EJPAU 2009. Jamiołkowska A. THE INFLUENCE OF BIO-PREPARATION BIOSEPT 33 SL ON FUNGI COLONIZING OF SWEET PEPPER PLANTS (<I>Capsicum</I> <I>annuum</I> L.) CULTIVATED IN THE FIELD

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.

2009
Volume 12
Issue 3

Topic:

Horticulture

ELECTRONIC
JOURNAL OF
POLISH
AGRICULTURAL
UNIVERSITIES

Jamiołkowska A. 2009. THE INFLUENCE OF BIO-PREPARATION BIOSEPT 33 SL ON FUNGI COLONIZING OF SWEET PEPPER PLANTS (Capsicum annuum L.) CULTIVATED IN THE FIELD, EJPAU 12(3), #13.
Available Online: http://www.ejpau.media.pl/volume12/issue3/art-13.html

THE INFLUENCE OF BIO-PREPARATION BIOSEPT 33 SL ON FUNGI COLONIZING OF SWEET PEPPER PLANTS (CAPSICUM ANNUUM L.) CULTIVATED IN THE FIELD

Agnieszka Jamiołkowska
Department of Plant Protection and Quarantine, Faculty of Horticulture and Landscape Architecture, University of Life Sciences in Lublin, Poland

ABSTRACT

The influence of Biosept 33 SL on healthiness of sweet pepper plants was examined. A field experiment was conducted in the years 2005, 2007, 2008 on a field in Zezulin. The object of study was sweet pepper plants of cultivar Ożarowska. Plants were sprayed and watered alternately using Biosept 33 SL. The mycological analyses showed that Biosept 33 SL influenced on the reduction of Fusarium oxysporum colony number and in part inhibited alternariosis on sweet pepper plants. Biosept 33 SL did not decreased the number of Fusarium equiseti and Colletotrichum coccodes on sweet pepper plants. The bio-preparation affected the Trichoderma spp. growth on roots and stem base at sweet pepper. The structural similarity between the fungal communities investigated was evaluated with SYNTAX 5.01.

Key words: pepper, Biosept 33 SL, bio-preparation, Fusarium spp., C. coccodes, A. alternata.

INTRODUCTION

One of the most popular method of plant control against pathogenic fungi is the chemical control with use of different fungicides [28,30]. Recently, the use of chemicals in plant production has been reduced. The methods of plant control against pests are the use of biological products – compounds of plant origin [5,9,12,16,18,21]. One of the bio-prepatations is Biosept 33 SL, containing 33% grapefruit extract. The preparation contains a spectrum of active substances (endogenous flavonoids) that have strong antibiotic properties, and at the same time, it does not invoke any side-effects. Inhibition of many disease-forming bacteria and fungi has been reported in studies upon grapefruit extracts [3,14]. Biosept 33 SL not only inhibits the development of harmful microorganisms, but also improves plant's immune system [14]. The literature provides with information on the effectiveness of Biosept 33 SL in the control vegetables, ornamental plants, both in vitro and in vivo conditions [2,3,11,15,18,19,20,26,27].

The aim of the study was to estimate the influence of grapefruit extract – Biosept 33 SL on fungal diseases of sweet pepper plants cultivated in the field.

MATERIAL AND METHODS

The studies were conducted in three years (2005, 2007, 2008) in Zezulin (Lubelski province). The object of study was sweet pepper plants (Capsicum annuum L.) of cultivar Ożarowska. Tillage was made in accordance to agricultural recommendations. Before plantation setting, while mineral fertilization was used in accordance to fertilization recommendations for Solanaceae plants on a base of earlier soil analyses with subsequent ploughing. The pepper plants were set into the field at the end of May in 35 × 67 cm spacing. The experimental combination consisted of 80 plants of each variety (20 plants in 4 replications). The experimental combination consisted of plants protected with bio-preparation Biosept 33 SL at 0.2% concentration. Plants without any protective treatment constituted the control. Two sweet pepper rows were sown around the experimental plants, and they were not used for strict study. No chemical operations were applied on the plantation; weed control was performed manually. Biosept 33 SL was used for spraying and for watering, alternately every 7 days. The first operation was done a month after plants setting in the field. In total, 8 operations were carried out.

