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Optimization of Mycobiosynthesis of Silver Nanoparticles by Using Fusarium 4f1 and Trichoderma Trs Isolates

Shital Pal
Department of Botany, Jagannath University, Dhaka-1100, Bangladesh

KS Hossain
Department of Botany, Jagannath University, Dhaka-1100, Bangladesh

Abstract/ Executive Summary:

Silver nanoparticles (Ag-NPs) by mixing silver nitrate (AgNO₃) with cell-free filtrate (CFF) of the two fungal isolates viz., Fusarium 4F1 and Trichoderma TrS were synthesized. pH, substrate concentration and incubation period for the production of better quality and quantity of Ag-NPs was optimized. The Ag-NPs by UV-vis spectroscopy were characterized. Between the two fungal isolates, pH levels, AgNO₃ concentrations and incubation periods studied, the highest number of spherical shaped, monodispersed and stable Ag-NPs were recorded from Fusarium 4F1 at pH 9, 2 mM AgNO₃ and 72 hrs of incubation.

Keywords: Mycobiosynthesis, Siolver nanoparticles, Fusarium, Trichoderma

Location: National Botanical Garden, Mirpur, Dhaka, Bangladesh

Start Date:

End Date:

Program Area: Crop-Soil-Water Management

Commodity: Soil Health

Objectives:

To synthesize Ag-NPs in laboratory scale by Bangladeshi fungal isolates obtained from soil and optimized pH level, substrate concentration and incubation period for the production of higher quality and quantity of Ag-NPs.

Methodology:

Soil samples were collected from the pine plantation of the National Botanical Garden, Mirpur, Dhaka, Bangladesh. Fungi were isolated in pure culture following serial dilution and pour plate method using potato dextrose agar (PDA) medium supplemented with 0.1 mg/ml streptomycin sulfate to exclude soil bacteria (Aneja 2003). The fungal isolates were identified following standard literatures on the basis of their morphological characteristics (Booth 1971, Bissett 1991). Among the isolated fungi, Fusarium 4F1 and Trichoderma TrS were selected for the biosynthesis of Ag-NPs because the genera are reported to be promising Ag-NPs producer (Vahabi et al. 2011, Birla et al. 2013, Korbekandi et al. 2013). To prepare biomass, Fusarium 4F1 and Trichoderma TrS were grown aerobically in potato sucrose and potato dextrose broth media (pH 7), respectively at 24 ± 2°C. The culture broth in the flasks were agitated at 170 rpm on a rotary shaker for 72 hrs. The biomass was harvested after incubation using a plastic sieve. After that, it was thoroughly washed with sterile distilled water to remove any other component from the biomass. To prepare Cell-Free Filtrate (CFF), approximately 20 g of fresh and clean biomass were taken in a 500 ml Erlenmeyer flask containing 150 ml of sterilized deionized water. The flask was incubated at 24 ± 2°C on a rotary shaker for 72 hrs and agitated at 170 rpm. After the incubation period, CFF was obtained by passing it through sterile Whatman filter paper No. 1. For the biosynthesis of Ag-NPs, the silver nitrate (AgNO₃) solution was mixed with ca. 50 ml fresh CFF of Fusarium 4F1 and Trichoderma TrS in several sets in 250 ml Erlenmeyer flasks. The final concentration of the substrate (AgNO₃) in the solutions was 1 mM and pH was adjusted to 7, 8, and 9 separately. The flasks containing CFF without AgNO₃ at pH 7, 8 and 9 were also maintained along with the treatment sets. These flasks were marked as control. Depending on the effects of pH on the quality and quantity of Ag-NPs, separate sets of experiments were run where final concentration of the AgNO₃ were 2 mM and pH were 9 for both fungal isolates following the above mentioned procedure. In every sets, 3 replications were maintained. All flasks were fully covered with thick black paper and incubated on a rotary shaker and agitated at 170 rpm at 24 ± 2°C in dark. Flasks were visually examined after every 24 hrs intervals up to 45 days to find any kind of change of CFF and AgNO₃ mixture color. In every observation, data on color change were recorded and photographs of the flasks were also taken with a digital camera. Reaction mixture was withdrawn every after 24, 48 and 72 hrs from each flask. UV-visible light absorption profile of the mixture was recorded to study the change in intensity of the brown color during incubation periods using a UV-Vis spectrophotometer (SPECORD 250 plus, Analytic Jena). The absorbance was measured from 300 - 600 nm at a resolution of 1 nm. The control sets were used as reference in spectroscopy. It is generally recognized that UV-Vis spectroscopy could be used to examine the quality and quantity of NPs in aqueous suspensions (Wiley et al. 2006). The results were analyzed statistically for the test of significance using WASP 2.0 software. The experiments were conducted in the laboratory at the department of Botany, Jagannath University, Dhaka.

Source of Information: Bangladesh Journal of Botany; 2079-9926; Vol 49 No 2 (2020), ISSN 0253-5416 eISSN

Article Link (Online Journal): https://doi.org/10.3329/bjb.v49i2.49315

Funding Source:

Budget:

Total Budget (Tk.) :

Achievement/Findings:

From this study, it may be inferred that pH 9, 72 hrs of incubation period and 2 mM concentration of AgNO₃ are optima for mycobiofabrication of spherical, monodispersed and stable Ag-NPs by both fungal isolates.

Knowledge Management: Journal