Discrimination of three trachurus species using both mitochondrial- and nuclear-based DNA approaches

A double-DNA approach was developed to discriminate the three Trachurus species that abide in European waters: T. trachurus, T. mediterraneus, and T. picturatus. The analysis aimed at both mitochondrial and nuclear loci. Polymerase Chain Reaction (PCR) amplification of the cytochrome b gene of mtDNA was followed by restriction analysis with three species-specific enzymes: NlaIII, NciI, and BsmAI. Digestion with these endonucleases yielded species-specific electrophoretic profiles. The universality of the results was verified by screening a large number of individuals from 12 geographical regions covering most of the distribution of the species. Additionally, the nuclear multicopy 5S rRNA gene was selected as an alternative candidate for the discrimination of the three Trachurusspecies. A simple agarose gel electrophoretic analysis of the amplicons proved to be capable of leading to unambiguous identification of the three Trachurus species. Thus, the double-DNA methodology presented here allows the accurate discrimination of Trachurus fish species and the detection of commercial fraud.     

Marine spatial plans focusing on biodiversity conservation: The case of the Aegean Sea

1. The expansion and intensification of marine uses have severe cumulative impacts on marine ecosystems and human well-being, unless they are properly managed with an ecosystem-based management approach.
2. A systematic conservation planning approach, using marxan with zones , was applied to generate alternative marine spatial plans for the Aegean Sea. Relevant human uses were included and their cumulative impact on a wide set of key biodiversity features was considered in the analysis. Different cost scenarios were developed to gain insight on the effects of the approaches used to assess socio-economic factors, and their potential impact on spatial plans.
3. The spatial plans generated differed greatly depending on the method used to estimate opportunity costs and evaluate human activities in monetary terms.
4. The vulnerability weights (the relative vulnerability of ecological features to specific human activities and their impacts) that were estimated based on a cumulative impact assessment, allowed the assessment of each zone in contributing to the achievement of conservation targets, through a transparent planning approach.
5. Results indicate that special care should be given to how socio-economic activities, their impact on the ecosystems, and related costs are incorporated into planning.
6. The proposed approach demonstrates how EU member states may effectively comply with the new Biodiversity Strategy 2030 targets, while planning for the sustainable use of their marine resources.

     

Greenhouse biofumigation with <Emphasis Type="Italic">Melia azedarach</Emphasis> controls <Emphasis Type="Italic">Meloidogyne</Emphasis> spp. and enhances soil biological activity

The need for environmentally friendly agricultural practices has led to the development of plant-based nematicides for root-knot nematode control. The efficacy of these botanicals has been tested primarily under laboratory and rarely under actual field conditions. Moreover, any side effects on non-target soil organisms that support soil biological activity are usually ignored. Herein, we evaluate the efficacy of Melia azedarach preparations against Meloidogyne spp. in a tomato greenhouse, by root gall examination and soil J2 enumeration. We also assessed side effects on soil microbes through PLFA analysis and microbivorous nematodes, and we quantified several plant growth parameters (e.g., fruit number and weight, root weight). Different treatments within the greenhouse included M. azedarach ripe fruit powder (MFP), ripe fruit water extract (MWE) and furfural, one of the principal active ingredients of M. azedarach and previously known to exhibit fumigant nematicidal activity. Results were compared to those obtained with the commercial nematicide oxamyl (Vydate^® 10 SL) and an untreated control. All treatments were repeated every 20 days throughout the cultivation period. MFP and MWE suppressed Meloidogyne spp. often at the same levels obtained by furfural and oxamyl treatments and enhanced soil biological activity, as indicated by the proliferation of soil microbes and microbial feeding nematodes. Furfural and oxamyl adversely affected the soil community, especially the free-living nematodes. Moreover, furfural was phytotoxic to tomato plants in spite of its natural origin.     

PIN1 auxin efflux carrier absence in Meloidogyne incognita-induced root-knots of tomato plants

The nematode species Meloidogyne incognita infects a large variety of cultivated crops and is one of nature’s most notorious pests. One cultivated plant which is prone to M. incognita infestation is the tomato. Knowing that in A. thaliana the PIN auxin efflux transporters distribution is being altered upon early invasion by M. incognita, the PIN1 allocation patterns in the giant cells of tomato plants after 15 and 25 days of infection were investigated. PIN1 was absent from the giant cells’ membrane in both assessment timings examined, indicating the maintenance of a local auxin maxima, which was also supported by IAA immunodetection. PIN1 distribution pattern could be attributed on either the nematodes nutritional needs throughout its life cycle or due to the tomato plants differential responses upon M. incognita infection.

     

Aliphatic ketones from Ruta chalepensis (Rutaceae) induce paralysis on root knot nematodes

This paper reports on the use of Ruta chalepensis L. extracts as a potential nematicide against root knot nematodes Meloidogyne incognita and Meloidogyne javanica . The essential oil (REO) and methanol extract (RME) of R. chalepensis were tested against second-stage juveniles, with REO inducing paralysis in both species (EC(50/1d) = 77.5 and 107.3 mg/L) and RME being selective for M. incognita (EC(50/1d) = 1001 mg/L). Chemical characterization of extracts was done by means of GC-MS and LC-MS, revealing mainly aliphatic ketones and coumarins, respectively. The first-ranking volatile nematicidal component in terms of individual activity against both species was 2-undecanone (EC(50) = 20.6 and 22.5 mg/L for M. incognita and M. javanica, respectively). This fact together with its high concentration in the most active extract found in this study, namely, REO (2926 mg/kg), categorizes 2-undecanone among the nematicidal principles of R. chalepensis. On the contrary, coumarins rutin and 8-methoxypsoralen were not found to be nematicidal at concentrations of ≤500 mg/L. Interestingly, M. incognita was found more sensitive than M. javanica.     

