Molecular Genotyping, 16S rRNA Sequencing, and Evolutionary Matrix of Streptomyces sp. Strain ANU-27 Isolated from Estuarine Mangrove Sediments

Anusha Rani Yavvari; Nagalakshmi Devamma Mundla

doi:10.37022/jis.v9i2.147

Anusha Rani Yavvari Department of Botany, SVU College of Sciences, Sri Venkateswara University, Tirupati-517502.
Nagalakshmi Devamma Mundla Department of Botany, SVU College of Sciences, Sri Venkateswara University, Tirupati-517502.

DOI https://doi.org/10.37022/jis.v9i2.147

Abstract

Estuarine mangrove environments are unique, highly dynamic habitats harboring a rich and unexplored diversity of bioactive actinobacteria. In this study, we highlight the extensive molecular identification, genetic sequencing, and evolutionary positioning of the bacterial isolate ANU-27, which exhibited superior broad-spectrum antimicrobial efficacy during initial phenotypic screenings. Genomic DNA was successfully extracted using the CTAB-lysozyme methodology, yielding634±32 µg/gof high-integrity DNA with a distinct genomic G+C content of 55.1% calculated via thermal denaturation midpoint (T_m= 92.3C).The 16S rRNA gene locus was targeted and amplified via PCR utilizing 27F and 1492R universal primers, generating a sharp ~1500 bp amplicon. Bidirectional Sanger sequencing resolved a definitive sequence spanning 1177 nucleotides with a high ribosomal G+C ratio 59.13%. Comprehensive homology mapping via NCBI BLASTn paired with a1000-replicate Maximum Parsimony cladogram clustered the strain with 100% sequence identity( E-Value = 0.0) within the genus Streptomyces, revealing an immediate phylogenetic node shared with Streptomyces maritimus strain SBK2-IR9 and Streptomyces rochei strain ABU8. These findings firmly clarify the taxonomic assignment of strain ANU-27, spotlighting its high secondary metabolic and biotechnological promise.

Keywords: Streptomyces sp, ANU-27, 16S rRNA Sequencing, Molecular Genotyping, Maximum Parsimony, Estuarine Actinomycetes

Author Biography

Nagalakshmi Devamma Mundla, Department of Botany, SVU College of Sciences, Sri Venkateswara University, Tirupati-517502.

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