Reuse of Wasted Bread as Soil Amendment: Bioprocessing, Effects on Alkaline Soil and Escarole (Cichorium endivia) Production

: In an era characterized by land degradation, climate change, and a growing population, ensuring high-yield productions with limited resources is of utmost importance. In this context, the use of novel soil amendments and the exploitation of plant growth-promoting microorganisms potential are considered promising tools for developing a more sustainable primary production. This study aimed at investigating the potential of bread, which represents a large portion of the global food waste, to be used as an organic soil amendment. A bioprocessed wasted bread, obtained by an enzymatic treatment coupled with fermentation, together with unprocessed wasted bread were used as amendments in a pot trial. An integrated analytical plan aimed at assessing i) the modification of the physicochemical properties of a typical Mediterranean alkaline agricultural soil, and ii) the plant growth-promoting effect on escarole (Cichorium endivia var. Cuartana), used as indicator crop, was carried out. Compared to the unamended soils, the use of biomasses raised the soil organic content (up to 37%) and total nitrogen content (up to 40%). Moreover, the lower pH and the higher organic acid content, especially in bioprocessed wasted bread, determined a major availability of Mn, Fe, and Cu in amended soils. The escaroles from pots amended with raw and bioprocessed bread had a number of leaves, 1.7- and 1.4-fold higher than plants cultivated on unamended pots, respectively, showing no apparent phytotoxicity and thus confirming the possible re-utilization of such residual biomasses as agriculture amendments.

Leaf nutrient content and transcriptomic analyses of endive (Cichorium endivia) stressed by downpour-induced waterlog reveal a gene network regulating kestose and inulin contents

Endive (Cichorium endivia L.), a vegetable consumed as fresh or packaged salads, is mostly cultivated outdoors and known to be sensitive to waterlogging in terms of yield and quality. Phenotypic, metabolic and transcriptomic analyses were used to study variations in curly- (‘Domari’, ‘Myrna’) and smooth-leafed (‘Flester’, ‘Confiance’) cultivars grown in short-term waterlog due to rainfall excess before harvest. After recording loss of head weights in all cultivars (6-35%), which was minimal in ‘Flester’, NMR untargeted profiling revealed variations as influenced by genotype, environment and interactions, and included drop of total carbohydrates (6–50%) and polyols (3–37%), gain of organic acids (2–30%) and phenylpropanoids (98–560%), and cultivar-specific fluctuations of amino acids (−37 to +15%). The analysis of differentially expressed genes showed GO term enrichment consistent with waterlog stress and included the carbohydrate metabolic process. The loss of sucrose, kestose and inulin recurred in all cultivars and the sucrose-inulin route was investigated by covering over 50 genes of sucrose branch and key inulin synthesis (fructosyltransferases) and catabolism (fructan exohydrolases) genes. The lowered expression of a sucrose gene subset together with that of SUCROSE:SUCROSE-1-FRUCTOSYLTRANSFERASE (1-SST) may have accounted for sucrose and kestose contents drop in the leaves of waterlogged plants. Two anti-correlated modules harbouring candidate hub-genes, including 1-SST, were identified by weighted gene correlation network analysis, and proposed to control positively and negatively kestose levels. In silico analysis further pointed at transcription factors of GATA, DOF, WRKY types as putative regulators of 1-SST.

Reduction of Nitrate Content in Baby-Leaf Lettuce and Cichorium endivia Through the Soilless Cultivation System, Electrical Conductivity and Management of Nutrient Solution

