Interactive impacts of microplastics and arsenic on agricultural soil and plant traits.

The ability of microplastics (MPs) to interact with environmental pollutants is currently of great concern due to the increasing use of plastic. Agricultural soils are sinks for multipollutants and the safety of biodegradable MPs in field conditions is questioned. However, still few studies have investigated the interactive effects between MPs and metals on the soil-plant system with agricultural soil and testing crops for human consumption. In this work, we tested the effect on soil and plant parameters of two common MPs, non-degradable plastic low-density polyethylene and biodegradable polymer polylactic acid at two different sizes (<250 µm and 250-300 µm) in association with arsenic (As). Lettuce (Lactuca sativa L.) was used as a model plant in a small-scale experiment lasting 60 days. Microplastics and As explained 12 % and 47 % of total variance, respectively, while their interaction explained 21 %, suggesting a higher toxic impact of As than MPs. Plant growth was promoted by MPs alone, especially when biodegradable MPs were added (+22 %). However, MPs did not affect nutrient concentrations in roots and leaves. The effect of MPs on enzyme activities was variable depending on the time of exposure (with larger effects immediately after exposure), the type and size of the MPs. On the contrary, the co-application of MP and As, although it did not change the amount of bioavailable As in soil in the short and medium term, it resulted in a significant decrease in lettuce biomass (-19 %) and root nutrient concentrations, especially when polylactic acid was applied. Generally, MPs in association with As determined the plant-soil toxicity. This work provides insights into the risk of copollution of MPs and As in agricultural soil and its phytotoxic effect for agricultural crops. However, the mechanisms of the joint effect of MP and As on plant toxicity need further investigation, especially under field conditions and in long-term experiments.

Effective bioremediation of clarithromycin and diclofenac in wastewater by microbes and Arundo donax L.

Bioremediation of pharmaceuticals has gained large research efforts, but there is still a need to improve the performance of bioremediation systems by selecting effective organisms. In this study, we characterized the capability to remove clarithromycin (CLA) and diclofenac (DCF) by the bacterium Streptomyces rochei, and the fungi Phanerochaete chrysosporium and Trametes versicolor. The macrolide antibiotic CLA and the non-steroid anti-inflammatory DCF were selected because these are two of the most frequently detected drugs in water bodies. Growth and content of the PhCs and a DCF metabolite (MET) by the energy crop Arundo donax L. were also evaluated under hydroponic conditions. The removal rate (RR) by S. rochei increased from 24 to 40% at 10 and 100 µg CLA L-1, respectively, averaged over incubation times. At 144 h, the RR by P. chrysosporium was 84%, while by T. versicolor was 70 and 45% at 10 and 100 CLA µg L-1. The RR by S. rochei did not exceed 30% at 1 mg DCF L-1 and reached 60% at 10 mg DCF L-1, whereas approached 95% and 63% by P. chrysosporium and T. versicolor, respectively, at both doses. Root biomass and length of A. donax were strongly affected at 100 µg CLA L-1. CLA concentration in roots and shoots increased with the increase of the dose and translocation factor (TF) was about 1. DCF severely affected both shoot fresh weight and root length at the highest dose and concentration in roots and shoots increased with the increase of the dose. DCF concentrations were 16-19 times higher in roots than in shoots, and TF was about 0.1. MET was detected only in roots and its proportion over the parent compound decreased with the increase of the DCF dose. This study highlights the potential contribution of A. donax and the tested microbial inoculants for improving the effectiveness of bioremediation systems for CLA and DCF removal.

Multiple Arbuscular Mycorrhizal Fungal Consortia Enhance Yield and Fatty Acids of Medicago sativa: A Two-Year Field Study on Agronomic Traits and Tracing of Fungal Persistence.

