Resource utilization and trophic position of nematodes and harpacticoid copepods in and adjacent to Zostera noltii beds

This study examines the resource use and trophic position of nematodes and harpacticoid copepods at the genus/species level in an estuarine food web in Zostera noltii beds and in adjacent bare sediments using the natural abundance of stable carbon and nitrogen isotopes. Microphytobenthos and/or epiphytes are among the main resources of most taxa, but seagrass detritus and sediment particulate organic matter contribute as well to meiobenthos nutrition, which are also available in deeper sediment layers and in unvegetated patches close to seagrass beds. A predominant dependence on chemoautotrophic bacteria was demonstrated for the nematode genus Terschellingia and the copepod family Cletodidae. A predatory feeding mode is illustrated for Paracomesoma and other Comesomatidae, which were previously considered first-level consumers (deposit feeders) according to their buccal morphology. The considerable variation found in both resource use and trophic level among nematode genera from the same feeding type, and even among congeneric nematode species, shows that the interpretation of nematode feeding ecology based purely on mouth morphology should be avoided

Tomato Response to Fusarium spp. Infection under Field Conditions: Study of Potential Genes Involved

Tomato is one of the most important horticultural crops in the world and is severely affected by Fusarium diseases. To successfully manage these diseases, new insights on the expression of plant– pathogen interaction genes involved in immunity responses to Fusarium spp. infection are required. The aim of this study was to assess the level of infection of Fusarium spp. in field tomato samples and to evaluate the differential expression of target genes involved in plant–pathogen interactions in groups presenting different infection levels. Our study was able to detect Fusarium spp. in 16 from a total of 20 samples, proving the effectiveness of the primer set designed in the ITS region for its detection, and allowed the identification of two main different species complexes: Fusarium oxysporum and Fusarium incarnatum-equiseti. Results demonstrated that the level of infection positively influenced the expression of the transcription factor WRKY41 and the CBEF (calcium-binding EF hand family protein) genes, involved in plant innate resistance to pathogens. To the best of our knowledge, this is the first time that the expression of tomato defense-related gene expression is studied in response to Fusarium infection under natural field conditions. We highlight the importance of these studies for the identification of candidate genes to incorporate new sources of resistance in tomato and achieve sustainable plant disease management.This research was supported by projects “Development of a new virus-based vector to control TSWV in tomato plants” with the references ALT20-03-0145-FEDER-028266 and PTDC/ASP-PLA/28266/2017, and “Control of olive anthracnose through gene silencing and gene ex- pression using a plant virus vector” with the references ALT20-03-0145-FEDER-028263 and PTDC/ASP- PLA/28263/2017, both projects co-financed by the European Union through the European Regional Development Fund, under the ALENTEJO 2020 (Regional Operational Program of the Alentejo), ALGARVE 2020 (Regional Operational Program of the Algarve) and through the Foundation for Science and Technology (FCT), in its national component. M.P. was supported by Portuguese National Funds through FCT/MCTES, under the PhD scholarship SFRH/BD/145321/2019, co-financed by the European Social Fund through the Regional Operational Program of the Alentejo. J.A.R. was supported by Portuguese National Funds through Project ALT20-03-0246-FEDER-000056, “BIOPRO- TOMATE: Bioproteção do tomateiro contra a fusariose—impacto das práticas agronómicas”, under scholarship BI_MESTRE_Uevora_CER_BIOPROTOMATE, co-financed by the European Regional Development Fund through Regional Operational Program Alentejo 2020

Defense Strategies: The Role of Transcription Factors in Tomato–Pathogen Interaction

