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Among foods protected by the European Union with the PGI (Protected Geographical Indication) mark, several ecotypes of “Fagioli di Sarconi” common beans, typical legumes of Basilicata Region, are included. This work aimed to conduct a survey of seed-borne pathogens isolated from “Ciuoto” and “Cannellino rosso”, ecotypes of “Fagioli di Sarconi” common beans, in two years and cultivation areas of the National Park of the Agri Valley, for identifying resistant and climatic changes well-adapted genotypes. Three validated methods were used for the seed-borne pathogens screening. Eighteen fungi were differently found for ecotype and year of observation by the washing test. Saprophyte contaminants pathogens isolated and detected by the blotter test were strongly reduced by 1% sodium hypochlorite treatment. Using the between paper test, specific for detecting Colletotrichum lindemuthianum, the presence of this pathogen for both ecotypes, years and cultivation areas, and also some bacteria were individuated. Therefore, area-, environment- and ecotype-dependent differences were revealed, probably also caused by a different polyphenolic content and thickness of integument of two ecotypes. This study represents a baseline information for further studies, development of forecasting models and management of seed-borne diseases associated with common beans.

The yield and fruit quality of processing tomatoes (Solanum lycopersicum L.) have increased markedly over the past decades. The aim of this work was to assess the effects of the organic (OFS) and conventional farming systems (CFS) on the main agronomic parameters involved in processing tomato yield components and fruit quality traits of heirloom and modern genotypes. Marketable yield increased from heirloom to modern genotypes, both in OFS and in CFS, showing a difference of ≈20 t per hectare in favor of CFS. Total fruit yield (TY) was not improved from heirloom to modern assessed genotypes, and a difference of ≈35 t per hectare was observed in favor of CFS. In both farming systems, the highest marketable yield of modern genotypes was due to a higher number of fruits per plant, harvest index, nitrogen agronomic efficiency (NAE), and fruit water productivity. Moreover, the main growth parameters involved in the yield differences between OFS and CFS were the number of leaves per plant, the average fruit weight, the normalized difference vegetation index (NDVI), and NAE. It is noteworthy that fruit quality improvement in terms of color and brix per hectare was paralleled by a decrease of tomato pH in both farming systems. According to our results, we conclude that to reduce the current yield gap between OFS and CFS, agronomic and breeding efforts should be undertaken to increase leaf area index, fruit number per plant, and NAE for better genotype adaptation to organic farming systems.

Currently, vegetable production systems have been changed to improve yield and quality, leading to an improvement in sustainability. In horticultural cultivation, one of the most important sectors is the seedling production in nurseries. In the past, horticultural seedlings were produced directly by farmers. Nowadays, industrialized nurseries provide seedlings characterized by a uniform growth and an early and contemporaneous development. In addition, consumer concern about the impact of food production on the environment is driving an increased demand for organic vegetables with a consequent increase of agricultural land cultivated by organic methods. Hence, there is a need to produce high-quality seedlings suitable to be cultivated in the organic farming system. Root zone management, for improving seedling quality of organic horticultural crops, remains largely unexploited, such that this review highlights some of the current research and future development priorities, providing useful information to nursery growers. In particular, we reviewed all the scientific and modern knowledge on the production of organic, healthy and vigorous seedlings including the use of: (1) compost and compost tea; (2) agroindustrial byproducts; (3) microbial and non-microbial plant biostimulants; (4) beneficial microorganisms.

This study evaluated, for the first time, the suitability of on-farm green compost (GC) from the fourth range sector for the production, in a controlled environment, of six baby leaf species (radish, lettuce, rocket, chard, sorrel and dill), characterized by short cultivation cycles. The pH and electrical conductivity of potting soil-compost mixtures were affected by the different amounts of GC added to the potting soil. Germination tests on Lactuca sativa seeds indicated no phytotoxic effects when GC was used to prepare aqueous extracts diluted at 50% and 25% (v/v). In the pot experiment, GC was used at four different dosages in the growing medium, inducing different impacts on growth parameters, depending on the species and applied dose. The greatest plant growth responses were determined by using less than 3.5 g of GC in 100 g of potting soil. Significant positive correlations between the dry weights of root and shoot (r = 0.77 **), seedling height (r = 0.51 **) and the number of true leaves number (r = 0.56 **) confirmed the close relationship between the root system biomass and the growth parameters of aboveground organs. The study highlights that on-farm green compost, used as partial growing substrate, provides a good opportunity to obtain baby leaf species with well-developed root systems.