Arbuscular mycorrhizal fungi are promoted as biofertilizers due to potential benefits in crop productivity, and macro- and microelement uptake. However, crop response to arbuscular mycorrhizal fungi (AMF) inoculation is context-dependent, and AMF diversity and field establishment and persistence of inoculants can greatly contribute to variation in outcomes. This study was designed to test the hypotheses that multiple and local AMF inoculants could enhance alfalfa yield and fatty acids (FA) compared to exotic isolates either single or in the mixture. We aimed also to verify the persistence of inoculated AMF, and which component of the AMF communities was the major driver of plant traits. Therefore, a field experiment of AMF inoculation of alfalfa (Medicago sativa L.) with three single foreign isolates, a mixture of the foreign isolates (FMix), and a highly diverse mixture of local AMF (LMix) was set up. We showed that AMF improved alfalfa yield (+ 68%), nutrient (+ 147% N content and + 182% P content in forage), and FA content (+ 105%). These positive effects persisted for at least 2 years post-inoculation and were associated with enhanced AMF abundance in roots. Consortia of AMF strains acted in synergy, and the mixture of foreign AMF isolates provided greater benefits compared to local consortia (+ 20% forage yield, + 36% forage N content, + 18% forage P content, + 20% total FA in forage). Foreign strains of Funneliformis mosseae and Rhizophagus irregularis persisted in the roots of alfalfa 2 years following inoculation, either as single inoculum or as a component of the mixture. Among inoculants, F. mosseae BEG12 and AZ225C and the FMix exerted a higher impact on the local AMF community compared with LMix and R. irregularis BEG141. Finally, the stimulation of the proliferation of a single-taxa (R. irregularis cluster1) induced by all inoculants was the main determinant of the host benefits. Crop productivity and quality as well as field persistence of inoculated AMF support the use of mixtures of foreign AMF. On the other hand, local mixtures showed a lower impact on native AMF. These results pave the way for extending the study on the effect of AMF mixtures for the production of high-quality forage for the animal diet.

Direct transfer of zinc between plants is channelled by common mycorrhizal network of arbuscular mycorrhizal fungi and evidenced by changes in expression of zinc transporter genes in fungus and plant.

The role that common mycorrhizal networks (CMNs) play in plant-to-plant transfer of zinc (Zn) has not yet been investigated, despite the proved functions of arbuscular mycorrhizal fungi (AMF) in crop Zn acquisition. Here, two autotrophic Medicago truncatula plants were linked by a CMN formed by Rhizophagus irregularis. Plants were grown in vitro in physically separated compartments (Donor-C and Receiver-C) and their connection ensured only by CMN. A symbiosis-defective mutant of M. truncatula was used as control in Receiver-C. Plants in both compartments were grown on Zn-free medium, and only the leaves of the donor plants were Zn fertilized. A direct transfer of Zn was demonstrated from donor leaves to receiver shoots mediated by CMN. Direct transfer of Zn was supported by changes in the expression of fungal genes, RiZRT1 and RiZnT1, and plant gene MtZIP2 in roots and MtNAS1 in roots and shoots of the receiver plants. Moreover, Zn transfer was supported by the change in expression of MtZIP14 gene in AM fungal colonized roots. This work is the first evidence of a direct Zn transfer from a donor to a receiver plant via CMN, and of a triggering of transcriptional regulation of fungal-plant genes involved in Zn transport-related processes.

Transcriptional Regulation of Genes Involved in Zinc Uptake, Sequestration and Redistribution Following Foliar Zinc Application to Medicago sativa.

