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 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.     

Changes in the content of steroidal glycoalkaloids in potato tubers grown in the field and in the greenhouse under different conditions of light, temperature and daylength

Differences in the content of steroid glycoalkaloids (SGA) were noted in the peels of potato tubers cultivars Alpha, Cara, Désirée and LT-7. Cultivar LT-7 had always the highest SGA content, Désirée was intermediate and Alpha and Cara had the lowest SGA content, regardless of growing conditions, tuber ageing and tuber treatments. Tubers grown during the summer developed the highest SGA content as compared to those grown in the spring and winter seasons, respectively. Tubers grown during winter showed an increase in SGA toward the end of the storage period. Similar results were obtained with tubers of LT-7 grown in a greenhouse under cool temperatures in combination with short days (which imitate the winter conditions in Israel). A decrease in SGA content was observed in tuber slices treated with a fungal cell-wall preparation fromPhytophthora citrophthora. This decrease was much stronger in the pith than in the peel, indicating a possible role for SGA in the defence mechanism because peels are the first part of the tuber to be invaded by pathogens. The above results were observed in tubers of all cultivars regardless of the growing season or tuber ageing.     

Evaluation of sampling methods and assessment of the sample size to estimate the weed seedbank in soil, taking into account spatial variability

As field sampling is time consuming, it is necessary to develop efficient sampling techniques to obtain accurate estimates of the weed seedbank in soil. The relative efficiency between sampling schemes depends on the spatial variability in weed seed density across agricultural fields. Spatial variability of the weed seed density was characterized by theoretical correlograms. A systematic sampling (square grill) scheme was considered and it was found that, taking into account spatial variability, this sampling scheme was more efficient than simple random sampling. As a result, the sample size can be reduced in comparison with that given in previous studies, where spatial correlation was ignored. The reduction depends on the correlation structure defined as a function of the ratio, τ, between the nugget effect and the sill of the variogram. The maximum reduction of the sample size, without loss of either precision or confidence level corresponds to the case where there is no nugget effect, τ = 0. The opposite extreme case, where the reduction is nil, corresponds to the case of a pure nugget effect τ = 1. The abaci based on given expressions are provided to determine the sample size in species whose spatial pattern can be fitted either to a Poisson or to a negative binomial distribution.     

Fireblight monitoring in Lithuania1

Lithuania has requested that its whole territory should be recognized by the EU as a protected zone for Erwinia amylovora . Fireblight monitoring was performed in 1998/2002 with the aim of detecting and identifying the bacterium, and of determining its distribution in the country. The study consisted of periodic surveys (at least twice a year) of nurseries, orchards, collective farms and host plants, growing individually or in small groups, as well as the surrounding zone within a radius of 250 m. Tests, under conditions of quality control, were applied to host plants with and without symptoms, using detection methods such as ELISA and immunofluorescence (with polyclonal antibodies), semi-selective plating and pathogenicity.     

Biology of a new virus isolated from Lupinus nootkatensis plants in Alaska

A new virus named Nootka lupine vein-clearing virus (NLVCV) was isolated from Lupinus nootkatensis plants that were confined to a relatively small area in the Talkeetna mountains of south-central Alaska. Annual surveys (2000–03) consistently found leaf symptoms of pronounced vein clearing and mosaic on 3- to 4-week-old plants in late June. Spherical particles ≈30 nm in diameter were isolated from these leaves. Virions contained a single-stranded RNA of ≈4·0–4·2 kb and one species of capsid protein estimated to be ≈40 kDa. The double-stranded RNA profile from naturally infected leaves consisted of three major bands ≈4·2, 1·9 and 1·5 kbp. Protein extractions from either sap or virions of diseased plants reacted to polyclonal antiserum made against the virions in Western blot assays. A predicted PCR product ≈500 bp was synthesized from virion RNA using primers specific to the carmovirus RNA-dependent RNA polymerase (RDRP) gene. The nucleotide sequence of the amplified DNA did not match any known virus, but contained short regions of identity to several carmoviruses. Only species belonging to the Fabaceae were susceptible to NLVCV by mechanical inoculation. Based on dsRNA profile, size of virion RNA genome and capsid protein, and similarity of the RDRP gene to that of other carmoviruses, it is suggested that NLVCV is a member of the family Tombusviridae , and tentatively of the genus Carmovirus . As the host range, RDRP gene and dsRNA profile of NLVCV are different from those of known viruses, this is a newly described plant virus.