Common bean (Phaseolus vulgaris L.) is a major grain legume consumed worldwide with high nutritional value. Seed-borne pathogens represent a serious problem for seeds, causing losses of yield and quality reduction. Basilicata Region (southern Italy) is a traditional, vocational area of bean production, in particular for "Fagioli di Sarconi" common beans, including different ecotypes protected by the European Union with the mark PGI (Protected Geographical Indication). The study aimed to determine the presence of seed-borne pathogens isolated from seeds of two ecotypes of "Fagioli di Sarconi" common beans, "Ciuoto" and "Cannellino rosso", collected from three different cultivation areas of Basilicata Region during the years 2018 and 2019, for identifying genotypes resistant to seed-borne pathogens and well adapted to climatic changes. The seeds were preliminary evaluated for their purity and germinability and treated or not with sodium hypochlorite for seed-borne pathogens screening, by using three validated seed health testing methods, according to the 2020 International Rules for seed testing: washing, blotter and between paper tests. The washing test identified 17 fungal pathogens, different for ecotype and year of observation. Furthermore, the 1% sodium hypochlorite treatment strongly reduced the contaminants maintaining the Rhizoctonia solani, Colletotrichum lindemuthianum, Fusarium oxysporum, Fusarium solani pathogens; instead, using the blotter test, Aspergillus spp., Cladosporium spp., Botrytis spp., Mucor spp., Rhizophus spp. saprophyte pathogens resulted. Finally, between paper test, specific for detecting the C. lindemuthianum, revealed the presence of this pathogen for the both ecotypes and years, in all areas. This assay individuated some bacteria, too.

In most countries the historical-cultural heritage also refers to the foods traditionally consumed. Southern Italy maintains a strong tradition of dishes based on wild plants, considered an important dietary foundation. Despite this, the nutritional properties and nutraceutical value of some of these plants are little known and, therefore, need to be investigated. Sixteen spontaneous species, traditionally used as food plants, were collected from their natural habitat in four different areas of Campania Region, and their phenolic content and nutritional value were determined according to European model of food labelling. The species analysed resulted quite homogeneous regarding the nutritional value, as demonstrated by cluster analyses. Energy values were always low. Minerals content (especially calcium and potassium) and phenols differed among the species, the sodium-potassium ratio was almost always

In the last few years, the aromatic plant market for fresh consumption has been growing, especially packaged herbs for Mass Market Retailers. Pruning and selection of aromatic plants during packaging leads to the accumulation of a large amounts of plant residues with consequent disposal costs. In a circular economy perspective, the aim of this study was to recover residues of three aromatic plants (Ocimum basilicum L., Rosmarinus officinalis L. and Salvia officinalis L.) by extracting essential oil and aromatic water and, subsequently, reusing oil-free biomasses for composting. In fact, these by-products are currently considered very interesting and sustainable. The essential oils are natural products that have a wide range of biological activities useful for pharmaceutical, medical, veterinary and agriculture innovative purposes. Hydrolates have a much softer scent than the corresponding essential oils. The compost can be successfully applied for the restoration and maintenance of soil fertility. Results of this study showed that yields of essential oils obtained from plant residues, were sufficiently higher, especially for sage and rosemary, also if they were collected well far from plant balsamic period. Analysis of composition of essential oils confirmed the presence of characteristic compounds for each species. The aromatic waters were found to be an easily usable product due to the favourable physicochemical characteristics and, in particular, the aromatic waters of basil showed high antioxidant activity. About compost, the main physicochemical (humidity, pH, electrical conductivity) and biological (basal respiration and hydrolase activity) properties, were considered. Phytotoxicity tests indicated that composts derived from the three species can be used in agriculture. This research demonstrates that it is possible to eliminate completely plant residues and recover new products from aromatic species, supporting the effectiveness of an eco-friendly model to recover and reuse all fresh aromatic plant residues.

The study of the biochemical and molecular mechanisms deriving from the host-pathogen-antagonist interaction is essential to understand the dynamics of infectious processes and can be useful for the development of new strategies to control phytopathogens, particularly viruses, against which chemical treatments have no effect. In this work, we demonstrate the ability of the rhizospheric fungus Trichoderma harzianum strain T-22 (T22) to induce defense responses in tomato (Solanum lycopersicum var. cerasiforme) against Cucumber mosaic virus (CMV, family Bromoviridae, genus Cucumovirus) strain Fny. A granule formulation containing T22 was used for treating the plants before, simultaneously or after the CMV inoculation, in order to study the molecular and biochemical aspects of the interaction between T22 and tomato against the virus. Reactive oxygen species (ROS) and the genes encoding for ROS scavenging enzymes were investigated. Histochemical analysis revealed a different increase in the superoxide anion ( $$ {\text{O}}_{2}^{ \bullet - } $$ ) and hydrogen peroxide (H2O2) content in plants infected by CMV alone or in the presence of T22, confirming the involvement of ROS in plant defense responses. Gene expression analysis suggested a definite improvement in oxidative stress when plants were treated with T22 after inoculation with CMV. In conclusion, our data indicate that Trichoderma harzianum T-22 stimulates the induction of tomato defense responses against CMV, an action that implies the involvement of ROS, pointing towards its use as a treatment rather than as a preventive measure.

