Abstract:
Quick and authentic identification of exotic and potentially invasive taxa with
capability
of causing high economic losses or detriments is essential prerequisite for
effective plant quarantine and biological control initiatives. The order Thysanoptera
includes several agricultural pest species that, not only because of their minute size but
als
o due to their cryptic behavior, incline to undetected transport through international
trade of plants. Identification of thrips, particularly at species level, is pretty demanding
and requires expertise in knowledge about Thysanoptera. Moreover, in most c
ases,
identification of larval Thysanoptera to species is impossible without presence of adults.
Hence, there is a great desire for a facile, accurate, and highly reliable technique for
thrips identification. The present study describes species
-
specific pr
imers for four pest
thrips species, and the use of a multiplex PCR assay to detect and to distinguish between
the four target species. Five primers were used to simultaneously amplify a specific region
of the mitochondrial DNA and produce species
-
specific
fragments. Results indicated that
the primers were capable of detecting these four species and amplifying uniquely sized,
species
-
specific PCR products. Furthermore, using a multiplex PCR assay, the primers
maintained specificity and sensitivity, and allow
ed detection of each of the four species in
a single reaction. The stringency of the method was tested using specimens of different
developmental stages and consistent results were obtained for all of the examined
samples. This method is simple enough to b
e implemented by non
-
experts and also can be
extended to any organism for which quick and reliable identification is needed.
Keywords:
Mitochondrial DNA, Multiplex polymerase chain reaction, Species
identification, Thripidae, Thysanoptera.