Soilless cultivation systems are efficient tools to control nitrates by managing nutrient solution (NS) salinity and nitrogen availability, however, these nitrate-lowering strategies require appropriate calibration based on species/genotype-specific responses interacting with climate and growing conditions. Three experiments were carried out on lettuce and Cichorium endivia grown in ebb-and-flow (EF) and floating (FL) systems at two levels of NS salinity (EC = 2.5 and 3.5 dS m−1) (EC2.5, EC3.5, respectively) under autumn and early-spring (lettuce) and winter and late-spring conditions (C. endivia). Nitrogen deprivation (NS withdrawal a few days before the harvest) was tested at EC2.5, in the autumn and winter cycles. The EF-system caused an increase in salinity in the substrate where roots mainly develop so it mimicked the effect of the EC3.5 treatment. In the winter-grown lettuce, the EF-system or EC3.5 treatment was effective in reducing the nitrate level without effects on yield, with the EF baby-leaf showing an improved quality (color, dry matter, chlorophylls, carotenoid, vitamin C, phenol). In both seasons, the EF/EC3.5 treatment resulted in a decline in productivity, despite a further reduction in nitrate content and a rise in product quality occurring. This response was strictly linked to the increasing salt-stress loaded by the EC3.5/EF as highlighted by the concurrent Cl− accumulation. In early-spring, the FL/EC3.5 combination may represent a trade-off between yield, nitrate content and product quality. In contrast, in winter-grown endive/escarole the EC3.5, EF and EC3.5/EF reduced the nitrate level with no effect on yield, product quality or Cl− uptake, thus proving them to be more salt-tolerant than lettuce. High temperatures during the late-spring cycle promoted nitrate and Cl− uptake, overcoming the nitrate-controlling effect of salinity charged by the EF system or EC3.5. The nitrate level decreased after 3 day-long (lettuce) or 6 day-long (C. endivia) NS withdrawal. In C. endivia and EF-grown lettuce, it provoked a decrease in yield, but a concurrent improvement in baby-leaf appearance and nutritional quality. More insights are needed to fine-tune the duration of the NS removal taking into account the soilless system used and species-specific characteristics.

First Report of Cichorium endivia (Asteraceae) as a Natural Host of Groundnut ringspot orthotospovirus in Brazil.

Endive (Cichorium endivia L.) is a very important cash crop for small farmers in Brazil. During inspections conducted in the summer season of 2019–2020, leaf samples of C. endivia ‘La Spezia’ seedlings exhibiting typical symptoms of orthotospoviruses infection (viz. concentric chlorotic spots and apical leaf deformation; ≈ 10%) were collected in commercial greenhouses in Brasília–DF, Central Brazil. Leaves of one healthy and three symptomatic plants were initially evaluated via double antibody sandwich enzyme-linked immunosorbent assay (DAS–ELISA) with polyclonal antibodies (produced at CNPH) raised against the nucleoprotein of the three major orthotospoviruses: tomato spotted wilt orthotospovirus (TSWV), groundnut ringspot orthotospovirus (GRSV) and tomato chlorotic spot orthotospovirus (TCSV). Strong serological reactions were observed only against GRSV antibodies exclusively in extracts from symptomatic samples. In order to confirm the causal agent of those symptoms, total RNA was extracted (Trizol®; Sigma) from infected leaf samples and used in a two-step reverse transcriptase polymerase chain reaction (RT–PCR) approach. Synthesis of the cDNA was carried out with the J13 primer (5’–CCC GGA TCC AGA GCA AT–3’) (Cortez et al., 2001) followed by PCR assays with the primer pair BR60 (5’–AGA GCA ATC GTG TCA–3`) and BR65 (5’–ATC AAG CCT TCT GAA AGT CAT–3’) (Eiras et al., 2001). This primer set amplifies a fragment of 453 bp including the untranslated region at the 3’ terminus of the small RNA and the protein N–coding gene of at least five orthotospoviruses: TSWV, GRSV, TCSV, chrysanthemum stem necrosis orthotospovirus (CSNV) and zucchini lethal chlorosis orthotospovirus (ZLCV) (Eiras et al., 2001). The obtained amplicons (≈ 432 bp) were subsequently subjected to Sanger dideoxy nucleotide sequencing at CNPH. BLASTn analysis showed >99% identity with a wide array of GRSV isolates available in the GenBank. The nucleotide sequence of Tospo #1 (MT215222) and Tospo #3 (MT215224) isolates displayed 100% identity between them, whereas the Tospo #2 (MT215223) isolate displayed one non–synonymous point mutation in the 3’ untranslated region in comparison with the former two isolates. Three plants of C. endivia, Capsicum annuum L. cv. Ikeda, tomato (Solanum lycopersicum L.) cv. Santa Clara and its isoline ‘LAM–147’ (with the Sw–5 resistance gene), Nicotiana rustica L., Lactuca sativa L. (‘Vanda’ and ‘PI-342444’) and Gomphrena globosa L. were mechanically inoculated individually with each GRSV isolate in order to confirm their pathogenicity. Chlorotic lesions and mosaic were observed seven days after inoculation of all plant materials, except the tomato inbred line ‘LAM–147’, which has the Sw-5 gene that confers broad–spectrum resistance to all Brazilian orthotospoviruses (Boiteux and Giordano, 1993). The GRSV infection was confirmed via DAS–ELISA and RT–PCR 15 days after inoculation, using the same set of antibodies and the primer pair BR60 / BR65. Transmission electron microscopy of ultrathin sections from symptomatic leaf tissues, both from field–infected and experimentally inoculated endive revealed the presence of typical orthotospovirus particles, within endoplasmic reticulum cisternae. Natural infection of endive by TSWV has been reported in Greece (Chatzivassiliou et al., 2000) and by TCSV in São Paulo State, Brazil and in Florida, USA (Subramanya Sastry et al., 2019). To our knowledge, it is the first report of GRSV naturally infecting this Asteraceae species in Brazil. Confirmation of GRSV infection of C. endivia plants is a relevant piece of information aiming to design effective disease management strategies. References: Boiteux, L.S. and Giordano, L. B. 1993. Euphytica 71: 151. Eiras, M. et al. 2001. Fitopatol. Bras. 26: 170. Chatzivassiliou, E.K. et al. 2000 Ann. Appl. Biol. 137: 127. Cortez, I., et al. 2001. Arch. Virol. 146: 265. Subramanya Sastry, K., et al. 2019. Encyclopedia of plant viruses and viroids. Springer, New Delhi. https://doi.org/10.1007/978-81-322-3912-3.

