IN THE RHIZOSPHERE OF FABACEAE PLANTS Elżbieta Patkowska, Danuta Pięta, Alina Pastucha Department of Plant Pathology, University of Agriculture in Lublin, Poland, 1. INTRODUCTION In recent years intensive studies have been conducted searching for new possibilities of plant protection with a view to reducing the chemicals, which contaminate agricultural and horticultural products as well as the environment. One of the methods used in the technology of plant cultivation is the application of biopreparations based on antagonistic microorganisms or organic compounds [1-8]. Biochikol 020 PC, based on chitosan belongs to them. It can have anti-virus, anti-bacterial and anti-fungal effects and it belongs to the stimulators of plants resistance [3, 7, 9-12]. Contrary to the typical fugicidal preparations, Biochikol 020 PC applied to the soil or the leaves stimulates the resistance mechanism of plants, besides having a direct effect on pathogenic factors [13]. Chitosan contained in Biochikol 020 PC, as an elicitor of resistance, accelerates the activity of the genes through the contact with a plant and the genes cause formation of biochemical compounds that have fungistatic or fungicidal effect, which is termed as plants immunization [14]. It follows from the studies conducted by the authors that Biochikol 020 PC can reduce the mycelium growth, sporulation and the spore-forming formations of fungi pathogenic towards papilionaceous plants [12]. Besides, pre-sowing dressing of the seeds of those plants with Biochikol 020 PC can protect the germinating seeds, and then the roots of the seedlings and older plants from infection by pathogenic soil-borne fungi [4, 15]. The purpose of the present studies was to determine the effect of Biochikol 020 PC on the formation of microorganism communities in the rhizosphere of papilionaceous plants. Polish Chitin Society, Monograph XI,

2 2. Material and methods E. Patkowska, D. Pięta, A. Pastucha The studies were carried out in the years at the Experimental Station of Czesławice near Nałęczów. The object of the studies was the rhizosphere soil of such plants as pea Sześciotygodniowy TOR cv., common bean Narew cv. and runner bean Westa cv., grown on the fields of monoculture with naturally accumulated infective material in the soil. Earlier studies [16, 17] showed that in this soil environment the following fungi considered to be pathogenic are present: Alternaria alternata, Botrytis cinerea, Fusarium culmorum, Fusarium oxysporum, Phoma exigua, Pythium irregulare, Rhizoctonia solani, and Sclerotinia sclerotiorum (unpublished results of pathogenicity test). The seeds were dressed with the 2.5% solution of Biochikol 020 PC before the sowing. Besides, for each plant species a combination with a chemical preparation, i.e. Zaprawa Oxafun T (active substance: carboxine 37.5% + tiuram 37.5%), was considered in the quantity 2g. kg -1 of seeds and a control combination, i.e. without any dressing. Each combination included 4 plots (4 repetitions), where 100 seeds were sown on each. The second treatment, which was spraying the plants with a biopreparations or Bravo Plus 500 SC was performed at the beginning of anthesis. In each year of the studies during the full anthesis of particular plant species, rhizosphere soil was subject to a microbiological analysis, which was conducted according to the method earlier described by Patkowska [16]. The bacteria Bacillus spp. and Pseudomonas spp. as well as fungi Gliocladium spp. and Trichoderma spp. obtained during those analyses were used to determine their antagonistic effect towards fungi pathogenic towards the examined Fabaceae plants such as B. cinerea, F. culmorum, F. oxysporum, R. solani, S. sclerotiorum according to the method described in the paper by Pięta and Patkowska [17]. The results concerning the microorganism populations obtained during the microbiological analysis of the rhizosphere soil were statistically analyzed and the significance of differences was determined on the basis of Tukey s confidence intervals [18]. 3. Results and discussion A laboratory microbiological analysis of the rhizosphere soil of the studied Fabaceae plants gave different numbers of bacteria and fungi (Table 1). The greatest total number of bacteria (from 5.06 x 10 6 to 6.45 x 10 6 cfu., depending on the plant species) occurred in 1 g of d.w. of the rhizosphere soil taken from the combination with Biochikol 020 PC, while the smallest number occurred in the control combinations (from 2.53 x 10 6 to 4.22 x 10 6 cfu.). 1 g of d.w. of the rhizosphere soil of particular plant species contained more bacteria from genus Bacillus as compared to the bacteria from genus Pseudomonas. Besides, the total number of fungi in 1 g of d.w. of the rhizosphere soil of the studied Fabaceae plants was the lowest in the combinations where Biochikol 020 PC was used and it ranged from x 10 3 to x 10 3 cfu.a slightly greater number of fungi was observed in the combinations with Zaprawa Oxafun T. On the 172 Polish Chitin Society, Monograph XI, 2006