At the start of fruity (second decade of August) and during full fruiting (second decade of September), 8 randomly selected pepper plants were sampled from each plot. Roots, stem and leaves of pepper plants were analyzed in laboratory according to the method described by Jamiołkowska [6]. For each experimental treatment 10 dishes with plant material, 10 plant fragments per each dish, were prepared and incubated in thermostat at 20-22°C for 7 days in darkness. Obtained colonies of fungi were transferred to potato-dextrose medium (PDA-Difco) and identified to the species with the keys and monographs. During fungi identification the available monographs were used [4,10,13,23,24,25].

The obtained fungal communities from each year were compared according to numerical classification method using statistical program SYN-TAX 5.01 [22]. In this study analysis of communities similarity was carried out basing on similarity coefficients i.e. percent differences showed on axis in 0-1 scale. In this case 0.1 percent difference between communities corresponds to 90% similarity of these communities [22]. Analysing the data, which describe studied communities, with SYN-TAX program dendrograms showing relation between compared communities was obtained.

RESULTS AND DISCUSSION

Meteorological conditions during 3-years experiment are present in Table 1 and 2. Vegetation season 2008 was warmer and more dry that 2005 and 2007 (Table 1, 2).

Table 1. Average monthly temperature, °C

Year	Month
Year	V	VI	VII	VIII	IX
2005	13.2	16.0	19.8	16.9	14.9
2007	15.0	18.1	19.2	18.4	13.0
2008	14.0	19.0	19.4	19.7	13.1
Average	14.0	17.7	19.4	18.3	13.6

Table 2. Precipitation in months, mm

Year	Month
Year	V	VI	VII	VIII	IX
2005	98.0	55.9	109.8	108.7	18.0
2007	81.5	87.8	87.0	37.6	129.8
2008	76.0	25.1	76.2	32.5	69.4
Average	85.1	56.2	91.0	59.6	72.4

Mycological analysis of above ground part of pepper plants resulted in 1027 isolates of fungi representing 17 species and micelia sterilia (Table 3). Mycological analysis roots and stem base of pepper resulted in 912 isolates of fungi representing 21 species and micelia sterilia (Table 4). The dominating pathogenic fungi were Alternaria alternata, Colletotrichum coccodes and Fusarium spp., represented by F. equiseti, F. oxysporum, F. avenaceum, F. culmorum, F. solani.

A. alternata made up from 48.3 to 84.0% fungi isolated from plants above ground parts, while its number varied during experimental years. Population of A. alternata was slightly lower in combination with Biosept 33 SL than in control. The exception occurred in 2008, particularly favorable year for the fungus development (Table 3). Efficiency of the bio-preparation in alternariosis reduction on potato leaves was confirmed by Lenc [11]. Here achieved results suggest that there is a possibility to partial reducing the A. alternata infection on sweet pepper above ground parts, but it also can be supposed that during warm and extremely wet seasons, the preparation would not efficiently protect plants against alternariosis.

Table 3. Fungi occurring the stem and leaves of sweet pepper plants of cultivar Ożarowska

Fungus species

Number of isolates/ absolute (%)

2005

2007

2008

Alternaria alternata Keiss.

76(76.6)

60(61.2)

55(63.2)

68(84.0)

53(62.3)

50(59.6)

43(48.3)

38(49.3)

50(63.3)

56(66.7)

51(67.1)

58(66.0)

Aureobasidium pullulans (de Bary) Arnoud

–

10(11.8)

5(5.9)

–

6(7.1)

–

Colletotrichum coccodes (Wallr.) Hughes

1(1.0)

–

2(2.2)

1(1.3)

3(3.5)

1(1.2)

–

Epicoccum purpurascens Ehr. ex Schl

–

4(4.9)

–

2(2.4)

–

9(11.4)

8(9.5)

1(1.3)

11(12.5)

Fusarium avenaceum
(Fr.) Sacc.

–

1(1.2)

–

4(4.5)

5(6.5)

2(2.5)

–

12(15.8)

6(8.8)

Fusarium culmorum (Smith) Sacc.

8(8.1)

–

1(1.1)

9(11.7)

3(3.8)

–

Fusarium equiseti (Corda) Sacc.

8(8.1)

26(26.5)

1(1.2)

4(4.9)

11(12.9)

5(5.9)

20(22.5)

1(1.3)

10(12.7)

7(8.3)

7(9.2)

10(11.4)

Fusarium oxysporum Schl.

