Identification of bacterial species from healthy wood of Aquilaria crassna using colony PCR

¹ Department of Biotechnology, Faculty of Applied Science, King Mongkut's University of Technology North Bangkok, Bangkok 10800, Thailand
² Water and Soil Environmental Research Unit, Mahidol University, Nakhonsawan Campus, Nakhonsawan 60130, Thailand
³ Aircraft Maintenance Engineering, Department of Mechanical and Industrial Engineering, Rajamangala University of Technology Krungthep, Bangkok 10120, Thailand
⁴ Department of Chemistry, Faculty of Science, University of New England, Australia
⁵ Biorefinery and Process Automation Engineering Center, Department of Chemical and Process Engineering, The Sirindhorn Thai-German International Graduate School of Engineering, King Mongkut's University of Technology North Bangkok, Thailand

^* Corresponding author: phusantisampan.t@gmail.com

Abstract

Agarwood is one of the most expensive, sacred and valuable woods used in the biotechnology industry. This natural raw material is in high demand as a commodity and can be used to manufacture a variety of products. Isolating a microbe from its environment is crucial because several bacterial strains can produce novel compounds for biotechnological applications. Exploration of bacterial strains, such as bacterial endophytes, has been associated with bioactive compounds of the agarwood plant. The purpose of this study was to isolate and identify bacteria from Agarwood trees in the Kaeng Hang Maeo Sub-district of Chanthaburi Province, Thailand by using molecular biology techniques. A colony PCR technique without prior DNA extraction for rapid and simple detection of bacteria was performed. After the DNA sequence analysis, the results showed that most populations in the normal layer are belonged to Pantoea dispersa (99%). Through analysis of phylogenetic tree by neighbor-joining method, the sequences of the 16s rRNA gene were analyzed to understand evolutionary relationships of bacteria found in agarwood. In conclusion, good amplification of the 16s rRNA sequence (1.5 kb) was detected with the specificity of the target sequences of the 16s RNA. In addition, the isolated bacterial strains are the most common species in agarwood, however, there has been no previous research on isolating these bacterial strains in Aquilaria crassna.