Marcello Guiducci

Use of MSAP Markers to Analyse the Effects of Salt Stress on DNA Methylation in Rapeseed (Brassica napus var. oleifera)

Excessive soil salinity is a major ecological and agronomical problem, the adverse effects of which are becoming a serious issue in regions where saline water is used for irrigation. Plants can employ regulatory strategies, such as DNA methylation, to enable relatively rapid adaptation to new conditions. In this regard, cytosine methylation might play an integral role in the regulation of gene expression at both the transcriptional and post-transcriptional levels. Rapeseed, which is the most important oilseed crop in Europe, is classified as being tolerant of salinity, although cultivars can vary substantially in their levels of tolerance. In this study, the Methylation Sensitive Amplified Polymorphism (MSAP) approach was used to assess the extent of cytosine methylation under salinity stress in salinity-tolerant (Exagone) and salinity-sensitive (Toccata) rapeseed cultivars. Our data show that salinity affected the level of DNA methylation. In particular methylation decreased in Exagone and increased in Toccata. Nineteen DNA fragments showing polymorphisms related to differences in methylation were sequenced. In particular, two of these were highly similar to genes involved in stress responses (Lacerata and trehalose-6-phosphatase synthase S4) and were chosen to further characterization. Bisulfite sequencing and quantitative RT-PCR analysis of selected MSAP loci showed that cytosine methylation changes under salinity as well as gene expression varied. In particular, our data show that salinity stress influences the expression of the two stress-related genes. Moreover, we quantified the level of trehalose in Exagone shoots and found that it was correlated to TPS4 expression and, therefore, to DNA methylation. In conclusion, we found that salinity could induce genome-wide changes in DNA methylation status, and that these changes, when averaged across different genotypes and developmental stages, accounted for 16.8% of the total site-specific methylation differences in the rapeseed genome, as detected by MSAP analysis.

Nitrogen Concentration Estimation in Tomato Leaves by VIS-NIR Non-Destructive Spectroscopy

Nitrogen concentration in plants is normally determined by expensive and time consuming chemical analyses. As an alternative, chlorophyll meter readings and N-NO3 concentration determination in petiole sap were proposed, but these assays are not always satisfactory. Spectral reflectance values of tomato leaves obtained by visible-near infrared spectrophotometry are reported to be a powerful tool for the diagnosis of plant nutritional status. The aim of the study was to evaluate the possibility and the accuracy of the estimation of tomato leaf nitrogen concentration performed through a rapid, portable and non-destructive system, in comparison with chemical standard analyses, chlorophyll meter readings and N-NO3 concentration in petiole sap. Mean reflectance leaf values were compared to each reference chemical value by partial least squares chemometric multivariate methods. The correlation between predicted values from spectral reflectance analysis and the observed chemical values showed in the independent test highly significant correlation coefficient (r = 0.94). The utilization of the proposed system, increasing efficiency, allows better knowledge of nutritional status of tomato plants, with more detailed and sharp information and on wider areas. More detailed information both in space and time is an essential tool to increase and stabilize crop quality levels and to optimize the nutrient use efficiency.

Actual N availability from winter catch crops used for green manuring in maize cultivation.

Field experiments were carried out in Central Italy on several green manure species (pure or mixed) and on succeeding grain maize to find practical relationships for predicting the N effect (N absorbed by maize that derives from the actual gain in soil available N due to green manure). Actually, little information is available for Mediterranean environments, where green manure species and strategies may be other than those of northern-temperate and tropical regions. Relationships were found for maize shooting (Neff = −3.61DW + 29.75N%; R2 = 0.67), flowering (Neff = −1.32DW + 27.47N%; R2 = 0.66) and physiological maturity (Neff = −4.84DW + 50.43N%; R2 = 0.74) (DW = incorporated biomass, N% = N concentration).

Phenolic compounds in grains, sprouts and wheatgrass of hulled and non-hulled wheat species.

