THE EFFECT OF BIOPREPARATIONS ON THE HEALTHINESS OF SOYBEAN CULTIVATED IN A GROWTH CHAMBER EXPERIMENT

Elżbieta Patkowska
Department of Plant Pathology, Agricultural University in Lublin, Poland

ABSTRACT

The purpose of a growth chamber experiment was to determine the effect of bipreparations (Biochikol 020 PC, Biosept 33 SL and Polyversum) and the chemical preparation (Zaprawa Oxafun T) applied for seed dressing on the number and healthiness of plants Glycine max (L.) Merrill. The seeds that were not dressed constituted the control. Soybean plants grew on the medium overgrown with the following pathogenic fungi: A. alternate 18, B. cinerea 31, F. culmorum 33, F. oxysporum f. sp. glycines 14, F. solani 6, P. sojae 51, P. irregulare 93, R. solani 14 and S. sclerotiorum 83.

The best emergencies were achieved after applying Zaprawa Oxafun T or Biosept 33 SL. The smallest number of plants grew in the control combination, i.e. without seed dressing. The greatest proportion of the infected plants was observed in the combinations with F. culmorum 33, F. oxysporum f. sp. glycines 14, F. solani 6 and P. sojae 51. Besides, the worst healthiness was characteristic of the plants growing in the control, while the best was characteristic of the plants grown from the seeds dressed with Polyversum or Biosept 33 SL. The plants grown from the seeds dressed with Polyversum or Zaprawa Oxafun T had the lowest mean value of the infection index, while the highest was found for the plants in the control.

Key words: soybean, Biochikol 020 PC, Biosept 33 SL, Polyversum, Zaprawa Oxafun T, seed dressing.

INTRODUCTION

Throughout the period of vegetation plants can be infected by such pathogenic soil-borne fungi as Botrytis cinerea Pers., Fusarium culmorum (W.G.Sm) Sacc., F. oxysporum Schl. F.sp. glycines Amst. Amst., F. solani (Mart.) Sacc., Rhizoctonia solani Kühn and Sclerotinia sclerotiorum (Lib.) de Bary [8, 13, 14, 18, 20, 22]. Those fungi first infect the germinating seeds and next the roots of the seedlings and older plants. They cause worse emergencies and healthiness. Besides, the species Phomopsis sojae Lehman has a negative effect on the size and quality of the soybean yield [8, 14, 25].

The chemical method, based on the application of fungicides for the dressing of the multiplication material or for the spraying of the above-ground parts of plants, is the most widespread method of protecting plants from plant pathogens. However, the tendency is to reduce the use of chemical preparations and to replace them with biopreparations based on antagonistic microorganisms, organic compounds or plants extracts [1, 10, 15, 16, 19, 21, 26, 29]. Biosept 33 SL, Biochikol 020 PC and Polyversum belong to the biopreparations that are effective in protecting the plants.

Biosept 33 SL, containing 33% grapefruit juice, acts directly on pathogenic factors and it induces plants’ resistance to certain pathogens [12]. 7-geranoxicumarin, found in grapefruit juice, can have this effect [10]. Besides, as observed by Caccioni et al. [2], aliphatic aldehydes, monoterpenes and nutkakon dominate among the many components of this extract. These compounds can show synergism in inhibiting the growth of a definite pathogenic factor or stimulate the germination of fungi sporules. The protective effect of Biosept 33 SL is also connected with the presence of endogenous flavonoids, glycosides, citrinian and limenon in this preparation [24].

Biochikol 020 PC, where chitosan is the active substance, can show an anti-viral, anti-bacterial and anti-fungal effect and it belongs to the inducers of plants’ resistance, towards both soil pathogens and epiphytic ones [1, 12, 15].

On the other hand, Polyversum, produced on the basis of the oospores of mycoparasite Pythium oligandrum acts directly on soil or phyllosphere pathogens and it can induce plants’ resistance to certain plant pathogens [10, 12, 16, 27].