Zinc (Zn) is an essential micronutrient for plants and animals, and Zn deficiency is a widespread problem for agricultural production. Although many studies have been performed on biofortification of staple crops with Zn, few studies have focused on forages. Here, the molecular mechanisms of Zn transport in alfalfa (Medicago sativa L.) were investigated following foliar Zn applications. Zinc uptake and redistribution between shoot and root were determined following application of six Zn doses to leaves. Twelve putative genes encoding proteins involved in Zn transport (MsZIP1-7, MsZIF1, MsMTP1, MsYSL1, MsHMA4, and MsNAS1) were identified and changes in their expression following Zn application were quantified using newly designed RT-qPCR assays. These assays are the first designed specifically for alfalfa and resulted in being more efficient than the ones already available for Medicago truncatula (i.e., MtZIP1-7 and MtMTP1). Shoot and root Zn concentration was increased following foliar Zn applications ≥ 0.1 mg plant-1. Increased expression of MsZIP2, MsHMA4, and MsNAS1 in shoots, and of MsZIP2 and MsHMA4 in roots was observed with the largest Zn dose (10 mg Zn plant-1). By contrast, MsZIP3 was downregulated in shoots at Zn doses ≥ 0.1 mg plant-1. Three functional gene modules, involved in Zn uptake by cells, vacuolar Zn sequestration, and Zn redistribution within the plant, were identified. These results will inform genetic engineering strategies aimed at increasing the efficiency of crop Zn biofortification.

HyLength: a semi-automated digital image analysis tool for measuring the length of roots and fungal hyphae of dense mycelia.

In plant-fungus phenotyping, determining fungal hyphal and plant root lengths by digital image analysis can reduce labour and increase data reproducibility. However, the degree of software sophistication is often prohibitive and manual measuring is still used, despite being very time-consuming. We developed the HyLength tool for measuring the lengths of hyphae and roots in in vivo and in vitro systems. The HyLength was successfully validated against manual measures of roots and fungal hyphae obtained from all systems. Compared with manual methods, the HyLength underestimated Medicago sativa roots in the in vivo system and Rhizophagus irregularis hyphae in the in vitro system by about 12 cm per m and allowed to save about 1 h for a single experimental unit. As regards hyphae of R. irregularis in the in vivo system, the HyLength overestimated the length by about 21 cm per m compared with manual measures, but time saving was up to 20.5 h per single experimental unit. Finally, with hyphae of Aspergillus oryzae, the underestimation was about 8 cm per m with a time saving of about 10 min for a single germinating spore. By benchmarking the HyLength against the AnaMorf plugin of the ImageJ/Fiji, we found that the HyLength performed better for dense fungal hyphae, also strongly reducing the measuring time. The HyLength can allow measuring the length over a whole experimental unit, eliminating the error due to sub-area selection by the user and allowing processing a high number of samples. Therefore, we propose the HyLength as a useful freeware tool for measuring fungal hyphae of dense mycelia.

Interaction Between Conservation Tillage and Nitrogen Fertilization Shapes Prokaryotic and Fungal Diversity at Different Soil Depths: Evidence From a 23-Year Field Experiment in the Mediterranean Area.

Soil biodiversity accomplishes key roles in agro-ecosystem services consisting in preserving and enhancing soil fertility and nutrient cycling, crop productivity and environmental protection. Thus, the improvement of knowledge on the effect of conservation practices, related to tillage and N fertilization, on soil microbial communities is critical to better understand the role and function of microorganisms in regulating agro-ecosystems. In the Mediterranean area, vulnerable to climate change and suffering for management-induced losses of soil fertility, the impact of conservation practices on soil microbial communities is of special interest for building mitigation and adaptation strategies to climate change. A long-term experiment, originally designed to investigate the effect of tillage and N fertilization on crop yield and soil organic carbon, was utilized to understand the effect of these management practices on soil prokaryotic and fungal community diversity. The majority of prokaryotic and fungal taxa were common to all treatments at both soil depths, whereas few bacterial taxa (Cloacimonates, Spirochaetia and Berkelbacteria) and a larger number of fungal taxa (i.e., Coniphoraceae, Debaryomycetaceae, Geastraceae, Cordicypitaceae and Steccherinaceae) were unique to specific management practices. Soil prokaryotic and fungal structure was heavily influenced by the interaction of tillage and N fertilization: the prokaryotic community structure of the fertilized conventional tillage system was remarkably different respect to the unfertilized conservation and conventional systems in the surface layer. In addition, the effect of N fertilization in shaping the fungal community structure of the surface layer was higher under conservation tillage systems than under conventional tillage systems. Soil microbial community was shaped by soil depth irrespective of the effect of plowing and N addition. Finally, chemical and enzymatic parameters of soil and crop yields were significantly related to fungal community structure along the soil profile. The findings of this study gave new insights on the identification of management practices supporting and suppressing beneficial and detrimental taxa, respectively. This highlights the importance of managing soil microbial diversity through agro-ecological intensified systems in the Mediterranean area.