Tomato, one of the most cultivated and economically important vegetable crops throughout the world, is affected by a panoply of different pathogens that reduce yield and affect product quality. The study of tomato–pathogen system arises as an ideal system for better understanding the molecular mechanisms underlying disease resistance, offering an opportunity of improving yield and quality of the products. Among several genes already identified in tomato response to pathogens, we highlight those encoding the transcription factors (TFs). TFs act as transcriptional activators or repressors of gene expression and are involved in large-scale biological phenomena. They are key regulators of central components of plant innate immune system and basal defense in diverse biological processes, including defense responses to pathogens. Here, we present an overview of recent studies of tomato TFs regarding defense responses to biotic stresses. Hence, we focus on different families of TFs, selected for their abundance, importance, and availability of functionally well-characterized members in response to pathogen attack. Tomato TFs’ roles and possibilities related to their use for engineering pathogen resistance in tomato are presented. With this review, we intend to provide new insights into the regulation of tomato defense mechanisms against invading pathogens in view of plant breeding.This work was funded by the projects “Development of a new virus-based vector to control TSWV in tomato plants” with the references ALT20-03-0145-FEDER-028266 and PTDC/ASP-PLA/28266/2017, and “Control of olive anthracnose through gene silencing and gene expression using a plant virus vector” with the references ALT20-03-0145-FEDER-028263 and PTDC/ASP-PLA/28263/2017, co-financed by the European Union through the European Regional Development Fund, under the ALENTEJO 2020 (Regional Operational Program of the Alentejo), ALGARVE 2020 (Regional Operational Program of the Algarve) and through the Foundation for Science and Technology (FCT), in its national component. M.P. was supported by Portuguese National Funds through FCT/MCTES, under the PhD scholarship SFRH/BD/145321/2019, co-financed by the European Social Fund through the Regional Operational Program of the Alentejo. This work was also supported by National Funds through FCT under the Project UIDB/05183/2020

Co infection of OMMV and OLV-1 enhances symptoms and increases both viruses accumulation and viral derived siRNAs in plants

Previous extensive field surveys in olive orchards have revealed high levels of Olive mild mosaic virus (OMMV) and Olive latent virus 1 (OLV-1), frequently appearing in mixed infections. These viruses belong to genus Alphanecrovirus and their RNA dependent RNA polymerase (RdRp),as well as their p6 and p8 amino acid sequences share over 87% identity

Metagenomic Assessment Unravels Fungal Microbiota Associated to Grapevine Trunk Diseases

Grapevine trunk diseases (GTDs) are among the most important problems that affect the longevity and productivity of vineyards in all the major growing regions of the world. They are slow-progression diseases caused by several wood-inhabiting fungi with similar life cycles and epidemiology. The simultaneous presence of multiple trunk pathogens in a single plant together with the inconsistent GTDs symptoms expression, their isolation in asymptomatic plants, and the absence of effective treatments make these diseases extremely complex to identify and eradicate. Aiming to gain a better knowledge of GTDs and search sustainable alternatives to limit their development, the present work studied the fungal community structure associated with GTDs symptomatic and asymptomatic grapevines, following a metagenomic approach. Two important cultivars from the Alentejo region with different levels of susceptibility to GTDs were selected, namely, ‘Alicante Bouschet’ and ‘Trincadeira’. Deep sequencing of fungal-directed ITS1 amplicon led to the detection of 258 taxa, including 10 fungi previously described as responsible for GTDs. Symptomatic plants exhibited a lower abundance of GTDs-associated fungi, although with significantly higher diversity of those pathogens. Our results demonstrated that trunk diseases symptoms are intensified by a set of multiple GTDs-associated fungi on the same plant. The composition of fungal endophytic communities was significantly different according to the symptomatology and it was not affected by the cultivar. This study opens new perspectives in the study of GTDs-associated fungi and their relation to the symptomatology in grapevines.This research was supported by Portuguese National Funds through FCT- MCTES under the PhD scholarship SFRH/BD/145321/2019, attributed to M.P., co-financed by the European Social Fund through the Regional Operational Program of the Alentejo. It was also supported by the project “Control of olive anthracnose through gene silencing and gene expression using a plant virus vector” (ALT20-03-0145-FEDER-028263 and PTDC/ASP-PLA/28263/2017) and by the project “Development of a new virus-based vector to control TSWV in tomato plants” (ALT20-03- 0145-FEDER-028266 and PTDC/ASP-PLA/28266/2017), both projects co-financed by the European Union through the European Regional Development Fund, under the Alentejo 2020, Algarve 2020 and through the FCT, in its national component

