The greening-causing agent alters the behavioral and electrophysiological responses of the Asian citrus psyllid to a putative sex pheromone.

The Asian Citrus Psyllid (ACP), Diaphorina citri, is a vector of the pathological bacterium Candidatus Liberibacter asiaticus (CLas), which causes the most devastating disease to the citrus industry worldwide, known as greening or huanglongbing (HLB). Earlier field tests with an acetic acid-based lure in greening-free, 'Valencia' citrus orange groves in California showed promising results. The same type of lures tested in São Paulo, Brazil, showed unsettling results. During the unsuccessful trials, we noticed a relatively large proportion of females in the field, ultimately leading us to test field-collected males and females for Wolbachia and CLas. The results showed high rates of Wolbachia and CLas infection in field populations. We then compared the olfactory responses of laboratory-raised, CLas-free, and CLas-infected males to acetic acid. As previously reported, CLas-uninfected males responded to acetic acid at 1 µg. Surprisingly, CLas-infected males required 50 × higher doses of the putative sex pheromone, thus explaining the failure to capture CLas-infected males in the field. CLas infection was also manifested in electrophysiological responses. Electroantennogram responses from CLas-infected ACP males were significantly higher than those obtained with uninfected males. To the best of our knowledge, this is the first report of a pathogen infection affecting a vector's response to a sex attractant.

Laboratory and field evaluation of acetic acid-based lures for male Asian citrus psyllid, Diaphorina citri.

The Asian citrus psyllid (ACP) is a vector of a pathogen associated with greening and thus a major problem in citriculture worldwide. Lures are much needed for improving ACP trapping systems for monitoring populations and surveillance. Previously, we have identified acetic acid as a putative sex pheromone and measured formic acid- and propionic acid-elicited robust electroantennographic responses. We have now thoroughly examined in indoor behavioral assays (4-way olfactometer) and field tests the feasibility of these three semiochemicals as potential lures for trapping ACP. Formic acid, acetic acid, and propionic acid at appropriate doses are male-specific attractants and suitable lures for ACP traps, but they do not act synergistically. An acetic acid-based homemade lure, prepared by impregnating the attractant in a polymer, was active for a day. A newly developed slow-release formulation had equal performance but lasted longer, thus leading to an important improvement in ACP trap capture at low population densities.

Putative sex pheromone of the Asian citrus psyllid, Diaphorina citri, breaks down into an attractant.

Under laboratory conditions, mating activity in Asian citrus psyllid (ACP) started 4 days after emergence, peaked at day 7, and showed a clear window of activity starting 8 h into the photophase and extending through the first hour of the scotophase. We confirmed that ACP males are attracted to emanations from conspecific females. Traps loaded with a candidate compound enriched with female extract, lignoceryl acetate (24Ac), at various doses were active only after being deployed for several weeks in the field, suggesting that a degradation product, not the test compound, was the active ingredient(s). Lignocerol, a possible product of 24Ac degradation, was not active, whereas acetic acid, another possible degradation product, was found in the airborne volatile collections from lures matured under field conditions and detected in higher amounts in volatiles collected from females at the peak of mating activity than in male samples. Acetic acid elicited dose-dependent electroantennographic responses and attracted ACP males, but not females, in Y-type and 4-way olfactometers. Field tests showed that acetic acid-baited traps captured significantly more males than control traps. Surprisingly, captures of females in acetic acid-baited traps were also higher than in control traps, possibly because of physical stimuli emitted by captured males.