The Major Antigenic Membrane Protein of “Candidatus Phytoplasma asteris” Selectively Interacts with ATP Synthase and Actin of Leafhopper Vectors

Phytoplasmas, uncultivable phloem-limited phytopathogenic wall-less bacteria, represent a major threat to agriculture worldwide. They are transmitted in a persistent, propagative manner by phloem-sucking Hemipteran insects. Phytoplasma membrane proteins are in direct contact with hosts and are presumably involved in determining vector specificity. Such a role has been proposed for phytoplasma transmembrane proteins encoded by circular extrachromosomal elements, at least one of which is a plasmid. Little is known about the interactions between major phytoplasma antigenic membrane protein (Amp) and insect vector proteins. The aims of our work were to identify vector proteins interacting with Amp and to investigate their role in transmission specificity. In controlled transmission experiments, four Hemipteran species were identified as vectors of “Candidatus Phytoplasma asteris”, the chrysanthemum yellows phytoplasmas (CYP) strain, and three others as non-vectors. Interactions between a labelled (recombinant) CYP Amp and insect proteins were analysed by far Western blots and affinity chromatography. Amp interacted specifically with a few proteins from vector species only. Among Amp-binding vector proteins, actin and both the α and β subunits of ATP synthase were identified by mass spectrometry and Western blots. Immunofluorescence confocal microscopy and Western blots of plasma membrane and mitochondrial fractions confirmed the localisation of ATP synthase, generally known as a mitochondrial protein, in plasma membranes of midgut and salivary gland cells in the vector Euscelidius variegatus. The vector-specific interaction between phytoplasma Amp and insect ATP synthase is demonstrated for the first time, and this work also supports the hypothesis that host actin is involved in the internalization and intracellular motility of phytoplasmas within their vectors. Phytoplasma Amp is hypothesized to play a crucial role in insect transmission specificity

Disruption of a Gene Predicted To Encode a Solute Binding Protein of an ABC Transporter Reduces Transmission of Spiroplasma citri by the Leafhopper Circulifer haematoceps

Spiroplasma citri is transmitted from plant to plant by phloem-feeding leafhoppers. In an attempt to identify mechanisms involved in transmission, mutants of S. citri affected in their transmission must be available. For this purpose, transposon (Tn4001) mutagenesis was used to produce mutants which have been screened for their ability to be transmitted by the leafhopper vector Circulifer haematoceps to periwinkle plants. With one mutant (G76) which multiplied in leafhoppers as efficiently as S. citri wild-type (wt) strain GII-3, the plants showed symptoms 4 to 5 weeks later than those infected with wt GII-3. Thirty to fifty percent of plants exposed to leafhoppers injected with G76 remained symptomless, whereas for wt GII-3, all plants exposed to the transmission showed severe symptoms. This suggests that the mutant G76 was injected into plants by the leafhoppers less efficiently than wt GII-3. To check this possibility, the number of spiroplasma cells injected by a leafhopper through a Parafilm membrane into SP4 medium was determined. Thirty times less mutant G76 than wt GII-3 was transmitted through the membrane. These results suggest that mutant G76 was affected either in its capacity to penetrate the salivary glands and/or to multiply within them. In mutant G76, transposon Tn4001 was shown to be inserted into a gene encoding a putative lipoprotein (Sc76) In the ABCdb database Sc76 protein was noted as a solute binding protein of an ABC transporter of the family S1_b. Functional complementation of the G76 mutant with the Sc76 gene restored the wild phenotype, showing that Sc76 protein is involved in S. citri transmission by the leafhopper vector C. haematoceps

Clay-clast aggregates: a new structural evidence for seismic sliding?

International audienceTo determine the processes responsible for slip-weakening in clayey gouge zones, rotary-shear experiments were conducted at seismic slip rates (equivalent to 0.9 and 1.3 m/s) at 0.6 MPa normal stress on a natural clayey gouge for saturated and non-saturated initial conditions. The mechanical behavior of the simulated faults shows a reproducible slip-weakening behavior, whatever initial moisture conditions. Examination of gouge obtained at the residual friction stage in saturated and non-saturated initial conditions allows the definition of two types of microstructures: a foliated type reflecting strain localization, and a non-foliated type composed of spherical aggregates. Friction experiments demonstrate that liquid-vapor transition of water within gouge due to frictional heating has a high capacity to explain the formation of spherical aggregates in the first meters of displacement. This result suggests that the occurrence of spherical aggregates in natural clayey fault gouges can constitute a new textural evidence for shallow depth pore water phase transition at seismic slip velocity and consequently for past seismic fault sliding

Pouvoir pathogène des mollicutes: du modèle Spiroplasma citri ... aux phytoplasmes

National audienc

Pouvoir pathogène des mollicutes: du modèle Spiroplasma citri ... aux phytoplasmes

National audienc

Late Quaternary co-seismic sedimentation in the Sea of Marmara's deep basins

25 pages, 15 figures, 1 tableauInternational audienceThe deep, northern, part of the Sea of Marmara (northwestern Turkey) is composed of several aligned, actively subsiding, basins, which are the direct structural and morphological expression of the North Anatolian Fault's northern branch. The last 20 kyr of their sedimentary fill (lacustrine before 12 kyr BP) have been investigated through giant piston coring onboard R/V MARION-DUFRESNE (MARMACORE Cruise, 2001) and chirp subbottom profiler onboard R/V ATALANTE during MARMARASCARPS Cruise (2002). Especially during the lacustrine stage, the infilling of the deep basins (Tekirda?, Central, Kumburgaz, and Çinarcic Basins; up to 1250 m depth) was dominated by turbidites (with coarse mixed siliciclastic and bioclastic basal part), intercalated in “hemipelagic-type” finegrained calcareous and slightly siliceous clays. Often – especially in the thickest ones – the turbidites show strong segregation and a sharp boundary between coarse part and suspendedload part. In the Central Basin, 8 m of a unique sedimentary event include a 5 to 8m-thick “homogenite” well imaged on seismic profiles. The latter is interpreted as related to a major – possibly triggered - tsunami effect, as described in the Eastern Mediterranean by Kastens and Cita (1981). In the marine (Holocene) upper part of the sedimentary fill, repeated to-and-from structures, affecting silt or fine sand, are evidencing seiche-like effects and, thus, earthquake triggering. Detailed correlations between two deep coring sites (1250 and 1200 m) indicate more than 100 % overthickening in the deepest one; this implies specific processes of distribution of terrigenous input by dense hyperpycnal currents (high kinetic energy, seiche effect, complex reflections on steep slopes). The peculiar sedimentary infilling of the Sea of Marmara's Central Basin (and, by extrapolation, of the whole set) is tentatively interpreted as a direct consequence of the strong seismic activity; the imprint of the latter is more obvious prior to the base of the Holocene, as environmental conditions favoured marginal accumulation (especially on the southern shelf) of large amounts of erosion products available for mass wasting