Fungi associated with esca disease in grapevine

Cross sections of woody stems of 309 diseased grapevines collected in France showed two kinds of necrosis typical of esca: a) A central light-colored necrosis of soft consistency, consisting of three zones, preceded by a centrally discolored wood, and b) a sectorial light-colored necrosis composed of two zones preceded by a sectorial brown necrosis. Isolations showed that different microflora was associated with each necrosis. Phaeoacremonium aleophilum and Phaeoacremonium chlamydosporum occurred in the discolored wood and the zones bordering the central decayed wood. Eutypa lata was the main fungus isolated from sectorial brown necrosis and the zones adjoining the decay wood. Phellinus punctatus was isolated from the sectorial and central decayed wood. Stereum hirsutum was present in decayed wood of 15 grapevines with esca symptoms not inhabited by P. punctatus. Wood decay tests and pathogenicity tests showed that S. hirsutum and P. punctatus were responsible for the decayed wood. Phaeoacremonium chlamydosporum and S. hirsutum produced a centrally discolored wood similar to that found in esca-affected vines. Phaeoacremonium aleophilum caused a sectorial brown necrosis of soft texture. From these studies, it was found that esca is a complex disease involving several microorganisms whose role in the process leading to wood degradation is discussed.     

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.     

Comparison of an in vitro and a damping-off assay to test soils for suppressiveness to Pythium aphanidermatum

Testing of soil samples in greenhouse assays for suppressiveness to soilborne plant pathogens requires a considerable investment in time and effort as well as large numbers of soil samples. To make it possible to process large numbers of samples efficiently, we compared an in vitro growth assay with a damping-off assay using Pythium aphanidermatum as the test organism on tomato seedlings. The in vitro test compares the radial growth or relative growth of the fungus in soil to that in autoclaved soil and reflects suppressiveness of soils to the pathogen. We used soils from a field experiment that had been farmed either organically or conventionally and into which a cover crop (oats and vetch in mixture) had been incorporated 0, 10, 21, and 35 days previously. We obtained a significant, positive correlation between damping-off severities of tomato seedlings in damping-off assays and both relative and radial growth in vitro. In addition, radial and relative growth of P. aphanidermatum in the in vitro assay were positively correlated with several carbon and nitrogen variables measured for soil and incorporated debris. We did not find differences between the two farming systems for either growth measures of P. aphanidermatum or disease severities on tomato at different stages of cover crop decomposition. The in vitro assay shows potential for use with any fungus that exhibits rapid saprophytic growth, and is most suitable for routine application in suppressiveness testing.     

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.     

Analytical models of weed biocontrol with sterilizing fungi: the consequences of differences in weed and pathogen life-histories

A model of the population dynamics of healthy weed plants, weed seeds in the soil, pathogen-infected weed plants and pathogen spores in the soil, was devised to investigate interactions that are important for the success of biocontrol with pathogens that prevent seed set. Three particular features of the host-pathogen interaction were examined in detail: the form of the density dependent relationship which determined seed and spore production, the host life stage at which infection could occur, and the relative competitive abilities of healthy and infected host plants. It was found that, when both weed and pathogen coexist, the equilibrium abundance of the weed in the presence of the pathogen was independent of the form of the relationship between plant density and seed or spore production. However, the form of this relationship did affect estimated equilibrium densities in the absence of biocontrol, and also affected the parametrizations under which both host and pathogen could coexist. Parameters derived from experiments with isolated host plants may therefore be sufficient to assess the biocontrol potential of new pathogens along with knowledge of densities achievable when the weed is uncontrolled. The form of the relationship used to control seed and spore production also had a marked influence on the range of parameter values over which the pathogen could persist. Other control measures were represented by changes to appropriate parameter values, e.g. weeding was represented by a change in weed death rate. With one exception, the use of additional control measures was not antagonistic to biocontrol. Often, however, the combined effect was less than additive, and the existence of synergy (where the combined effects are more than additive) was critically dependent on the form of the relationship of the rate of seed production per plant with density and the efficiency of the other control measures.     

