Isolation and Identification of Endophytic Fungi from Banana Leaves (Musa sp.)

Mohan, Anand (2020) Isolation and Identification of Endophytic Fungi from Banana Leaves (Musa sp.). Final Year Project (Bachelor), Tunku Abdul Rahman University College.

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Abstract

Mutualistic microbial endophytes are present in the cells of the stem, leaves and roots of all plants. The banana plant has had a history with fungus such as when the Panama Disease took down the globally exported banana variety called Gros Michel. The objective of this study was to isolate and identify endophytic fungus communities present in banana leaves. Young and mature banana leaves were collected from four different locations. Leaf sections were sterilized and inoculated on Potato Dextrose Agar to allow fungal isolation. Morphological identification by staining and microscopy was done where colony pigmentation, colony texture, presence of septa and conidia were observed. To further identify the endophytic fungus, molecular identification was carried out on four isolates of distinct morphological characteristics. Three different genomic DNA extraction methods were carried out which were the CTAB method, thermolysis method and microwave method. Polymerase chain reaction was carried out using ITS (Internal Transcribed Spacer region) sequences as primers. The amplified DNA showed single bands of approximately 500bp. An endophytic fungus was identified as Mucor sp. In addition, other endophytic microorganisms such as Bacillus subtilis and Sphingomonas sp. was also identified. Keywords: Morphological Identification, DNA Extraction, CTAB method, Thermolysis method, Polymerase Chain Reaction (PCR), DNA Sequencing.