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Title: A rapid method for preparation of nucleic acid extracts from potato psyllids for detection of 'Candidatus Liberibacter solancearum' and molecular analysis

Author
item Crosslin, James
item Swisher Grimm, Kylie
item Cimrhakl, Launa

Submitted to: Southwestern Entomologist
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/8/2013
Publication Date: 4/1/2013
Citation: Crosslin, J., Swisher, K.D., Hamlin, L.L. 2013. A rapid method for preparation of nucleic acid extracts from potato psyllids for detection of 'Candidatus Liberibacter solancearum' and molecular analysis. Southwestern Entomologist. 38:41-48.

Interpretive Summary: Potato zebra chip disease (ZC) is caused by a bacterium transmitted to potatoes by the potato psyllid. The disease has caused severe economic damage in Mexico, Central America, New Zealand, the south-central United States. The ZC disease first appeared in the major potato producing region of Washington, Oregon, and Idaho in 2011. Since the ZC bacterium is transmitted by the potato psyllid, testing large numbers of the potato psyllids is necessary for studying the epidemiology of ZC. We have developed a method for testing psyllids for the bacterium that does not require organic solvents or lengthy DNA isolation techniques. The technique is rapid, reproducible, and useful for detection of the bacterium in individual psyllids and for genetic analysis of the potato psyllids.

Technical Abstract: A rapid method has been developed and validated for PCR analysis of potato psyllids for Candidatus Liberibacter solanacearum (Lso), the causal agent of zebra chip disease of potatoes. The method is also suitable for PCR amplification and high resolution melting analysis of the cytochrome oxidase I gene of potato psyllids which is useful for psyllid population studies. The new method takes less than 30 minutes, requires only common laboratory reagents, uses one inexpensive enzyme, no organic solvents, and no lengthy centrifugation steps are necessary. The method should be suitable for rapid testing of large numbers of potato psyllids for studies on zebra chip disease epidemiology and genetic analysis of psyllid populations.