Analysis of the olive fruit fly Bactrocera oleae transcriptome and phylogenetic classification of the major detoxification gene families

he olive fruit fly Bactrocera oleae has a unique ability to cope with olive flesh, and is the most destructive pest of olives worldwide. Its control has been largely based on the use of chemical insecticides, however, the selection of insecticide resistance against several insecticides has evolved. The study of detoxification mechanisms, which allow the olive fruit fly to defend against insecticides, and/or phytotoxins possibly present in the mesocarp, has been hampered by the lack of genomic information in this species. In the NCBI database less than 1,000 nucleotide sequences have been deposited, with less than 10 detoxification gene homologues in total. We used 454 pyrosequencing to produce, for the first time, a large transcriptome dataset for B. oleae. A total of 482,790 reads were assembled into 14,204 contigs. More than 60% of those contigs (8,630) were larger than 500 base pairs, and almost half of them matched with genes of the order of the Diptera. Analysis of the Gene Ontology (GO) distribution of unique contigs, suggests that, compared to other insects, the assembly is broadly representative for the B. oleae transcriptome. Furthermore, the transcriptome was found to contain 55 P450, 43 GST-, 15 CCE- and 18 ABC transporter-genes. Several of those detoxification genes, may putatively be involved in the ability of the olive fruit fly to deal with xenobiotics, such as plant phytotoxins and insecticides. In summary, our study has generated new data and genomic resources, which will substantially facilitate molecular studies in B. oleae, including elucidation of detoxification mechanisms of xenobiotic, as well as other important aspects of olive fruit fly biology

Effects of Salinity on Tagetes Growth, Physiology, and Shelf Life of Edible Flowers Stored in Passive Modified Atmosphere Packaging or Treated With Ethanol

Irrigation with saline water causes significant crop yield loss. However, short-term saline application might cause less negative effects on yield yet at the same time improve quality aspects of edible products. Tagetes (Tagetes patula L.) plants were subjected to salinity (0, 50, and 100 mM NaCl) and harvested flowers were stored up to 14 days in passive modified atmosphere packaging (with or without ethanol application). Salinity of 100 mM NaCl decreased plant biomass and plant size (i.e., height) and had a negative effect on physiological processes such as stomatal closure and chlorophylls content decrease. Salinity increased flower polyphenols, antioxidant activities, and total carotenoids but decreased anthocyanins, and greater impacts were found at salinity of 100 mM NaCl, providing higher antioxidant value of the edible flowers. Short-term saline exposure of tagetes plants activated metabolic processes and as a result there was an accumulation of minerals such as N, P, Na, and Zn on edible flowers. During storage, salinity maintained but ethanol application increased the flower CO2 production. Ethanol application decreased the decay of flowers subjected to 100 mM NaCl. Flower weight losses and marketability accelerated at salinity of 100 mM NaCl after 14 days of storage. Tagetes flowers demonstrated induction in both non-enzymatic (i.e., proline content) and enzymatic mechanisms (catalase) to overcome stress caused by salinity during harvest stage and/or ethanol at storage. Our results have shown that short-term exposure to salinity and/or ethanol is able to achieve higher carotenoids and anthocyanins levels and these compounds can be considered as a new source of nutraceuticals

Effect of phosphorus application rate on Mentha spicata L. grown in deep flow technique (DFT)

The present study evaluated the impact of phosphorus application rate on plant growth and physiological parameters, antioxidant activity, chemical composition, and essential oil yield and composition of hydroponically grown spearmint plants. Increased P levels resulted in high dry matter content of the aerial part. Antioxidant activity of spearmint leaves was significantly higher at the highest P levels. Although essential oil yield was not affected, essential oil composition varied among the studied P levels, especially carvone content. Total and individual organic acids content was higher when 50 mg/L P were added in the nutrient solution. Rosmarinic acid was the main detected phenolic compound, while the highest total phenolic compounds and rosmarinic acid content was observed at 50 and 70 mg/L of P, respectively. In conclusion, phosphorus application rate may affect spearmint growth and development, as well as chemical composition and essential oil composition.The authors are grateful to the project AgroLabs that has been developed under the Programme Interreg V-B Balkan – Mediterranean 2014-2020, co-funded by the European Union and National Funds of the participating countries. Also, the authors are grateful to the Foundation for Science and Technology (, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2013), A. Fernandes (SFRH/BPD/114753/2016) and L. Barros contract; also to FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E.info:eu-repo/semantics/publishedVersio

