Construction of a cDNA library of Bemisia tabaci for use in the ''yeast two-hybrid screen'' method

Bemisia tabaci was reported for the first time in the Mediterranean part of Croatia in 2000. It was found in glasshouses in the agricultural area between the towns of Trogir and Omis, on the following crops: Euphorbia pulcherrima , Thunbergia grandiflora , Cucumis sativus (cucumber), Solanum melongena (aubergine), Phaseolus spp. (beans), Ficus carica (fig), Rubus spp. and several weeds of the families Asteraceae and Solanaceae . In 2001, monitoring for the pest was organized all over the country, in each of the 21 counties. In each county, there were several monitoring points so that all the major vegetable and flower producers were included. A special effort was made to record the spread of B. tabaci in the region where it was first found, bearing in mind that optimal conditions for outdoor spread exist along the Adriatic coast. Yellow sticky traps and visual inspection are used to monitor for B. tabaci . Eradication measures are being applied, and regulatory measures have been taken to prevent further spread of B. tabaci to continental parts of Croatia.     

Genetic analyses of resistance of the wheat differential cultivars Carstens V and Spaldings Prolific to two races of Puccinia striiformis

Genetic analysis of resistance of wheat seedlings to two races of Puccinia striiformis was conducted on F₁, F₂ and F₃ generations from crosses Carstens V (CV) × Lee, Spaldings Prolific (SPA) × Lee and CV × SPA. F₂ generations from crosses of CV and SPA with Strubes Dickkopf (SD) were also studied. The plants were classified into six resistance classes and analysed by factorial correspondence analysis and nonhierarchical classification. The two P. striiformis isolates tested were a French isolate of race 43E138 and a Lebanese isolate of race 2E16, selected for the differences in their virulence spectra for the common differential cultivars Strubes Dickkopf and Nord Desprez. Resistance of CV and SPA was recessive and dominant to races 43E138 and 2E16, respectively. CV possessed three or four resistance genes, one of them being expressed with both races. Two genes of CV had a cumulative effect for resistance to 43E138 and two or three gave dominant resistance to 2E16. SPA had three resistance genes, all of which gave resistance to 2E16 and two of which also gave resistance to 43E138. SPA had one gene in common with CV for resistance to both races. Furthermore, the gene for resistance to race 2E16 in CV and SPA was allelic with a gene in SD, and was probably Yr25 .     

Tomato yellow leaf curl Sardinia virus and its vector Bemisia tabaci in Sicilia (Italy): present status and control possibilities*

Tomato yellow leaf curl Sardinia begomovirus (TYLCSV) appeared in Sicilia (IT) in 1988, creating great threats to agriculture and causing huge losses, especially in south‐eastern areas of the island, where protected tomato cultivation is widespread. Towards the mid‐1990s, a reduction occurred in the virus epidemics, probably due to new approaches which have been applied to rational control of its vector, the whitefly Bemisia tabaci. More recently, TYLCSV has increased again, creating great concern among local tomato producers and stimulating new research. Besides studies on natural enemies of the vector, aiming to investigate their role and distribution, the main current research lines in Sicilia concern the possibility of reducing both whitefly activity, using photoselective plastics as covers, and virus damage, by growing tolerant tomato genotypes.     

Orobanche species and population discrimination using intersimple sequence repeat (ISSR)

Summary Orobanche species are commonly identified using morphological characteristics. In many cases, the distinction of closely related species is difficult, and a molecular tool is more suitable to differentiate them. In this study, genomic polymorphism between morphologically distinct species was investigated through amplification by polymerase chain reaction (PCR) of intersimple sequence repeat (ISSR) regions. Five primers were used to study genetic variation in the morphologically distinct species O. hederae and O. amethystea, as well as the closely related species O. cernua and O. cumana. For the first two species, all the primers detected genetic polymorphism. Anchored primers allowed the identification of more specific molecular markers than non‐anchored tri‐ and tetranucleotide primers. Genetic polymorphism was investigated among three O. hederae populations using the two types of primer. One non‐anchored and two anchored primers detected intraspecific variation, which was not correlated with the geographical location of those populations. The primer (GATA)₄ detected polymorphism between five specimens each of O. cernua and O. cumana species collected from different countries, permitting these two closely related species to be clearly differentiated. This study demonstrated that ISSR markers can be highly reliable for precise identification of Orobanche species.     

