Assessment of bacterial and fungal (hemi)cellulose-degrading enzymes in saccharification of ammonia fibre expansion-pretreated Arundo donax.

This study reports enzymatic hydrolysis of the biomass of the giant reed (Arundo donax L.) after ammonia fibre expansion (AFEX) pretreatment. In particular, the capacity of the arabinofuranosidase from the fungus Pleurotus ostreatus recombinantly expressed in Pichia pastoris rPoAbf, its evolved mutant rPoAbf F435Y/Y446F and the endo-cellulase from Streptomyces sp. G12 CelStrep recombinantly expressed in Escherichia coli to enhance the hydrolysis of AFEX-treated A. donax was investigated, using the corn stover as reference feedstock. The investigated enzymes were assayed using a mixture of purified cellulases (CBHI, CBHII, EGI and ßG), endoxylanases (LX3, LX4) and accessory hemicellulases (LarbF and LßX) as reference enzyme mixture and substituting EGI with rCelStrep and LarbF with rPoAbf or rPoAbf F435Y/Y446F. The use of rPoAbf F435Y/Y446F in the substitution of LarbF led to improvements in sugar conversion, giving a glucan, xylan and arabinan conversion after 72 h of around 62, 63 and 80 %, respectively, similar or higher than those (44, 66 and 55 %) achieved by 72 h hydrolysis with commercial enzymes Novozymes Cellic®, Ctec3 and Htec3. The enzymes rPoAbf, rPoAbf F435Y/Y446F and rCelStrep were also investigated for their effect on hydrolysis of AFEX-pretreated A. donax by addition to commercial enzyme mixture Novozymes Cellic®, Ctec3 and Htec3, and it was shown that the addition of rPoAbf and its evolved mutant rPoAbf F435Y/Y446F enhanced both xylan and arabinan conversions, which achieved 80 % after 6 days of saccharification with rPoAbf F435Y/Y446F.

Infrared Thermography Monitoring of Durum and Common Wheat for Adaptability Assessing and Yield Performance Prediction.

Wheat is one of the most cultivated cereals thanks to both its nutritional value and its versatility to technological transformation. Nevertheless, the growth and yield of wheat, as well as of the other food crops, can be strongly limited by many abiotic and biotic stress factors. To face this need, new methodological approaches are required to optimize wheat cultivation from both a qualitative and quantitative point of view. In this context, crop analysis based on imaging techniques has become an important tool in agriculture. Thermography is an appealing method that represents an outstanding approach in crop monitoring, as it is well suited to the emerging needs of the precision agriculture management strategies. In this work, we performed an on-field infrared monitoring of several durum and common wheat varieties to evaluate their adaptability to the internal Mediterranean area chosen for cultivation. Two new indices based on the thermal data useful to estimate the agronomical response of wheat subjected to natural stress conditions during different phenological stages of growth have been introduced. The comparison with some productive parameters collected at harvest highlighted the correlation of the indices with the wheat yield (ranging between p < 0.001 and p < 0.05), providing interesting information for their early prediction.

Integrating Smart Greenhouse Cover, Reduced Nitrogen Dose and Biostimulant Application as a Strategy for Sustainable Cultivation of Cherry Tomato.

Fruit yield and quality of greenhouse tomatoes are strongly influenced by light conditions and nitrogen (N) availability, however, the interaction between these factors is still unclear. We evaluated the effects on cherry tomatoes of two tunnel plastic covers with different optical properties and three N doses, also in combination with a biostimulant treatment. We compared a diffuse light film (Film1) and a conventional clear film (Film2), and three N levels, corresponding to 50% (N50), 75% (N75) and 100% (N100) of the optimal dose, with and without a microbial plus a protein hydrolysed biostimulant, compared to a non-treated control. The three experimental treatments significantly interacted on several yield and quality parameters. In control plants (untreated with biostimulants), the early yield was higher at reduced N doses compared to N100, with greater increments under the diffusive Film1 compared to the clear Film2 (+57.7% and +37.0% vs. +31.7% and +16.0%, in N50 and N75 respectively). Film1 boosted the total fruit production at all the N rates and with or without biostimulants, compared to Film2, with stronger effects under sub-optimal N (+29.4% in N50, +21.2% in N75, and +7.8% in N100, in plants untreated with biostimulant). Total yield decreased with decreasing N levels, while it always increased with the application of biostimulants, which counterbalanced the detrimental effects of N shortage. Quality traits were mainly affected by the cover film and the biostimulant treatment. The diffusive film increased the content of carotenoids, lycopene and total phenols compared to the clear one, and the biostimulants increased texture, soluble solids, phenols and ascorbic acid compared to the untreated control. It is worth noting that in plants fertilized at 75% of the reference N dose, the biostimulants determined higher yield than the N100 untreated control, under both the covers (+48% in Film1 and +20% in Film2). In conclusion, the diffusive film improved the fruit yield and quality of greenhouse tomatoes in the spring-summer period, presumably avoiding plant stress due to high-intensity direct light. Reduced N rates limited the plant productivity, however, the biostimulant application was effective in compensating for the detrimental effects of sub-optimal supply of N synthetic fertilizers.