Magnesium and manganese induced changes on chemical, nutritional, antioxidant and antimicrobial properties of the pansy and Viola edible flowers

composition was determined using standard methods. Free sugars, fatty acids, organic acids, tocopherols, and phenolic compounds were analyzed using various HPLC and GC devises. The extract’s antimicrobial, antioxidant, cytotoxicity, and anti-inflammatory activity were assessed. The results indicated that Mg enrichment negatively affected plant growth and mineral accumulation but improved photosynthetic performance. The edible flowers contained significant amounts of protein, low levels of fat, and varying sugar contents, such as glucose and fructose. Various fatty acids and phenolic compounds were identified, with different concentrations depending on the treatment. The flowers exhibited antioxidant potential, antimicrobial activity, cytotoxic effects, and antiinflammatory properties. The correlations between the investigated parameters not only expand knowledge on Mg and Mn interaction but also catalyze significant advancements in sustainable agriculture and food health, fostering a healthier and more conscious future.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES (PIDDAC) to CIMO (UIDB/00690/2020 and UIDP/00690/ 2020) and SusTEC (LA/P/0007/2020). L. Barros, Ricardo C. Calhelha, S. A. Heleno and T. C. Finimundy thank the national funding by FCT through the institutional scientific employment program-contract for her contract, while M. Carocho thanks FCT through the individual scientific employment program-contract (CEECIND/00831/2018). I. Oliveira thanks FCT for her PhD grant (BD/06017/2020).info:eu-repo/semantics/publishedVersio

The effects of biostimulants, biofertilizers and water-stress on nutritional value and chemical composition of two spinach genotypes (Spinacia oleracea L.)

In the present study, the e ect of biostimulants application on the nutritional quality and bioactive properties of spinach cultivated in protected environment under water stress conditions was evaluated. For this purpose, four commercially available biostimulant products (Megafol (MEG), Aminovert (AM), Veramin Ca (V), Twin Antistress (TA), and two spinach genotypes (Fuji F1 and Viroflay) were tested under two irrigation regimes (normal irrigation (W+), and water-holding (W–). Fat and carbohydrates content was favored by water stress when Megafol (MEGW+) and Veramin (VW+) were applied on Fuji plants, while calorific value was also increased by MEGW+ treatment. In contrast, protein and ash content increased when AMW– and TAW+ were applied on Viroflay plants. Ra nose and glucose were the most abundant sugars, followed by sucrose and fructose, with the highest contents recorded for Fuji plants when AMW+ (fructose, glucose and total carbohydrates), CW– (sucrose), and TAW– (ra nose) treatments were applied. Regarding organic acids, oxalic and malic acid which had the highest contents for the TAW– (Viroflay plants) and AMW– (Fuji plants) treatments, respectively. - and -tocopherol were the only isoforms detected with MEGW– and VW– inducing the biosynthesis of -tocopherol, while AMW+ increased -tocopherol content in Fuji plants. The main fatty acids were -linolenic and linoleic acids which were detected in the highest amounts in AMW–, AMW+, and TAW+ the former and in AMW–, VW–, and CW+ the latter. Regarding phenolic compounds content, peak 12 (5,30,40-Trihydroxy-3-methoxy-6:7-methylenedioxyflavone-40-glucuronide) was the most abundant compound, especially in Viroflay plants under normal irrigation and no biostimulants added (CW–). The antioxidant and cytotoxic activity of the tested samples did not show promising results when compared with the positive controls, while a variable antibacterial activity was recorded depending on the tested biostimulant, irrigation regime and genotype. In conclusion, a variable e ect of the tested biostimulants and irrigation regimes was observed on bioactive properties and chemical composition of both spinach genotypes which highlights the need for further research in order to make profound conclusions regarding the positive effects of biostimulants under water stress conditions.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2019); national funding by FCT, P.I., through the institutional scientific employment program-contract for M.I.D., R.C.C. and L.B.; C.P. contract though the celebration of program-contract foreseen in No. 4, 5 and 6 of article 23º of Decree-Law No. 57/2016, of 29th August, amended by Law No. 57/2017, of 19th July; to FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E and TRANSCoLAB 0612_TRANS_CO_LAB_2_P.info:eu-repo/semantics/publishedVersio

The sustainable use of cotton, hazelnut and ground peanut waste in vegetable crop production