Benthic nematode assemblage composition and diversity during a natural recovery process of Zostera noltii seagrass beds,

In 2008, the stable seagrass beds of the Mira estuary (SW Portugal) disappeared completely; however, during 2009, they have begun to present early symptoms of natural recovery, characterised by a strongly heterogeneous distribution. This study was designed to investigate the spatial and temporal variability patterns of species composition, densities and trophic composition of the benthic nematode assemblages in this early recovery process, at two sampling sites with three stations each and at five sampling occasions. Because of the erratic and highly patchy seagrass recovery and the high environmental similarity of the two sampling sites, we expected within-site variability in nematode assemblages to exceed between-site variability. However, contrary to that expectation, whilst nematode genus composition was broadly similar between sites, nematode densities differed significantly between sites, and this between-site variability exceeded within-site variability. This may be linked to differences in the Zostera recovery patterns between both sites. In addition, no clear temporal patterns of nematode density, trophic composition and diversity were evident

Agricultura sustentável e as novas técnicas genómicas

As Novas Técnicas Genómicas (NGTs) são técnicas desenvolvidas a partir de 2001, após a adoção da legislação sobre organismos gene ticamente modificados (OGM). Estas incluem a mutagénese dirigida, cisgénese, intragénese e transgénese. As NGTs permitem mutações genómicas precisas num organismo, podendo faze lo sem inserção de material genético estranho (mutagénese dirigida); ou com a inserção de material genético, diretamente (cisgénese) ou de uma cópia reorganizada (intragénese), proveniente de um organismo com o qual se poderia cruzar na natureza; ou ainda, por outro lado, com inserção de material genético de um organismo não cruzável na natureza (transgénese). Independentemente destas diferenças, atualmente todas as NGTs estão enquadradas na legislação dos OGMs, pelo que a sua aplicação em agricultura é bastante restritiva. Recentemente, e como já se verifica em vários outros países, a Europa tem mostrado abertura para excluir algumas NGTs da legislação dos OGM. Este artigo descreve as principais NGTs e o seu papel potencial para responder aos desafios atuais da agricultura. São ainda mostrados os avanços realizados na Europa para criação de uma legislação própria para algumas NGTs, assim como as vantagens que tal poderia trazer para a sustentabilidade da agricultura e a competitividade da economia europeia.Fundação para a Ciência e a Tecnologiainfo:eu-repo/semantics/publishedVersio

Enzymatic activity on sediments and nematode assemblage responses during seagrass beds habitat recovery following the disturbance of the traditional digging activity of bivalves harvesting.

Sediment digging is an anthropogenic activity connected to the exploitation of living resources in estuarine and marine environments. The knowledge on the functional responses of the benthic assemblages to the physical disturbance is an important baseline to understand the ecological processes of the habitat recovery and restoration and to develop tools for the management of the harvesting activities. To investigate the effects of the digging activity of the bivalves on Zostera noltii seagrass beds a manipulative field experiment was conducted that included the enzymatic activity of sediments and the associated nematode assemblages. Four plots (two undisturbed serving as control and two dug to collect bivalves - treatment) with 18 subplots were randomly located at seagrass beds in the Mira estuary at the SW coast of Portugal. Samples were randomly and unrepeatably collected from three subplots of each plot in five different occasions, before sediment digging (T0) up to six months after disturbance (T5). Microbial activity in sediments was assess by determining the extracelular enzymatic activity of six hydrolytic enzymes (sulfatase, phosphatase, b -N-acetilglucosaminidase, b-glucosidase, urease, protease) and two oxidoreductases (phenol oxidase and peroxidase). The microbial community status was also assessed through the measurement of dehydrogenase, which reflects microbial respiration. The nematode assemblages composition, biodiversity and trophic composition at different sampling occasions were also analyzed. The fluorometric and biochemical parameters analysed of the Z. noltii plants during the experimental period showed a recovery of the seagrass beds, and it was detected an increase of the enzymatic activity of the sediments after disturbance. The nematodes assemblages were similar in all sampling occasions. The seagrass beds and the nematodes assemblages associated showed a high resilience to the stress caused by the traditional bivalves digging activity. The obtained results allow the development of a management programme for the commercial fishing activity to maintain the good environmental status and minimized the secondary environmental effects on marine and estuarine habitats through the establishment of a baseline for the regulation of the harvesting frequency