The purpose of the present studies was to determine the effect of biopreparations (Biochikol 020 PC, Biosept 33 SL and Polyversum) and fungicide (Zaprawa Oxafun T), applied for seed dressing, on the number and healthiness of soybean plants.

MATERIAL AND METHODS

The experiment was conducted in a growth chamber. It used the seeds of soybean ‘Mazovia’ cv. and the garden earth overgrown with the fungi species pathogenic towards soybean: Alternaria alternate 18, Botrytis cinerea 31, Fusarium culmorum 33, Fusarium oxysporum f. sp. glycines 14, Fusarium solani 6, Phomopsis sojae 51, Pythium irregulare 93, Rhizoctonia solani 14 and Sclerotinia sclerotiorum 83. Isolates of those fungi were obtained from the infected plants of soybean cultivated in a field experiment, and their pathogenic character was confirmed in earlier studies. A special form of F. oxysporum was established using the method described by Łacicowa [7]. The infection mixture of the pathogenic fungi used in the experiment was prepared according to Noll’s method described by Łacicowa [6]. To this aim, the compost earth with a 5% addition of barley grain was placed in Erlenmayer’s flasks with the volume of 3000 cm³. The flasks with the mixture were sterilized three times for 2 hours in an autoclave at the pressure of 1 atmosphere and the temperature of 121°C. After sterilization, 14-day-old cultures of the enumerated pathogenic fungi growing on PDA medium were inoculated into the earth in the flasks, considering 1 flask for each fungus isolate. Next, the flasks with the inoculum were kept at temperature 23°C for three weeks so that the fungus overgrew the earth, which could be acknowledged macroscopically.

Plastic pots, where 25 seeds of soybean were sown into each, were filled with this medium. Before the sowing, the seeds were dressed with the following biopreparations: 0.2% Biosept 33 SL (33% grapefruit extract), 2.5% Biochikol 020 PC (containing 1.88% s.a) and Polyversum (containing 10⁶ oospores in 1 g), using 1 g preparation · 100 g^-1 seeds. For a comparison, chemical preparation Zaprawa Oxafun T (s.a.: carboxin 37.5% + tiuram 37.5%) was used in the quantity 2 g·kg^-1. The seeds that were not dressed constituted the control. 4 replications (4 pots) were used for each experimental combination. The conditions for the seed germination and the growth of plants in the growth chamber were the same as in an earlier experiment described by Patkowska and Pięta [15].

Five weeks after the sowing of soybean seeds the number of seedlings was determined and the proportion of plants with disease symptoms on the roots and the stem base was established. The healthiness of plants was evaluated according to a five-grade scale described by Pięta [17], and the infection index was calculated according to the formula applied by Sadowski and Rzekanowski [23].

The results were statistically analyzed and the significance of differences was determined on the basis of Tukey’s confidence intervals.

RESULTS AND DISCUSSION

The results obtained from the studies pointed to the differences in the number and healthiness of plants in particular experimental combinations (fig. 1, photo 1). The number of soybean plants grown from the seeds dressed with particular biopreparations, i.e. Biosept 33 SL, Polyversum and Biochikol 020 PC, was close to the number of plants obtained in the combination with Zaprawa Oxafun T. The biggest number of seedlings grew from the seeds of soybean dressed with fungicide Zaprawa Oxafun T or biopreparation Biosept 33 SL (respectively, from 64 to 84 and from 63 to 84 seedlings), depending on the species of pathogenic fungus present in the medium, but with an exception of F. oxysporum f. s. glycines 14, Rhizoctonia solani 14 and Sclerotinia sclerotiorum 83. In the combinations with F.oxysporum f.sp. glycines 14 or R.solani 14 the best emergencies were achieved after applying biopreparation Polyversum or Zaprawa Oxafun T. On the other hand, the presence of S. sclerotiorum 83 in the medium caused that the biggest number of plants grew from the seeds dressed with Biosept 33 SL (83 seedlings) or Biochikol 020 PC (81 seedlings). The smallest number of plants grew in control combinations, i.e. without seed dressing (from 46 to 68 seedlings, depending on the species of pathogenic fungus).

