Beth Grafton-Cardwell and her team at Lindcove - Citrus Research ...

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24 Citrograph July/August 2011 CRB Funded Research Reports 2010 Research Project Progress Report Identification of odor-based lures and repellents for the Asian citrus psyllid The Asian citrus psyllid (ACP) is known to transmit a bacterium called Candidatus Liberibacter asicaticus, which causes the deadly citrus greening disease (also known as huanglongbing (HLB)) around the world. This psylllid has recently been found in areas of Southern California, which is a major cause of concern for the citrus industry here. The adult psyllids feed on citrus plants, and females gather in numbers and lay eggs on the young leaves. In order for a psyllid to spread the disease, it needs to first find and feed on an infected plant to pick up the bacteria and subsequently find and feed on a second plant, thereby inoculating the tree. Like many other insects, the ACP relies heavily on the sense of smell to find the plants. The olfactory system provides an ideal target to design methods to interfere with the ability of the insect to smell and therefore disrupt its ability Anandasankar Ray, Lisa Forster and Robert Luck to find host plants and subsequently transmit the bacterium. In our project we are using sophisticated methods to study the olfactory system in ACP to find odors that can be used to block the insect from finding citrus plants. These methods allow us to precisely measure the signals generated in the nose of the psyllid, called the antenna, when it comes in contact with plant odors. In the first part of the project, we are interested in finding a highly effective odor lure that can be used to attract ACP effectively to traps for surveillance or to larger traps for population control. In the second part of the project, we are interested in finding odors that ACP avoid strongly and can therefore be effective in repelling ACP from citrus plants and groves. All of our experiments are currently being performed in the confines of the secure quarantine facility at the Uni- Fig. 1. Photograph of a single-unit electrophysiology apparatus that is used to measure activity of the ACP antennal cells. versity of California Riverside. We have established a disease-free rearing facility inside the quarantine facility that supplies us with healthy adult ACP to perform our experiments. Inside this facility we take several precautions to contain the insects securely, such as cages with double walls, tightly sealed containers, and careful handling of insects by highly trained personnel. After setting up a healthy colony, we embarked upon the process of setting up the sensitive instrumentation inside the quarantine facility, which would enable us to identify the various lures and repellent odors for ACP. Specialized equipment has been designed to perform behavioral testing of psyllids at high-throughput to test for attraction and repellency. These assays will enable us to quickly and reliably identify odors that can act as attractants or repellents. We believe that ours is the first and only single-unit electrophysiology instrument (Figure 1.) that is currently used to study the ACP antenna in close detail. Once the instrumentation was set up and an appropriate protocol was established, we started testing compounds that are potential candidates for attractants. Tiny hairs which house sensory cells called neurons cover the surface of the psyllid antenna. Along with the hairs, the antennae have pit-like structures that also house sensory cells (Figure 2). Odor-detecting cells are found inside every pit-like structure, however only a few hair-like structures contain them. Since very little is known about the different types of sensory cells, we first asked which of these cells on the antenna are involved in the detection of odors and act as sensors for the insect. We were able to identify several cells on the antenna that specifically respond
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