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Detection by PCR of Psyllid, Diaphorina citri Kuwayama, Vector of Citrus Huanglongbing Disease
The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is the most important citrus pest, because it serves as a vector of “Candidatus Liberibacter” species that cause huanglongbing disease. Thus, when exporting Rutaceae (citrus), exporting countries put them into quarantine. However, as the size of an imago is so small that only skilled experts can identify it using morphology-based species identification. A PCR-based assay was developed for monitoring psyllids using a rapid, using the DNA extraction from psyllid bodies and PCR amplification. Thus, this study aims to develop a DNA marker system with high species specificity and discrimination power that helps identifying psyllid. We analyzed the base sequence of mitocondria DNA. Based on this sequence, restriction sites were determined and a species particularity primer was made. Several polymorphic regions of mitochondrial DNA of both species were sequenced and used for developing specific restriction sites and polymerase chain reaction (PCR) primers. Also, species-specific PCR primers were devised to develop diagnostic PCR method for identifying the internal feeders. Base sequence results from PCR, which used psyllid species-specific primers, was analyzed by the Mealign program of the CLC Main Workbench analysis software, and used to develop a phylogenic tree. After analyzing this, Hemiptera proved to be allied species.
