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Electronic Journal of Polish Agricultural Universities (EJPAU) founded by all Polish Agriculture Universities presents original papers and review articles relevant to all aspects of agricultural sciences. It is target for persons working both in science and industry,regulatory agencies or teaching in agricultural sector. Covered by IFIS Publishing (Food Science and Technology Abstracts), ELSEVIER Science - Food Science and Technology Program, CAS USA (Chemical Abstracts), CABI Publishing UK and ALPSP (Association of Learned and Professional Society Publisher - full membership). Presented in the Master List of Thomson ISI.

Volume 9
Issue 1
Topic:
ELECTRONIC
JOURNAL OF
POLISH
AGRICULTURAL
UNIVERSITIES
. , EJPAU 9(1), #19.
Available Online: http://www.ejpau.media.pl/volume9/issue1/art-19.html


 

ABSTRACT

A field experiment was conducted in the years 2003-2004 on a field of 10-years’ soybean monoculture, with naturally accumulated infection material in the soil. Soybean of ‘Mazovia’ was the object of studies. Before the sowing the seeds were dressed, and at the beginning of anthesis the plants were sprayed with biopreparations Polyversum, Biosept 33 SL and Biochikol 020 PC. The experiment considered the combination with dressing the seeds by means of Zaprawa Oxafun T (a.s. carboxine 37.5% + tiuram 37.5%) and spraying the plants with fungicide Bravo Plus 500 S.C. (a.s.: chlorotalonile 50%). The seeds that were not dressed constituted the control. During the vegetation the number and healthiness of soybean plants were determined twice (in the seedling phase and at anthesis), at the same time a laboratory mycological analysis was carried out. After the harvest the studies established the size and quality of the seed yield of soybean growing in particular experimental combinations.

The best number of plants was obtained in the combination with Biosept 33 SL, slightly weaker after applying Polyversum or Zaprawa Oxafun T, and the worst in the control combination. Biosept 33 SL and the fungicide were the most effective in protecting the plants from infection by soil-borne fungi. Polyversum and Biochikol 020 PC also showed a good protective effect. The greatest seed yield was collected from soybean plants after applying Biosept 33 SL, while the lowest – from control plants. Soybean plants were mainly infected by Fusarium spp.,A. alternate, P. exigua, R. solani and P. irregulare. Those fungi were most frequently isolated from control plants, and the least frequently after applying Biosept 33 SL or Polyversum. The proportion of pathogenic fungi (P. sojae, Fusarium spp., S. sclerotiorum, P. exigua) in seed infection of soybean was the smallest in the combinations with Biosept 33 SL or Zaprawa Oxafun T, while the biggest in the control.

Key words: .

INTRODUCTION

One of the most popular method of plant control against pathogenic fungi is the chemical method, which means pre-sowing dressing of the seeds and bulbs as well as spraying the plants with different fungicides [7,11,15,16,22,31,32,33]. In recent years more and more often measures are taken, especially in food production, which makes it possible to reduce the use of chemicals. The methods of plant control against pests are improved for example through the elimination of pesticides and the use of biological means containing antagonistic microorganisms or compounds of plant origin [2,5,7,10,13,20,26,28,29]. According to Nuti and Sirsi [18], about 70% of the biopreparations used per about 30 mln ha is made up of bioinsecticides based on Bacillus thuringiensis. A large proportion of the biopreparations used in practice is taken by the preparations produced on the basis of other species from the genus of Bacillus B. mycoides, B. subtilis as well as Pseudomonas spp. and Trichoderma spp. [8,17,30]. The effect of preparations containing organic compounds and plant extracts is also commonly known, for example Biochikol 020 PC, based on chitosan [1,3,20,23], Bioczos BR, containing garlic pulp [6,12] and Biosept 33 SL, 33% grapefruit extract [3,20,21,23,26,27].

