Molecular Identification of Some Wood-decay Fungi of Some Softwood Timber by Using Internal Transcribed Spacer Primers in Akwa Ibom State, Nigeria
Ajefu, Monica I. *
Department of Botany and Ecological Studies. Faculty of Biological Sciences. University of Uyo, Uyo, Nigeria.
I. N. Bassey
Department of Botany and Ecological Studies. Faculty of Biological Sciences. University of Uyo, Uyo, Nigeria.
Asuquo, Terrance J.
Department of Botany and Ecological Studies. Faculty of Biological Sciences. University of Uyo, Uyo, Nigeria.
*Author to whom correspondence should be addressed.
Abstract
Aims: This research focuses on molecular identification of some wood-decay fungi of some softwood timber in some parts of Akwa Ibom State.
Study Design: The study was conducted in the laboratory using culture vessels to maintain isolated fungal culture from decaying wood samples.
Place and Duration of Study: Samples were obtained from Idu timber market, Uruan, LGA, Akwa Ibom State which were then taken Botany and Microbiology laboratories at the University of Uyo for culturing. Molecular identification was done in Inqaba South Africa. The entire work was done in summer seasons of 2024.
Methodology: The isolates were inoculated on Sabouraud Dextrose Agar (SDA) plates. The plates were incubated at 27oC for seven days. Then molecular identification and sequencing were carried out. Extraction was done using a ZR fungal/bacterial DNA mini prep extraction kit. The extracted genomic DNA was quantified using the Nanodrop 1000 spectrophotometer. The Internal Transcribed Spacer 1(ITS) as forward and ITS 4 as reverse primers. Sequencing was done using the BigDye Terminator kit on a 3510 ABI sequencer. Obtained sequences were trimmed or filtered using bioinformatics algorithm. Similar sequences were downloaded from the National Center for Biotechnology Information (NCBI) database using BLASTN. The accession number of amplified nucleotides sequences for fungal isolate and morphological characteristics were obtained. Finally, percentage distribution of fungi isolated were calculated.
Results: The DNA extractions reveal the following fungi that occur in the three wood samples: Candida parapislosis, Aspergillus flavus, A. terreus, A. niger and Cyberlindnera jadinii. The percentage distribution of fungi isolated in wood sample shows that Aspergillus genus had the highest percentage (60%) of occurrence, while C. jadinii and C. parapislosis had the lowest occurrence with 20% each. Percentage distribution of fungi isolated from different timber samples showed that Atarabang species produced the highest percentage of fungi distribution of (60%) while Ntufiak and Ufu had the lowest percentage of distribution with 20% each.
Conclusion: The results of this work show that fungi are present in all the sampled locations because of negligence in wood treatment and favourable environmental conditions, and as such, adequate procedures should be put in place by relevant authorities to establish strict regulations for timber treatment and the use of high-quality wood for construction.
Keywords: Molecular identification, softwood; fungi, DNA extraction, ITS region