Genetic variability and virulence among Verticillium albo-atrum isolates from hop

Verticillium wilt, caused by Verticillium albo-atrum or V. dahliae, is an important disease of many worldwide crop species. In Europe, V. albo-atrum isolates infecting hop express different levels of virulence, inducing mild or lethal disease syndromes, and it is therefore an attractive model for studying the virulence of this pathogen. In this work, eleven amplified fragment length polymorphism (AFLP) primer combinations were used to analyze genetic variability among 55 V. albo-atrum hop isolates from four European hop growing regions, as well as isolates from other hosts and V. dahliae isolates. Cluster analysis divided V. albo-atrum and V. dahliae isolates into two well-separated groups. Within the V. dahliae cluster, isolates were separated without host specific grouping, although no host adapted isolates were included. In V. albo-atrum, the alfalfa isolates were distinct from isolates of other hosts, where a high association with virulence was observed in hop and tomato isolates. All lethal hop isolates were genetically different from mild hop isolates. The lethal hop isolates from England and Slovenia expressed the same virulence phenotype, although they showed a different AFLP pattern. The mild hop isolates formed two subgroups, to which isolates clustered irrespective of geographical location. These data suggest multiple origins of V. albo-atrum hop isolates, and the possible appearance of new virulent isolates in the future in other hop growing regions.     

Multi Criteria Assessment of Zero Residue Apple Production

During the 2008–2010 growing seasons, an alternative ‘zero residue apple production system’ was compared with integrated apple production with cvs ‘Idared’, ‘Golden Delicious’, ‘Jonagold’ and ‘Gala’ in commercial orchards at several locations throughout Slovenia, using data envelopment analysis (DEA), and multi criteria assessment by an analytical hierarchical process (AHP). The principle of the ‘zero residue apple production’ is a combination of integrated (IP) and organic apple production. During the first 3 months of the growing season (1 April–30 June), pesticides used in IP with rapid degradation (8–10 applications) were used to control pests and diseases. During the second part of the season from 1 July to harvest, organic products (6–12 applications) were employed compared with 19–25 applications overall in IP. The goal of the alternative system was to reduce the amount of applied conventional pesticides by 40 % and to minimize pesticide residues in fruits to below the limit of 0.5 % of the legal maximum residue level (MRL) or below the residue concentrations of 0.005–0.01 mg/kg and to retain the high long-term level of yield, fruit quality, and net income per hectare. The number of pesticide residues was reduced from 4.2–5.5 in IP to 1.8–3.4 in zero residue cultivation, while 3 year average yields (class 1 fruit) were 4–9 % lower than in IP. The break even prices ranged from € 0.31 for Idared in IP, € 0.34 for ‘Elstar’ of both production systems to € 0.35/kg for zero residue cultivated ‘Golden Delicious’. Overall, a price increase of just € 0.02/kg for residue free apples would make this new ‘zero residue apple production’ profitable then representing a realistic alternative to the standard integrated apple production system.     

Molecular diversity of ‘Candidatus Phytoplasma pyri’ isolates in Slovenia

A European quarantine organism ‘Candidatus Phytoplasma pyri’ causing devastating pear decline disease has been reported to affect pear trees in several European countries. In this study a multilocus sequence analysis was successfully used to gain detailed insight into the molecular diversity of thirty closely related ‘Candidatus Phytoplasma pyri’ isolates from different orchards in Slovenia. Among three genomic regions analyzed, the 16S/23S rRNA intergenic spacer region was the most conserved among Slovenian isolates with 99.7 % sequence identity, yielding only three distinct genotypes. On the other hand, five different genotypes were detected when analyzing secY and aceF genomic regions that shared sequence identity of 94.8 and 97.2 %, respectively. Six of the detected genotypes, specifically four in the secY region and one in each of the two other analyzed genomic regions, were unique for Slovenia. At least eight different haplotypes were found with multilocus sequence analysis, indicating high molecular diversity among Slovenian ‘Ca. P. pyri’ isolates. Haplotypes were clustered into two major clusters, separated by at least 45 mutations. No connection was established between haplotype occurrence and cultivar type.     

Manipulating behaviour with substrate‐borne vibrations – potential for insect pest control

This review presents an overview of the potential use of substrate‐borne vibrations for the purpose of achieving insect pest control in the context of integrated pest management. Although the importance of mechanical vibrations in the life of insects has been fairly well established, the effect of substrate‐borne vibrations has historically been understudied, in contrast to sound sensu stricto. Consequently, the idea of using substrate‐borne vibrations for pest control is still in its infancy. This review therefore focuses on the theoretical background, using it to highlight potential applications in a field environment, and lists the few preliminary studies that have been or are being performed. Conceptual similarities to the use of sound, as well as limitations inherent in this approach, are also noted. © 2014 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Identification of QTLs for alpha acid content and yield in hop (Humulus Lupulus L.)

