Development of single locus DNA microsatellite markers in Oryctes rhinoceros (Linnaeus) using 5′ anchored RAMs-PCR method

Abstract

Oryctes rhinoceros, commonly known as rhinoceros beetle, is an important pest in oil palm plantations. The presence of this pest in replanting sites as early as six months after replanting has alarmed planters due to the possibility of increased crop damage (Samsuddin et al. 1993; Kamarudin and Wahid 1997). Being a nocturnal animal with a destructive feeding habit, it is difficult to eliminate this pest (Young 1986). Pheromone trapping using a speciesspecific-aggregation pheromone is commonly used to trap O. rhinoceros in replanting sites (Hallet et al. 1995). However, not all population of O. rhinoceros in the field were observed to be attracted by it. This suggests the possibility of a cryptic species complex occurrence in this insect. To investigate this notion, a thorough study utilizing molecular markers is necessary. An example of a powerful marker with proven capability for resolving such issues is the single-locus DNA microsatellite marker. Being codominant, multiallelic and highly polymorphic, microsatellites are a powerful and promising genetic marker, suitable for precise discrimination of closely related individuals (Smouse and Chevillon 1998). A previous study on the genetic variation of O. rhinoceros had highlighted the necessity of further studying this pest species using single-locus microsatellite DNA markers as O. rhinoceros were reported to exhibit possible occurrence in two groups (Manjeri et al. 2011). Therefore, with interest to further study the population genetic structure of O. rhinoceros; this study was carried out to isolate sufficiently novel single-locus microsatellite markers for the insect. The isolation was carried out based on the 5′-anchored polymerase chain reaction (PCR) technique (Fisher et al. 1996)