3 The effect of Biochikol 020 PC on microorganism communities in the rhizosphere of... Table 1. The number of bacteria and fungi in the rhizosphere soil of Fabaceae plants (mean from the years ). Experimental combination Total number of bacteria, mln g -1 d. w. of soil Total number of Bacillus spp., mln g -1 d. w. of soil Total number of Pseudomonas spp., mln g -1 d. w. of soil Total number of fungi, thous. g -1 d. w. of soil Pea Biochikol 020 PC 5,06 c 2,49 a 0,49 a 43,04 a Zaprawa Oxafun T 4,07b 2,51a 0,70b 60,46b Control 3,67a 2,50a 0,32a 62,24c Common bean Biochikol 020 PC 6,45c 2,97c 0,52a 22,70a Zaprawa Oxafun T 5,87b 2,35b 1,05b 33,72b Control 4,22a 1,34a 0,91b 34,07b Runner bean Biochikol 020 PC 5,06b 1,82c 1,95c 16,48a Zaprawa Oxafun T 2,70a 1,06a 0,99a 28,35b Control 2,53a 0,92a 1,37b 35,77c * Mean values in the columns for plant species differ significantly (P 0,05), if they are not marked with the same letter. other hand, the greatest number of fungi was found in the rhizosphere soil taken from control combinations, i.e. without any seed dressing (from x 10 3 to x 10 3 cfu., depending on the plant species) (Table 1). The fact that changes occurred in the qualitative and qualitative composition of particular microorganism populations can be explained by the modifying effect of chitosan contained in Biochikol 020 PC or the chemical elements in Zaprawa Oxafun T [19-21]. Besides, Myśków [22] stated that definite proportions exist between the populations of microorganisms, i.e. intensive multiplication of bacteria corresponds to poor multiplication of fungi and vice versa. As stated by Pospieszny [23], chitosan may inhibit the development of some pathogenic bacteria only with a long and direct, and not a short-lasting, contact with those bacteria. Besides, the studies conducted by Atabekov et al. [24] showed that chitosan caused an increase of the population of Bacillus thuringiensis and Escherichia coli bacteria and it protected them from the infection by bacteriophagous species. It should probably be supposed that in the present studies the increase of the number of bacteria from the genera of Bacillus and Pseudomonas after the application of chitosan was also possible. The species composition of fungi isolated from the rhizosphere of pea, common bean and runner bean, growing in particular combinations of the experiment, was varied (Table 2). Totally, 2471 fungi isolates were obtained, which belonged to 25 species. The proportion of pathogenic fungi obtained from the rhizosphere of the studied plants was 30.3% and it was always the highest in control combinations, i.e. without any seed dressing. The dominant pathogenic fungi were Fusarium oxysporum, F. solani, Pythium irregulare and Rhizoctonia solani (unpublished results of pathogenicity test). The most frequently isolated saprophytic fungi were Gliocladium spp., Penicillium pp. and Polish Chitin Society, Monograph XI,

4 E. Patkowska, D. Pięta, A. Pastucha Table 2. Fungi isolated from rhizosphere of Fabaceae plants (total from the years ). Experimental combination Number of isolates Fungus species Pea Common bean Runner bean B Z K B Z K B Z K Total Alternaria alternata (Fr.) Keissler Aspergillus flavus Link Cladosporium cladosporioides Fres de Vries Fusarium oxysporum Schl Fusarium solani (Mart.) Sacc Gliocladium catenulatum Gilman Abbott Gliocladium roseum Bainier Humicola grisea Domsch Mucor hiemalis Bainier Mucor mucedo Fresenius Papulaspora irregularis Hotson Penicillium canescens Scopp Penicillium fellutanum Biourge Penicillium nigricans (Bain.) Than Penicillium verrucosum Dierckx var. cyclopium (West.) Penicillium verrucosum Dierckx var. verrucosum Samson, Stolk et Hadlok Polish Chitin Society, Monograph XI, 2006

5 The effect of Biochikol 020 PC on microorganism communities in the rhizosphere of... Table 2a. Fungi isolated from rhizosphere of Fabaceae plants (total from the years ). Experimental combination Number of isolates Fungus species Pea Common bean Runner bean B Z K B Z K B Z K Total Pythium irregulare Buisman Rhizopus nigricans Ehrenberg Rhizoctonia solani Kűhn Talaromyces flavus (Ben.) emend. Stolk et Samson Trichoderma aureoviride Rifai Trichoderma harzianum Rifai Trichoderma koningii Oud Trichoderma polysporum (Link ex Pers.) Rifai Trichoderma viride Pers. ex S. F. Gray Total B Biochikol 020 PC; Z Zaprawa Oxafun T; K Control Polish Chitin Society, Monograph XI,

