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Efficacy of Akanda (calotropis Gigantea L.) Against Some Phytopathogenic Bacteria

M.M. Hasan
Department of Plant Pathology, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh.

A. khatun
Department of Plant Pathology, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh.

M.A.A. Bachchu
Department of Entomology, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh.

M.M.H. Bhuyain
Department of Entomology, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh.

M.A. Hossain
Department of Entomology, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh.

Abstract/ Executive Summary:

The antibacterial activities of ethyl alcohol, ethyl acetate, chloroform, acetone, n-hexane and dichloromethane extracts from the leaf, stem and root of akanda (Calotropis gigantea L.) were investigated against two phytopathogenic bacteria: Xanthomonas oryzae pv. oryzae and Ralstonia solanacearum, and a symbiotic bacterium Rhizobium sp. Leaf and root extracts of ethyl alcohol, ethyl acetate and chloroform; and stem extracts of ethyl alcohol and ethyl acetate showed antibacterial activities against all the test organisms. The maximum inhibition zone of all the test organisms were found with the leaf extracts followed by the root and the stem extracts. Among all the extracts, the maximum zone of inhibition of X. oryzae pv. oryzae have been observed with the leaf extracts of ethyl acetate followed by the stem and the root extracts of acetone, respectively. The maximum zone of inhibition of R. solanacearum and Rhizobium sp. was found with the ethyl alcohol leaf extract followed by the stem and the root extracts. The filter paper disc diffusion method and the broth serial dilution technique were applied to screen the antibacterial efficacies of the extracts and to determine the minimum inhibitory values, respectively. The minimum inhibitory concentration (MIC) of all the extracts ranged from 2.0 - 8.0 mg/mL. These results revealed that C. gigantea L. has antimicrobial activities against the three tested bacteria.

Keywords: Calotropis gigantea L., Antibacterial activity, Xanthomonas oryzae pv. oryzae, Ralstonia solanacearum, Rhizobium sp, MIC

Location: Department of Plant Pathology, Hajee Mohammad Danesh Science and Technology University (HSTU), Dinajpur

Start Date: 00-00-2011

End Date: 00-12-2011

Program Area: Pest Management

Commodity: Akanda

Objectives:

1. The purpose of the present study was aided to prepare different extracts from leaf, stem and root bark of C. gigantea L. to assess their antibacterial activities against two plant pathogenic bacteria namely Xanthomonas oryzae pv. oryzae and Ralstonia solanacearum, and a symbiotic bacterium, Rhizobium sp.

Methodology:

The experiment on antibacterial activities of Akanda (Calotropis gigantea L.) against three bacteria species was conducted at the laboratory, Department of Plant Pathology, Hajee Mohammad Danesh Science and Technology University (HSTU), Dinajpur, Bangladesh. Plant materials: The fresh leaves, stems and roots of C. gigantea L. were collected from the HSTU campus. At first, fresh specimens were washed thoroughly 2-3 times with running tap water followed by sterile water and shade-dried. After that the dried plant parts were made powder and used for extraction. Test microorganisms: The test bacteria (Xanthomonas oryzae pv. oryzae, Ralstonia solanacearum) and a symbiotic bacterium (Rhizobium sp). were isolated from the bacterial leaf blight disease of rice, wilt of potato and root nodules of pulse crops. After isolation, the bacteria were cultured on nutrient agar medium and subsequently pure cultures of all bacteria were made at the laboratory. Preparation of solvent extractions: The powdered plant materials (25 g) were extracted with ethyl alcohol, ethyl acetate, chloroform, n-hexane, acetone and dichloromethane solvents using a Soxhlet extractor for 48 hours and the extraction process was performed four times. The extracts were filtered through Whatman No. 1 filter paper. The filtrates were concentrated with a rotary evaporator under reduced pressure at 60⁰C to have the crude extracts. After complete evaporation, each of those extracts were weighed and preserved at room temperature in airtight bottles until further use. One gram of each solvent residue was dissolved in 10.0 ml of respective solvents and used as the test extracts for antimicrobial activity assay. Antibacterial activity assay: Antibacterial activity of the aqueous and solvent extracts was determined by the disc diffusion method on nutrient agar medium. Sterile Whatman filter discs (6 mm diameter) were made in the nutrient agar plate using sterile cork borer (5 mm). To study the antibacterial activities, standard kanamycin disc (30 µg/disc) and blank disc impregnated with the respective solvents were used as the positive and the negative controls, respectively. The antibacterial activities of ethyl alcohol, ethyl acetate, chloroform, n-hexane, acetone and dichloromethane were tested at concentrations of 30 µg/disc. The plates were incubated for 72 hours at 28⁰C. At the end of the incubation period, the clear zone of inhibition around the disc was measured in millimeter (mm). Determination of Minimum Inhibition Concentrations (MIC): The MIC values for the six solvents were determined by the serial tube dilution technique against X. oryzae pv. oryzae, R. solanacearum and Rhizobium sp.

Source of Information: J. Agrofor. Environ. 5 (1): 15-18, 2011; ISSN 1995-6983

Article Link (Online Journal):

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

The maximum inhibition zone of all the test organisms were found with the leaf extracts followed by the root and the stem extracts. Among all the extracts, the maximum zone of inhibition of X. oryzae pv. oryzae have been observed with the leaf extracts of ethyl acetate followed by the stem and the root extracts of acetone, respectively. The maximum zone of inhibition of R. solanacearum and Rhizobium sp. was found with the ethyl alcohol leaf extract followed by the stem and the root extracts. The filter paper disc diffusion method and the broth serial dilution technique were applied to screen the antibacterial efficacies of the extracts and to determine the minimum inhibitory values, respectively. The minimum inhibitory concentration (MIC) of all the extracts ranged from 2.0 - 8.0 mg/mL. These results revealed that C. gigantea L. has antimicrobial activities against the three tested bacteria.

Knowledge Management: Journal