A model of plant virus disease epidemics in asynchronously-planted cropping systems

A mathematical model was developed of the dynamics of a plant virus disease within a spatially-referenced lattice of fields of a host crop. The model can be applied to crops in continuous, contiguous cultivation such as tropical irrigated rice. Disease progress in each field of the host crop was assumed to be logistic and determined by incidence within the field itself as well as incidence in neighbouring fields, depending on the gradient of disease spread. The frequency distribution of planting dates (represented by the proportion of the total number of fields planted in successive months) was assumed to follow a normal distribution and the variance of planting date was used as a measure of cropping asynchrony. Analysis of the model revealed that disease incidence within the lattice (i.e. mean incidence over all fields) depended upon the infection efficiency, the slope of the dispersal gradient, and the variance in planting date. Disease endemicity depended mainly on planting date variance and disease persisted in the lattice if this variance exceeded a certain threshold. Above the threshold for persistence, the response of mean disease incidence to planting date variance was non-linear and the region of greatest sensitivity was closest to the threshold. Thus, disease systems that show moderate rather than high cropping asynchrony are more likely to be influenced by changes in the variance of planting date. Implications for the area-wide management of rice tungro virus disease are discussed.     

Role of acquisition and inoculation time in the expression of clonal variation for BYDV-PAV transmission in the aphid species Rhopalosiphum padi

Vector efficiency of 20 Rhopalosiphum padi clones, originating from Europe, North America and North Africa and exhibiting different types of life cycle, was evaluated by transmitting a French BYDV-PAV isolate to barley plants under five different acquisition/inoculation sequences (AAP/IAP). Differences between clones in transmission efficiency were found only when a short AAP was followed by a long IAP (6 h/120 h) and, to some extent, when a long AAP (48 h) was followed by a short IAP (6 h), but no differences were found when the conditions for virus transmission were optimal, i.e. long AAP followed by long IAP (48 h/120 h). There were no differences in transmission rates by clones of different geographical origins and with different life cycles. As a consequence, clonal variation is probably of little importance in the vector aspects of the epidemiology of PAV serotypes transmitted by R. padi , but the availability of a range of clones exhibiting transmission differences under limiting AAP or IAP conditions could be of interest for studies of virus–vector relationships.     

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.     

The effect of component number on Rhynchosporium secalis infection and yield in mixtures of winter barley cultivars

Mixtures of winter barley cultivars containing up to six components were grown over three years with and without fungicide treatment. Yield increases were recorded for mixtures compared with the mean of their monoculture components and there was a significant trend towards greater benefit from increased number of components. These benefits were partially attributable to a corresponding increase in control of Rhynchosporium secalis as component number increased. The potential for exploitation of mixtures in cereals for control of splash-dispersed pathogens is discussed.     

Transfer from Erwinia herbicola to Escherichia coli of a plasmid associated with biocontrol of fire blight

Strains of Erwinia herbicola effective in the biocontrol of fire blight of hawthorn were used to investigate the possibility that the antagonistic activity is coded by plasmid-born genes. Agarose gel electrophoresis of isolated plasmids from four antagonistic Erw. herbicola strains showed a band of a supercoiled 12 kb plasmid in each strain, with a second band greater than 16.2 kb consistently seen in two strains. Erw. herbicola strains showed resistance to penicillin-G, which could be conferred on penicillin-G sensitive Escherichia coli TG1 by transformation with a pure Erw. herbicola plasmid preparation. Transformed strains of Esc. coli appeared to contain the Erw. herbicola 12 kb plasmid, but not the > 16.2 kb plasmid. In an agar plate assay, Esc. coli transformants produced an inhibition zone against Erw. amylovora similar to those produced by the original Erw. herbicola strains. In two biocontrol assays, the transformed Esc. coli strains had a suppressive effect on disease development on infected pear fruit slices and hawthorn blossoms.