The mycorrhizal pathway of zinc uptake contributes to zinc accumulation in barley and wheat grain.

BACKGROUND: Increasing zinc (Zn) concentrations in crops is important for alleviation of human Zn deficiency. Arbuscular mycorrhizal fungi (AMF) contribute to plant Zn uptake, but their contribution to Zn in the edible portion of crops has not yet been investigated. This study aimed to quantify the mycorrhizal pathway of Zn uptake into grain of wheat and barley under varying soil Zn availabilities. Bread wheat (Triticum aestivum) and barley (Hordeum vulgare) were grown in pots with a hyphal compartment containing 65Zn. Plants were inoculated with Rhizophagus irregularis and grown at three soil Zn concentrations. Radioactive Zn in grain and straw was measured and the contribution of AMF to Zn uptake was calculated. RESULTS: The mycorrhizal pathway of Zn uptake contributed up to 24.3% of total above-ground Zn in wheat, and up to 12.7% of that Zn in barley. The greatest contribution by the mycorrhizal pathway was observed in barley at the lowest Zn addition, and in wheat at the highest one. In addition, grain yield of bread wheat was increased by AMF. CONCLUSIONS: These results suggest that AMF have a substantial role in uptake of Zn into cereals, and the proportional contribution by the MPU is dependent on plant species, as well as available soil Zn.

Forage Rotations Conserve Diversity of Arbuscular Mycorrhizal Fungi and Soil Fertility.

In the Mediterranean, long-term impact of typical land uses on soil fertility have not been quantified yet on replicated mixed crop-livestock farms and considering the variability of soil texture. Here, we report the effects, after 15 years of practice, of two legume-winter cereal rotations, olive orchards and vineyards on microbiological and chemical indicators of soil fertility and the communities of arbuscular mycorrhizal fungi (AMF). We compare the changes among these four agricultural land-use types to woodland reference sites. Root colonization by AMF of English ryegrass (Lolium perenne L.), a grass that occurred under all land use types, was only half as heavy in biannual berseem clover (Trifolium alexandrinum L.)-winter cereal rotations than in 4-year alfalfa (Medicago sativa L.)-winter cereal rotations. In olive (Olea europaea L.) orchards and vineyards (Vitis vinifera L.), where weeds are controlled by frequent surface tillage, the AMF root colonization of ryegrass was again much lower than in the legume-cereal rotations and at the woodland reference sites. All the microbial parameters and soil organic carbon correlated most strongly with differences in occurrence and relative abundance (ß-diversity) of AMF genera in soil. The soil pH and mineral nutrients in soil strongly correlated with differences in AMF root colonization and AMF genus richness (α-diversity) in soil. Diversity of AMF was much less affected by soil texture than land use, while the opposite was true for microbial and chemical soil fertility indicators. Land uses that guaranteed a continuous ground cover of herbaceous plants and that involved only infrequent tillage, such as multiyear alfalfa-winter cereal rotation, allowed members of the AMF genus Scutellospora to persist and remain abundant. On the contrary, under land uses accompanied by frequent tillage and hence discontinuous presence of herbaceous plants, such as tilled olive orchard and vineyard, members of the genus Funneliformis dominated. These results suggest that multiyear alfalfa-winter cereal rotation with active plant growth throughout the year is the least detrimental agricultural land use in soil carbon and AMF abundance and diversity, relative to the woodland reference.

