Identifikasi Aktinomiset Endofit Asal Tanaman Obat Berdasarkan 16S rRNA

Abstract

Indigenous actinomycetes are known to have high biodiversity and chance to acquire a novel species. Various plants bioactive compounds with many functions may also be produced by endophytic microbes including actinomycetes. Many actinomycetes have been isolated from various medicinal plants. The research aimed to identify selected endophytic actinomycets from Curcuma xanthorrhiza (CX 10.1), Guazuma ulmifolia Lamk (AJB 4(1)), and Centella asiatica (AEP-2) based on 16S rRNA. Selected isolates of endophytic actinomycetes were grown in Yeast Starch Agar (YSA). DNA genome was isolated from CX 10.1, AJB 4(1), and AEP-2 isolates respectively, using Genomic DNA Mini Kit followed by amplification of 16S rRNA with PCR. PCR product was sequenced and analyzed by bio-informatics software to construct phylogenetic tree. The morphology colony of CX 10.1 did not produce aerial mycelium and spore chain type. Based on 16S rRNA gene sequence, CX 10.1 isolate was closed related with Micromonospora aurantiaca strain DSM 43813, Micromonospora coxensis strain 2-30-b(28) and Micromonospora krabiensis strain MA-2 with maximum identity 97%, 96% and 96% respectively. Meanwhile, morphology colony of AJB 4(1) and AEP-2 produced brown grey and white aerial mycelium, respectively and both of isolates produced Spiral spore chain type. The 16S rRNA sequence of AJB 4(1) showed to have the close relationship with Streptomyces misionensis strain NRRL B- 3230, Streptomyces africanus strain CPJVR-H, and Streptomyces eurythermus strain ATCC 14975, with maximum identity 98%, 97% and 97% respectively. The 16S rRNA sequence of AEP-2 showed close related with Streptomyces galilaeus strain JCM 4757,with 96% of maximum identity, while the close relatedness with both Streptomyces flavogriseus strain CBS 101.34, and S. polyantibioticus strain SPR; DSM 44925 was 95%. The maximum identity of AEP-2 was less then 97% and with further identification approaches may indicate a novel species.