Functional analysis of the 3′ end of avrBs3/pthA genes from two Xanthomonas species

The phytopathogens Xanthomonas oryzae pathovar (pv.) oryzae and Xanthomonas axonopodis pv. citri each contain several avrBs3/pthA family genes. Structural features of these genes important for avirulence and/or virulence functions include a central region of multiple direct repeats and three nuclear localization signals (NLSs) and an acidic activation domain (AAD) at the 3′ end. To identify other regions critical to function in the 3′ ends of these genes, we constructed several chimeras using apl1 and apl2 from X. axonopodis pv. citri and avrXa10 and avrXa7 from X. oryzae pv. oryzae and evaluated their functions by inoculation to citrus and rice. The apl1 and avrXa7 genes are major virulence determinants in citrus and rice, respectively, while the contributions of apl2 and avrXa10 to virulence are negligible or not measurable. Constructs that contained a 417 bp HincII-SphI fragment from the 3′ end of apl1 in combination with the repeats from avrXa7, avrXa10, and apl1 caused a canker phenotype on citrus. Interchange of the HincII-SphI fragment between avrXa7 and avrXa10 abolishes avrXa7 avirulence function and reduces its virulence but it does not affect avrXa10 avirulence function in rice. avrXa7 caused a hypersensitive response (HR) in citrus and replacement of it's 3′ end with that of apl1 resulted in loss of canker and induction of HR. Thus, the HincII-SphI fragment of the avrBs3/pthA gene family is important for avirulence and virulence functions in two different plant species, Oryza sativa and Citrus natsudaidai HAYATA.     

The organophosphorous insecticide chlorpyrifos affects the neuroepithelial junction, the bioelectric parameters of the skin of the frog Caudiverbera caudiverbera, and the structure of model cell membranes

The current study examines the acute effects of the organophosphorus pesticide chlorpyrifos on a sympathetic synapse of the frog Caudiverbera caudiverbera. Nerve stimulation was followed immediately by a transient increase in the short-circuit current (SCC) and in the potential difference (PD), which consisted of a rapid and then a slow component. Chlorpyrifos concentrations from 5 μM to 1.0 mM caused a dose-dependent block of both components to a 10% of their control values, which was reversed by washout. The pesticide blockade did not affect the skin response to noradrenaline. X-ray diffraction and fluorescence spectroscopy studies on membrane models showed marked phospholipid perturbation, which favors changes in ion channel conformation and interferes with receptor proteins, thus altering noradrenergic transmission and Cl⁻ secretion in the mucous glands. The foregoing results may be interpreted as a reversible inhibition of the neuroepithelial synapse to nerve stimulation, possibly due to non-specific lip μd-protein perturbation, interference with synaptic transmission, and transient Cl⁻ channel inactivation.     

Decision support systems used in the Netherlands for reduction in the input of active substances in agriculture

Since the middle of the 1980s, Dutch farmers have been using decision support systems (DSS) as an aid in the control of pests. This started with EPIPRE, then weather-related potato blight warning systems were developed (Prophy and Plant-Plus). In the 1990s, many weather-based DSS were developed against pests of orchards, flower bulbs, arable crops and field-produced vegetables. Also, a DSS was developed to predict and check the effect of meteorological conditions on the effectiveness of application timing of plant protection products (GEWIS). The use of these systems resulted in more sustainable crop protection: sustainable because the use of DSS led to a lower risk of crop damage and, in many cases, to a lower input of active substances, by optimization of the product and dose to actual phytosanitary and meteorological conditions. The use of GEWIS to ensure application at the right time of day further reduced the input of active substances and increased efficacy.     

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.     

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.     

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.     

Studies on the potential use of Pasteuria penetrans as a biocontrol agent of root-knot nematodes (Meloidogyne spp.)

Some aspects of the interaction of the bacterial parasite Pasteuria penetrans and the root-knot nematode ( Meloidogyne spp.) were investigated in laboratory and pot experiments. The variable spore attachment on juveniles exposed to water suspensions of the bacterium is probably attributed to differential susceptibility of biotypes within a heterogeneous Meloidogyne population. The relationship between spore concentration and attachment level is not linear over a range of spore dosages, indicating that even at very high spore concentrations the number of spores capable of attachment may not be present in excess and it is difficult to ensure sufficient numbers of spores to ensure infection will attach to all nematodes. Attempts to apply the bacterium in conditions such as might occur in seedbeds did not suppress nematode multiplication after transplanting in nematode-infested soil, indicating that the only effective application method is a thorough spore distribution in the planting sites. Two major constraints were revealed: high levels of spore attachment to juveniles does not always guarantee a significant reduction of egg laying and this is greatly influenced by the Meloidogyne biotype. Furthermore, the cumulative effect of the parasite in reducing Meloidogyne populations over several crop cycles was less than expected as the bacterium reduced intra-specific competition for the food supply and the less damaged root enabled many nematodes to survive.     

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.     

The effects of ultraviolet-B (UV-B: 290–320 nm) radiation on blister blight disease of tea (Camellia sinensis)

Sunlight is known to reduce the incidence of blister blight disease of tea, caused by Exobasidium vexans . The effects of the ultraviolet-B (UV-B: 290–320 nm) component of solar radiation on the disease and the development of E. vexans were studied in the field in Sri Lanka using UV-screening filter materials held over a commercial crop. Exclusion of UV-B radiation by Polyester, which reduces fluxes in the UV-B region by approximately 75–85%, increased both the number of translucent spots (immature sites of infection) and number of sporulating blisters (mature sites of infection), but it had little or no effect on the sporulation of the pathogen. When basidiospores were artificially inoculated on leaves, and were exposed to full or filtered solar radiation, their survival and germination increased when UV-B wavelengths were removed; significant differences were found 62 h after inoculation. However, UV-B did not affect the extension of germ tubes. This study indicates that the UV-B component of solar radiation plays an important role in the natural regulation of blister blight disease in the field.