Utjecaj metoda ispitivanja na energiju klijanja i klijavost sjemena endivije (Cichorium endivia L.)

Endivija (Cichorium endivia L.) je povrtna vrsta čije se potrebite količine sjemena u Hrvatskoj svake godine povećavaju zbog sve većeg broja vrtlara hobista. Posljednjih nekoliko godina u laboratorijima za kontrolu kvalitete poljoprivrednog sjemena zapaženo je da sjeme endivije neovisno o sorti ima nisku energiju klijanja i klijavost. Cilj ovog istraživanja je ispitati navedene pokazatelje kvalitete sjemena endivije različitim metodama koje se koriste kod ispitivanja klijavosti povrtnih kultura. Korištena je metoda na filtar- papiru s četiri tretmana 1) vlaženje podloge destiliranom vodom, 2) vlaženje podloge 0,2%-tnom otopinom KNO3, 3) prethodno hlađenje + vlaženje podloge destiliranom vodom, 4) prethodno hlađenje + vlaženje podloge 0,2%-tnom otopinom KNO3. Ispitivanje je provedeno u laboratoriju za kontrolu kvalitete poljoprivrednog reprodukcijskog materijala Visokog gospodarskoga učilišta u Križevcima. Nakon provedenog ispitivanja rezultati pokazuju da su najvišu energiju klijanja i klijavost sjemena imali uzorci kod kojih je korišten predtretman prethodno hlađenje i destilirana voda, a značajno nižu energiju i klijavost sjemena imali su uzorci kod kojih je korištena metoda vlaženja podloge s 0,2%-tnom otopinom KNO3.

Genotyping by RAD Sequencing Analysis Assessed the Genetic Distinctiveness of Experimental Lines and Narrowed down the Genomic Region Responsible for Leaf Shape in Endive (Cichorium endivia L.)

The characterization of genetic diversity in elite breeding stocks is crucial for the registration and protection of new varieties. Moreover, experimental population structure analysis and information about the genetic distinctiveness of commercial materials are essential for crop breeding programs. The purpose of our research was to assess the genetic relationships of 32 endive (Cichorium endivia L.) breeding lines, 18 from var. latifolium (escarole) and 14 from var. crispum (curly), using heterologous Cichorium intybus-derived simple sequence repeats (SSR) markers and single-nucleotide polymorphisms (SNP) markers. We found that 14 out of 29 SSR markers were successfully amplified, but only 8 of them were related to polymorphic loci. To overcome the limitation of the low number of informative SSR marker loci, an alternative SNP-based approach was employed. The 4621 SNPs produced by a restriction site-associated DNA marker sequencing approach were able to fully discriminate the 32 endive accessions; most importantly, as many as 50 marker loci were found to distinguish the curly group from the escarole group. Interestingly, 24 of the marker loci mapped within a peripheral segment of chromosome 8 of lettuce (Lactuca sativa L.), spanning a chromosomal region of 49.6 Mb. Following Sanger sequencing-based validation, three genes were determined to carry nonsynonymous SNPs, and one of them matched a putative ortholog of AtELP1, subunit 1 of the Elongator complex. Considering that several previously characterized Elongator complex subunit mutants exhibited elongated and/or curly leaf phenotypes, this gene should be taken into consideration for a better understanding of the underlying mechanism controlling leaf shape in endive.

