Molecular Identification and Phylogenetic Analysis of Fungi Contaminants Associated with In Vitro Cultured Banana Based on ITS Region Sequence

Abstract

This study characterized, identified and conducted phylogenetic analysis on fungi contaminants in vitro bananas based on the sequence of inter-space (ITS) regions. Genomic DNA was extracted from the pure culture of fungi contaminants, amplified and sequenced using ITS1 and ITS4 markers. Analysis of the sequences using MEGA 7 Software at higher similarity sequence identified five Aspergillus spp., three Penicillium spp., one each of Fusarium, Trichoderma and Cladosporium as the contaminants. The genetic distance between the fungi species was 0.205, which suggests a homogeneous substitution between the sequences, and thiamine was the most stable. The fungi clustered in three major groups at 0.10 genetic distance, subdivided into five clusters. A cluster and sub-cluster consisting of five Aspergillus strains; a major cluster of three Penicillium strains; a cluster comprising of Fusarium chlamydosporum and Trichoderma viride; and a sole fungi Cladosporium tenuissimum. The Aspergillus group were phylogenetically related to A. flavus and A. parvissclerotigenus, the identified Penicillium spp. were closely related to Penicillium citrinum while the detected Cladosporium aligned with Cladosporium tenuissium and Phoma multirostrata. The information provided by this study could be utilized to develop a specific and compelling sterilization protocol to minimize the rate of contamination during in vitro culture procedures.

Managing Lethal Browning and Microbial Contamination in Musa spp. Tissue Culture: Synthesis and Perspectives

“…Tissue culture technology is useful for providing disease-free and consistent planting materials that can be made available throughout the year [45]. Thus, the technique is crucial for banana and plantain micropropagation, ensuring the availability of genetically identical and disease-free plantlets for commercial production of the crop, which would significantly improve food security and economic benefits for farmers and consumers around the world [10]. Despite the multiple benefits of tissue culture technologies to banana and plantain production and agricultural growth, contamination has been a serious impediment to their use in crop micropropagation [46].…”

“…Accurate identification of common contaminants is required to effectively combat the threat of fungal contamination. Animasaun et al [10] isolated and identified eleven fungi strains of five genera, Aspergillus, Cladosporium, Fusarium, Penicillium, and Trichoderma as the significant contaminants of in vitro banana culture. In early studies, Odutayo et al [60] and Rahman et al [61] identified four and five genera of fungi contaminants in tissue culture materials, respectively.…”

“…This method is able to produce diseasefree plants, high-quality banana plants, and high-quality planting material, and the rapid production of many uniform plants in a short time, which can support strong plant growth during the subsequent growth cycle [6,7]. However, explant browning and contamination are the main obstacles that often occur in the micropropagation of banana and plantain explants using tissue culture techniques [8][9][10].…”

Elucidation of the composition, antioxidant, and antimicrobial properties of essential oil and extract from Citrus aurantifolia (Christm.) Swingle peel

“…Meanwhile, fungal contaminants that often contribute to contamination include Aspergillus flavus , A. niger , Fusarium spp., and Trichoderma spp. ( Agbadje et al, 2021 , Animasaun et al, 2022 , Kithaku et al, 2019 , Mokbel et al, 2017 ). Inadequate treatment of explant contamination can result in less regeneration capacity, decreased callus formation, and suppressed adventitious shoot growth ( Cobrado and Fernandez, 2016 , El-Banna et al, 2021 ).…”