1(1.0)

2(2.0)

15(17.2)

4(4.9)

–

2(2.4)

6(6.8)

–

5(6.3)

–

Fusarium solani (Mart.) Sacc.

–

12(15.6)

–

Gilmaniella humicola Barron

–

1(1.2)

–

2(2.4)

3(3.6)

–

Mucor hiemalis Wehm.

5(5.1)

4(4.1)

–

2(2.4)

1(1.1)

2(2.6)

–

1(1.3)

–

Penicillium expansum Link

–

3(3.4)

–

7(8.3)

–

4(5.3)

1(1.1)

Penicillium jensenii Zaleski

–

6(7.8)

–

Penicillium purpurogenum Stoll

–

8(9.2)

–

2(2.4)

–

Slerotinia sclerotiorum (Lib.) de Bary

–

14(15.7)

–

Trichoderma harzianum Rifai

–

3(3.1)

–

6(7.1)

5(5.9)

–

4(5.2)

–

Trichoderma koningii Oud.

–

3(3.1)

1(1.2)

–

2(2.4)

–

1(1.1)

Micelia sterilia

–

5(6.0)

–

1(1.1)

Total

99(100)

98(100)

87(100)

81(100)

85(100)

84(100)

89(100)

77(100)

79(100)

84(100)

76(100)

88(100)

I – start of fruity, II – full of fruity
C – control, B – Biosept 33SL

Within Fusarium genus, F. equiseti was isolated from above ground parts of sweet pepper most numerously. It made up from 1.2 to 22.5% fungi isolated from studied experimental combinations. However, its population was much higher on plants protected with the bio-preparation, than in control, except from 2007 (Table 3). On a base of achieved data, it can be supposed that Biosept 33 SL does not reduce the F. equiseti population, but it stimulates its growth. Due to the bio-preparation, number of F. oxysporum on sweet pepper above ground parts was much lower than in the control. And similarly on roots and stem base, the examined bio-preparation reduced the population of F. oxysporum (Table 4). That fact was confirmed earlier by Pięta [19] indicating the decrease of F. oxysporum populations on soybean seedlings sprayed with Biosept. Andrzejak [1] also reported inhibiting effects of Biosept on fusariosis at asparagus in the pot experiments. Besides fungi isolated from pepper plants Trichoderma spp. were often isolated from the combination witch Biosept, that control (Table 3, 4). According to many authors [7,8,9] the number of F. oxysporum colonies decreases with abundant occurrence of. Trichoderma spp. This fact was confirmed by results of the mycological analysis of the pepper plants. Antagonistic Trichoderma spp. colonizing the rhizosphere of pepper, can constitute a protective barrier against pathogens [17].

Table 4. Fungi occurring the roots and stem base of sweet pepper plants of cultivar Ożarowska

Fungus species

Number of isolates/ absolute (%)

2005

2007

2008

Alternaria alternata Keiss.

12(14.8)

6(7.1)

–

1(1.1)

1(2.0)

–

7(10.0)

Aureobasidium pullulans
(de Bary) Arnoud

–

1(1.7)

–

7(8.8)

1(1.4)

5(6.7)

10(14.3)

Colletotrichum coccodes (Wallr.) Hughes

18(22.2)

31(36.5)

54(62.1)

77(84.6)

–

2(3.5)

2(2.3)

–

23(28.7)

–

11(14.9)

–

Epicoccum purpurascens Ehr. ex Schl

–

1(1.1)

–

7(8.8)

1(1.4)

3(4.1)

6(8.6)

Fusarium avenaceum
(Fr.) Sacc.

–

3(3.8)

1(1.4)

6(8.1)

7(10.0)

Fusarium culmorum (Smith) Sacc.

1(1.2)

7(8.2)

–

4(4.8)

2(2.5)

–

Fusarium equiseti (Corda) Sacc.

4(5.0)

1(1.2)

–

7(13.7)

5(8.7)

23(27.4)

–

1(1.3)

2(2.8)

Fusarium oxysporum Schl.

14(17.3)

3(3.5)

12(13.8)

4(4.4)

1(2.0)

1(1.7)

2(2.3)

10(12.5)

28(38.9)

22(29.7)

10(14.3)

Fusarium solani
(Mart.) Sacc.