Spatial dependence and the relationship of soil organic carbon and soil moisture in the Luquillo Experimental Forest,Puerto Rico

We used geo-spatial statistical techniques to examine the spatial variation and relationship of soil organic carbon (SOC) and soil moisture (SM) in the Luquillo Experimental Forest (LEF), Puerto Rico, in order to test the hypothesis that mountainous terrain introduces spatial autocorrelation and crosscorrelation in ecosystem and soil properties. Soil samples (n = 100) were collected from the LEF in the summer of 1998 and analyzed for SOC, SM, and bulk density (BD). A global positioning system was used to georeference the location of each sampling site. At each site, elevation, slope and aspect were recorded. We calculated the isotropic and anisotropic semivariograms of soil and topographic properties, as well as the cross-variograms between SOC and SM, and between SOC and elevation. Then we used four models (random, linear, spherical and wave/hole) to test the semi-variances of SOC, SM, BD, elevation, slope and aspect for spatial dependence. Our results indicate that all the studied properties except slope angle exhibit spatial dependence within the scale of sampling (200 – 1000 m sampling interval). The spatially structured variance (the variance due to the location of sampling sites) accounted for a large proportion of the sample variance for elevation (99%), BD (90%), SOC (68%), aspect (56%) and SM (44%). The ranges of spatial dependence (the distances within which parameters are spatially dependent) for aspect, SOC, elevation, SM, and BD were 9810 m, 3070 m, 1120 m, 930 m and 430 m, respectively. Cross correlograms indicate that SOC varies closely with elevation and SM depending on the distances between samples. The correlation can shift from positive to negative as the separation distance increases. Larger ranges of spatial dependence of SOC, aspect and elevation indicate that the distribution of SOC in the LEF is determined by a combination of biotic (e.g., litterfall) and abiotic factors (e.g., microclimate and topographic features) related to elevation and aspect. This demonstrates the importance of both elevation and topographic gradients in controlling climate, vegetation distribution and soil properties as well as the associated biogeochemical processes in the LEF.This revised version was published online in May 2005 with corrections to the Cover Date.     

Analysis of the association among three viruses infecting hop in Australia

Associations among Hop latent virus (HpLV), Hop mosaic virus (HpMV), and Apple mosaic virus (ApMV) were assessed in five hop cultivars at four commercial hop-growing regions in Victoria and Tasmania, Australia. The presence or absence of each virus was confirmed by double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). Spatial patterns of virus-infected plants were characterized using the Spatial Analysis by Distance IndicEs ( sadie ) system of pattern analysis. The association among viruses (occurrence and covariation) was assessed using the Jaccard similarity index, Spearman's rank correlation coefficient, and sadie . The spatial pattern of plants infected by HpLV and HpMV ranged from random to highly aggregated depending upon the cultivar infected and the mean disease incidence. The spatial pattern of plants infected by ApMV was aggregated in six of the seven plots where ApMV was present. A strong positive association between HpLV and HpMV was found in all cultivars at all locations. This association may be the result of the viruses sharing a common aphid vector species, the presence of one virus enhancing the ability of the aphid vector to acquire the other virus either through transencapsidation or influences on virus titre, or mixed infections within source plants. Significant associations, positive or negative, were found less frequently between HpLV and ApMV, and HpMV and ApMV.     

Characterization of phytoplasmas detected in Chinaberry trees with symptoms of leaf yellowing and decline in Bolivia

Polymerase chain reaction (PCR) assays were used to detect phytoplasmas in foliage samples from Chinaberry ( Melia azedarach ) trees displaying symptoms of yellowing, little leaf and dieback in Bolivia. A ribosomal coding nuclear DNA (rDNA) product (1·8 kb) was amplified from one or more samples from seven of 17 affected trees by PCR employing phytoplasma-universal rRNA primer pair P1/P7. When P1/P7 products were reamplified using nested rRNA primer pair R16F2n/R16R2, phytoplasmas were detected in at least one sample from 13 of 17 trees with symptoms. Restriction fragment length polymorphism (RFLP) analysis of P1/P7 products indicated that trees CbY1 and CbY17 harboured Mexican periwinkle virescence (16SrXIII)-group and X-disease (16SrIII)-group phytoplasmas, respectively. Identification of two different phytoplasma types was supported by reamplification of P1/P7 products by nested PCR employing X-disease-group-specific rRNA primer pair R16mF2/WXint or stolbur-group-related primer pair fSTOL/rSTOL. These assays selectively amplified rDNA products of 1656 and 579 bp from nine and five trees with symptoms, respectively, of which two trees were coinfected with both phytoplasma types. Phylogenetic analysis of 16S rDNA sequences revealed Chinaberry yellows phytoplasma strain CbY17 to be most similar to the chayote witches'-broom (ChWBIII-Ch10) agent, a previously classified 16SrIII-J subgroup phytoplasma. Strain CbY1 resembled the Mexican periwinkle virescence phytoplasma, a 16SrXIII-group member. The latter strain varied from all known phytoplasmas composing group 16SrXIII. On this basis, strain CbY1 was assigned to a new subgroup, 16SrXIII-C.