Author Correction: Impact of genetically engineered maize on agronomic, environmental and toxicological traits: a meta-analysis of 21 years of field data.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has not been fixed in the paper.

Impact of genetically engineered maize on agronomic, environmental and toxicological traits: a meta-analysis of 21 years of field data.

Despite the extensive cultivation of genetically engineered (GE) maize and considerable number of scientific reports on its agro-environmental impact, the risks and benefits of GE maize are still being debated and concerns about safety remain. This meta-analysis aimed at increasing knowledge on agronomic, environmental and toxicological traits of GE maize by analyzing the peer-reviewed literature (from 1996 to 2016) on yield, grain quality, non-target organisms (NTOs), target organisms (TOs) and soil biomass decomposition. Results provided strong evidence that GE maize performed better than its near isogenic line: grain yield was 5.6 to 24.5% higher with lower concentrations of mycotoxins (-28.8%), fumonisin (-30.6%) and thricotecens (-36.5%). The NTOs analyzed were not affected by GE maize, except for Braconidae, represented by a parasitoid of European corn borer, the target of Lepidoptera active Bt maize. Biogeochemical cycle parameters such as lignin content in stalks and leaves did not vary, whereas biomass decomposition was higher in GE maize. The results support the cultivation of GE maize, mainly due to enhanced grain quality and reduction of human exposure to mycotoxins. Furthermore, the reduction of the parasitoid of the target and the lack of consistent effects on other NTOs are confirmed.

Biofortification with Iron and Zinc Improves Nutritional and Nutraceutical Properties of Common Wheat Flour and Bread.

The effect of field foliar Fe and Zn biofortification on concentration and potential bioavailability of Fe and Zn and health-promoting compounds was studied in wholemeal flour of two common wheat varieties (old vs modern). Moreover, the effect of milling and bread making was studied. Biofortification increased the concentration of Zn (+78%) and its bioavailability (+48%) in the flour of the old variety, whereas it was ineffective in increasing Fe concentration in both varieties. However, the old variety showed higher concentration (+41%) and bioavailability (+26%) of Fe than the modern one. As regard milling, wholemeal flour had higher Fe, Zn concentration and health-promoting compounds compared to white flour. Bread making slightly change Fe and Zn concentration but greatly increased their bioavailability (77 and 70%, respectively). All these results are of great support for developing a production chain of enriched functional bread having a protective role against chronic cardio-vascular diseases.

Investigation of a Staphylococcal Food Poisoning Outbreak from a Chantilly Cream Dessert, in Umbria (Italy).

On August 28, 2015, a staphylococcal food poisoning outbreak occurred in Umbria, Italy, affecting 24 of the 42 customers who had dinner at a local restaurant. About 3 h after ingesting a variety of foods, the customers manifested gastrointestinal symptoms. Within 24 h of notification from the hospital emergency department, Sanitary Inspectors of the local Public Health Unit performed an epidemiological investigation. A retrospective cohort study was conducted among the customers. Food and environmental samples were collected. Due to the rapid onset of symptoms (vomiting, diarrhea), the food samples were analyzed for the presence of toxigenic bacteria and their toxins; nasopharyngeal swabs were collected from the waiters and cooks. Among the food tested, high levels of coagulase-positive staphylococci (CPS) (3.4 × 108 CFU/g) and staphylococcal enterotoxins (2.12 ng SEA/g) were only detected in the Chantilly cream dessert. CPS were also detected on the surface of a kitchen table (10 CFU/swab), and five food handlers were positive for Staphylococcus aureus. In total, five enterotoxigenic S. aureus isolates were recovered from three food handlers, a kitchen surface, and the Chantilly cream dessert. These isolates were further characterized by biotyping, pulsed-field gel electrophoresis, and multiplex polymerase chain reaction assays for the detection of eleven enterotoxin encoding genes (sea, seb, sec, sed, see, seg, seh, sei, sej, sep, and ser) and three genes involved in antibiotic resistance (mecA, mecC, and mupA). Three sea-positive strains, isolated from the dessert, environment, and one of the cooks, had the same pulsed-field gel electrophoresis profile and belonged to the human biotype, suggesting that the contamination causing the outbreak most likely originated from a food handler. Moreover, improper storage of the dessert, at room temperature for about 5 h, permitted microbial growth and SEA production. This study underlines the importance of both laboratory evidence and epidemiological data for outbreak investigation.