Biological control of plants also makes use of preparations that act directly on pathogens. This group contains Polyversum and Contans, based on mycoparasitic fungi. The first of the enumerated preparations contains oospores of Pythium oligandrum and besides a direct effect on soil or leaf phytopathogens, defensive reactions can be also induced in a plant against pathogenic factors [20,21,23]. On the other hand, preparation Contans contains the spores of fungus Coniothyrium minitans. As stated by Benuzzi and Albonetti [2] and Grendene and Marciano [9], this fungus shows considerable effectiveness in plant control against Sclerotinia sclerotiorum, S. minor and S. trifolium. It colonizes the sclerotia of these pathogens, causing their decomposition and a decrease of their number in the soil, at the same time inhibiting the development of apothecia. Besides, according to Orlikowski et al. [21], the introduction of this preparation to the medium infected by S. sclerotiorum significantly limits the development of white mould on pyrethrum and marguerite.

The literature provides information, for example, on the effectiveness of Polyversum, Biosept 33 SL, Biochikol 020 PC in the control of cereals, vegetables or ornamental plants, both in in vitro and in vivo conditions [1,3,6,20,21,26,27]. Because there is no reference to the possibility of using these preparations in the protection of Glycine max, the present studies have been undertaken.

MATERIALS AND METHODS

A field experiment was conducted in the years 2003-2004 at the Experimental Station at Czesławice near Nałęczów on a plot of a 10-year-old monoculture of soybean with naturally accumulated infection material in the soil. The seeds of soybean ‘Mazovia’ cv. were sown during the first 10 days of May. The same substances were used as seed dressing preparations as those that were used in an earlier growth-chamber experiment (i.e. Polyversum in the quantity of 1 g × 100 g-1 seeds, 0.2% Biosept 33 SL, 2.5% Biochikol 020 PC and Zaprawa Oxafun in the quantity of 1 g × 100 g-1 seeds) [23]. The seeds that were not dressed constituted the control. Each experimental combination included 4 plots with the area of 1.25 m2 each, where 100 seeds were sown. At the beginning of anthesis the seeds were sprayed with the same biopreparations that had been used for seed spraying. In the case of the combination with Zaprawa Oxafun T (a.s.: carboxine 37.5% + tiuram 37.5%), the plants were sprayed with Bravo Plus S.C. (a.s.: chlorotalonile 50%). Observations were performed twice in the period of vegetation (in the seedling phase and at anthesis of soybean plants) and they established the number and healthiness of plants on particular plots. At that time plants with clear disease symptoms were also sampled from each experimental combination in order to conduct a laboratory mycological analysis [14]. After the harvest the studies determined the seed yield and the proportion of seeds with necrotic spots on the seed cover.

The results concerning the number, healthiness and yielding were subjected to statistical analysis and the significance of differences was established on the basis of Tukey’s confidence intervals [19].

RESULTS AND DISCUSSION

The use of biopreparations and a fungicide for the pre-sowing dressing of seeds and spraying the plants of Glycine max had a positive effect on the emergencies, the number, healthiness and yielding of the species in question (fig. 1, tab. 1).

Figure 1. The number and healthiness of soybean plants (means from the years 2003-2004):
1 – Polyversum, 2 – Biosept 33 SL, 3 – Biochikol 020PC, 4 – Zaprawa Oxafun T, 5 – Control

Table 1. Weight and quality of soybean seeds yield

Experimental combination

Yield of seeds (g) on the plot

Participation of infected seeds (%)

2003

2004

mean

2003

2004

mean

Polyversum

385cd

327c

356c

4.0a

4.0ab

4.0bc

Biosept 33SL

400d

334c

367c

2.0a

1.6a

1.8a

Biochikol 020 PC

327bc

323c

325c

2.0a

2.0a

2.0ab

Zaprawa Oxafun T

253ab

228b

240.5b

3.0a

6.0bc

4.5c

Control

200a

103a

151.5a

9.0b

7.4c

8.2d

* means in columns fellowed by the same letter do not differ significantly at P ≤ 0.05