BACKGROUND The use of sprouts and young plantlets in human nutrition is increasing because they often contain phytochemicals and other high value nutrients. This is also the case for wheat, although there is no literature for hulled wheat species. Thus we determined total polyphenols, phenolic acids (PAs), fibre and minerals in grains, 5-day-old sprouts and 12-day-old wheatgrass of einkorn (cv. Monlis), emmer (cvs Augeo, Rosso Rubino, Zefiro), spelt (cvs Pietro, Giuseppe), durum wheat (cv. Creso) and soft wheat (cv. Orso). RESULTS Grains of einkorn and emmer contained twice bound PAs as compared to soft and durum wheat and spelt, with p-coumaric acid accounting for about 50% of total bound PAs. In wheatgrass, differences between species for bound PAs decreased due to a decrease in einkorn and emmer and an increase in soft and durum wheat. In all species, total phenols and free PAs increased passing from grains to sprouts and wheatgrass. Neutral and acid detergent fibre content increased with sprouting only in einkorn and emmer. CONCLUSION Our evidence suggests that the grains of einkorn and emmer and the sprouts and wheatgrass of all Triticum species might potentially be valuable for the development of functional foods.

Crop Rotation as a System Approach for Soil Fertility Management in Vegetables

This paper reviews the recent literature on crop rotation as a tool to manage soil fertility specifically for vegetable production. All of the aspects dealing with soil fertility management, i.e. mineral and organic fertilisation, crop residue s management, cover cropping and green manuring, and intercrop ping, are examined in the frame of crop rotations in conventional and organic systems for both specialised and non-specialised vegetable production. A focus is given on conservation tillage practices to manage green manures and vegetable crop residues. The design and modelling of vegetable rotations are described under the viewpoint of increasing the nutrient use efficiency and the self-sufficiency of the system. Some long-term experiments including vegetables are described which evaluate cumulated effects of rotations on soil fertility and vegetable production. It is concluded that only integrating all the available techniques of soil fertility management at a whole rotation scale it is possible to contribute to the productive, economic and environmental sustainability of the system. For example, little supplementation of mineral or fast-release organic fertilisers delivered with rational fertilisation techniques (e.g. starter, split, and localised fertilisation; fertigation) may help compensate the temporal and spatial lack of matching between nitrogen release from slow-release organic sources and crop nitrogen demand. This would help modulate nutrient supply in a more flexible way and improve crop nutrient uptake, so allowing more constant yields across years and limited risks of nutrient loss to the environment.

RELIABILITY OF NDVI DERIVED BY HIGH RESOLUTION SATELLITE AND UAV COMPARED TO IN-FIELD METHODS FOR THE EVALUATION OF EARLY CROP N STATUS AND GRAIN YIELD IN WHEAT

This study was aimed at comparing in-field parameters and remote sensing NDVI (normalized difference vegetation index) by both satellite (SAT) and unmanned aerial vehicle (UAV) for the assessment of early nitrogen (N) status and prediction of yield in winter wheat (Triticum aestivum L.). Six increasing N rates, i.e., 0, 40, 80, 120, 160, 200 kg N ha−1 were applied, half at tillering and half at shooting. Thus, when the crop N status was monitored between the two N applications, consecutive N treatments differentiated from each other by just 20 kg N ha−1. The following in-field and remote sensed parameters were compared as indicators of crop vegetative and N status: plant N% (w:w) concentration; crop N uptake (Nupt); ratio between transmitted and incident photosynthetically active radiation (PARt/PARi); leaf SPAD values, an indirect index for chlorophyll content; SAT and UAV derived NDVI. As reliable indicators of wheat N availability, in-field parameters were ranked as follows: PARt/PARi ≅ Nupt > SPAD ≅ N%. The PARt/PARi, Nupt and SPAD resulted quite strongly correlated to each other. At all crop stages, the NDVI was strongly correlated with PARt/PARi and Nupt. It is of relevance that NDVI correlated quite strongly to in-field parameters and grain yield at shooting, i.e., before the second N application, when the N rate can still be adjusted. The SAT and UAV NDVIs were strongly correlated to each other, which means they can be used alternatively depending on the context.