title: DNA barcoding, species-specific PCR and real-time PCR techniques for the identification of six Tribolium pests of stored products.
abstract: Flour beetles of the genus Tribolium Macleay (Coleoptera: Tenebrionidae) are important stored product pests in China and worldwide. They are often found or are intercepted in grain depots, flour mills, and entry-exit ports, etc. Traditionally, Tribolium species are identified according to the morphological characteristics of the adult. However, it is almost impossible to rapidly identify adult fragments and non-adult stages based on external morphological characteristics. Molecular techniques for the rapid and accurate identification of Tribolium species are required, particularly for pest monitoring and the quarantine of stored products pests. Here, we establish DNA barcoding, species-specific PCR, and real-time PCR techniques for the identification of six stored-product pest Tribolium species including T. castaneum, T. confusum, T. destructor, T. madens, T. freemani and T. brevicornis. We detected the mitochondrial DNA cytochrome oxidase subunit I (COI) barcodes for Tribolium from 18 geographic populations and 101 individuals, built a Tribolium DNA barcode library, and designed species-specific primers and TaqMan probes for the above six Tribolium species. The three techniques were applied to identify Tribolium collected from stored samples and samples captured from quarantine ports. The results demonstrated that three techniques were all able to identify the six species of Tribolium both rapidly and accurately.
authors.affiliation: College of Plant Protection, China Agricultural University, Beijing 100193, China.|Academy of State Administration of Grain, Beijing 100037, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Zhang
authors.affiliation: College of Plant Protection, China Agricultural University, Beijing 100193, China.
authors.forename: Yi-Jiao
authors.initials: YJ
authors.lastname: Wang
authors.affiliation: Chinese Academy of Inspection and Quarantine, Beijing 100176, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Guo
authors.affiliation: College of Plant Protection, China Agricultural University, Beijing 100193, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Luo
authors.affiliation: Academy of State Administration of Grain, Beijing 100037, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Wu
authors.affiliation: Crop Research Institute, Drnovská 507, 161 06 Prague 6, Czech Republic.
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Kučerová
authors.affiliation: Crop Research Institute, Drnovská 507, 161 06 Prague 6, Czech Republic.
authors.forename: Václav
authors.initials: V
authors.lastname: Stejskal
authors.affiliation: Department of Entomology and Plant Pathology, 127 Noble Research Center, Oklahoma State University, Stillwater, OK 74078, USA.
authors.forename: George
authors.initials: G
authors.lastname: Opit
authors.affiliation: Academy of State Administration of Grain, Beijing 100037, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Cao
authors.affiliation: Academy of State Administration of Grain, Beijing 100037, China.
authors.forename: Fu-Jun
authors.initials: FJ
authors.lastname: Li
authors.affiliation: College of Plant Protection, China Agricultural University, Beijing 100193, China.
authors.forename: Zhi-Hong
authors.initials: ZH
authors.lastname: Li
mesh_terms: D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D058893:DNA Barcoding, Taxonomic; D017931:DNA Primers / Q000378:metabolism; D003576:Electron Transport Complex IV / Q000737:chemistry / Q000235:genetics; D010802:Phylogeny; D060888:Real-Time Polymerase Chain Reaction; D016415:Sequence Alignment; D013045:Species Specificity; D014231:Tribolium / Q000145:classification* / Q000235:genetics*
chemical_list: D017931:DNA Primers; D004247:DNA; D003576:Electron Transport Complex IV