Observations conducted both in the seedling phase and at anthesis of soybean showed that the number of plants grown out of the seeds dressed with the tested biopreparations was close to the number of plants obtained after using the fungicide Zaprawa Oxafun T. The best emergencies were achieved on the plots sown with the seeds dressed with Biosept 33 SL (88 seedlings, on average). A little smaller number of plants grew from the seeds dressed with Polyversum or Zaprawa Oxafun T (85 seedlings from each), while the smallest number was obtained in the control combination (68 seedlings, on average) (Fig. 1). Besides, Biosept 33 SL and Zaprawa Oxafun T were most effective in protecting the germinating seeds, and then the roots and the stem base from the infection by soil-borne fungi, since the proportion of the infected seedlings in those combinations was 2.2% and 2.8%, respectively. Polyversum and Biochikol 020 PC also showed a good protective effect (the proportion of the infected seedlings was 3.8% and 4.9%, respectively). The greatest number of infected seedlings grew from the seeds that had not been dressed (11.8%) (Fig. 1).

A similar relation was found out during the second observation, i.e. at soybean anthesis. Then a slight loss of plants and an increase of infected plants were observed on particular plots. The greatest proportion of infected plants at anthesis was found in the control combination and it was 12.2% (Fig. 1).

The obtained results confirmed the effect of biopreparations applied as dressing for soybean in an earlier growth chamber experiment [23]. Then the best emergencies were obtained after dressing the soybean seeds with Biosept 33 SL or Zaprawa Oxafun T, while the worst were observed for the control, i.e. without any dressing. That was similar to the results of the present studies. Besides, the best protective effect (both in a growth chamber experiment and a field one) among all the tested preparations was found for grapefruit extract, since the proportion of infected plants in this combination was the smallest. High effectiveness of preparations in the control of vegetables and ornamental plants was achieved in the studies by for example Baturo [1], Borkowski [3], Dłużniewska [6], Orlikowski and Skrzypczak [20], Orlikowski et al. [21]. Chitosan contained in Biochikol 020 PC and used for seed dressing protected the plants of spring barley from infection by Bipolaris sorokiniana [1], and wheat plants from Fusarium graminearum [25]. According to Borkowski [3], Biochikol 020 PC or Biosept 33 SL, used for spraying the plants of tomato ‘Corindo F1’ cv. protected them from Oidium lycopersici. Laboratory studies showed that those biopreparations had a similar cytological-anatomical mechanism of strengthening the resistance of tomato plants. This was especially well visible when the colour of the nerves of leaves treated with Biosept 33 SL and Biochikol 020 PC changed, which proves the formation of hydrogen peroxide in the tissues and points to a systemic stimulation of resistance [4].

The size and quality of soybean seeds yield obtained after the harvest was proportional to the number and healthiness of plants on the plots of particular experimental combinations (Table 1). The highest yield (367 g from a plot, on average) was obtained from soybean after using Biosept 33 SL. A good yielding was also found out for the plants on combinations with Polyversum and Biochikol 020 PC (356 g and 325 g from a plot, on average). The smallest yield (151.5 g from a plot, on average) with the greatest proportion of infected seeds (8.2%, on average) was obtained from control plants. The smallest number of seeds (1.8%, on average) with necrotic spots on the seeds cover was observed after using Biosept 33 SL (Table 1).

A mycological analysis of infected roots and the stem base of seedlings and older plants of soybean in all experimental combinations isolated 1664 fungi isolates belonging to 17 genera (Table 2). The dominating pathogenic fungi were Fusarium spp., represented by F. culmorum, F. oxysporum and F. solani. The total proportion of those fungi isolated from soybean seedlings constituted 25.2%, while the proportion of plants at anthesis was 32.4% of all isolations. Besides, Alternaria alternate, Phoma exigua, Rhizoctonia solani and Pythium irregulare were often isolated from the infected plants (but only from the seedlings). The enumerated pathogenic fungi were the least frequent to obtain from soybean plants after the application of Biosept 33 SL or Polyversum, and the most frequent – from control plants. A reverse relationship was observed in the case of Gliocladium spp., Penicillium spp. and Trichoderma spp. Besides, species from the genera of Acremonium, Cladosporium, Humicola, Mucor and Rhizopus were isolated from the infected plants of soybean (tab. 2).