Arabidopsis thaliana L. is a model plant but little information is available about morphological root changes as part of a phytohormonal common response against both biotic and abiotic stressors. For this purpose, two-week-old Arabidopsis seedlings were treated with 10 mu M CdSO4 or infected with CMV. After 12 days the entire aerial parts and the root system were analyzed, and the presence of CMV or the accumulation of Cd were detected. Microscopic analysis revealed that both CMV and Cd influenced root morphology by a marked development in the length of root hairs and an intense root branching if compared to controls. Among the three treatments, Cd-treated seedlings showed a shorter root axis length and doubled their lateral root diameter, while the lateral roots of CMV-infected seedlings were the longest. The root growth patterns were accompanied by significant changes in the levels of indole-3-acetic acid, trans-zeatin riboside, dihydrozeatin riboside, as a probable consequence of the regulation of some genes involved in their biosynthesis/degradation. The opposite role on root development played by the phythormones studied is discussed in detail. The results obtained could provide insights into novel strategies for plant defense against pathogens and plant protection against pollutants.

Understanding the induction of plant defenses against viruses using biocontrol agents is essential for developing new strategies against these pathogens, given the ineffectiveness of chemical treatments. The ability of Trichoderma harzianum, strain T-22 (T22) to control Cucumber mosaic virus (CMV) in Solanum lycopersicum var. cerasiforme plants and the changes in the physiology of tomato treated/infected with T22/CMV were examined. Plant growth-promoting effects, photosynthetic performance, reactive oxygen species (ROS) scavenging enzymes, and phytohormones were investigated. T22 improved tomato growth in terms of plant height and improved photosynthesis, total chlorophyll content and plant gas exchange. In contrast, CMV induced a negative effect on dry matter accumulation and inhibited the photosynthetic capacity. The analysis of plant hormones demonstrated that treating with T22 before or simultaneously to CMV infection, led to a systemic resistance by jasmonic acid/ethylene and salicylic acid signaling pathways. Conversely, systemic resistance was abscissic acid-dependent when T22 treatment was administered after the CMV infection. In conclusion, the data reported here indicate that the T22-based strategy may be the most effective measure against CMV.

Nowadays, crop production is at risk due to global warming, especially in Mediterranean areas where the increase of air temperature and/or reduction of precipitation is relevant. Climate changes that are occurring can severely prejudice plant defensive mechanisms during host-pathogen interactions by modifying growth and physiology of the host plant. In particular, viral diseases cause serious economic losses destroying crops and reducing agronomic productivity, and, in some cases such as tomato crops, they become the limiting factor production of both open field and under greenhouse cultivation systems. This is because plant viruses are obligate parasites and require living tissue for their multiplication and spread. Therefore, they are able to interfere with plant metabolism and compete for host plant resources, so determining a decrease of plant growth and productivity. Severe outbreaks of Cucumber mosaic virus (CMV) and other viruses caused disruption of tomato plants in the Mediterranean region and in Southern Italy since the 1970s. In such a scenario, it is necessary to introduce new strategies for controlling plant pathogens and parasites in order to help maintain ecosystems and to boost sustainable agriculture. The aim of this work is to give an up-to-date overview on the recent breakthroughs in the use of microorganisms on plants for improving crop yields, quality and plant tolerance against pathogens. In particular, here we report a case study regarding an innovative strategy to control a viral disease (CMV) in tomato, based on the use of rhizosphere microorganism (Trichoderma harzianum, strain T-22) as an antagonist biocontrol agent (BCA).

In this work, specific concentrations of cadmium, copper and zinc in double combination, were supplied for 12 days to growing seedlings of the model species Arabidopsis thaliana. Metal accumulation was measured in roots and shoots. Microscopic analyses revealed that root morphology was affected by metals, and that the root and shoot levels of indole-3-acetic acid, trans-zeatin riboside and dihydrozeatin riboside varied accordingly. Minor modifications in gibberellic acid levels occurred in the Zinc treatments, whereas abscisic acid level did not change after the exposition to metals. Reverse transcription polymerase chain reaction analysis of some genes involved in auxin and cytokinin synthesis (AtAAO, AtNIT and AtIPT) revealed that their expression were not affected by metal treatments. The root morphological alterations that resulted in an increased surface area, due to the formation of root hairs and lateral roots, could be signs of the response to metal stress in terms of a functionally-addressed reorientation of root growth. The root system plasticity observed could be important for better understanding the manner in which the root architecture is shaped by environmental and hormonal stimuli.

