Phylogenetic reconstruction of endophytic fungal isolates using internal transcribed spacer 2 (ITS2) region |
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Authors: | Kathamuthu GokulRaj Natesan Sundaresan Enthai Jagan Ganeshan Pandi Rajapriya Johnpaul Muthumary Jayavel Sridhar Mohan Pandi |
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Institution: | 1.Department of Molecular Microbiology, School of Biotechnology, Madurai Kamaraj University, Madurai – 21, Tamil Nadu, India;2.Department of Microbial Technology, School of Biological Sciences, Madurai Kamaraj University, Madurai – 21, Tamil Nadu,India;3.Centre for Advanced Studies in Botany, University of Madras, Guindy Campus, Chennai – 25, Tamil Nadu, India;4.Department of Biotechnology (DDE), Madurai Kamaraj University, Madurai – 21, Tamil Nadu, India |
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Abstract: | Endophytic fungi are inhabitants of plants, living most part of their lifecycle asymptomatically which mainly confer protection and
ecological advantages to the host plant. In this present study, 48 endophytic fungi were isolated from the leaves of three medicinal
plants and characterized based on ITS2 sequence – secondary structure analysis. ITS2 secondary structures were elucidated with
minimum free energy method (MFOLD version 3.1) and consensus structure of each genus was generated by 4SALE. ProfDistS
was used to generate ITS2 sequence structure based phylogenetic tree respectively. Our elucidated isolates were belonging to
Ascomycetes family, representing 5 orders and 6 genera. Colletotrichum/Glomerella spp., Diaporthae/Phomopsis spp., and Alternaria
spp., were predominantly observed while Cochliobolus sp., Cladosporium sp., and Emericella sp., were represented by singletons. The
constructed phylogenetic tree has well resolved monophyletic groups with >50% bootstrap value support. Secondary structures
based fungal systematics improves not only the stability; it also increases the precision of phylogenetic inference. Above ITS2 based
phylogenetic analysis was performed for our 48 isolates along with sequences of known ex-types taken from GenBank which
confirms the efficiency of the proposed method. Further, we propose it as superlative marker for reconstructing phylogenetic
relationships at different taxonomic levels due to their lesser length. |
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Keywords: | Endophytes ITS MEGA 5 1 ProfDistS Sequence-secondary structure 4SALE |
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