Análises parasitológicas de hortaliças comercializadas na feira livre do Ver-o-Peso, Belém – PA

Objetivo: Avaliar a ocorrência de diferentes formas parasitárias em amostras de Lactuca sativa, Brassica oleracea, Coriandrum sativum, Petroselinum crispum, Allium schoenoprasum, Ocimum basilicum, Ocimum pilosum, Cichorium endivia e Mentha spicata provenientes do mercado Ver-o-Peso. Métodos: Estudo transversal, descritivo e quantitativo. Foram coletadas 5 amostras de cada hortaliça, sendo processadas pelos métodos de sedimentação espontânea e centrifugação, seguida de análise e identificação de parasitas. Resultados: Todas as amostras estavam contaminadas por parasitas. A hortaliça Lactuca sativa apresentou a maior frequência, entre os helmintos (82,24%), os ovos de Ancylostoma sp e Ascaris sp., destacaram-se em ambos os métodos, entretanto, nos protozoários (60,05%), o Balantidium sp (cistos) foi predominante no método de Hoffman e, no de Ritchie, foram os cistos de Entamoeba sp e Larvas de Nematodos. Comparando a detecção entre métodos, não houve diferença significativa, entretanto, ressalta importância das duas metodologias concomitantes para detecção de parasitas. Conclusão: A presença de parasitas em hortaliças significa risco na sua transmissão, além de criar um alerta quanto à provável subnotificação de parasitoses intestinais. A conscientização da correta higienização de alimentos é fundamental para redução das parasitoses, devendo ser acompanhada da efetiva atuação da vigilância sanitária nas feiras livres.

Impact of climatic factors on growth and development of Cichorium Endivia in greenhouse in Leningrad region, Russia

The paper discusses the results of the study of the effect of climatic factors on the yield and biometric indicators of various varieties of endive chicory (Cichorium endivia (L.)) over a two-year study grown in the summer-autumn period in the plastic greenhouses on the experimental plots of the St. Petersburg Agrarian University, Pushkin, Russia. The plants of the genus Chicory are known and valued for their unique medicinal and nutritional properties. Total of 14 varieties of domestic and foreign Cichorium endivia breeding were studied. The study was carried out in 2015 and 2016years that were sharply differed in the amount of precipitation and temperature. The climate of Leningrad region is characterised with humid and long-lasting cold season. The aim of the research was to identify the variety of endive chicory that is most suitable and tolerant for the climatic conditions of Leningrad region, Russia. During the growing season the plant biometric parameters such as plant height (cm), rosette diameter of leaves (cm), and number of leaves (pieces) were measured. During harvesting, additional parameters such as biomass of plants and leaves, the area of the assimilation surface were determined. The decrease in plant biometric parameters during the phenological observations in September 2016 was associated with an increased amount of precipitation in July-August. The results showed that the variety "Broad Betavian fullhearted" was the most sensitive to the adverse effects of weather, showing the largest variation in biometrics over the studied years. In plants of this variety the mass of leaves decreased by 9.2 times, the area of the assimilation surface was reduced by 11.6 times, while the crop shortfall in 2016 was almost 90%. The varieties "Green curled" and "Scarola bionda" were the most tolerant to weather conditions. These varieties were characterized by the maximum yield in both years of research: "Green curled"-7.90 kg/m 2 and 2.87 kg/m 2 , "Scarola bionda"-8.06 kg/m 2 and 2.84 kg/m 2 in 2015 and 2016, respectively. Generally, the result of the study showed that most of the studied varieties were sensitive to the amount of precipitation and the number of sunny days. The study made it possible to identify the most sensitive and most tolerant varieties of endive chicory for growing in the Leningrad region.