Ascorbate peroxidase and catalase activities and their genetic regulation in plants subjected to drought and salinity stresses

Hydrogen peroxide (H2O2), an important relatively stable non-radical reactive oxygen species (ROS) is produced by normal aerobic metabolism in plants. At low concentrations, H2O2 acts as a signal molecule involved in the regulation of specific biological/physiological processes (photosynthetic functions, cell cycle, growth and development, plant responses to biotic and abiotic stresses). Oxidative stress and eventual cell death in plants can be caused by excess H2O2 accumulation. Since stress factors provoke enhanced production of H2O2 in plants, severe damage to biomolecules can be possible due to elevated and non-metabolized cellular H2O2. Plants are endowed with H2O2-metabolizing enzymes such as catalases (CAT), ascorbate peroxidases (APX), some peroxiredoxins, glutathione/thioredoxin peroxidases, and glutathione sulfo-transferases. However, the most notably distinguished enzymes are CAT and APX since the former mainly occurs in peroxisomes and does not require a reductant for catalyzing a dismutation reaction. In particular, APX has a higher affinity for H2O2 and reduces it to H2O in chloroplasts, cytosol, mitochondria and peroxisomes, as well as in the apoplastic space, utilizing ascorbate as specific electron donor. Based on recent reports, this review highlights the role of H2O2 in plants experiencing water deficit and salinity and synthesizes major outcomes of studies on CAT and APX activity and genetic regulation in droughtand salt-stressed plants

Synergistic effect of Trichoderma and chitosan application in tomato for the control of Cucumber mosaic virus infection

Soil and plant environment is characterized by a wide range of microorganisms able to interact with host plants and, in some cases, to differentially induce susceptibility or resistance to pathogens. Plant viruses cause chlorosis and necrosis, so decreasing plant growth and productivity. Differently, many fungi are able to trigger a beneficial relationship with plants. Trichoderma spp. are endophytic symbionts able to modify plants metabolism, increasing nutrient uptake by plants and photosynthetic efficiency, and protecting them from pathogens. A biopolymer able to elicit plant-immunity is chitosan, derived by the deacetylation of chitin, a component of some fungal cell walls. Chitosan improves the host hypersensitive response by the expression of pathogenesis-related proteins and the synthesis of secondary metabolites. Trichoderma harzianum T-22 (T22) induces defense responses against Cucumber mosaic virus Fny (CMV) in Solanum lycopersicum. On this basis, the aim of this work was to determine if the combination T22-chitosan has an antiviral activity against CMV in tomato plants. Plant physiological parameters (gas exchange, chlorophyll content and fluorescence) were followed throughout the experiment. Furthermore, ELISA test was employed to detect CMV. Results indicate that plants treated with T22 and chitosan had a strong attenuation of viral load, a higher chlorophyll content and a better photosynthetic performance compared to the untreated plants. Further investigations are in progress to determine plant antioxidant responses. In conclusion, combined treatment based on T22 and chitosan represents a highly effective strategy against CMV, embracing the criteria of sustainable agricultural practice and public health protection

Hormonal response and root architecture in Arabidopsis thaliana subjected to heavy metals

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

Response of Two Local Common Bean Ecotypes of “Fagioli di Sarconi” PGI (Phaseolus vulgaris L.) to Seed-Borne Pathogens and Environmental Change

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

Influence of Cultivation Areas on the Seed-Borne Pathogens on Two Local Common Bean Ecotypes of “Fagioli di Sarconi” PGI (Phaseolus vulgaris L.).

Abstract The “Fagioli di Sarconi” common beans, typical of Basilicata Region (Southern Italy), include 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 two ecotypes of “Fagioli di Sarconi” common beans, “Ciuoto” and “Cannellino rosso”, in two different cultivation areas during the years 2018 and 2019, for identifying genotypes resistant and well adapted to climatic changes. The seeds were evaluated for seed-borne pathogens screening by using three validated seed health testing methods, according to the 2020 International Rules for seed testing. The washing test identified 18 fungal pathogens, different for ecotype and year of observation; the 1% sodium hypochlorite treatment strongly reduced the contaminants. With the blotter test, several saprophyte pathogens were found. Between paper test, specific for detecting the C. lindemuthianum, revealed the presence of this pathogen for both ecotypes and years, in all areas, and individuated some bacteria, too. In conclusion, this work highlighted differences by the two PGI common bean ecotypes in response to seed-borne pathogens resistance and environmental change due probably to their different thickness and polyphenolic content of integument

Beneficial effects of Trichoderma harzianum T-22 in tomato seedlings infected by Cucumber mosaic virus (CMV)

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 (O2 ) 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

