Phytoplasmas are plant pathogenic bacteria transmitted by hemipteran phloem sucking insects in a persistent, propagative manner. Recent studies on 'Candidatus Phytoplasma asteris' showed that the major antigenic membrane protein (Amp) specifically interacts with vector proteins, both both cytosolic (actin and myosin) and membrane (ATP synthase) localization (Suzuki et al., 2006; Galetto et al., 2011). Following acquisition, phytoplasmas must cross the gut epithelium and colonize salivary glands for successful transmission. The gut epithelium and salivary glands are physical barriers that must be overcome by the phytoplasma during vector colonization. Specific interactions of Amp with vector membrane proteins occur in vitro at both sites. Aim of this work was to develop a system to study the effects of phytoplasma membrane proteins on vector acquisition and transmission capabilities. To investigate the biological effects of the interactions of CYP Amp at the gut epithelium, vectors of two species (Macrosteles quadripunctulatus Kirschbaum, and Euscelidius variegatus Kirschbaum), before acquisition on CYP infected plants, were fed on a medium containing i) a fusion construct of the protein, ii) a fusion protein of ArtI, a membrane protein not involved in specific interaction with vectors, iii and iv) antibodies raised against Amp and ArtI, v) no proteins. After latency, insects were singly caged on healthy plants for an inoculation period after which they were collected and assayed in nested PCR to detect the presence of CYP. Phytoplasma titre in insects was also quantified by RT-PCR. To determine the role of CYP membrane proteins at the salivary gland level, abdominal microinjection experiments with a suspension of CY phytoplasmas added with Amp fusion protein or Amp and its specific antibodies are in progress. The results of preliminary experiments are presented and discussed.
Trasmissione dei fitoplasmi mediante insetti vettori: metodi per lo studio in vivo del ruolo delle proteine di memebrana di chrysanthemum yellows phytoplasma
Rashidi M;Galetto L;Bosco D
2013
Abstract
Phytoplasmas are plant pathogenic bacteria transmitted by hemipteran phloem sucking insects in a persistent, propagative manner. Recent studies on 'Candidatus Phytoplasma asteris' showed that the major antigenic membrane protein (Amp) specifically interacts with vector proteins, both both cytosolic (actin and myosin) and membrane (ATP synthase) localization (Suzuki et al., 2006; Galetto et al., 2011). Following acquisition, phytoplasmas must cross the gut epithelium and colonize salivary glands for successful transmission. The gut epithelium and salivary glands are physical barriers that must be overcome by the phytoplasma during vector colonization. Specific interactions of Amp with vector membrane proteins occur in vitro at both sites. Aim of this work was to develop a system to study the effects of phytoplasma membrane proteins on vector acquisition and transmission capabilities. To investigate the biological effects of the interactions of CYP Amp at the gut epithelium, vectors of two species (Macrosteles quadripunctulatus Kirschbaum, and Euscelidius variegatus Kirschbaum), before acquisition on CYP infected plants, were fed on a medium containing i) a fusion construct of the protein, ii) a fusion protein of ArtI, a membrane protein not involved in specific interaction with vectors, iii and iv) antibodies raised against Amp and ArtI, v) no proteins. After latency, insects were singly caged on healthy plants for an inoculation period after which they were collected and assayed in nested PCR to detect the presence of CYP. Phytoplasma titre in insects was also quantified by RT-PCR. To determine the role of CYP membrane proteins at the salivary gland level, abdominal microinjection experiments with a suspension of CY phytoplasmas added with Amp fusion protein or Amp and its specific antibodies are in progress. The results of preliminary experiments are presented and discussed.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
