Teofilo Vamerali

Biochar addition to an arsenic contaminated soil increases arsenic concentrations in the pore water but reduces uptake to tomato plants (Solanum lycopersicum L.).

Arsenic (As) concentrations in soil, soil pore water and plant tissues were evaluated in a pot experiment following the transplantation of tomato (Solanum lycopersicum L.) plantlets to a heavily As contaminated mine soil (~6000 mg kg(-1) pseudo-total As) receiving an orchard prune residue biochar amendment, with and without NPK fertiliser. An in-vitro test was also performed to establish if tomato seeds were able to germinate in various proportions of biochar added to nutrient solution (MS). Biochar significantly increased arsenic concentrations in pore water (500 μg L(-1)-2000 μg L(-1)) whilst root and shoot concentrations were significantly reduced compared to the control without biochar. Fruit As concentrations were very low (<3 μg kg(-1)), indicating minimal toxicity and transfer risk. Fertilisation was required to significantly increase plant biomass above the control after biochar addition whilst plants transplanted to biochar only were heavily stunted and chlorotic. Given that increasing the amount of biochar added to nutrient solution in-vitro reduced seed germination by up to 40%, a lack of balanced nutrient provision from biochar could be concluded. In summary, solubility and mobility of As were increased by biochar addition to this soil, but uptake to plant was reduced, and toxicity-transfer risk was negligible. Therefore leaching rather than food chain transfer appears the most probable immediate consequence of biochar addition to As contaminated soils.

Phytoremediation trials on metal- and arsenic-contaminated pyrite wastes (Torviscosa, Italy).

At a site in Udine, Italy, a 0.7m layer of As, Co, Cu, Pb and Zn contaminated wastes derived from mineral roasting for sulphur extraction had been covered with an unpolluted 0.15m layer of gravelly soil. This study investigates whether woody biomass phytoremediation is a realistic management option. Comparing ploughing and subsoiling (0.35m depth), the growth of Populus and Salix and trace element uptake were investigated in both pot and field trials. Species differences were marginal and species selection was not critical. Impaired above-ground productivity and low translocation of trace elements showed that bioavailable contaminant stripping was not feasible. The most significant finding was of coarse and fine roots proliferation in surface layers that provided a significant sink for trace elements. We conclude that phytostabilisation and effective immobilisation of metals and As could be achieved at the site by soil amelioration combined with woody species establishment. Confidence to achieve a long-term and sustainable remediation requires a more complete quantification of root dynamics and a better understanding of rhizosphere processes.

A comparison of root characteristics in relation to nutrient and water stress in two maize hybrids

Root responses of maize (Zea mays L.) to limited nutrients and water availability were evaluated in two highly productive full-season hybrids, DK585 and Santos (Dekalb — Monsanto), in laboratory, pot and field tests. In the laboratory, under optimal nutrient and water supply, seedlings of DK585 had higher growth (leaves and roots). Under nitrate or sulphate deprivation, DK585 showed better ability in adapting its root/shoot ratio to stress conditions, whereas Santos showed less plastic behaviour. This morphological trait of DK585 was associated with higher sulphate and constitutive nitrate influxes. In pot trials (plants with four to five leaves), DK585 maintained a high transpiration level to very low values (around 0.2) of FTSW (fraction of transpirable soil water), whereas Santos showed a higher response to soil drying. The latter reduced the rate of transpiration starting from a FTSW of about 0.6. In the open field (trial in 2000, Legnaro, NE Italy), in conditions of fluctuating combined water and nitrogen stress, DK585 at flowering reached greater root length density (RLD) than Santos in deep layers (50–100-cm interval depth) of positions further from the plant. However, in these conditions, the yield of DK585 was found to be only slightly higher than that of Santos (8.88 vs. 8.49 t ha.−1 d.w.). An overall evaluation of the two hybrids indicates the more conservative strategy towards limited water and nutrient resources in Santos, and a greater tendency towards stress avoidance in DK585.

Oil crops for biodiesel production in Italy

There is an energy advantage in methylester production using sunflower, rapeseed and soybean as crop suppliers of energy, the process being practicable in terms of technique and energy gain. The three oilseed crops considered in this work are suitable for considerable reduction of inputs with consequent energy savings and low environmental impact. Nevertheless, because of its low arable surface area, the number of available hectares being static, Italy is oriented towards intensive management. As a consequence, optimisation of cropping techniques is the main strategy to be pursued for higher crop yields for methylester production.

An approach to minirhizotron root image analysis

Minirhizotrons speed up research on root demography, but image quality often hampers standardization of the image processing method. A simple procedure working on the blue band of colour images was tested on fibrous roots of sugarbeet (Beta vulgaris var. saccharifera). With respect to green and red, the blue band allows better detection of roots when their luminance is very similar to that of the background. The method makes use of an exponential algorithm of contrast stretching, which takes luminance frequency distribution into account. Based on a single threshold level, the procedure includes skeletonization. A minimum segment length was adopted to discriminate roots from extraneous objects. Although a specific minimum root length (MRL) value was calculated for each soil type, results show that a single value can be applied, indicating that this method can be profitably used for processing large samples of images.