Amplified fragment length polymorphism (AFLP) and microsatellite (SSR) markers were applied to a segregation population of 111 genotypes derived from a pseudo-testcross of hop (Humulus lupulus L.) in order to detect quantitative trait loci (QTLs) for alpha-acid content and yield traits. A total of 199 markers (150 AFLPs, 43 SSRs, one hypothetical sex marker, five chs genes) were located on the 20 linkage groups (LGs) of the maternal and paternal maps, covering 706 and 616 cM, respectively. Due to the presence of 16 common biparental SSR markers, homology of seven LGs between parental maps could be inferred. The progeny segregated quantitatively for alpha-acid content and yield determined in the years from 2002–2006. A total of 13 putative QTLs for alpha acid content, 13 for dry cone weight and 18 for harvest index were identified on the two maps across years. Possible homologies between the detected QTLs on the two maps as well as in different years were established for all three traits. The most promising QTL for alpha acid content was identified on LG03 flanked by two AFLP markers (E-ACC-M-CAA103F*/P-ACA-M-CAC412F). From 13.80 to 36.64% higher content of alpha acids than the averages obtained in different years was observed in plants having both flanking markers. The candidate region for further characterization of QTLs for yield traits was located on LG01 where the putative QTLs for harvest index were detected on both maps in each of the 5 years. The QTLs identified represent an important improvement in alpha acids MAS and the first step towards marker-assisted breeding for hop yield.     

Assessment of genetic variability of olive varieties by microsatellite and AFLP markers

Fourteen developed microsatellite markers were characterized for their use in genotyping and diversity studies of olive varieties. After optimisation of microsatellite assay and allele sizing, ninety-six alleles were found in nineteen varieties, with an average of 6.8 alleles per locus. The characteristics of the microsatellite markers were used to identify markers that can be reliably applied for variety genotyping. Such features were the generation of complex banding patterns supported by underlying allele sequences, `short allele dominance', an unstable repeat structure and a low number of alleles. AFLP analysis was performed on the same set of olive varieties using eight primer pair combinations. The genetic relationships among nineteen olive varieties were compared on the basis of microsatellite and AFLP polymorphisms. Genetic distances between all pairwise combinations of the varieties were calculated using Jaccard's coefficient of similarity and dendrograms were constructed by the UPGMA method. The results of clustering analysis with both molecular systems showed the common genetic background of Tuscan varieties, and genetic divergence within Slovene olive germplasm. Slovenian varieties ‘Buga’, ‘Štorta’ and ‘Samo’ might represent regionally selected olives, while ‘Zelenjak’ and ‘Črnica’ are probably derived from the Central Italian region. The predominant local ‘Istrska belica’ was introduced to Slovenia independently from the other regional varieties and showed the lowest genetic similarity with the other regional varieties. This revised version was published online in August 2006 with corrections to the Cover Date.     

Kinetics of abamectin disposition in blood plasma and milk of lactating dairy sheep and suckling lambs

Abamectin (ABM) has been used worldwide as an anthelmintic drug in veterinary medicine and as an agricultural pesticide. Its pharmacokinetics and permeation into milk was evaluated in dairy sheep after subcutaneous administration. ABM elimination half-lives and mean residence times were 1.7 and 3.7 days for blood plasma and 1.9 and 3.8 days for milk, respectively. The ABM milk to plasma concentration ratio (0.89) primarily depends on milk fat content. Transfer of ABM residues to suckling lambs was evaluated by determination of ABM concentration time courses in lambs' plasma. Mean maximal concentration in lambs was 1.6 microg L(-1) at 3.3 days, and elimination half-life was 2.7 days. In ewes' plasma and milk, ABM was detected up to 23 days. Because of different pharmacokinetics, ABM exposure in lambs was almost 10% of the exposure in ewes, although the amount excreted in milk was only 1.0% of the dose.     

Expression of estrogen receptor 1 and progesterone receptor in primary goat mammary epithelial cells

The exact role and sensitivity of cells to estrogen and progesterone mediated through the steroid receptors during lactation is not known. Expression of estrogen receptor 1 (ESR1) and progesterone receptor (PGR) was quantified in mammary tissue‐derived primary goat mammary epithelial cells (pgMECs) to determine the influence of donor tissue physiology (lactating and juvenile) and cell culture growth conditions (basal and lactogenic) on ESR1 and PGR expression in the derived cells. Relative messenger RNA (mRNA) levels for both receptors were the highest in cell lines derived from mammary tissue of juvenile goats. Maintaining pgMECs in lactogenic conditions resulted in up‐regulation of ESR1 (1.36‐ to 12.35‐fold) and in down‐regulation of PGR (‐2.53‐ to ‐3.62‐fold), compared to basal conditions. Based on Western blotting analysis we suggest that the differences in mRNA expression are translated to the protein level. We suggest that differential expression in lactating conditions is correlated with terminal differentiation of the pgMECs. Double immunostainings showed that estrogen receptor alpha (ER‐α) positive cells do not exclusively belong to the luminal lineage and that ER‐α and PGR can be expressed individually or co‐expressed in the pgMECs. The derived primary cultures/lines in early passages are hormone‐responsive and represent a useful surrogate for mammary tissue in research experiments.