Table 2. Fungi isolated from infected plants of soybean (sums from the years 2003-2004)

Fungus species

Experimental combination / Number of isolates

Seedlings

Plants at anthesis

Total

Pol.

Bios.

Bioch.

Z. Ox.

K

Mean

Pol.

Bios.

Bioch.

Z. Ox.

K

Mean

Acremonium roseum (Oud.)

2

-

-

5

-

7

12

9

6

3

1

31

38

Alternaria alternata (Fr.) Keissler

11

2

8

13

9

43

4

2

7

6

3

22

65

Aspergillus niger van Tiegh

1

-

-

-

-

1

-

1

-

-

-

1

2

Aureobasidium pullulans (de Bary) Arnoud.

16

4

12

10

-

42

11

5

6

2

1

25

67

Cladosporium cladosporioides (Fres.) de Vries

3

3

5

9

-

20

2

3

4

-

1

10

30

Epicoccum purpurascens (Ehr. ex. Schl.)

6

4

2

9

11

32

-

-

1

-

-

1

33

Fusarium culmorum (W. G. Sm.) Sacc.

9

5

7

20

33

74

7

5

8

26

38

84

158

Fusarium oxysporum Schl.

17

15

21

16

49

118

19

16

25

38

32

130

248

Fusarium solani (Mart.) Sacc.

5

4

11

7

14

41

2

-

5

6

13

26

67

Gliocladium catenulatum Gilman Abbott

-

-

-

-

-

-

4

6

3

-

2

15

15

Gliocladium fimbriatum Gilman et Abbott

16

13

20

2

2

53

-

3

-

-

-

3

56

Gliocladium roseum Bainier

19

17

20

3

3

62

5

9

4

-

-

18

80

Humicola grisea Domsch

1

-

-

-

-

1

-

-

3

-

1

4

5

Mucor hiemalis Wehmer

-

-

2

8

1

11

-

-

8

1

-

9

20

Mucor mucedo Fresenius

3

6

3

14

4

30

-

-

-

1

-

1

31

Papulaspora irregularis Hotson

-

-

-

-

-

-

-

-

-

1

4

5

5

Penicillium canescens Thom

3

1

3

6

11

24

-

-

2

-

3

5

29

Penicillium chrysogenum Thom

-

-

-

5

6

11

-

-

2

-

-

2

13

Penicillium notatum Westling

-

-

-

-

-

-

2

2

4

4

-

12

12

Penicillium nigricans (Bain.) Thom

-

-

-

-

-

-

1

3

3

4

1

12

12

Penicillium purpurogenum Stoll

-

7

2

6

7

22

-

-

1

5

-

6

28

Penicillium verrucosum Dierckx var.
cyclopium (West.) Samson, Stolk et Hadlok


3


-


2


18


10


33


3


8


6


2


-


19


52

Penicillium verrucosum Dierckx var. verrucosum Samson, Stolk et Hadlok


2


-


-


-


-


2


2


4


-


8


12


26


28

Phoma eupyrena Sacc.

1

-

-

-

-

1

-

-

-

-

3

3

4

Phoma exigua Desm.

5

2

1

4

8

20

1

-

2

6

1

10

30

Pythium irregulare Buisman

2

-

-

7

4

13

-

-

-

-

-

-

13

Rhizoctonia solani Kühn

4

2

6

16

24

52

1

-

5

4

8

18

70

Rhizopus nigricans Ehrenberg

2

7

4

19

8

40

1

-

3

1

-

5

45

Trichoderma hamatum (Bonord.) Bain.

