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Title: Polymerase Chain Reaction- Restriction Fragment Length Polymorphism method to distinguish three mealybug clades within the Planococcus citri-P. minor species complex

Author
item RUNG, A. - DEPT. OF FOOD & AGR., CA
item MILLER, D. - USDA,ARS,SEL (RETIRED)
item Scheffer, Sonja

Submitted to: Journal of Economic Entomology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/27/2008
Publication Date: 2/1/2009
Citation: Rung, A., Miller, D.R., Scheffer, S.J. 2009. Polymerase Chain Reaction- Restriction Fragment Length Polymorphism method to distinguish three mealybug clades within the Planococcus citri-P. minor species complex. Journal of Economic Entomology. 102:1-5.

Interpretive Summary: Mealybugs are serious agricultural pests and are common invasive species in the U.S. This research provides a molecular identification tool that distinguishes between two mealybug species (citrus mealybug and minor mealybug). These species are difficult or impossible to determine using morphological characters. The molecular tool is especially important to U.S. quarantine because the minor mealybug does not occur in the U.S. but is frequently intercepted at U.S. ports-of-entry. This research provides a tool for quarantine personnel so that they can accurately distinguish these species and prevent the minor mealybug from becoming a pest in the U.S.

Technical Abstract: The mealybug species Planococcus citri (Risso) and P. minor (Maskell) have special significance to U.S. quarantine and U.S. agriculture. These two species, commonly intercepted at U.S. ports-of-entry, are difficult to identify based on morphological characters. This study presents a molecular method for distinguishing P. citri, P. minor, and a genetically distinct clade that is morphologically identical to P. citri, from Hawaii. This method employs polymerase chain reaction (PCR) followed by restriction fragment polymorphism analysis (RFLP) using the restriction enzymes BspH1, BsmH1, HpH1. The resulting band patterns can be visualized in a 2% agarose gel and are sufficient to differentiate between the three entities mentioned above. PCR-RFLP diagnostics can be used for all life stages and is cheaper than DNA sequencing.