Detection of 'Candidatus liberibacter asiaticus' in cacopsylla (Psylla) citrisuga (hemiptera: Psyllidae)

Abstract

Nymphs of the pomelo psyllid, Cacopsylla (Psylla) citrisuga Yang & Li, were collected from huanglongbing (HLB) symptomatic lemon trees, Citrus limon (L.) in Yunnan Province, China. DNA samples extracted from groups of 210 pomelo psyllid nymphs of all stages and from leaves of lemon plants were analyzed with nested-PCR in order to detect 'Candidatus Liberibacter asiaticus' (Las). The results showed that 24 out of the 49 lemon trees were Las-positive. Also psyllid nymphs collected from 10 out of the 24 Las-positive plants were Las-positive. DNA extracted from individual late stage nymphs (3rd to 5th instars) collected from Las-infected trees were then subjected to nested-PCR trials. Twelve out of the 29 nymphs proved to be Las positive. All nymphs collected from Las-negative lemon trees were also Las-negative. Most psyllid samples which showed positive in nested-PCR were also positive in conventional-PCR detection. The amplified fragment of the 16S rRNA gene of 'Ca. Liberibacter spp.' from positive psyllid samples was 99% similar to those of Las strain psy62 in GenBank. These results demonstrate that Cacopsylla (Psylla) citrisuga is another Las carrier insect. Transmission studies are underway to determine whether Las-positive psyllids can transmit Las to healthy citrus.