Multiplex-PCR for identification of two Hishimonus species (Hemiptera: Cicadellidae) in jujube orchards and detection of jujube witches' broom (JWB) phytoplasma in their bodies

Hishimonus sellatus (Uhler) transmits jujube witches' broom (JWB). This study aims to develop and validate a rapid and inexpensive molecular method to discriminate between H. sellatus and H. lamellatus occurring in jujube orchards and to detect JWB phytoplasma in their bodies meantime. A multiplex PCR method was designed for identification of two species and detection of JWB phytoplasma in their bodies. The results showed that the multiplex PCR method established in this study could identify species of H. sellatus and H. lamellatus accurately, and could detect JWB phytoplasma in H. sellatus or H. lamellatus sensitively. The validation results with 48 field collections of male individuals pre-checked by morphological observation showed that there was no mismatch between multiplex PCR and morphological observation in all samples, indicating the 100% accuracy of the multiplex PCR for identification of H. sellatu and H. lamellatus.