Biodegradable Mulch Films and Bioformulations Based on Trichoderma sp. and Seaweed Extract Differentially Affect the Metabolome of Industrial Tomato Plants.

The use of biostimulants and biofilms in agriculture is constantly increasing, as they may support plant growth and productivity by improving nutrient absorption, increasing stress resilience and providing sustainable alternatives to chemical management practices. In this work, two commercial products based on Trichoderma afroharzianum strain T22 (Trianum P®) and a seaweed extract from Ascophyllum nodosum (Phylgreen®) were tested on industrial tomato plants (Solanum lycopersicum var. Heinz 5108F1) in a field experiment. The effects of single and combined applications of microbial and plant biostimulants on plants grown on two different biodegradable mulch films were evaluated in terms of changes in the metabolic profiles of leaves and berries. Untargeted metabolomics analysis by LC-MS Q-TOF revealed the presence of several significantly accumulated compounds, depending on the biostimulant treatment, the mulch biofilm and the tissue examined. Among the differential compounds identified, some metabolites, belonging to alkaloids, flavonoids and their derivatives, were more abundant in tomato berries and leaves upon application of Trichoderma-based product. Interestingly, the biostimulants, when applied alone, similarly affected the plant metabolome compared to control or combined treatments, while significant differences were observed according to the mulch biofilm applied.

Assessing Yield and Quality of Melon (Cucumis melo L.) Improved by Biodegradable Mulching Film.

Low-density polyethylene (LDPE) plastic mulching films have an important function, but at the end of their lifetime pose an economic and environmental problem in terms of their removal and disposal. Biodegradable mulching films represent an alternative to LDPE with the potential to avoid these environmental issues. In this preliminary study, we employed a biodegradable film based on Mater-Bi® (MB) in comparison with low-density polyethylene to assess their effect on the yield and particular quality traits (organoleptic and nutraceutical composition of the fruits) of muskmelon (cv Pregiato) grown on soils with different textures (clay-loam-CL and sandy loam-SL) in two private farms in South Italy. Soil temperature under the mulch was also measured. During the monitored periods, mean soil temperature under LDPE was higher (about 1.3 °C) than that under the biodegradable film and was higher in SL soil than in CL soil, at 25.5° and 24.2 °C, respectively. However, the biodegradable film was able to limit the daily temperature fluctuation, which was 1.7 °C in both soils compared with 2.3 °C recorded for LDPE. Fruit yields were higher with MB film than LDPE (+9.5%), irrespective of soil texture. MaterBi® also elicited increases in total soluble solids, polyphenols, flavonoids, and antioxidant activity compared with LDPE films: 13.3%, 22.4%, 27.2%, and 24.6%, respectively. Color parameters of flesh, namely brightness, chroma, and hue angle were better in fruits grown on LDPE. Our findings suggest that Mater-Bi® based biodegradable mulching film is a potentially valid alternative to traditional LDPE, particularly for obtaining the agronomical benefits outlined above and for promoting environmental sustainability due to its favourable biodegradable properties.

Biodegradable Mulching Film vs. Traditional Polyethylene: Effects on Yield and Quality of San Marzano Tomato Fruits.

Mulching is a common practice for improving crop yield and obtaining an out-of-season production, but when made using plastic materials it can bring environmental problems due to the management and the disposal of films at the end of the cropping seasons. To increase the sustainability of this practice, recently, mulching films made with biodegradable organic materials have become more widely used. Our aim was to evaluate the effect of a biodegradable mulching film on yield and qualitative traits of the San Marzano tomato fruits over two years (2014 and 2015). Two different types of mulching were tested: (i) black biodegradable film (MB12) and (ii) black low-density polyethylene (LDPE) were compared to bare soil (BS). Both mulching films elicited a 25% increase in yield, mainly due to the significantly higher number of fruits per square meter, compared to BS. Both mulching films also elicited a 9.9% increase in total soluble solids and a 57% increase in carotenoid content, while firmness showed the highest value in BS fruits. MB12 determined the highest value of the Hunter color ratio a/b of tomato fruits, followed by LDPE, while the lowest value was recorded in BS fruits. Both mulching films elicited an increase of 9.6%, 26.0%, and 11.7% for flavonoids, polyphenols, and AsA, respectively. In 2014, the MB12 degradation started at 71 days after transplant (DAT); in 2015, at 104 DAT. Therefore, replacing polyethylene with biodegradable film would seem to be an agronomically efficient and environmentally sustainable practice.

Bioactive Compounds and Antioxidant Properties of Wild Rocket (Diplotaxis Tenuifolia L.) Grown under Different Plastic Films and with Different UV-B Radiation Postharvest Treatments.