Nematicidal activity of (E,E)-2,4-decadienal and (E)-2-decenal from Ailanthus altissima against Meloidogyne javanica

Methanol extracts of various plant parts of Ailanthus altissima were tested against the root knot nematode Meloidogyne javanica . Extracts of bark (ABE), wood (AWE), roots (ARE), and leaves (ALE) from A. altissima were investigated against freshly hatched second-stage juveniles (J(2)). AWE was the most active extract, with EC(50/3d) of 58.9 mg/L, while ALE, ARE, and ABE did not show nematicidal activity. The chemical composition of the extracts of A. altissima was determined by gas chromatography-mass spectrometry, and (E,E)-2,4-decadienal, (E)-2-undecenal, (E)-2-decenal, hexanal, nonanal, and furfural were the most prominent constituents. (E,E)-2,4-Decadienal, (E)-2-decenal, and furfural showed the highest nematicidal activity against M. javanica , with EC(50/1d) = 11.7, 20.43, and 21.79 mg/L, respectively, while the other compounds were inactive at the concentrations tested. The results obtained showed that AWE and its constituents (E,E)-2,4-decadienal and (E)-2-decenal could be considered as potent botanical nematicidal agents.     

Synergistic and antagonistic interactions of terpenes against Meloidogyne incognita and the nematicidal activity of essential oils from seven plants indigenous to Greece

BACKGROUND: Biorational means for phytonematode control were studied within the context of an increasingly ecofriendly pest management global approach. The nematicidal activity and the chemical composition of essential oils (EOs) isolated from seven plants grown in Greece and ten selected compounds extracted from them against second‐stage juveniles (J2) of Meloidogyne incognita (Kof. & White) Chitwood were evaluated using juvenile paralysis experiments. Additionally, synergistic and antagonistic interactions between nematicidal terpenes were studied using an effect addition model, with the comparison made at one concentration level. RESULTS: The 96 h EC₅₀ values of Foeniculum vulgare Mill., Pimpinella anisum L., Eucalyptus meliodora A Cunn ex Schauer and Pistacia terebinthus L. were 231, 269, 807 and 1116 µg mL^?1, respectively, in an immersion bioassay. Benzaldehyde (9 µg mL^?1) was the most toxic compound, followed by γ‐eudesmol (50 µg mL^?1) and estragole (180 µg mL^?1), based on 96 h EC₅₀ values. The most potent terpene pairs between which synergistic actions were found, in decreasing order, were: trans‐anethole/geraniol, trans‐anethole/eugenol, carvacrol/eugenol and geraniol/carvacrol. CONCLUSION: This is the first report on the activity of F. vulgare, P. anisum, E. meliodora and P. terebinthus, and additionally on synergistic/antagonistic nematicidal terpene interactions, against M. incognita, providing alternative methods for nematode control. Copyright © 2010 Society of Chemical Industry     

Genetic identification and phylogenetic inferences in different Mugilidae species using 5S rDNA markers

A DNA methodology was developed to discriminate fry of six Mugilidae species found in the Mediterranean, namely Mugil cephalus, Mugil so‐iuy, Chelon labrosus, Liza aurata, Liza ramada and Liza saliens. Polymerase chain reaction (PCR) amplification of the 5S rDNA gene was used for the identification of the above six species. PCR products of two species showed different patterns on EtBr‐stained agarose gels; M. so‐iuy gave a pattern of three bands, while L. saliens gave a pattern of one band. Mugil cephalus, C. labrosus, L. aurata and L. ramada gave a pattern of two bands. Subsequent sequencing analyses revealed unique haplotypes for each of the remaining four species. This is a genetic technique that could be applied in hatcheries, for identification of fry mullet's species. Furthermore, 5S rDNA sequences of each of five of the studied species (i.e. M. cephalus, C. labrosus, L. aurata, L. ramada and L. saliens), as well as a sequence of Oedalechilus labeo that belongs to the Mugilidae family, were aligned and used for the examination of the phylogenetic relationships among them. Phylogenetic trees produced in our study are in general agreement with those presented in the literature. This is the first study to use a nuclear marker to elucidate the phylogeny of Mugilidae species.     

Metabolites from Marine-Derived Fungi as Potential Antimicrobial Adjuvants

Marine-derived fungi constitute an interesting source of bioactive compounds, several of which exhibit antibacterial activity. These acquire special importance, considering that antimicrobial resistance is becoming more widespread. The overexpression of efflux pumps, capable of expelling antimicrobials out of bacterial cells, is one of the most worrisome mechanisms. There has been an ongoing effort to find not only new antimicrobials, but also compounds that can block resistance mechanisms which can be used in combination with approved antimicrobial drugs. In this work, a library of nineteen marine natural products, isolated from marine-derived fungi of the genera Neosartorya and Aspergillus, was evaluated for their potential as bacterial efflux pump inhibitors as well as the antimicrobial-related mechanisms, such as inhibition of biofilm formation and quorum-sensing. Docking studies were performed to predict their efflux pump action. These compounds were also tested for their cytotoxicity in mouse fibroblast cell line NIH/3T3. The results obtained suggest that the marine-derived fungal metabolites are a promising source of compounds with potential to revert antimicrobial resistance and serve as an inspiration for the synthesis of new antimicrobial drugs.