Seedlings with inhibited growth, with symptoms of necrosis on the roots and the stem base, occurred in each experimental combination (photo 2). The highest proportion of infected plants of soybean was observed in the experimental combinations with such plant pathogens as Fusarium culmorum 33, F. oxysporum f.sp. glycines 14, F. solani 6 and Phomopsis sojae 51 (fig. 1). The germinating seeds as well as the roots and the stem base of soybean were the most effectively protected by Polyversum and Biosept 33 SL, since the proportion of the infected plants, depending on the species of pathogenic fungus, ranged from 5.8% to 14.8% and from 5.7% to 16.9%, respectively. A similar protective effect was shown by Biochikol 020 PC and Zaprawa Oxafun T. The proportion of infected plants of soybean grown from the seeds dressed with biopreparation Biochikol 020 PC ranged from 6.8% to 19.1%, while that of the seeds dressed with fungicide Zaprawa Oxafun T ranged from 6.1% to 11.6%. The worst healthiness was characteristic of control plants, since the proportion of infected plants ranged from 18.6% to 31.1% (fig. 1).

Considerable effectiveness of biopreparations in the protection of ornamental plants and vegetables was confirmed in the studies by Kurzawińska and Galda [5], Orlikowski et al. [11, 12], Borkowski et al. [1], Saniewska [24], Waksmundzka and Mazur [27] and Wojdyła [28, 29]. Biosept 33 SL added to the medium considerably reduced the number of Phytophthora spp., Pythium spp. and the special forms of Fusarium oxysporum [10, 11, 12]. When used for spraying, it protected plants from the fungi causing spots, rust and powdery mildew [12, 29].

Biochikol 020 PC, when applied to the soil for the dressing of bulbs and onions or for spraying, effectively protected them from Phytophthora spp., special forms of F. oxysporum. It also protected roses from Sphaerotheca pannosa var. rosae, Peronospora sparsa and Botrytis spp. [10, 28]. Besides, through its contact with a plant, chitosan contained in Biochikol 020 PC, as an elicitor of resistance enhances the activity of genes, which start the formation of biochemical compounds with fungistatic or fungicidal effects.

The mean value of the infection index of soybean plants, calculated on the basis of a five-grade scale, ranged from 4.7 and 78.2 in particular experimental combinations (fig. 2). The highest values of the infection index were found in the same combinations of the experiment as in the case of the greatest proportion of infected plants, i.e. with the following plant pathogens present in the medium: F. oxysporum f. s. glycines 14, F. culmorum 33, F. solani 6 or Phomopsis sojae 51.The lowest value of the infection index was characteristic of soybean plants grown from the seeds dressed with Polyversum (from 4.7 to 12.6) or Zaprawa Oxafun T (from 4.9 to 18.4). Slightly higher and similar to each other values of the infection index were obtained in the combinations with dressing the seeds with Biosept 33 SL or Biochikol 020 PC (respectively, from 12.6 to 38.4 and from 10.5 to 27.3). On the other hand, the highest value of the infection index was calculated for plants grown from the seeds that were not dressed (from 54.7 to 78.2, depending on the species of pathogenic fungus used in the experiment). A similar effect was obtained in an earlier experiment [15]. Then, the smallest value of the infection index was characteristic of runner bean plants grown from the seeds dressed with Polyversum. Besides, as observed by Eaksmundzka and Mazur [27], the application of Polyversum in the concentration 0.1% for the protection of sweet basil protects most plants from Fusatrium oxysporum and Cladosporium cladosporoides. On the other hand, according to Kurzawińska and Gajda [5] this biopreparation used for the dressing of tomato seeds stimulated the plants’ growth and inhibited the development of F. oxysporum f. sp. radicis-lycopersici. The protective effect of Polyversum can be based on the formation of auxin-like elements, for example trypamine (TNH2) as well as on the ability to colonize the root system of plants and to stay in the rhizosphere throughout the period of vegetation, in this way making it difficult for pathogens to colonize this zone [3].

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