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A one‐step, single tube, duplex PCR to detect predation by native predators on invasive B emisia tabaci MEAM 1 and F rankliniella occidentalis
Author(s) -
Zhang GuiFen,
Wu Xia,
Zhou ZhiXiang,
Meng XiangQin,
Wan FangHao
Publication year - 2014
Publication title -
entomologia experimentalis et applicata
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.765
H-Index - 83
eISSN - 1570-7458
pISSN - 0013-8703
DOI - 10.1111/eea.12134
Subject(s) - biology , predation , duplex (building) , predator , polymerase chain reaction , gene , genetics , dna , ecology
B emisia tabaci ( G ennadius) MEAM 1 ( H emiptera: A leyrodidae) and F rankliniella occidentalis ( P ergande) ( T hysanoptera: T hripidae), two important invasive species, are serious agricultural pests. In this study, a one‐step, single tube, duplex polymerase chain reaction ( PCR ) procedure was developed to allow rapid, specific, and sensitive identification of B . tabaci MEAM 1 and F . occidentalis in predator guts. The system and conditions used for the duplex PCR were optimized. The species specificity of the duplex PCR determined by comparison against non‐targets that might interact with B . tabaci MEAM 1 and F . occidentalis showed that oligonucleotide primers amplified nuclear gene target sequences present only in B . tabaci MEAM 1 or F . occidentalis . The limits of detection were 9.53 ng μl −1 for B . tabaci MEAM 1 and 8.94 ng μl −1 for F . occidentalis . Within a field cage study, in which predators H armonia axyridis ( P allas) ( C oleoptera: C occinellidae) and O rius sauteri ( P oppius) ( H emiptera: A nthocoridae) were allowed to feed on B . tabaci MEAM 1 and F . occidentalis for 10 h, the B . tabaci MEAM 1 DNA was detectable in 100% of H . axyridis and O . sauteri , and F . occidentalis DNA was detectable in 80% of H . axyridis and 90% of O . sauteri ; this implicated that B . tabaci MEAM 1 and F . occidentalis remains could be detected in native predator guts simultaneously. The accuracy and reliability of the assay suggested strongly that the duplex PCR optimized for B . tabaci MEAM 1 and F . occidentalis is sensitive and specific for both invasive insects and is therefore useful in early diagnosis and monitoring of B . tabaci MEAM 1 and F . occidentalis infections, and can be used to identify domestic predator species and food web relationships.