–

4(4.8)

13(16.6)

–

Gliocladium catenulatum Gilman et Abbott

–

2(2.8)

Humicola fuscoatra Traaen

–

1(1.2)

–

Mucor hiemalis Wehm.

5(6.2)

1(1.2)

–

1(1.7)

1(1.2)

2(2.5)

–

Mucor mucedo Mich.
ex St.-Am.

–

2(3.4)

–

1(1.2)

–

1(1.5)

Penicillium expansum Link

–

17(19.5)

–

Penicillium janczewskii Zaleski

–

8(10.0)

14(19.4)

–

Penicillium janthinellum Biourge

–

1(1.7)

–

2(2.8)

Rhizoctonia solani Kühn

–

39(76.5)

10(17.2)

22(26.2)

–

4(5.4)

2(2.8)

Slerotinia sclerotiorum (Lib.) de Bary

–

25(29.8)

–

Trichoderma hamatum (Bonord.) Bain.

15(18.5)

21(24.7)

–

10(17.2)

–

31(39.2)

6(7.5)

–

19(25.7)

13(18.6)

Trichoderma harzianum Rifai

12(14.8)

15(17.6)

–

3(5.8)

25(43.2)

–

31(39.2)

14(17.5)

26(36.1)

–

8(11.5)

Trichoderma koningii Oud.

–

4(4.6)

8(8.8)

–

Micelia sterilia

–

1(1.2)

1(1.4)

3(4.1)

–

Total

81(100)

85(100)

87(100)

91(100)

51(100)

58(100)

84(100)

79(100)

80(100)

72(100)

74(100)

70(100)

I – start of fruity, II – full of fruity
C – control, B – Biosept 33SL

Mycological analysis of roots and stem base revealed that among pathogenic species colonizing the root system, C. coccodes was numerously isolated (Table 4). It made up from 2.3% to 84.6% of fungal communities obtained. In combinations with Biosept, it was isolated more frequently than from control sample, except from 2008 (Table 4). C. coccodes is the important pathogens occurring under ground parts of pepper plants [7]. Achieved results suggest that Biosept 33 SL cannot be recommended for protecting the sweet pepper against root and stem base anthracnosis. Similar results were reported by Stomp-Chrzan [29] who confirmed that Biosept 33 SL did not ensure an appropriate protection of bean against infection and development of C. lindemuthianum neither under field and laboratory conditions.

Analysis of fungal communities performed by means of numerical classification using SYN-TAX software allowed for determining the similarity level between studied communities. Fungal communities isolated from leaves and stem of pepper plants protected by Biosept 33 SL in 2005 and 2007 (similarity 67%) were the most similar (Fig. 1).

Fig. 1. Dendrogram showing the classification of fungal communities from stems and leaves of pepper plants cultivar Ożarowska; 2005: 1 – control, 2 – Biosept 33 SL; 2007:
3 – control, 4 – Biosept 33 SL; 2008: 5 – control, 6 – Biosept 33 SL

Fig. 2. Dendrogram showing the classification of fungal communities from roots and stem base of pepper plants cultivar Ożarowska; 2005: 1 – control, 2 – Biosept 33 SL; 2007:
3 – control, 4 – Biosept 33 SL; 2008: 5 – control, 6 – Biosept 33 SL

Fungal communities isolated from roots and stem base of pepper plants protected by Biosept 33 SL in 2005 and 2007 (similarity 56%) were the most similar (Fig. 2).

Acknowledgments to Cintamani Poland – Piaseczno for supporting the bio-preparation Biosept 33 SL for scientific purposes.

CONCLUSIONS

Biosept 33 SL poorly reduces alternariosis at sweet pepper plants.
Biosept 33 SL contributes to the reduction in Fusarium oxysporum population at sweet pepper.
Biosept 33 SL does not reduce the number of Fusarium equiseti and Colletotrichum coccodes at sweet pepper.
The biofungicide affects the growth of Trichoderma spp. colonies on roots and stem base at sweet pepper.

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Accepted for print: 26.08.2009

Agnieszka Jamiołkowska
Department of Plant Protection and Quarantine, Faculty of Horticulture and Landscape Architecture, University of Life Sciences in Lublin, Poland
phone: (+48 81) 532-30-47
7 Leszczynskiego Street
20-069 Lublin
Poland
email: aguto@wp.pl

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