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Bio-priming of Cucurbits and Okra Seeds with Culture Filtrates of Trichoderma Harzianum for Controlling Seed-borne Fungi

Md. Naimur Rahman
Department of Plant Pathology, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh

Farzana Haque Tumpa
Department of Plant Pathology, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh

A.K.M. Sahfiqul Islam
Babylon Agri Science Limited, Dhaka, Bangladesh

Md. Atiqur Rahman Khokon
Department of Plant Pathology, Bangladesh Agricultural University, Mymensingh 2202, Bangladesh

Abstract/ Executive Summary:

Culture filtrates collected from Trichoderma harzianum were assayed to control seed-borne fungi of selected vegetables viz. bottle gourd, wax gourd, sweet gourd, snake gourd and okra. Three fungal genera comprising of four species viz. Aspergillus niger, Aspergillus flavus, Penicillium spp. and Fusarium oxysporum were found to be associated with most of the vegetable seeds. Among the seed borne fungi the highest incidence was recorded in case of A. niger (49.2%) followed by A. flavus (40.6%) and F. oxysporum (40.2%). Five treatments viz. 1%, 3%, 5%, 7% aqueous solution of culture filtrates and control (without culture filtrate) were assayed against individual seed-borne fungal pathogen following poison food technique. Among the treatments, 7% aqueous solution of culture filtrate showed superior performance by suppressing mycelial growth of all fungal genera and this treatment was further considered to treat the seeds to control seed borne fungi. Prevalence of seedborne A. niger, A. flavus, Penicillium spp. and F. oxysporum were reduced significantly by seed treatment with 7% aqueous solution of culture filtrates. The findings of the study suggested that seed priming by aqueous solution of culture filtrates of Trichoderma (7%) can be used for vegetable seed treatment for controlling seed- borne fungal infection.

Keywords: Culture filtrate, Trichoderma harziamum, Seed-borne fungi, Vegetables

Location: Department of Plant Pathology and Seed Pathology Centre (SPC), Bangladesh Agricultural University, Mymensingh, Bangladesh

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Okra, Cucurbit

Objectives:

To assess the in-vitro antagonistic ability of culture filtrates of Trichoderma harzianum against seed borne fungi of cucrbits and okra.

Methodology:

The experiments were conducted in the Laboratory of Bioactive Compounds, Bio-formulation and Biosignaling, Plant Disease Diagnostic Clinic (PDDC), Department of Plant Pathology and Seed Pathology Centre (SPC), Bangladesh Agricultural University. Seeds of bottle gourd, sweet gourd, snake gourd, wax gourd and okra were collected from the seed traders of Notun Bazar, Mymensingh. Blotter method was followed according to ISTA rules for seed health testing (ISTA, 2010). After 7 days of incubation, individual seed was observed under stereo-binocular microscope (Zesis) at 10x and 40x magnifications in order to record the association of seed borne fungi. For proper identification of fungi, temporary slides were prepared from the fungal colony and observed under compound microscope and identified with the help of keys suggested by Neergard (1979). Trichoderma harzianum was collected from IPM Laboratory, Department of Plant Pathology, Bangladesh Agricultural University, and Mymensingh. The effect of culture filtrate of T. harzianum on tested pathogens was studied by following methods, developed by Maheswari et al. (2001). Five discs (9 mm diameter) of T. harzianum were cut from vigorously growing margin of 5 days old cultures and inoculated separately into 100 ml sterile Potato Dextrose Broth (pH 5.6) (Singla, 2018). Flasks were incubated at 28°C with 100 rpm on a mechanical shaker for 10 days so as to fragment the hyphal mats and to maintain homogeneous growth in liquid medium. After incubation, the cultures were filtered first through Whatmann filter paper No.4 and finally through Millipore filter (0.45 μm) to obtain sterile culture filtrate. The culture filtrate was adjusted to pH 5.6 by using 0.1N HCl and / or 0.1N NaOH before use. From the stock solution 1%, 3%, 5% and 7% aqueous solutions of culture filtrate were prepared by adding required amount of double distilled water. In vitro efficacy of aqueous solution of culture filtrates To determine the growth inhibitory effects of the treatments against the test fungi A. niger, F. oxysporum and Penicillium spp were isolated from infected jute seeds. Different concentration of aqueous solution of culture filtrates viz. 1%, 3%, 5% and 7% were added to PDA medium and stirred for 10 minutes before pouring on petridishes. Growth inhibition of A. niger, F. oxysporum and Penicillium spp. were measured on the culture filtrates treated PDA medium. Ten mm mycelial disc of each fungus was placed in the centre of the each petridish and incubated at 24º C. Percent inhibition was measured by using this formula (Singh et al., 2010): I= [(C₂−C₁)/C₂)] ×100. Here, I = Percent inhibition of radial mycelial growth, C₂ = Radial mycelial growth of pathogen in control plates, C₁ = Radial mycelial growth of pathogen in treated plates. Seed priming with aqueous solution of culture filtrates Four hundred seeds from each seed sample were immersed in aqueous solution of culture filtrates. Then, the flask was shaken in rotary shaker for 15 minutes to give uniform coating of culture filtrates on the seeds. Afterwards, the health of seeds was examined following incubation test using blotting paper according to ISTA rules (ISTA, 2010). Statistical Analysis: The collected data on different parameters were analyzed statistically by using MSTAT C package program. The means for all the treatments were compared by DMRT (Duncan Multiple Range Test). The differences among the means were compared by LSD test (Least Significance Difference) (Gomez and Gomez, 1984).

Source of Information: J Bangladesh Agril Univ 18(1): 12–16, 2020

Article Link (Online Journal): https://doi.org/10.5455/JBAU.94724

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

Conclusion: Aqueous solution of culture filtrates of Trichoderma harzianum is potential compounds for priming of seed to control seed-borne fungi of different vegetables. Trichoderma is a well-known bio-agent and also beneficial for environment and completely safe for human health. Therefore, application of Trichoderma culture filtrates having antagonistic metabolites can be utilized as seed treating bio-primer in place of chemical pesticides. Further intensive researches are necessary to develop effective commercial formulation using different kinds of organic solvents.

Knowledge Management: Journal