Analysis of root images from auger sampling with a fast procedure: a case of application to sugar beet

Manual line-intersect methods for estimating root length are being progressively replaced by faster and more accurate image analysis procedures. These methods even allow the estimation of some more root parameters (e.g., diameter), but still require preliminary labour-intensive operations. Through a task-specific macro function written in a general-purpose image analysis programme (KS 300 — Zeiss), the processing time of root images was greatly reduced with respect to skeletonisation methods by using a high-precision algorithm (Fibrelength). This has been previously proposed by other authors, and estimates length as a function of perimeter and area of the digital image of roots. One-bit binary images were acquired, aiming at large savings in computer memory, and automatic discrimination of roots against extraneous objects based on their elongation index (perimeter2/area), was performed successfully. Of four tested spatial resolutions (2.9, 5.9, 8.8, 11.8 pixel mm−1), in clean samples good accuracy in root length estimation was achieved at 11.8 pixel mm−1, up to a root density of 5 cm cm−2 on the scanner bed. This resolution is theoretically suitable for representing roots at least 85 Am wide. When dealing with uncleaned samples, a thick layer of water was useful in speeding up spreading of roots on the scanner bed and avoiding underestimation of their length due to overlaps with organic debris. A set of fibrous root samples of sugar beet (Beta vulgaris var. saccharifera L.) collected at harvest over two years at Legnaro (NE Italy) was analysed by applying the above procedure. Fertilisation with 100 kg ha−1 of nitrogen led to higher RLD (root length density in soil) in shallow layers with respect to unfertilised controls, whereas thicker roots were found deeper than 80 cm of soil without nitrogen.

Culturable endophytic bacteria enhance Ni translocation in the hyperaccumulator Noccaea caerulescens

In this work, both culture-dependent and independent approaches were used to identify and isolate endophytic bacteria from roots of the Ni hyperaccumulator Noccaea caerulescens. A total of 17 isolates were cultured from root samples, selected for tolerance to 6mM Ni and grouped by restriction analysis of 16S rDNA. Bacterial species cultivated from roots belonged to seven genera, Microbacterium, Arthrobacter, Agreia, Bacillus, Sthenotrophomonas, Kocuria and Variovorax. The culture-independent approach confirmed the presence of Microbacterium and Arthrobacter while only other five clones corresponding to different amplified ribosomal DNA restriction patterns were detected. Five selected highly Ni-resistant bacteria showing also plant growth promoting activities, were inoculated into seeds of N. caerulescens, and in vivo microscopic analysis showed rapid root colonisation. Inoculated plants showed increased shoot biomass, root length and root-to-shoot Ni translocation. Root colonisation was also evident, but not effective, in the non-hyperaccumulating Thlaspi perfoliatum. Seed inoculation with selected Ni-resistant endophytic bacteria may represent a powerful tool in phytotechnologies, although transferring it to biomass species still requires further studies and screening.

Fibrous root turnover and growth in sugar beet (Beta vulgaris var. saccharifera) as affected by nitrogen shortage

The root system of plants is subject to fast cycles of renewal and decay within the growing season. In water and/or nutrient stress conditions, this turnover may become strategic for plant survival and productivity, but knowledge about its mechanisms is still insufficient. In order to investigate the effects of nitrogen fertilization on growth and turnover of sugar beet roots, an experiment was carried out over two growing seasons in northern Italy with two levels of N supply (0, 100 kg ha−1). Biomass production and partitioning were followed during growth, and fibrous root dynamics were inspected by means of computer-aided procedures applied to minirhizotron images.

In situ phytoremediation of arsenic- and metal-polluted pyrite waste with field crops: Effects of soil management

Sunflower, alfalfa, fodder radish and Italian ryegrass were cultivated in severely As-Cd-Co-Cu-Pb-Zn-contaminated pyrite waste discharged in the past and capped with 0.15m of unpolluted soil at Torviscosa (Italy). Plant growth and trace element uptake were compared under ploughing and subsoiling tillages (0.3m depth), the former yielding higher contamination (∼30%) in top soil. Tillage choice was not critical for phytoextraction, but subsoiling enhanced above-ground productivity, whereas ploughing increased trace element concentrations in plants. Fodder radish and sunflower had the greatest aerial biomass, and fodder radish the best trace element uptake, perhaps due to its lower root sensitivity to pollution. Above-ground removals were generally poor (maximum of 33mgm(-2) of various trace elements), with Zn (62%) and Cu (18%) as main harvested contaminants. The most significant finding was of fine roots proliferation in shallow layers that represented a huge sink for trace element phytostabilisation. It is concluded that phytoextraction is generally far from being an efficient management option in pyrite waste. Sustainable remediation requires significant improvements of the vegetation cover to stabilise the site mechanically and chemically, and provide precise quantification of root turnover.

Field release of genetically marked Azospirillum brasilense in association with Sorghum bicolor L.

The agronomic impact of genetically tagged azospirilla (Azospirillum brasilense)was assessed in open field and their fluctuation were monitored in the soil/rhizosphere. Strain performance, upon inoculation of sorghum, was evaluated over a two-years period; agronomic treatments included nitrogen application (0, 80, 160 kg ha−1), and types of inoculant (Sp245 lacZ, Sp6 gusA, Sp6 IAA++gusA). Grain yield was higher for inoculated seed plots than in non-inoculated ones, whereas nitrogen content, biomass of plant residues and nitrogen in plant residues gave values that were not statistically different. Root length density (RLD) of sorghum at the end of the stem elongation stage was affected only by the indole-3-acetic acid (IAA) overproducer Azospirillum strain (A. brasilense Sp6 IAA++gusA) with respect to the normal IAA producer (A. brasilense Sp6 gusA), being higher in the first 40 cm of depth, notwithstanding the level of nitrogen fertilization. The traceability of the released genetically modified strains enabled to monitor their ability to colonise soil and roots. Moreover, the genetic modification per se vs. the non-modified counterpart, did not affect the culturable aerobic population in soil, microfungi, streptomycetes, fluorescent pseudomonads, soil microbial biomass, or some microbial activities, all selected as important indicators.