Managing outbreaks of the citrus long-horned beetle Anoplophora chinensis (Forster) in Europe: molecular diagnosis of plant infestation.

BACKGROUND Anoplophora chinensis (Forster) (Coleoptera Cerambycidae) is a polyphagous long-horned beetle native to Eastern Asia that infests a wide range of broadleaved plants, causing disruption of vascular tissue, structural weakness and tree death. As a result, A. chinensis is a quarantine pest for the European Union. In order to confirm its infestations on plants, it is necessary to obtain adult or larval specimens on which morphological or molecular analysis can be performed. However, obtaining such specimens from infested plants can be a demanding and difficult task. Therefore, a diagnostic tool that is non-invasive for the plant and able to be performed in the absence of any insect stages may be useful to confirm infestations of A. chinensis on plants. RESULTS A protocol is presented that is based on polymerase chain reaction amplification using DNA extracted from A. chinensis faecal material collected in the field. CONCLUSION Results obtained in the present work show that the non-invasive approach is a reliable and accurate alternative diagnostic tool in phytosanitary surveys.

[1]  M. Quesada,et al.  A Simple and Rapid Method for DNA Isolation from Xylophagous Insects , 2010, International journal of molecular sciences.

[2]  P. Roversi,et al.  Feeding and oviposition of Anoplophora chinensis on ornamental and forest trees , 2010, Phytoparasitica.

[3]  P. Roversi,et al.  Evaluation of eradication measures against Anoplophora chinensis in early stage infestations in Europe. , 2010 .

[4]  S. Jarman,et al.  A molecular approach to identify prey of the southern rock lobster , 2008, Bulletin of Entomological Research.

[5]  C. Chapman,et al.  Plant DNA sequences from feces: potential means for assessing diets of wild primates , 2007, American journal of primatology.

[6]  M. Ciampitti,et al.  Anoplophora species in Europe : infestations and management processes , 2006 .

[7]  S. Regnaut,et al.  DNA degradation in avian faecal samples and feasibility of non-invasive genetic studies of threatened capercaillie populations , 2006, Conservation Genetics.

[8]  M. Bon,et al.  High through-put characterization of insect morphocryptic entities by a non-invasive method using direct-PCR of fecal DNA. , 2005, Journal of biotechnology.

[9]  T. Ladd,et al.  Development of SCAR markers for the DNA-based detection of the Asian long-horned beetle, Anoplophora glabripennis (Motschulsky). , 2003, Archives of insect biochemistry and physiology.

[10]  S. Wasser,et al.  Techniques for application of faecal DNA methods to field studies of Ursids , 1997, Molecular ecology.

[11]  E. Paxinos,et al.  A noninvasive method for distinguishing among canid species: amplification and enzyme restriction of DNA from dung , 1997, Molecular ecology.

[12]  B. Crespi,et al.  Evolution, weighting, and phylogenetic utility of mitochondrial gene sequences and a compilation of conserved polymerase chain reaction primers , 1994 .

[13]  R. Vrijenhoek,et al.  DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. , 1994, Molecular marine biology and biotechnology.

[14]  I. Adachi.  Population studies ofanoplophora malasiaca adults (Coleoptera: Cerambycidae) in a citrus grove , 1990, Researches on Population Ecology.

[15]  I. Adachi.  Reproductive biology of the white-spotted longicorn beetle, Anoplophora malasiaca Thomson (Coleoptera: Cerambycidae), in citrus trees. , 1988 .

[16]  Jean J Turgeon,et al.  Managing invasive populations of Asian longhorned beetle and citrus longhorned beetle: a worldwide perspective. , 2010, Annual review of entomology.

[17]  Julie Feinstein.  DNA sequence from butterfly frass and exuviae , 2004, Conservation Genetics.

[18]  M. P. Piggott.  Effect of sample age and season of collection on the reliability of microsatellite genotyping of faecal DNA , 2004 .