Land-use intensity and host plant simultaneously shape the composition of arbuscular mycorrhizal fungal communities in a Mediterranean drained peatland.

Land-use change is known to be a major threat to biodiversity and ecosystem services in Mediterranean areas. However, the potential for different host plants to modulate the effect of land-use intensification on community composition of arbuscular mycorrhizal fungi (AMF) is still poorly understood. To test the hypothesis that low land-use intensity promotes AMF diversity at different taxonomic scales and to determine whether any response is dependent upon host plant species identity, we characterised AMF communities in the roots of 10 plant species across four land use types of differing intensity in a Mediterranean peatland system. AMF were identified using 454 pyrosequencing. This revealed an overall low level of AMF richness in the peaty soils; lowest AMF richness in the intense cropping system at both virtual taxa and family level; strong modulation by the host plant of the impact of land-use intensification on AMF communities at the virtual taxa level; and a significant effect of land-use intensification on AMF communities at the family level. These findings have implications for understanding ecosystem stability and productivity and should be considered when developing soil-improvement strategies in fragile ecosystems, such as Mediterranean peatlands.

Prevalence and characteristics of verotoxigenic Escherichia coli strains isolated from pigs and pork products in Umbria and Marche regions of Italy.

In total 1095 samples from 675 pork products, 210 swine colon contents, and 210 swine carcass sponge swabs were collected in Umbria and Marche regions of Italy and examined for the presence of Shiga toxin-producing Escherichia coli (STEC), also known as Verotoxin-producing E. coli (VTEC). After an enrichment step, each sample was analysed by real-time PCR to detect the stx1, stx2, and eae genes. stx-Positive samples were further tested for the "top five" serogroup markers (O157, O26, O103, O111, O145) and cultured onto selective media. The isolates were assigned to stx subtypes and tested for the presence of aaiC and aggR genes. Out of 420 swine samples, 38.6% faecal samples and 13.8% carcass sponge swabs were stx-positive. In total, 33 E. coli STEC isolates were obtained from 30 samples (4 carcasses and 26 colon contents) indicating a culture-positive rate of 7.1%. A higher culture-positive rate was observed in faecal samples (12.4%) than in carcass sponge swabs (1.9%). Out of 675 pork samples, 19 (2.8%) were stx-positive. No STEC strains were isolated from stx-positive pork products. We concluded that STEC isolation from foodstuffs remains difficult, despite the application of ISO/TS 13136:2012. Furthermore, in accordance with the results of studies conducted in other countries, we observed that most of swine STEC strains carried stx2e gene and lacked of virulence genes, such as eae, aaiC and aggR, indicative of potential pathogenic characteristics for humans. Although the majority of STEC isolates did not express virulence factors correlating with severe human diseases, the association between swine STEC strains and human illness requires further investigations.

Role of Verocytotoxigenic Escherichia Coli in the Swine Production Chain.

