Seasonal progression of sex ratio and phytoplasma infection in Scaphoideus titanus Ball (Hemiptera: Cicadellidae).

The differences between the seasonal occurrence and likelihood of being infected by FD phytoplasmas, of male and female Scaphoideus titanus Ball, were investigated. Sex ratio (male: female) was calculated by counting males and females sampled by means of yellow sticky traps and sweep-nets and from adults derived from hatched eggs in field-collected grapevine wood. PCR essays were performed to test differences in infection between genders. Overall, the sex ratio on sticky traps was significantly more male biased (1.99:1) if compared to net sweeping (0.62:1) and laboratory rearing (0.60:1). The peak of male presence was recorded in the middle of July in laboratory rearing and sweep net, and in the middle of August on sticky traps; the maximum presence of females was detected at the end of July in laboratory rearing, and at the end of August in sweep net samplings and on sticky traps. The seasonal sex ratio was more male biased at the beginning in laboratory rearing (1.50:1) and sticky traps (9:1), and then decreased in favor of females at the end of the sampling period, both in laboratory rearing (0.17:1) and in sticky traps (0.07:1). This trend was significantly less skewed, although similar, in sweep net samplings that recorded a sex ratio of 1:1 and 0.16:1 at the beginning and at the end of the sampling period, respectively. Concerning phytoplasma detection, an interaction between gender and sampling period was observed, the males showing a peak of infected individuals later in the season (35%). Some possible behavioral explanations of the data obtained are given.

Multiple symbiosis in the leafhopper Scaphoideus titanus (Hemiptera: Cicadellidae): details of transovarial transmission of Cardinium sp. and yeast-like endosymbionts.

Scaphoideus titanus is the insect vector of flavescence dorée (FD), a yellow disease of grapevines. Observations on adult females and nymphs of S. titanus showed that this insect is associated with a complex microbial community. Ultrastructural analysis showed that the fat body, salivary glands and ovary of the insect harbour microorganisms showing the brush-like structure typically observed in the genus Cardinium. In particular, it has been shown that these symbiotic bacteria are present both in the follicular cells and in the eggs. In addition, cells resembling bacteriocytes, harbouring numerous Cardinium symbionts in the cytoplasm, were observed in the apical portion of the ovary in adult females. These cells are likely responsible for bacterial transmission to the ovary. Optical microscopy showed that the fat body harbours an enormous population of yeast-like symbionts (YLSs). Ultrastructural observations showed that these symbionts are enclosed within specialized cells of the fat body and are also present in the ovary, where they are found in both the follicular cells and the eggs. There is thus evidence that both Cardinium and the YLSs are transovarially transmitted to the offspring. To our knowledge, S. titanus is the sole insect known to transmit two different kinds of symbionts to the eggs, a prokaryote and an eukaryote. Gene sequence analysis and in situ hybridization led to the identification of YLSs as members of the class Sordariomycetes (=Pyrenomycetes). Finally, ultrastructural observation of the midgut content revealed the presence, in both adult females and nymphs, of a complex microbial community, which include a phytoplasma-like microorganism, likely the agent of FD.

Ultrastructure of a novel Cardinium sp. symbiont in Scaphoideus titanus (Hemiptera: Cicadellidae).

An ultrastructural study of the novel symbiont Cardinium sp. was performed with particular attention to the description of the structure and organization of highly elaborated cytoplasmic complexes containing microtubule-like elements (MLC). Three major components were observed. The first was a system of microtubule-like elements (ML) arranged in parallel array extending from the plasma membrane into the cytosol of the bacterium. The second, an fibrous electrondense plaque (FEP), approximately 8 nm thick, located 7.5 nm away from the plasma membrane and parallel to it. The third component, not previously reported, was described for the first time in this paper. This consisted of a set of regularly distributed 8 nm electron-dense structures (ES), with a center-to-center spacing of about 12 nm, adhering to the outer leaflet of the plasma membrane. Often, the ES created a close connection between the plasma membrane and the outer membrane, so that in this area they became straight and stiff. The first and second component of these structures are compared to previously described microtubules and microfilaments.