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Formulation of Trichoderma Based Biopesticide for Controlling Damping Off Pathogen of Eggplant Seedling

Md. Jannatul Adan
Department of Plant pathology, Sher-e-Bangla Agricultural University, Bangladesh

Afsana Jahan
Department of Plant pathology, Sher-e-Bangla Agricultural University, Bangladesh

Md. Rafiqul Islam
Department of Plant pathology, Sher-e-Bangla Agricultural University, Bangladesh

Md. Mahfuzar Rahman
Department of Agronomy, Sher-e-Bangla Agricultural University, Bangladesh

Md. Abdullahil Baque
Department of Agronomy, Sher-e-Bangla Agricultural University, Bangladesh

Abstract/ Executive Summary:

The effect of eight Trichoderma based substrates viz. rice bran, wheat bran, lentil bran, gram bran, black gram bran, mustard oil cake, grass pea bran and saw dust in mixing with peat soil and water were used in the formulation, these were evaluated for sporulations of Trichoderma harzianum and acting against Sclerotium rolfsii for the management of damping off of eggplant seedlings with control. The effect of the treatments varied significantly in terms of production of Trichoderma spore and reducing damping off and tip over increasing germination percentage, plant height, seedling vigor and fresh weight of eggplant seedlings in comparison to control. Among the treatments soil application with T₅ (Trichoderma + Black gram bran + Peat soil + Water), T₇ (Trichoderma + Grass Pea bran + Peat soil + Water) and T₄ (Trichoderma + Gram bran + Peat soil + Water) showed the promising effect in controlling pre-emergence damping off, post-emergence damping off, tip over and increasing germination percentage, plant height, vigor index and fresh weight of seedlings. The highest germination percentage (78.00%) was observed in treatment T₅ at 16 days after sowing (DAS). The lowest percentage (6.33%) post- emergence damping off was observed in T₅ in eggplant seedling at 16 DAS. The lowest pre- emergence damping off and tip over were observed in T₅ in eggplant (2.33% and 1.33%).

Keywords: Trichoderma, Biopesticide, Damping off Pathogen, Eggplant Seedling

Location: The experiments were conducted at the central laboratory as well as in the nursery house Department of Plant Pathology, Sher-e-Bangla Agricultural University, Dhaka, 1207.

Start Date: 00-12-2013

End Date: 00-09-2014

Program Area: Pest Management

Commodity: Brinjal

Objectives:

To isolate Trichoderma harzianum from rhizosphere soil,
To formulate Trichoderma based biopesticide using different grain brans and peat soil based substrates,
To find out the efficacy of biopesticide against damping off of eggplant seedlings and
To determine the sustainability of Trichoderma in different formulations.

Methodology:

Plastic tray or soil pots were used as unit plot. The experiments were carried out during the period from December 2013 to September 2014. Trichoderma sp. was isolated from rhizosphere soil from four different Districts (Gazipur, Dhaka, comilla and Rangpur) in Bangladesh by soil dilution technique. Peat soil was collected from Tungipara, Gopalgonj, Bangladesh. Substrates were collected from local shops of Kawran Bazar, Dhaka. Substrates were kept in 4⁰C until use. Eggplant (BARI Brinjal-1) seeds were collected from Bangladesh Agricultural Development Corporation (BADC), Gabtoli, Dhaka. All together a peat soil based substrates (substrate : peat soil : water = 1:1:2) along with a control were explored in the experiment T₁ (Trichoderma + Rice bran + Peat soil + Water), T₂ (Trichoderma + Wheat bran + Peat soil + Water), T₃ (Trichoderma + Lentil bran + Peat soil + Water), T₄ (Trichoderma + Gram bran + Peat soil + Water), T₅ (Trichoderma + Black gram bran + Peat soil + Water), T6(Trichoderma + Mustard oil cake + Peat soil + Water), T₇ ( Trichoderma + Grass Pea bran + Peat soil + Water), T₈ ( Trichoderma + Saw Dust + Peat soil + Water). The requisite amount of materials for each substrates were thoroughly mixed in a 1000 ml Erlenmeyer flask and autoclaved at 121⁰C for 15 minutes for sterilization. The sterilized substrates allowed to cool down and then inoculated with 5 mm dia mycelia disc of 7 days old Trichoderma culture. Seven discs for each flask were used for inoculation. Inoculated flasks were then incubated at room temperature (25±2)⁰C. After incubation for 25 days, the contents were taken out from the flasks, air dried in laminar airflow cabinet and grinded in a blender. The grinded materials were kept in polythene bag with labeling and treated as formulated Trichoderma. The spores of Trichoderma per gram of formulated products were measured by Haemocytometer. Number of spores/conidia per ml spread is counted with the help of Haemocytometer following the procedure of Ashrafuzzaman. The pathogen was obtained from naturally infected tomato plant grown in the experimental field of the Department of Plant Pathology, SAU, Dhaka. The typical collar rot symptoms of tomato plant showed a rot with dry black to brown black lesions around the stem at collar region. The plant was still alive with pale green, and reduced sized leaves. Numbers of round brown to black sclerotia were found. The infected tissue of the collar region of the plant was collected and repeatedly washed in fresh water and surface was sterilized with 10% Clorox for 1 minute followed by three times washing in distilled water. Then the pieces of infected tissue were placed on PDA acidified with one drop of 5% lactic acid and inoculated at 22± 2ºC for 7 days. After incubation, white mycelia and sclerotia were formed. The pathogen was purified and multiplied subsequently through hyphal tip culture on PDA, for preparation of inocula. Barley culture method was followed to culture and multiply Sclerotium rolfsii. Inocula of Sclerotium rolfsii were prepared in barley culture. Barley grains collected from market were thoroughly washed in water and kept soaked in fresh water for 24hrs. After decantation, barley grains were taken in 500 ml Erlenmeyer flask at the rate of 200g in each. The flasks were plugged with cotton followed by wrapping the mouth with brown paper. The flasks containing moist barley grains were sterilized in autoclave at 121ºC under 15lbs pressure for 15 minutes. The sterilized barley grains in the flask were cooled and inoculated aseptically with mycelial blocks (5mm) of pure culture of Sclerotium rolfsii on PDA and inoculated at room temperature for 7-8 days. The flasks were shaken periodically with hand for proper distribution of fungal mycelium throughout the entire mass of the inoculated barley grains. The mycelial growth of the fungus covered entire barley mass in the flask when small round white sclerotia started to form. It was taken out of the flask after fifteen days. The entire mass was spread on brown paper and air dried at room temperature. The colonized dried barley grains were used as inocula for inoculation of plants Soil and cow dung were mixed in (2:1) ratio and kept for 15 days and then the soil sterilized with 5 ml formalin (40%) diluted with 20 ml water for 4 kg soil and the prepared soil was heaped in square block. Soil heap was covered by polyethylene sheet for 48 hr. After 4 days of treatment pots were filled up with the sterilized soil. Formulated Trichoderma were mixed with the soil (except control) @ of 20g/kg soil. The treated soil was incubated for 7 days maintaining proper soil moisture. After that, soil was inoculated with barley grain colonized by Sclerotium rolfsii @ 20g/kg of soil. Inoculated soil was incubated for 7 days maintaining proper soil moisture. One hundred seeds of eggplant were sown in each soil pot after 7 days of application of formulated Trichoderma and Sclerotium rolfsii. The data were recorded on the following parameters-% of seed germination, % pre-emergence damping off, % post-emergence damping, % tip over, seedling height, vigor index and fresh weight of seedling. The collected data were compiled and analyzed statistically using the analysis of variance (ANOVA) technique with the help of a computer package program MSTAT-C and the mean differences were adjusted by Duncan’s Multiple Range Test (DMRT).

Source of Information: Universal Journal of Agricultural Research 3(3): 106-113, 2015

Article Link (Online Journal): http://www.hrpub.org DOI: 10.13189/ujar.2015.030305

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Trichoderma formulation based on black gram bran proved the best among eight substrate combinations in reducing the damping off, pre-emergence - and post emergence death of seedlings and tip over. The formulation also proved to enhance growth of eggplant seedlings.

Knowledge Management: Journal