6 E. Patkowska, D. Pięta, A. Pastucha Trichoderma spp. The genus of Gliocladium was represented by G. catenulatum and G. roseum, while genus Trichoderma was represented by T. aureoviride, T. harzianum, T. koningii, T. polysporum and T. viride. The greatest number of the abovementioned saprophytic fungi occurred in the rhizosphere of Fabaceae plants in the experimental combinations with Biochikol 020 PC, a little smaller number occurred after the application of Zaprawa Oxafun T, and the smallest number in control combinations (Table 2). It should be supposed that chitosan stimulated the growth and development of Gliocladium spp. and Trichoderma spp. The species of those fungi are considered to be antagonists that can influence the reduction of the number of pathogenic fungi. This fact is confirmed in numerous items of literature [1, 25]. Besides, as stated by Transmo [26], some natural antagonists, including the fungi of Trichoderma spp., show a considerable increase of biomass and increased production of chitinolytic enzymes in the environment containing chitosan. That is why it should be supposed that in the present studies the use of chitosan could have also contributed to the increased population of fungi from Trichoderma genus. The rhizosphere soil of the studied plants contained bacteria (Bacillus spp. and Pseudomonas spp.) and fungi (Gliocladium spp. and Trichoderma spp.) distinguished by their antagonistic effect towards phytopathogens. The greatest number of antagonistic microorganisms (both bacteria and fungi) was obtained from the combination with the application of Biochikol 020 PC, a little smaller number was achieved from the combination with Zaprawa Oxafun T, and the smallest from control combinations and that was irrespective of the species of the studied Fabaceae plant (Table 3). It seems that chitosan contained in Biochikol 020 PC could have had a positive effect of the development of antagonistic microorganisms. It should be supposed that only a big number of antagonists may effectively limit the development of pathogenic fungi, which is confirmed in earlier studies [5, 17]. As stated by Orlikowski et al. [19] and Wojdyła [27] this compound, when applied to the soil for the dressing of bulbs and onions of ornamental plants or for the spraying, effectively protects the plants from Phytophthora spp., special forms of Fusarium oxysporum. Besides, it protects roses from Sphaerothe- Table 3. The number of antagonistic bacteria and fungi in the rhizosphere soil of Fabaceae plants (total from the years ). Antagonistic bacteria and fungi Experimental combination / Number of isolates Pea Common bean Runner bean B Z K B Z K B Z K Bacillus spp Pseudomonas spp Mean Gliocladium spp Trichoderma spp Mean Total B Biochikol 020 PC; Z Zaprawa Oxafun T; K - Control 176 Polish Chitin Society, Monograph XI, 2006