A plant bioreactor has enormous capability as a system that supports many biological activities, that is, production of plant bodies, virus-like particles (VLPs), and vaccines. Foreign gene expression is an efficient mechanism for getting protein vaccines against different human viral and nonviral diseases. Plants make it easy to deal with safe, inexpensive, and provide trouble-free storage. The broad spectrum of safe gene promoters is being used to avoid risk assessments. Engineered virus-based vectors have no side effect. The process can be manipulated as follows: (a) retrieve and select gene encoding, use an antigenic protein from GenBank and/or from a viral-genome sequence, (b) design and construct hybrid-virus vectors (viral vector with a gene of interest) eventually flanked by plant-specific genetic regulatory elements for constitutive expression for obtaining chimeric virus, (c) gene transformation and/or transfection, for transient expression, into a plant-host model, that is, tobacco, to get protocols processed positively, and then moving into edible host plants, (d) confirmation of protein expression by bioassay, PCR-associated tests (RT-PCR), Northern and Western blotting analysis, and serological assay (ELISA), (e) expression for adjuvant recombinant protein seeking better antigenicity, (f) extraction and purification of expressed protein for identification and dosing, (g) antigenicity capability evaluated using parental or oral delivery in animal models (mice and/or rabbit immunization), and (h) growing of construct-treated edible crops in protective green houses. Some successful cases of heterologous gene-expressed protein, as edible vaccine, are being discussed, that is, hepatitis C virus (HCV). R9 mimotope, also named hypervariable region 1 (HVR1), was derived from the HVR1 of HCV. It was used as a potential neutralizing epitope of HCV. The mimotope was expressed using cucumber mosaic virus coat protein (CP), alfalfa mosaic virus CP P3/RNA3, and tobacco mosaic virus (TMV) CP-tobacco mild green mosaic virus (TMGMV) CP as expression vectors into tobacco plants. Expressed recombinant protein has not only been confirmed as a therapeutic but also as a diagnostic tool. Herpes simplex virus 2 (HSV-2), HSV-2 gD, and HSV-2 VP16 subunits were transfected into tobacco plants, using TMV CP-TMGMV CP expression vectors.

A primary therapeutic goal in Alzheimer’s disease (AD) is to reduce the quantity of amyloid b protein (A b) present in the brain. To develop an effective, safe system for vaccination against Alzheimer’s disease, the plant virus Cucumber mosaic virus (CMV) was engineered genetically to express A b -derived fragments that stimulate mainly humoral immune responses. Six chimeric constructs, bearing the A b 1–15 or the A b 4–15 sequence in positions 248, 392 or 529 of the CMV coat protein (CP) gene, were created. Viral products proved to be able to replicate in their natural host. However, only chimeric A b 1–15-CMVs were detected by A b 1–42 antiserum in Western blot analysis. Experimental evidence of Immunoelectron microscopy revealed a complete decoration of A b 1–15-CMV248 and A b 1–5-CMV392 following incubation with either anti-A b 1–15 or anti-A b 1–42 polyclonal antibodies. These two chimeric CMVs appear to be endowed with features making them possible candidates for vaccination against Alzheimer’s disease.

The Cucumber mosaic virus (CMV) is an isodiametric plant virus with an extremely wide host range, present worldwide. CMV chimeric particles (R9-CMV), engineered to express a 27-aa synthetic peptide derived from Hepatitis C virus (HCV), were demonstrated to be stable under simulated gastric and intestinal conditions. Then the possibility of inducing a humoral immune response in rabbits fed with R9-CMV infected lettuce plants was demonstrated, suggesting that this system could function as a confirming tool of a bioreactor for the production of a stable edible vaccine against HCV.

Chimeric plant viruses are emerging as promising vectors for use in innovative vaccination strategies. In this context, cucumber mosaic virus (CMV) has proven to be a suitable carrier of the hepatitis C virus (HCV)-derived R9 mimotope. In the present work, a new chimeric CMV, expressing on its surface the HCV-derived R10 mimotope, was produced but lost the insert after the first passage on tobacco. A comparative analysis between R10- and R9-CMV properties indicated that R9-CMV stability was related to structural features typical of the foreign insert. Thus, in order to combine high virus viability with strong immuno-stimulating activity, we doubled R9 copies on each of the 180 coat protein (CP) subunits of CMV. One of the chimeras produced by this approach (2R9-CMV) was shown to systemically infect the host, stably maintaining both inserts. Notably, it was strongly recognized by sera of HCV-infected patients and, as compared with R9-CMV, displayed an enhanced ability to stimulate lymphocyte IFN-gamma production. The high immunogen levels achievable in plants or fruits infected with 2R9-CMV suggest that this chimeric form of CMV may be useful in the development of oral vaccines against HCV.