Epidemiological survey of Phthorimaea operculella granulosis virus in Tunisia

In Tunisia, Phthorimaea operculella is an important pest in traditional, unrefrigerated potato stores. An integrated control approach including the use of biological control agents and specifically insect viruses is being developed. From this perspective, we carried out a screening and an evaluation of local Phthorimaea operculella granulosis virus naturally present in Tunisian pest populations. On the basis of symptomatology, P. operculella larvae were collected from different regions (North, Cap Bon, Sahel). Indirect ELISA using a serum raised against the purified Tunisian isolate of the virus was used to establish the geographical distribution of the virus. Comparison of the two approaches showed that symptomatology is not so virus-specific, as morphologically healthy insects were sometimes ELISA-positive. The serological technique therefore gives better detection of the virus.     

Estimating the potential development of Diaporthe helianthi epidemics in Italy*

Diaporthe helianthi is the causal agent of a severe sunflower disease but, in Italy, disease outbreaks are sporadic with no significant losses. The present work investigates the role of meteorological conditions on the potential development of D. helianthi epidemics in Italy, using the French model Asphodel, which simulates the effect of air temperature, relative humidity and rainfall on ascospore maturation and dispersal, infection establishment, disease onset and severity during the period of host susceptibility. Meteorological data measured in eight stations distributed from north to south Italy, over a 5‐year period (1995–99), was used as model input. Results showed that meteorological conditions in Italy are frequently favourable for D. helianthi infections on sunflower, and severe epidemics are possible. Therefore, climatic conditions are not a limiting factor for disease development in the Italian sunflower‐growing areas. The lack of disease epidemics in Italy may be related to differences in the pathogen populations compared with the French ones.     

A coat protein transgene from a Scottish isolate of potato mop-top virus mediates strong resistance against Scandinavian isolates which have similar coat protein genes

Resistance tests were made on seedlings of transformed lines of Nicotiana benthamiana which contain a transgene encoding the coat protein (CP) gene of a Scottish isolate of potato mop-top virus (PMTV). This transgene has been reported to confer strong resistance to the PMTV isolate from which the transgene sequence was derived and also to a second Scottish isolate. Plants of lines of the transgenic N. benthamiana were as resistant to two Swedish and two Danish PMTV isolates as to a Scottish isolate, and of five lines tested, greater than 93.5% of transgenic plants were immune. The coat protein gene sequences of these four Scandinavian isolates were very similar to those of the two Scottish isolates. The greatest divergence between the isolates was three amino acid changes and there was less than 2% change in CP gene nucleotide sequence. It is concluded that the PMTV CP transgene used in these experiments could confer resistance against isolates from different geographical areas because it is becoming apparent that the CP genes of PMTV isolates are highly conserved.     

Production and characterization of a monoclonal antibody specific to the M serotype of plum pox potyvirus

A monoclonal antibody to an Albanian isolate of plum pox potyvirus (PPV) was obtained (MAbAL), that specifically recognized strain M of this virus. The specificity of MAbAL, assessed by comparative ELISA on 130 PPV isolates of different geographical origin, 22 of which were also tested by comparative IC-PCR, gave consistent and highly reproducible results. MAbAL seems to be elicited by a stable surface determinant that makes it particularly suitable for successful use under a wide range of conditions. MAbAL is an useful addition to the panel of PPV-specific MAbs available to date.     

Non-structural plum pox potyvirus proteins detected by immunogold labelling

Plum pox potyvirus (PPV) induces in infected Nicotiana clevelandii cells characteristic crystalline inclusions known as nuclear inclusions (NI) when located in the nucleus and as dense material (Dm) when located in the cytoplasm. Crystalline inclusions contain protease (NIa) and RNA-dependent RNA polymerase (NIb) proteins. It is now well established for all potyviruses that cylindrical inclusions contain CI helicase ATPase protein (Martin et al., 1992). The intracellular location of other non-structural PPV proteins remains unknown. Using Escherichia coli expression vectors, specific antibodies were obtained against P1, P3, 6K2 and NIb PPV proteins for which antibodies were not yet available. As expected, NIb antiserum labelled crystalline inclusions. P1, P3 and 6K2 proteins were present in both types of crystalline inclusions found in the nucleus and in the cytoplasm of PPV-infected leaves of N. clevelandii, suggesting that nuclear inclusions and dense material were composed of the same proteins. This composition is discussed.     

Frequency of transmission of artichoke Italian latent nepovirus by Longidorus fasciatus (Nematoda: Longidoridae) from artichoke fields in the Iria and Kandia areas of Argolis in northeast Peloponnesus, Greece

Artichoke Italian latent nepovirus (AILV) transmitted by Longidorus fasciatus is a causative agent of artichoke patchy chlorotic stunting (APCS) in northeast Peloponnesus in southern Greece. Populations of L. fasciatus collected from the Iria and Kandia areas of Argolis in northeast Peloponnesus were used in laboratory experiments to determine the frequency of transmission of the virus by its natural vector. One tenth to almost one half of the L. fasciatus specimens recovered from soil collected in two artichoke fields showing APCS transmitted AILV. Allowing nematodes access for 4 wk to Nicotiana clevelandii mechanically infected with AILV did not increase the number of individual specimens able to transmit virus. The total number of specimens transmitting virus in an experiment did not exceed fifty percent of the individuals tested. Virus-like particles were only observed adsorbed to the inner surface of the odontostyle and it is suggested that the high frequency of transmission of AILV by L. fasciatus is a result of efficient dissociation of virus particles from the specific sites of retention in the vector.