Bisifusarium lunatum causing cladode soft rot in pear cactus (Nopalea cochenillifera) in Brazil

Cactus pear variety Miuda (Nopalea cochenillifera L. Salm-Dyck) is an important crop for the Northeast region of Brazil, composing one of the main sources of animal feed. By April 2021, cladode rot caused death of several cactus pear plants in a production area located in Itaporanga, Paraíba State, Brazil (7°21'55.35'S, 38°11'38.68'W). The infected cladodes showed brown circular necrotic spots and soft rot with perforations that extended throughout the cladode, followed by tipping over and death of the infected plants. The incidence of the disease ranged from 10 to 30% of the plants. Bisifusarium strains were isolated and cultured on potato dextrose agar (PDA) and synthetic nutrient-poor agar (SNA). The colonies were purple in color on PDA. On SNA, macroconidia (n = 100) were abundant, hyaline, slightly falcate, and three-septate, measuring 11.0 to 23.1 × 2.3 to 4.1 µm. Microconidia (n = 100) were oval and generally aseptate, measuring 4.1 to 8.7 × 2.3 to 3.0 µm. Conidiogenic cells formed into short monophialides. Chlamydospores were not observed. According to these morphological features, the pathogen was initially identified as Bisifusarium lunatum (Gryzenhout et al. 2017). For further confirmation of the identification, the partial sequences of translation elongation factor 1-alpha (TEF1-a) and the second largest subunit of RNA polymerase II (RPB2) genes were sequenced for a representative isolate (CMA 34; GenBank accession nos. OR536502 for TEF1-a and OR553509 for RPB2) and compared with other Bisifusarium species from the GenBank database. Subsequently, it was subjected to a phylogenetic analysis of maximum likelihood, including previously published sequences. According to BLAST searches, the TEF1-a and RPB2 sequences were 99% (637/640 nt) and 100% (312/312 nt) similar to B. lunatum (COUFAL0213: TEF1-a [MK640219] and RPB2 [MK301291]), respectively. The isolate was also clustered in a clade containing the ex-type of B. lunatum with 100% support (SH-aLRT and UFboot), being confidently assigned to this species. The pathogenicity test was performed as described by Medeiros et al. (2015) by using healthy 2-month-old cactus pear seedlings (n = 10) cultivated in a greenhouse. Sterile toothpicks were distributed over colonies of the representative isolate grown on PDA at 25 ± 2°C for 7 days. Seedling cladodes were stuck with the toothpicks, moistened with sterile water, and covered with transparent plastic bags for 24 h, thus simulating a humid chamber. Following 3 months, all control plants (stuck with sterile toothpicks) remained healthy, whereas those inoculated with the representative isolate exhibited rot symptoms. This test was performed two times. B. lunatum was reisolated from symptomatic cladodes and identified as previously described, thus fulfilling Koch’s postulates. To our best knowledge, this is the first report of B. lunatum causing soft rot on N. cochenillifera in Brazil. Besides N. cochenillifera, this species was also reported on Opuntia ficus-indica in India (Gryzenhout et al. 2017), which raises concern regarding its ability to infect other forage sources for cattle feed in Brazilian semiarid regions. The present study highlights that the precise identification of B. lunatum is a key factor to adjust control strategies and management of the disease to prevent the spread of this disease to other crops.Instituto Nacional do Semiárido (INSA); Fundação de Apoio à Pesquisa do Estado da Paraíba (FAPESQ); Fundação para a Ciência e a Tecnologiainfo:eu-repo/semantics/publishedVersio