Application of molecular tools for detection of plant viruses

Grapevine leafroll disease (GLRD) is one of the most important virus diseases of grapevines worldwide, causing major economical impact. The disease has a complex aetiology and currently eleven phloem-limited viruses, termed in general Grapevine leafroll-associated virus (GLRaVs), have been identified. Two of the GLRaVs, GLRaV-1 and GLRaV-3, are included in the European certification scheme of propagation material. However, the flawed notion that GLRaV-3 is more frequent than GLRaV-1 and that all other GLRaVs are possibly not as relevant for GLRD, has until now precluded the development of specific serological and molecular detection assays and limited the scope of molecular characterization of the viruses known to be associated with the disease. Hence, few studies have addressed the phylodynamics of GLRaVs or even characterized the genetic structure of their natural populations. This generalized lack of molecular information, in turn underlie the deficient capacity to detect the viruses. The phylogenetic analyses were conducted on the basis of the heat shock protein 70 homologue (HSP70h) and the coat protein (CP) genes for GLRaV-1 and the HSP70h, the heat shock protein 90 homologue (HSP90h) and the CP genes for GLRaV-5. The data obtained for GLRaV-1 contributed 83 new CP sequences. This information was combined with previous analysis by other authors and used for the production of new polyclonal IgG, capable of detecting CP variants from all the phylogroups observed. Successful testing of this new tool included tissue print immunoblotting (TPIB) and in situ immunoassay (ISIA). The data obtained for GLRaV-5, contributed 61 new CP and 28 new HSP90h gene sequences. Eight phylogenetic groups were identified on the basis of the CP. Characterization of the genetic structure of the isolates revealed a higher diversity than previously reported and allowed the identification of dominant virus variants. For both GLRaV-1 and GLRaV-5, the effect of vegetative propagation on the virus transmission dynamics was addressed.

Assessment of catches, landings and fishing effort as useful tools for MPA management

Marine protected areas (MPAs) have been widely recognized as a tool to achieve both fisheries management and conservation goals. Simultaneously achieving these multiple goals is difficult due to conflicts between conservation (often long-term) and economic (often short-term) objectives. MPA implementation often includes additional control measures on fisheries (e.g. vessel size restrictions, gear exclusion, catch controls) that in the short-term may have impacts on local fishers' communities. Thus, monitoring fisheries catches before, during and after MPA implementation is essential to document changes in fisheries activities and to evaluate the impact of MPAs in fishers' communities. Remarkably, in contrast with standard fisheries-independent biological surveys, these data are rarely measured at appropriate spatial scales following MPA implementation. Here, the effects of MPA implementation on local fisheries are assessed in a temperate MPA (Arrabida Marine Park, Portugal), using fisheries monitoring methods combining spatial distribution of fishing effort, on-board observations and official landings statistics at scales appropriate to the Marine Park. Fisheries spatial distribution, fishing effort, on-board data collection and official landings registered for the same vessels over time were analysed between 2004 and 2010. The applicability and reliability of using landings statistics alone was tested (i.e. when no sampling data are available) and we conclude that landings data alone only allow the identification of general patterns. The combination of landings information (which is known to be unreliable in many coastal communities) with other methods, provides an effective tool to evaluate fisheries dynamics in response to MPA implementation. As resources for monitoring socio-ecological responses to MPAs are frequently scarce, the use of landings data calibrated with fisheries information (from vessels, gear distribution and on-board data) is a valuable tool applicable to many worldwide coastal small-scale fisheries. (C) 2015 Elsevier B.V. All rights reserved.