In vitro nematicidal activity of naphthoquinones against the root-lesion nematode Pratylenchus thornei

The root-lesion nematode Pratylenchus thornei is a widely distributed and important parasite of cereals and legumes. As migratory endoparasites, P. thornei are difficult to manage  because they enter and leave host roots and may remain quiescent inside roots or in soil when conditions are unfavourable for plant growth. The number of available nematicides to manage these nematodes is restricted, so new, effective and eco-friendly sustainable management strategies are needed. The effects of naphthoquinones (juglone, 1,4-naphtoquinone and plumbagin) produced by some plants species, including walnut (Juglandaceae), were assessed against P. thornei. An additional treatment of a mixture of juglone and 1,4-naphtoquinone (2:1, w/w), was included because these compounds are frequently found at these propotion in walnut extracts. Juveniles and adult nematodes were exposed to different concentrations of each naphthoquinone and nematode mortality was assessed. Juglone and 1,4-naphthoquinone (at 500 ppm) were more effective than plumbagin, and gave 100% mortality after 24 h of exposure. A synergistic effect was not detected when juglone and 1,4-naphthoquinone (2:1, w/w) were combined. Estimated lethal concentrations causing 50% P. thornei mortality (LC50s) (72 h exposure) were: 134.7 ppm for juglone, 161.2 ppm for 1,4-naphthoquinone, 207.6 ppm for juglone + 1,4-naphthoquinone (2:1, w/w), and 178.8 ppm for plumbagin. This study has demonstrated the nematicidal potential of these naphthoquinones against P. thornei, and has shown that walnut residues may be valuable sources for extraction of these compounds

Transcriptome analysis reveals the high ribosomal inhibitory action of 1,4-naphthoquinone on Meloidogyne luci infective second-stage juveniles

The root-knot nematode (RKN) Meloidogyne luci presents a threat to the production of several important crops. This nematode species was added to the European Plant Protection Organization Alert list in 2017. The scarce availability of efficient nematicides to control RKN and the phasing out of nematicides from the market have intensified the search for alternatives, such as phytochemicals with bionematicidal properties. The nematicidal activity of 1,4-naphthoquinone (1,4-NTQ) against M. luci has been demonstrated; however, knowledge of the potential mode(s) of action of this compound is still scarce. In this study, the transcriptome profile of M. luci second-stage juveniles (J2), the infective stage, in response to 1,4-NTQ exposure was determined by RNA-seq to identify genes and pathways that might be involved in 1,4-NTQ’s mode(s) of action. Control treatments, consisting of nematodes exposed to Tween® 80 (1,4-NTQ solvent) and to water, were included in the analysis. A large set of differentially expressed genes (DEGs) was found among the three tested conditions, and a high number of downregulated genes were found between 1,4-NTQ treatment and water control, reflecting the inhibitory effect of this compound on M. luci, with a great impact on processes related to translation (ribosome pathway). Several other nematode gene networks and metabolic pathways affected by 1,4-NTQ were also identified, clarifying the possible mode of action of this promising bionematicide

Music and heritage in the context of Musical Sciences studies

Relatório de estágio de mestrado em Ensino de MúsicaO presente relatório diz respeito ao Projeto de Estágio - Música e Património no contexto da aprendizagem das Ciências Musicais, implementado no Conservatório de Música de Paredes, no ano letivo 2014-2015, na turma de História da Cultura e das Artes. O Conservatório de Música situa-se numa zona de grande riqueza patrimonial. O Património Histórico edificado, constituído pela Rota do Românico do Vale do Sousa, foi o ponto de partida para a realização deste projeto pedagógico e de investigação. Em particular, o Projeto de Estágio foi dedicado às práticas musicais na Rota do Românico, especificamente, no Mosteiro de Santa Maria de Pombeiro, em Felgueiras. A aplicação do projeto foi administrada em três momentos distintos. A primeira em contexto de sala de aula (pela apresentação da Rota do Românico, do Mosteiro de Santa Maria de Pombeiro e da prática musical, realizada pela congregação dos monges beneditinos); a segunda, a concretização de uma visita guiada ao Mosteiro; e por fim, a apresentação de um trabalho executado pelas alunas. A investigação efetuada e a intervenção pedagógica resultaram numa maior valorização do património histórico, na sua preservação e fruição e no conhecimento das práticas musicais, praticadas nos mosteiros Beneditinos, nos séculos XVII e XVIII. A aplicação de estratégias diversificadas, nomeadamente, os princípios da pedagogia de projeto, conduziu a aprendizagens significativas no campo das Ciências Musicais.This report concerns the Project internship – Music and Heritage in the context of Musical Sciences studies, implemented at the Music conservatory of Paredes in the lecture year of 2014- 2015, in the class of Cultural History and Arts. The Music Conservatory is located in an area of great patrimonial wealth. The existing Historic Patrimony, composed of the Romanic Route of Vale do Sousa, was the starting point for the present pedagogical project and research. In particular, the internship project focused on the musical practices in the Romanic Route, specifically at the Monastery of Santa Maria de Pombeiro, in Felgueiras. The application of the project was conducted at three distinct time points: the first in the context of a classroom (through the presentation of the Romanic Route, of the monastery of Santa Maria de Pombeiro and musical practices conducted by the congregation of the Benedictine monks at that time); secondly, the achievement of a tour guide of the Monastery; and last, presentations of assignments performed by the students. The performed research and pedagogical intervention resulted in an increase in appreciation of Historic Heritage value, of its preservation and fruition, and in the knowledge of musical practices conducted at the Benedictine monasteries in the XVII and XVIII centuries. The application of diversified strategies, namely principles of project teaching, led to significant increases in learning in the field of Musical Sciences

