Does aphid salivation affect phloem sieve element occlusion in vivo?

To protect against loss of photo-assimilate-rich phloem sap, plants have evolved several mechanisms to plug phloem sieve tubes in response to damage. In many Fabaceae, each sieve element contains a discrete proteinaceous body called a forisome, which, in response to damage, rapidly transforms from a condensed configuration that does not impede the flow of sap to a dispersed configuration that plugs the sieve element. Aphids and other specialized phloem sap feeders can ingest phloem sap from a single sieve element for hours or days, and to do this, they must be able to suppress or reverse phloem plugging. A recent study provided in vitro evidence that aphid saliva can reverse forisome plugs. The present study tested this hypothesis in vivo by inducing forisome plugs which triggered aphids to switch behaviour from phloem sap ingestion to salivation into the sieve element. After salivating into the sieve element for various periods of time, the aphids were instantaneously cryofixed (freeze fixed) in situ on their leaf. The state of the forisome was then determined in the penetrated sieve element and in nearby non-penetrated sieve elements which served as controls for sieve elements not subjected to direct aphid salivation. Forisomes were almost always in close contact with the stylet tips and thus came into direct contact with the saliva. Nonetheless, forisome plugs in the penetrated sieve element did not revert back to a non-plugging state any faster than those in neighbouring sieve elements that were not subjected to direct aphid salivation.