The environmental burden from crop production byproducts is gradually increasing and necessitates the sustainable management of waste towards a circular economy approach. In the present study, three byproducts (cotton ginning waste (CGW), ground hazelnut husks (GHH) and ground peanut husks (GPH)) were evaluated in lettuce cultivation. For this purpose, the tested materials were incorporated in soil at two different rates (25% and 50% of total substrate volume) while a control treatment (no addition of byproducts) was also considered. Fresh weight per plant and total yield was the highest for the GHH50% treatment. The highest fat, protein, carbohydrates and energy content were observed for the CGW25% treatment. Chemical composition also differed among the tested byproducts where CGW25% treatment had the highest total tocopherols, sugars (sucrose, fructose, trehalose and total sugars) and organic acids content. The most abundant fatty acids were α-linolenic, linoleic and palmitic acid in all the tested treatments, while the highest antioxidant activity was observed for the GHH50% treatment. Regarding polyphenols, phenolic acids content was the highest in the GHH treatments, whereas flavonoids were the highest for the CGW25% treatment. No cytotoxicity against the PLP2 non-tumor cell line was observed, whereas only the GPH50% treatment showed moderate efficacy against HeLa, HepG2 and MCF-7 cell lines. The tested extracts also showed moderate antibacterial activities and only the extracts from the CGW50% treatment were more effective than the positive control against Trichoderma viride. In conclusion, the present results showed the great potential of using the tested byproducts as soil amendments for vegetable crops production, since they may improve the nutritional parameters, the chemical profile and the bioactivities of the final product. The suggested alternative use of the tested byproducts not only will increase the added value of crops but will also alleviate the environmental burden from bulky agroindustry byproducts.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support by national funds FCT/MCTES to CIMO (UIDB/00690/2020) and national funding by FCT, P.I., through the institutional scientific employment program-contract for L. Barros, A. Fernandes, M. I. Dias and R. Calhelha. C. Pereira though the celebration of program-contract foreseen in No. 4, 5 and 6 of article 23 of Decree-Law No. 57/2016, of 29 August, mended by Law No. 57/2017, of 19 July. The authors are grateful to the FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E; and to the Ministry of Education, Science and Technological Development of Republic of Serbia (451-03-68/2020-14/200007).info:eu-repo/semantics/publishedVersio

Narrativas dos profissionais dos ensinos pré-escolar e primeiro ciclo face às crianças com necessidades educativas especiais: o outro lado da história

[Resumo] As debilidades na formação dos educadores e professores quanto as NEE sao uma importante variável influenciadora na e para a construção de urna equidade educativa. Registe-se a afirmação proferida na Declaras;ao de Salamanca: "a preparação adequada de todo o pessoal educativo é o factor chave na promosção das escolas inclusivas." Toma-se imprescindível conhecermos as formas de pensar, de actuar, a diversidade de sentimentos e estratégias que os profissionais poderao demonstrar como resultado de lidarem com uma criança com NEE, para que possamos melhor compreender e interpretar a direcs;ao para a construção de um diálogo efectivo de parceria educacional. Pretende-se, assim, através de urna metodologia qualitativa - análise de narrativas - o desenvolvimento de um programa de forma<;ao que promova desenvolvimento de atitudes positivas face a escola inclusiva, a aquisis;ao de novas competencias de ensino, que permita aos profissionais serem mais responsivos as necessidades educativas das criança

Comparison between different extraction methods in the recovery of bioactive molecules from Melissa officinalis L. under sustainable cultivation: chemical and bioactive characterization