Susceptibility of crop plants to the root-knot nematode Meloidogyne luci, a threat to agricultural productivity

The root-knot nematode (RKN) Meloidogyne luci is included in the Alert List of the European Plant Protection Organization, because it has potential negative impacts on economically important crops. Identification of plant species/cultivars resistant to M. luci is important for its management. Susceptibility of 35 commercial plant species/cultivars, from nine families to a M. luci isolate from Portugal was evaluated in pot assays, assessing root gall index (GI) and reproduction factor (Rf) 60 d after inoculation, with tomato ‘Coração-de-Boi’ used as the positive susceptible experimental control. Presence/absence of RKN resistance genes was also determined in the tomato and pepper cultivars. One cultivar of cabbage, three of lettuce, ten of pepper, one of sugar beet, and all the cultivars of Cucurbitaceae (five), Fabaceae (two) and Poaceae (one) were susceptible to M. luci (GI = 4-5; Rf = 2.1-152.3). One cultivar each of carrot, passion fruit, lettuce ‘Cocktail’, cabbage ‘Bacalan’, ‘Coração’ and ‘Lombarda’, and spinach ‘Tayto’ were resistant/hypersensitive (Rf 2). The tomato ‘Actimino’, ‘Briomino’, ‘Veinal’ and ‘Vimeiro’, which carried at least one copy of the Mi-1.2 gene, were resistant to the nematode (GI = 1-2; 0.0 < Rf < 0.1). These results indicate that the tomato cultivars have potential to contribute to reduction of M. luci populations in agro-ecosystems and improve the crop yields

Commercial Potato Cultivars Exhibit Distinct Susceptibility to the Root Lesion Nematode Pratylenchus penetrans

The root lesion nematode Pratylenchus penetrans is an important plant-parasitic nematode of potato. In this study, the susceptibility of commercial potato cultivars to P. penetrans was assessed. Nematode penetration was evaluated in cultivars Agria, Camel, Kennebec, Laura, Royata, and Stemster at 1, 3, 7, and 15 days after inoculation (DAI) with 750 nematodes/plant, and an egression assay at 3 DAI with 1000 nematodes/plant. Reproduction assays of cultivars Agata, Agria, Camel, Désirée, Dirosso, Kennebec, Laura, Picasso, Royata, and Stemster were performed in 2 L pots inoculated with four P. penetrans/g soil and quantified at 60 DAI. Tenue or moderate root cell browning to advanced necrotic areas were observed after nematode penetration, and the number of nematodes/g of root gradually increased with time of infection. A lower number of deposited eggs and nematodes were observed within the roots of cultivar Laura in all assays comparatively to other cultivars. The susceptibility index (SI) was significantly lower in cultivar Laura (0.4–0.6), followed by cultivars Camel and Picasso (0.8–0.9). All remaining cultivars showed SI values above 1. Although the potato susceptibility to the nematode varied among cultivars, no differences on the average number or weight of tubers produced by each plant of inoculated versus non-inoculated plants were detected. Our data reveals that these cultivars have a distinct ability to support the reproduction of P. penetrans

Species-Specific Molecular Detection of the Root Knot Nematode Meloidogyne luci

Meloidogyne luci has been identified in various countries around the world parasitizing economically important crops and, due to its potential to cause serious damage to agriculture, was included in the European and Mediterranean Plant Protection Organization Alert List in 2017. This species shares morphological and molecular similarities with M. ethiopica and M. inornata, and a M. ethiopica group was therefore established. Although specific primers for the DNA amplification of species belonging to the M. ethiopica group have been developed previously, the primers were not species-specific, so molecular markers for the specific detection of M. luci are still needed. The objective of this study was to develop a SCAR marker for the detection of M. luci and the discrimination from other Meloidogyne spp. based on the intraspecific variability found in RAPD markers. RAPD screening of M. luci and M. ethiopica genome was used for the identification of a specific amplification product on M. luci, which was cloned, sequenced and converted into a SCAR marker. The specificity of the designed primers (Mlf/r) was tested and produced a fragment (771 bp) for all nine M. luci isolates with no amplification for the other nine Meloidogyne spp., including M. ethiopica and M. inornata. Additionally, the proper amplification of the M. luci SCAR-marker was also successful with DNA from galls of M. luci infected tomato roots. The results obtained in this study reveal that the specific molecular detection of M. luci was achieved and that the developed methodology can be used for routine diagnosis purposes, which are essential to monitoring the distribution and spread of M. luci in order to implement future effective and integrated nematode pest management programs

Pratylenchus penetrans Parasitizing Potato Crops: Morphometric and Genetic Variability of Portuguese Isolates

The root lesion Pratylenchus penetrans is an economically important pest affecting a wide range of plants. The morphometry of five P. penetrans isolates, parasitizing potato roots in Portugal, was compared and variability within and between isolates was observed. Of the 15 characters assessed, vulva position (V%) in females and the stylet length in both females/males showed the lowest coefficient of intra and inter-isolate variability. Moreover, DNA sequencing of the internal transcribed spacers (ITS) genomic region and cytochrome c oxidase subunit 1 (COI) gene was performed, in order to evaluate the intraspecific genetic variability of this species. ITS revealed higher isolate genetic diversity than the COI gene, with 15 and 7 different haplotypes from the 15 ITS and 14 COI sequences, respectively. Intra- and inter-isolate genetic diversity was found considering both genomic regions. The differentiation of these isolates was not related with their geographical origin. In spite of the high intraspecific variability, phylogenetic analyses revealed that both ITS region and COI gene separate P. penetrans from other related species. Our findings contribute to increasing the understanding of P. penetrans variability