Abstract
Rice (Oryza sativa L.) is the main cereal crop in many Asian countries. The Asian rice planthoppers, Nilaparvata lugens (Stål) (brown planthopper), Sogatella furcifera (Horváth) (white-backed planthopper), and Laodelphax striatellus (Fallén) (small brown planthopper) (Hemiptera: Delphacidae), are the most economically important pests of rice. These three rice planthopper species often co-occur in the same paddy field. Traditionally, species identification of individuals of the three rice planthopper species has relied on morphological characters, but accurate discrimination of early-instar nymphs is very difficult, even for expert researchers. In this study, we developed a rapid one-step multiplex PCR assay using conserved and species-specific 5.8S-ITS2 rDNA gene primers for simultaneous identification of individuals of the three rice planthopper species. The multiplex PCR results showed that the three rice planthopper species could be identified accurately based on the length of the resultant amplicon, regardless of the individual developmental stage. Furthermore, we applied this assay for the first accurate quantification of early-instar nymphs of each rice planthopper species in paddy fields. Notably, we found that the species composition of early-instar nymphs cannot be extrapolated from that of adults. Thus, the multiplex PCR assay developed here facilitates detection of each rice planthopper species at the beginning of outbreaks in paddy fields.
Highlights
Numerous insects migrate long distances between countries and continents [1, 2]
The 5.8SITS2-28S rDNA gene sequences obtained in this study were deposited in the DDBJ/EMBL/ GenBank nucleotide sequence databases under the accession numbers MT950726–MT950755
This study focused only on the three rice planthopper species, this multiplex PCR method could be useful for detecting individuals of the three rice planthopper species from samples containing other planthopper species by redesigning the species-specific primers as needed
Summary
Numerous insects migrate long distances between countries and continents [1, 2]. Some of them are serious agricultural pests, such as locusts, armyworms, and planthoppers [3,4,5]. The timing and magnitude of migration of these transboundary pests vary from year to year because their long-distance migrations occur under specific weather conditions [3,4,5]. It is necessary to monitor the migration and subsequent population growth of such insect pests continuously for their management [6, 7]. Molecular methods, including PCR-based techniques and DNA sequencing, are useful for identifying species, strains, and source.
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