Melissa officinalis L., from the Lamiaceae family, is one of the most important medicinal and aromatic plants with potential in the market. With the passing of time, the use of medicinal plants in the treatment of some illness has gone from the simplest forms of local treatment to the industrial manufacture of phytotherapics. In addition to their medicinal effect, they can also be used in the form of infusions and decoctions and in various food preparations. In this sense, the objective of this work was to compare three different extraction methods: infusion (100% water), maceration (80:20 ethanol: water v:v) and ultrasound-assisted extraction (UAE) under previous optimized extraction conditions (33.0 +- 3.2 min, 371.7 +- 19.3Wand 39.9 +- 1.4% ethanol), in plants grown under sustainable cultivation under full irrigation in June. The parameters studied included bioactive evaluation by antioxidant (thiobarbituric acid reactive substances—TBARS), cytotoxicity (sulforhodamine B) and anti-inflammatory (RAWcells) assays. The composition of phenolic compounds and organic acids was also studied by GC-MS and HPLC-DAD, respectively. According to the obtained results, eight phenolic compounds were identified and quantified, being rosmarinic acid the main one for the three extraction methods (infusion: 107.1 +- 0.9 mg/g extract; maceration: 155.7 +- 0.3 mg/g extract; UAE: 118.7 0.6 mg/g extract). For Lithospermic acid A isomer (25.25 +- 0.01 mg/g) and Hydroxylsalvianolic E (111.70 +- 2.20 mg/g), the UAE revealed the lowest content of individual polyphenols, whereas maceration recorded the highest extractability. On the other hand, the content of six of the eight polyphenols detected for the ultrasound-assisted extraction was similar to the infusion and maceration methods. In terms of antioxidant activity, the infusions showed the highest capacity (3.00 +- 0.14 +-g/mL), followed by maceration (5.33 +- 0.30 +-g/mL) and UAE (12 +- 0.15 +-g/mL). The highest anti-inflammatory activity was verified for the infusion (244 +- 11 +-g/mL), followed by UAE (305 +- 9 +-g/mL), with no activity recorded for the maceration extract (>400 +-g/mL). The antitumor properties were evaluated in five cell lines, with the best results being recorded for infusion, except AGS (24 +- 1 +-g/mL) whereThe authors are grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020). L. Barros and M.I. Dias thank the national funding by FCT through the institutional scientific employment program-contract for her contract, while M. Carocho and S. Heleno thank FCT through the indiviual scientific employment program-contracts (CEECIND/00831/2018 and CEECIND/03040/2017). I. Oliveira thanks FCT for her PhD grant (BD/06017/2020). To FEDER-Interreg España-Portugal programme for financial support through the project TRANSCoLAB 0612_TRANS_CO_LAB_2_P; to ERDF through the Regional Operational ProgramNorth 2020, within the scope of Project GreenHealth— Norte-01-0145-FEDER-000042.info:eu-repo/semantics/publishedVersio

Phytotoxic and insecticidal activity of industrial hemp (Cannabis sativa L.) extracts against Plodia interpunctella Hübner—A potential sunflower grain protectant

The biological activity (contact and contact-digestive toxicity, repellent and fumigant effects, effect on the insect’s development and life cycle parameters) of industrial hemp (Cannabis sativa L.) ethanolic extract was assessed against Plodia interpunctella, the most destructive storage pest of sunflower. Additionally, the study aimed to examine the phytotoxic activity of the extract in order to assess its potential as a sunflower grain protectant. Phytotoxicity assessment was based on the effect on germination energy and seed germination and the activity of antioxidative enzymes, enzymes of the polyphenolic metabolism, and the intensity of lipid peroxidation in sunflower seedlings. The antioxidant capacity and content of phenolic compounds (total phenolics and total tannins) were also measured in seedlings. In the experiments, 70% ethanolic extract of dried flowering buds of industrial hemp (variety Helena) was applied at 0.5%, 1.0% and 2.0% concentrations directly on sunflower seeds. Ethanol solution (70%) was the control. The hemp extract (1% and 2%) revealed medium repellence for P. interpunctella larvae (L3–4) while at 2% concentration it caused 42.5% larval mortality after 72 h. Moreover, the insect’s development was prolonged and fecundity significantly reduced in hemp treatments. The extract did not exhibit fumigant activity. Germination energy and germination of sunflower seeds were stimulated in treatment with 2% hemp extract, while most biochemical parameters of the seedlings were not significantly affected by the hemp extract

Effects on Lettuce Yield Parameters and Toxicological Safety Assessment of a Plant-Derived Formulation Based on Rosemary and Eucalyptus Essential Oils

Essential oils from medicinal and aromatic plants are increasingly recognized as a promising class of green molecules for use in crop production. In many cases, the beneficial aspects of a substance are not supported by sufficient toxicological safety testing, even though recent reports suggest that some compounds may be toxic to terrestrial or aquatic non-target species. It is, therefore, essential to investigate the possibility of adverse effects on non-target animals and humans exposed to these substances through the consumption of fruit and/or vegetables. The present study aims to examine the potential effects on yield and quality parameters and investigate the level of in vitro and in vivo toxicity of an Eco-product (EP) based on rosemary and eucalyptus essential oils, to provide a measure for safe use in the agricultural sector. The product was evaluated in lettuce crop production and indicated that one-time application of the EP formula increases yield, activating various secondary metabolism pathways of the plant to cope with oxidative stress. Cytotoxicity assays and in vivo acute oral and dermal toxicity studies suggest that the tested compound does not pose any significant health hazard, and the dissolved product can be classified in Category 5, according to the Globally Harmonized System of Classification and Labelling of Chemicals (GHS)