Physico-Chemical Characterization and Biological Activities of a Digestate and a More Stabilized Digestate-Derived Compost from Agro-Waste

The excessive use of agricultural soils and the reduction in their organic matter, following circular economy and environmental sustainability concepts, determined a strong attention in considering composting as a preferred method for municipalities and industries to recycle organic by-products. Microorganisms degrade organic matter for producing CO2, water and energy, originating stable humus named compost. The current study analyzed the chemical composition of a cow slurry on-farm digestate and a more stabilized digestate-derived compost (DdC), along with their phytotoxic, genotoxic and antifungal activities. The chemical analysis showed that digestate cannot be an ideal amendment due to some non-acceptable characteristics. Biological assays showed that the digestate had phytotoxicity on the tested plants, whereas DdC did not induce a phytotoxic effect in both plants at the lowest dilution; hence, the latter was considered in subsequent analyses. The digestate and DdC induced significant antifungal activity against some tested fungi. DdC did not show genotoxic effect on Vicia faba using a micronuclei test. Soil treated with DdC (5 and 10%) induced damping-off suppression caused by Fusarium solani in tomato plants. The eco-physiological data indicated that DdC at 5–10% could increase the growth of tomato plants. In conclusion, DdC is eligible as a soil amendment and to strengthen the natural soil suppressiveness against F. solani

Cucumber mosaic virus Is Unable to Self-Assemble in Tobacco Plants When Transmitted by Seed

Cucumber mosaic virus (CMV), which has great impact on agronomic production worldwide, is both aphid and seed transmitted. Although the mechanisms of aphid transmission have been widely studied, those underlying the ability of CMV to survive and remain infectious during the passage from one generation to the next through the seeds are still to be clarified. Moreover, the viral determinants of seed transmission rate are poorly understood. Three viral genotypes produced from same RNA 1 and 2 components of CMV-Fny but differing in RNA 3 (the wild type CMV-Fny, a pseudorecombinant CMV-Fny/CMV-S and a chimeric CMV previously obtained by our group, named F, FS and CS, respectively) were propagated in Nicotiana tabacum cv Xanthi plants in order to assess differences in tobacco seed transmission rate and persistence through plant generations in the absence of aphid transmission. Seed-growth tests revealed CMV infection in the embryos, but not in the integuments. Seedlings from seed-growth tests showed the presence of all considered viruses but at different rates: from 4% (F, FS) to 16% (CS). Electron microscopy revealed absence (CS) of viral particles or virions without the typical central hole (F and FS). In agreement, structural characteristics of purified CMV particles, assessed by circular dichroism spectroscopy, showed anomalous spectra of nucleic acids rather than the expected nucleoproteins. These alterations resulted in no seed transmission beyond the first plant generation. Altogether, the results show for the first time that correct virion assembly is needed for seed infection from the mother plant but not to seedling invasion from the seed. We propose that incorrect virion formation, self-assembly and architecture stability might be explained if during the first stages of germination and seedling development some tobacco seed factors target viral regions responsible for protein-RNA interactions

Trichoderma harzianum T-22 induces systemic esistance in tomato Infected by Cucumber mosaic virus

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 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

Correlation between hormonal homeostasis and morphogenic responses in Arabidopsis thaliana seedlings growing in a Cd/Cu/Zn multi-pollution context

To date, almost no information is available in roots and shoots of the model plant Arabidopsis thaliana about the hierarchic relationship between metal accumulation, phytohormone levels, and glutathione/phytochelatin content, and how this relation affects root development. For this purpose, specific concentrations of cadmium, copper and zinc, alone or in triple combination, were supplied for 12 days to in vitro growing seedlings. The accumulation of these metals was measured in roots and shoots, and a significant competition in metal uptake was observed. Microscopic analyses revealed that root morphology was affected by metal exposure, and that the levels of trans-zeatin riboside, dihydrozeatin riboside, indole-3-acetic acid, and the auxin/cytokinin ratio varied accordingly. By contrast, under metal treatments, minor modifications in gibberellic acid and abscisic acid levels occurred. RT-PCR analysis of some genes involved in auxin and cytokinin synthesis (e.g., AtNIT in roots and AtIPT in shoots) showed on average a metal up-regulated transcription. The production of thiol-peptides was induced by all the metals, alone or in combination, and the expression of the genes involved in thiol-peptide synthesis (AtGSH1, AtGSH2, AtPCS1 and AtPCS2) was not stimulated by the metals, suggesting a full post-transcriptional control. Results show that the Cd/Cu/Zn-induced changes in root morphology are caused by a hormonal unbalance, mainly governed by the auxin/cytokinin ratio