Shiga toxin-producing Escherichia coli (STEC) can cause severe clinical diseases in humans, such as haemorrhagic colitis (HC) and haemolytic-uremic syndrome (HUS). Although ruminants, primarily cattle, have been suggested as typical reservoirs of STEC, many food products of other origins, including pork products, have been confirmed as vehicles for STEC transmission. Only in rare cases, pork consumption is associated with severe clinical symptoms caused by high pathogenic STEC strains. However, in these outbreaks, it is unknown whether the contamination of food products occurs during swine processing or via cross-contamination from foodstuffs of different sources. In swine, STEC plays an important role in the pathogenesis of oedema disease. In particular a Shiga toxin subtype, named stx2e, it is considered as a key factor involved in the damage of swine endothelial cells. On the contrary, stx2e-producing Escherichia coli has rarely been isolated in humans, and usually only from asymptomatic carriers or from patients with mild symptoms, such as uncomplicated diarrhoea. In fact, the presence of gene stx2e, encoding for stx2e, has rarely been reported in STEC strains that cause HUS. Moreover, stx2e-producing STEC isolated from humans and pigs were found to differ in serogroup, their virulence profile and interaction with intestinal epithelial cells. Because of the limited epidemiologic data of STEC in swine and the increasing role of non-O157 STEC in human illnesses, the relationship between swine STEC and human disease needs to be further investigated.

The response of durum wheat to the preceding crop in a Mediterranean environment.

Crop sequence is an important management practice that may affect durum wheat (Triticum durum Desf.) production. Field research was conducted in 2007-2008 and 2008-2009 seasons in a rain-fed cold Mediterranean environment to examine the impact of the preceding crops alfalfa (Medicago sativa L.), maize (Zea mays L.), sunflower (Helianthus annuus L.), and bread wheat (Triticum aestivum L.) on yield and N uptake of four durum wheat varieties. The response of grain yield of durum wheat to the preceding crop was high in 2007-2008 and was absent in the 2008-2009 season, because of the heavy rainfall that negatively impacted establishment, vegetative growth, and grain yield of durum wheat due to waterlogging. In the first season, durum wheat grain yield was highest following alfalfa, and was 33% lower following wheat. The yield increase of durum wheat following alfalfa was mainly due to an increased number of spikes per unit area and number of kernels per spike, while the yield decrease following wheat was mainly due to a reduction of spike number per unit area. Variety growth habit and performance did not affect the response to preceding crop and varieties ranked in the order Levante > Saragolla = Svevo > Normanno.

Coordination between plant and apex development in Hordeum vulgare ssp. distichum.

Developmental scales for cereals describe apex and plant morphology separately. In order to link crucial steps of internal and external development, in three varieties of Hordeum vulgare spp. distichum L., sown in autumn and in spring, we recorded plant, leaf and apex stage, following the scales of Zadoks, Haun, and Banerjee and Wienhues, the number of primordia, culm and spike length, and the final number of leaves and spikelets. Primordia initiation was coordinated with leaf appearance and the relative rate was constant for the initiation of productive primordia. The maximum number of primordia was achieved just before the first node became detectable, but development was completed only by those initiated before floret differentiation and internode distension started. The first spikelet was initiated when the third leaf tip became visible, and the last one when plants were at the pseudo stem erection stage and five leaves had still to appear.

The stucco work of Giacomo Serpotta and the Serpotta School. Constitutive materials, degradation and decay.

Giacomo Serpotta (1656-1732) was the major figure of a school of stucco work which was growing and developing in Palermo in the course of several decades towards the end of the 17th century. This paper presents some preliminary results of a wide and systematic comparative study, aimed at investigation of the compositional and textural characteristics of the stucco, and of the techniques of the modeling and finishing of its surfaces, by means both of measurements of physico-chemical properties of the materials and of their mineralogical and petrographic characterization. The opportunity for such a study was made available by the launching of the "Progetto Serpotta" by the Soprintendenza dei Beni Ambientali e Culturali di Palermo. This project is aimed at the restoration and conservation of most of the Serpotta school stucco works. The project offers a unique opportunity to extract samplings from the entire cycle of works prior to restoration. Moreover, the study of the stucco's material composition serves as an aid in better understanding the nature and the causes of degradation and decay, which in many cases have seriously damaged the works.