7 The effect of Biochikol 020 PC on microorganism communities in the rhizosphere of... ca pannosa var. rosae, Peronospora sparsa and Botrytis spp. Moreover, according to Borkowski et al. [10], Biochikol 010 PC can effectively protect tomato plants from infection by Oidium lycopersicum. The effect of Biochikol 020 PC consists of inducing the formation of hydrogen peroxide (H 2 O 2 ) in the plant tissues, which hampers the germination of spores and the development of mycelium of Oidium lycopersicum [10]. Basing on the obtained results and the information from literature, Biochikol 020 PC can be included among the biopreparations with a protective effect on phytopathogens and with a positive influence on the development of antagonists. 4. REFERENCES 1. McQuilken M. P., Gemmell J., Lahdenpera M. L.: Gliocladium catenulatum as a potential biological control agent of damping off in bedding plants. J. Phytopath. 149, ľ, 2001, Orlikowski L. B., Skrzypczak Cz., Harmaj I.: Biological activity of grapefruit extract in the control of forme speciales of Fusarium oxysporum. J. Plant Prot. Res. 41 (4), 2001, Orlikowski L. B., Skrzypczak Cz.: Biocides in the control of soil-borne and leaf pathogens. Hortic. Veget. Grow. 22 (3), 2003, Patkowska E., Pięta D.: Introductory studies on the use of biopreparations and organic compounds for seed derssing of runner bean (Phaseolus coccineus L.). Folia Univ. Agric. Stetin., Agricultura 239 (95), 2004, Pięta D., Pastucha A., Patkowska E.: The use of antaginistic microorganisms in biological control of bean diseases. Hortic. Veget. Grow. 22 (3), 2003, Singh G., Mukhopadhyay A.N.: Biocontrol potential of mutants of Gliocladium virens for wilt complex of lentil. Legume Res. 23 (2), 2000, Sobiczewski P., Berczyński S., Kordyla-Bronka M.: Integrowana ochrona roślin przed zarazą ogniową (Erwinia amylovora). Prog. Plant Prot./Postepy Ochr. Rośl. 44 (1), 2004, Wojdyła A.: Wyciąg z grejpfruta w ochronie chryzantem i wierzby przed rdzą. Prog. Plant Prot./Postepy Ochr. Rośl. 44 (2), 2004, Borkowski J., Dyki B.: Wpływ chitozanu, Tytanitu i innych preparatów na ograniczanie rozwoju mączniaka prawdziwego na pomidorach w szklarni. Folia Hortic. Suplement 2003/1, 2003, Borkowski J., Dyki B., Niekraszewicz A., Struszczyk H.: Effect of the preparations Biochikol 020 PC, Tytanit, Biosept 33 SL and others on the healthiness of tomato plants and their fruiting in glasshouse. Progress on Chemistry and Application of Chitin and Its Derivatives Monograph. X, 2004, H. Struszczyk (ed.). Polish Chitin Society, Łódź. 11. Jarosz A., Gołębniak B.: Wpływ środków biologicznych na rozwój skórzastej zgnilizny owoców truskawki (Phytophthora cactorum). Mat. XLV Sesji Nauk. IOR, 2005, Pięta D., Patkowska E., Pastucha A.: Oddziaływanie biopreparatów na wzrost i rozwój niektórych grzybów chorobotwórczych dla roślin motylkowatych. Acta Sci. Pol., Hortorum Cultus 3(2), 2004, Orlikowski L. B., Skrzypczak Cz., Wojdyła A., Jaworska-Marosz A.: Wyciągi roślinne i mikroorganizmy w ochronie roślin przed chorobami. Zesz. Nauk. AR Kraków 387 (82), 2002, Kuć L.: Plant immunization and its applicability for disease control. Innovative Apptoaches to Plant Dis., 1987, Pastucha A., Patkowska E.: Skuteczność biopreparatów w ochronie grochu (Pisum sativum L.) przed grzybami chorobotwórczymi. Folia Univ. Agric. Stetin., Agricultura 239 (95), 2004, Patkowska E.: The effect of Oxafun T on the healthiness of pea (Pisum sativum L.) and on the formation of communities of rhizosphere microorganisms of this plant. Ecological Chemistry and Engineering 12, 2005, Polish Chitin Society, Monograph XI,

8 E. Patkowska, D. Pięta, A. Pastucha 17. Pięta D., Patkowska E.: The role of antagonistic fungi and bacteria limiting the occurrence of some phytopathogens inhabiting the soybean soil environment. EJPAU, Horticulture, v. 6, Issue 2, 2003, Oktaba W.: Metody statystyki matematycznej w doświadczalnictwie. PWN Warszawa (1987) p Orlikowski L. B., Skrzypczak Cz., Wojdyła A.: Biological activity of plant extracts and chitosan towards soil-borne and leaf pathogens. Botanica Lithuanica 3, 1999, Pospieszny H., Struszczyk H.: Chitozan potencjalny biopreparat przeciwko patogenom roślin. Mat. XXXIV Sesji Nauk. IOR, 1994, Borecki Z.: Fungicydy stosowane w ochronie roślin. PWRiL, Warszawa Myśków W.: Związek między aktywnością biologiczną gleby a jej żyznością i urodzajnością. Biologiczne metody podnoszenia żyzności urodzajności gleb. Mat. Szkol., Puławy, 1989, Pospieszny H.: Niektóre aspekty stosowania chitozanu w ochronie roślin. Prog. Plant Prot./Postępy Ochr. Rośl. 37(1), 1997, Atabekov J. G., Koczkina Z. M., Pospieszny H., Chirkov C. N.: Sposob podowlenia fagoliza pri kultirowani mikroorganizmow. (1993). Patent rosyjski Nr Kredics L. D., Doczi I., Antol Z., Manczinger L.: Effect of heavy metals on growth and extracellular enzyme activities of mycoparasitic Trichoderma strains. Biulletin of Environmental Contamination and Toxicology 66, 2, 2000, Transmo A., Skaugrud O., Harman G. E.: Use of chitin and chitosan in biological control of plant disease. In,,Chitin Enzymology. R. A. A. Muzzarelli (ed.). I, 1993, Wojdyła A. T.: Chitosan in the control of rose diseases 6 year trials. Bull. Pol. Ac. Sci., Biol. Sci. 49 (3), 2001, Acknowledgment The studies in the year 2004 were financed by the Ministry of Science and Information within the project No 3 PO Polish Chitin Society, Monograph XI, 2006