-

-

-

-

-

-

3

5

1

4

-

13

13

Trichoderma harzianum Rifai

25

29

12

3

-

69

27

37

3

1

-

68

137

Trichoderma koningii Oud.

22

28

14

5

1

70

10

20

8

2

6

46

116

Trichoderma polysporum (Link ex. Pers.) Rifai

1

3

-

-

-

4

9

12

8

8

-

37

41

Trichoderma viride Pers. ex. S. F. Gray

9

11

5

3

-

28

28

34

6

2

3

73

101

Total

188

163

160

208

205

924

154

184

134

135

133

740

1664

Pol. – Polyversum, Bios. – Biosept 33 SL, Bioch. – Biochikol 020 PC, Z. Ox. – Zaprawa Oxafun T, K – Control

The positive effect of Biosept 33 SL and Polyversum consists in limiting the growth and development of, for example, Phoma narcisiI [26], Fusarium oxysporum f.sp. tulipae [20], Phytophthora cinnamoni and Pyhtium ultimum [21]. As stated by Orlikowski et al. [21], spraying rose shrubs infected by Sphaerotheca pannosa var. rosae with grapefruit extract caused dehydration of the cytoplasm of mycelium cells and conidial spores, followed by inhibited growth of the pathogen. Also, watering the peat substrate with this extract contributed to a decreased population of P. cryptogea by about 70%, and, besides, it considerably improved the healthiness of Lawson falsecypress plants in its protection from P. cinnamoni [21]. On the other hand, introducing the oospores of P. oligandrum in the form of Polyversum into the soil considerably decreased the population of F. oxysporum f.sp. cyclaminis, Phytophthora spp. and Pythium spp. [21].

Figure 2. Participation of fungi isolated from soybean seeds (means from the years 2003-2004):
1 – A. alternata, 2 – Fusarium spp., 3 – Ph. exigua, 4 – Ph. sojae, 5 – R. solani, 6 – S. sclerotiorum,7 – Penicillium spp.,
8 – Trichoderma spp., 9 – other saprophytic fungi

Almost four times as few fungi as from the seeds with spots on the cover were obtained from normally coloured seeds as a result of mycological analysis. Totally, 666 fungi isolates belonging to 27 species were isolated from the seeds of all experimental combinations. Such fungi as Phomopsis sojae, where the proportion of this species ranged from 10.3% to 35.8%, Fusarium spp. – from 8.4% to 21.2%, and S. sclerotiorum – from 5.0% to 12.1%, depending on the experimental combination, proved to be dominating on the seeds with spots (Fig. 2). Besides, R. solani, P. exigua, A. alternate and different species from the genera Penicillium and Trichoderma were often isolated. The participation of the enumerated pathogenic fungi in infecting soybean seeds was the smallest after applying Biosept 33 SL or Zaprawa Oxafun T, slightly higher in the combinations with Polyversum and Biochikol 020 PC, and the highest in the control combination. On the other hand, the smallest proportion of saprophytic fungi Penicillium spp. and Trichoderma spp. was observed in the case of seeds obtained from control plants (Fig. 2). Within the group of other saprophytic fungi, different species from the genera of Acremonium, Aspergillus, Cladosporium, Epicoccum, Mucor and Torula were isolated from soybean seeds. According to Pięta and Pastucha [24], chitosan and Biospept 33 SL, used to spray common bean plants with at the beginning of anthesis, protected the pods and seeds from the infection by pathogenic fungi. Besides, dressing the seeds of Callistephus chinesis with grapefruit extract considerably eliminated the fungi colonizing the seeds and had a good effect on their germination [27].

It can be stated on the basis of the obtained results and information from the literature of the subject that the tested biopreparations can be recommended for the biological control of soybean plants against the soil-borne fungi.

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  34. The studies conducted in 2004 were financed by the Ministry of Science and Informatisation, grant no. 3PO6 03425.



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