Actinobacterial community and diversity in rhizosphere soils of Aquilaria crassna Pierre ex Lec assessed by RT-PCR and PCR-DGGE

The actinobacterial community in rhizospheres of eaglewood (Aquilaria crassna Pierre ex Lec) was analyzed using culture-independent methods of RT-PCR and PCR DGGE of 16S rRNA gene. We conducted the experiments to investigate the difference in diversity and community structure of actinobacteria with respect to sampling sites and seasons and to determine effect of plant species on selection of rhizosphere community from different sampling sites. Total genomic DNA and RNA were extracted from rhizosphere soils collected from two plantations in Phetchabun province and one plantation in each Nakhonnayok province, Rayong province and Chiang Mai province of Thailand during dry and rainy seasons. The UPGMA dendrogram generated from DGGE fingerprints showed that the actinobacterial community was separated corresponding to sampling sites, suggesting sampling sites effect. The shift in community and diversity between two seasons was detected in all sampling sites. RNA-based analyses showed that several actinobacterial groups appeared to be ubiquitous but different in metabolic activity in different environments. Species diversity (S) and simple indexes (I) indicate the increase in species diversity of actinobacteria from all sampling sites in rainy season. Cloning and sequencing of 16S rRNA gene fragments obtained from DGGE bands revealed that 14 of 40 dominant species of actinobacteria in the rhizospheres of this plant belonged to uncultured actinobacteria. Besides the uncultured actinobacteria, Nocardioides sp., Streptomyces sp., Mycobacterium sp., Rhodococcus sp. and Actinoplanes sp. were indentified and frequently found more than other genera.