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.     

Viruses in the northern potato-producing regions of Saudi Arabia

Agar double diffusion tests and, later, ELISAs were used to detect viruses associated with potato in 242 samples collected in 16 trips to Tabuk and Hail, northern regions of Saudi Arabia, in four consecutive growing seasons (autumn 1989, spring and autumn 1990 and spring 1991). Eleven different viruses were detected in Tabuk and 12 in Hail. The viruses detected in Tabuk were alfalfa mosaic (AMV), cucumber mosaic (CMV), tobacco mosaic (TMV), potato leaf roll (PLRV), tomato spotted wilt (TSWV), tobacco ringspot (TRSV) and potato A, M, S, X and Y. The same viruses, plus potato yellow dwarf (PYDV), were detected in Hail. AMV was most frequently and CMV least frequently detected in Tabuk, whereas in Hail the most and least common were PVA and PLRV respectively.     

Preliminary investigation of the causal agent(s) of a disease causing leaf curl of tobacco in South Africa

A high incidence (86%) of potyvirus infection was noted in tobacco plants exhibiting a form of leaf curl in South Africa. Despite leaf curl being reported in the literature to be of geminiviral aetiology, no geminiviruses were detected. Furthermore, no other virus particles were detected by virus purification, TEM and serology. Twelve species of dsRNA were consistently isolated from these tobacco plants, but were absent from other forms of leaf curl-affected and healthy tobacco. Aphid and mechanical inoculation demonstrated that the purified potyvirus(es) did not cause leaf curl symptoms, but rather mild mottle and mosaic symptoms in tobacco. Partial characterization of the potyvirus preparation showed a possible relationship to a South African strain of potato virus Y. Because potyvirus-inoculated plants did not manifest leaf curl symptoms, and because leaf curl symptoms were noted in some plants not infected with a potyvirus, it was concluded that the potyvirus is not involved in the leaf curl aetiology, but causes a latent infection, the symptoms of which are masked. The pattern of the dsRNA banding, induction of enations and lack of mechanical and seed transmission are common to plant reoviruses. The possibility of a phytoreovirus involvement in this form of leaf curl is currently being investigated. The results from this study suggest that tobacco leaf curl disease worldwide, with regard to geminiviruses, be re-evaluated.     

Effect of soil moisture content on the suppression of Rhizoctonia stem canker on potato by the nematode Aphelenchus avenae and the springtail Folsomia fimetaria

The effect of soil moisture content on the suppression of Rhizoctonia stem canker on potato by mycophagous soil animals was studied in growth chambers. Three soil moisture levels were established in two bioassays, in which potato sprouts grew through a 15-cm soil layer inoculated with sclerotia of Rhizoctonia solani (AG-3). In one experiment two levels of R. solani inoculum were applied. The effect on plant disease of mycophagous soil fauna was assessed by adding the springtail Folsomia fimetaria and/or the nematode Aphelenchus avenae to the soil. In the absence of mycophagous organisms, Rhizoctonia disease severity on potato stems was highest in dry soil. A. avenae and F. fimetaria reduced Rhizoctonia stem canker when applied at populations found in the field. They were effective over a broad range of soil moistures. The stimulatory effect of dry soil conditions on Rhizoctonia stem canker was counteracted by a greater efficacy of the mycophagous soil fauna under these conditions. Mild drought stress did not seem to be a limiting factor in the biological control of stem canker by these two organisms.     

Development of a powder formulation of Pseudomonas fluorescens for control of rice blast

Pseudomonas fluorescens strain Pf1, inhibitory to the growth of the rice blast pathogen Pyricularia oryzae in vitro , was developed as a talc-based powder formulation. When rice seeds were treated with this formulation, the bacteria spread to roots, stems and leaves of the plants and protected against leaf infection by P. oryzae . When applied as a foliar spray, the bacteria survived on the leaves. The powder formulation controlled leaf blast under greenhouse conditions. In tests as a seed treatment and foliar spray in four field trials it effectively controlled the disease and increased grain yield.     

The effect of solar heating of soil on natural and inoculated agrobacteria

Solarization trials were carried out over 3 years and in two countries to control crown gall disease on fruit trees and eliminate Agrobacterium . In 1992, agrobacteria in naturally infested soils of two Italian nurseries were monitored before and after solarization. Agrobacteria populations decreased by 99% and 92% after the treatment; however, crown gall incidence did not decrease. In 1993 and 1994 solarization was tested in Oregon in fields artificially infested with two marked strains of A. tumefaciens . In sandy loam soil, the target bacteria were eliminated in 4 weeks, while in silty clay soil the populations were markedly reduced after 2 months of treatment. Crown gall incidence on cherry rootstocks transplanted to the field at the end of 1993 was 3.7% in the sandy loam soil control plots, while no tumours were observed on plants from solarized plots. The use of solarization in combination with reduced doses of metham-sodium was also evaluated.     

Transmission of grapevine leafroll-associated closteroviruses by Pseudococcus longispinus and P. calceolariae

The transmission of two closteroviruses associated with grapevine leafroll, GLRaV-1 and GLRaV-3, from grapevine to grapevine by the mealybugs, Pseudococcus longispinus and P. calceolariae (Homoptera: Pseudococcidae) was studied. Controlled transmission experiments using the first and third instars of each insect were conducted twice during the 1993–94 growing season to investigate the consequence of virus accumulation within the donor vine leaf tissue on the incidence of virus transmission to healthy recipient vines. Transmission of GLRaV-1 and GLRaV-3 was determined by ELISA testing recipient vines in July 1994 and March 1995. GLRaV-3 was transmitted to recipient vines by P. longispinus and P. calceolariae first instars only. An increase in virus titre within the season did not significantly alter the transmission rate of GLRaV-3 by either P. longispinus or P. calceolariae first instars. P. longispinus and P. calceolariae failed to transmit GLRaV-1 to recipient vines.