Rocket species are rich in nutrients with well-known bioactive activity, but their content depends on several factors, such as plant-UV radiation interaction. In this work, we measured the production of nutritional elements in wild rocket (Diplotaxis tenuifolia L.) leaves as a function of exposure to UV-B radiation by adopting a combined approach. The wild rocket plants were grown under three greenhouse cover films (A, B, and C) having different transmittivity to UV-B and the fresh-cut leaves were exposed to UV-B in postharvest for 45, 150, 330, and 660 s. The content of chlorophyll, carotenoids, phenolic compounds, ascorbic acid, and the antioxidant activity were determined. Chlorophyll, carotenoids, and total phenolic content were significantly increased by the combination of Film C and treatment with UV-B for 45 s. The predominant phenolic compounds were kaempferol, isorhamnetin, and quercetin. Film C also elicited an increase in ascorbic acid (the most abundant antioxidant compound in the range 374-1199 per 100 g of dry matter) and antioxidant activity. These findings highlighted an increase in bioactive compound content in the wild rocket when it was cultivated under Film C (diffused light film with a tailored UV-B transmission dose) and treated with UV-B radiation for 45 s postharvest, corresponding to an energy dose of 0.2 KJ m-2.

Plant-Based Biostimulants Influence the Agronomical, Physiological, and Qualitative Responses of Baby Rocket Leaves under Diverse Nitrogen Conditions.

Nitrogen is the primary technical means responsible for food production increase, but on the other hand, wise management is needed because its excessive use can have a negative impact on the environment and on green leafy vegetable quality, such as that rocket. Rocket has the characteristics of accumulating nitrate in leaves with possible impacts on human health. In order to overcome this issue, researchers are focusing their attention on the use of alternative means, such as plant biostimulant application. The scope of this study was to assess the effect of legume-derived protein hydrolysate(LDPH) and tropical plant extract(TPE), combined with various doses of nitrogen (0 kg ha-1 non-fertilized; N0); 60 kg ha-1 (sub-optimal; N1); 80 kg ha-1 (optimal; N2); and 100 kg ha-1 (supra-optimal; N3)), in order to reduce nitrogen use, boost yield, and enhance the chemical and nutritional value of leaves without significantly accumulating nitrate. Both vegetal-based plant biostimulants enhanced plant growth, boosted the marketable yield (especially at N0 and N1 levels, by 38.2% and 28.2%, respectively, compared to the non-treated control), and increased the SPAD (Soil Plant Analysis Development) index and leaf pigments content, such as chlorophyll and carotenoids, especially in treated-LDPH rocket. The plant-based biostimulants also produced a major amplification in lipophilic antioxidant activity (+ 48%) and total ascorbic acid content (average + 95.6%), especially at low nitrogen fertilization levels, and maintained nitrate content under the legal European Comission limits.

Comparative assessment of autochthonous bacterial and fungal communities and microbial biomarkers of polluted agricultural soils of the Terra dei Fuochi.

Organic and inorganic xenobiotic compounds can affect the potential ecological function of the soil, altering its biodiversity. Therefore, the response of microbial communities to environmental pollution is a critical issue in soil ecology. Here, a high-throughput sequencing approach was used to investigate the indigenous bacterial and fungal community structure as well as the impact of pollutants on their diversity and richness in contaminated and noncontaminated soils of a National Interest Priority Site of Campania Region (Italy) called "Terra dei Fuochi". The microbial populations shifted in the polluted soils via their mechanism of adaptation to contamination, establishing a new balance among prokaryotic and eukaryotic populations. Statistical analyses showed that the indigenous microbial communities were most strongly affected by contamination rather than by site of origin. Overabundant taxa and Actinobacteria were identified as sensitive biomarkers for assessing soil pollution and could provide general information on the health of the environment. This study has important implications for microbial ecology in contaminated environments, increasing our knowledge of the capacity of natural ecosystems to develop microbiota adapted to polluted soil in sites with high agricultural potential and providing a possible approach for modeling pollution indicators for bioremediation purposes.

Degradative actions of microbial xylanolytic activities on hemicelluloses from rhizome of Arundo donax.

Polysaccharidases from extremophiles are remarkable for specific action, resistance to different reaction conditions and other biotechnologically interesting features. In this article the action of crude extracts of thermophilic microorganisms (Thermotoga neapolitana, Geobacillus thermantarcticus and Thermoanaerobacterium thermostercoris) is studied using as substrate hemicellulose from one of the most interesting biomass crops, the giant reed (Arundo donax L.). This biomass can be cultivated without competition and a huge amount of rhizomes remains in the soil at the end of cropping cycle (10-15 years) representing a further source of useful molecules. Optimization of the procedure for preparation of the hemicellulose fraction from rhizomes of Arundo donax, is studied. Polysaccharidases from crude extracts of thermophilic microorganisms revealed to be suitable for total degradative action and/or production of small useful oligosaccharides from hemicelluloses from A. donax. Xylobiose and interesting tetra- and pentasaccharide are obtained by enzymatic action in different conditions. Convenient amount of raw material was processed per mg of crude enzymes. Raw hemicelluloses and pretreated material show antioxidant activity unlike isolated tetra- and pentasaccharide. The body of results suggest that rhizomes represent a useful raw material for the production of valuable industrial products, thus allowing to increase the economic efficiency of A. donax cultivation.