Wild cereals from Holocene central Sahara archaeological sites: aDNA and archaeobotany from the Takarkori rockshelter

Both pollen and macroremains recovered from archaeological sites point to a long-time exploitation of wild cereals and to the prevalence of these plants among those selected and transported to shelters and caves of central Sahara (southwestern Libya)(1). Spikelets, florets and grains of Panicoideae (Brachiaria, Cenchrus, Echinochloa, Panicum, Setaria, Urochloa, Sorghum) are the most abundant plant remains in these sites. The archaeological excavation of the Takarkori rockshelter by the Italian-Libyan Archaeological Mission in the Acacus and Messak (Central Sahara), Sapienza University of Rome (directed by S. di Lernia), exposed a surface of c. 120 m2 in extent. The 1.6 m thick deposit includes stone structures, fireplaces, plant accumulations, dung layers and a burial area. Layers were deposited during hunter-gatherer and later pastoralist occupational phases. Chronology ranged from c. 9000 to 4200 BP, and was based on stratigraphy, radiocarbon dates, and archaeological materials (2). Systematic morphological analysis was carried out on 200 spikelets/florets selected as representative of different chronological contexts. The records of Panicum, Echinochloa and Sorghum showed homogeneous typology and fairly uniform size in each genus. aDNA extraction confirming the morphological identifications of the three taxa was obtained by Olmi et al. (3). aDNA was successfully performed using several methods (4, 5) and then assayed using PCR with a primer set for the rbcL gene. New aDNA extraction was obtained from Panicum spikelets and the analysis of matK, trnH-psbA and trnL barcode regions may help to identify the records at species level

Infected pancreatic necrosis: outcomes and clinical predictors of mortality. A post hoc analysis of the MANCTRA-1 international study

: The identification of high-risk patients in the early stages of infected pancreatic necrosis (IPN) is critical, because it could help the clinicians to adopt more effective management strategies. We conducted a post hoc analysis of the MANCTRA-1 international study to assess the association between clinical risk factors and mortality among adult patients with IPN. Univariable and multivariable logistic regression models were used to identify prognostic factors of mortality. We identified 247 consecutive patients with IPN hospitalised between January 2019 and December 2020. History of uncontrolled arterial hypertension (p = 0.032; 95% CI 1.135-15.882; aOR 4.245), qSOFA (p = 0.005; 95% CI 1.359-5.879; aOR 2.828), renal failure (p = 0.022; 95% CI 1.138-5.442; aOR 2.489), and haemodynamic failure (p = 0.018; 95% CI 1.184-5.978; aOR 2.661), were identified as independent predictors of mortality in IPN patients. Cholangitis (p = 0.003; 95% CI 1.598-9.930; aOR 3.983), abdominal compartment syndrome (p = 0.032; 95% CI 1.090-6.967; aOR 2.735), and gastrointestinal/intra-abdominal bleeding (p = 0.009; 95% CI 1.286-5.712; aOR 2.710) were independently associated with the risk of mortality. Upfront open surgical necrosectomy was strongly associated with the risk of mortality (p < 0.001; 95% CI 1.912-7.442; aOR 3.772), whereas endoscopic drainage of pancreatic necrosis (p = 0.018; 95% CI 0.138-0.834; aOR 0.339) and enteral nutrition (p = 0.003; 95% CI 0.143-0.716; aOR 0.320) were found as protective factors. Organ failure, acute cholangitis, and upfront open surgical necrosectomy were the most significant predictors of mortality. Our study confirmed that, even in a subgroup of particularly ill patients such as those with IPN, upfront open surgery should be avoided as much as possible. Study protocol registered in ClinicalTrials.Gov (I.D. Number NCT04747990)

“Banca del Germoplasma” per la conservazione della biodiversità dei cereali: progetto e prospettive future

A partire dal 2011 è iniziato presso il Centro Interdipartimentale “Biogest Siteia” dell‟Università degli Studi di Modena e Reggio Emilia un progetto per la realizzazione di una Banca del Germoplasma per la conservazione e la valorizzazione di specie di interesse agrario, in particolare di varietà di frumento tenero (Triticum aestivum L.). Nell‟ambito del progetto sono previste la caratterizzazione morfologica di spighe e cariossidi, sulla base di descrittori standard, e la caratterizzazione biochimica di ciascuna varietà, basata sulla determinazione quali-quantitativa di metaboliti primari e secondari di interesse nutrizionale

“Banca del Germoplasma” per la conservazione della biodiversità dei cereali: progetto e prospettive future

La conservazione di germoplasma, di corrente o potenziale valore, si profila come un sicuro investimento per poter disporre, anche in futuro, di preziose risorse genetiche per le specie di interesse agrario. Si assiste spesso alla scomparsa di antiche varietà locali e di specie selvatiche parentali di nuove cultivars, le quali presentano elevata variabilità genetica e buona adattabilità alle condizioni ambientali del luogo di origine e/o di coltivazione. La severa e continua selezione di sempre nuove varietà infatti, se da un lato ha consentito di disporre di prodotti di elevata qualità, ha inevitabilmente comportato una severa erosione delle risorse genetiche delle piante coltivate, con una diminuzione della resistenza a stress di natura abiotica e biotica. Risultano quindi di fondamentale importanza le attività di conservazione ex situ, come ad esempio la costituzione di banche del germoplasma vegetale che, se affiancate a vivai di campo per il periodico rinnovo del materiale conservato, possono garantire sopravvivenza ai pool genici vegetali. Presso il Centro Interdipartimentale Biogest-Siteia è stato sviluppato, a partire dal 2011, un progetto che prevede la realizzazione di una banca del germoplasma per la conservazione e la valorizzazione di cereali, in particolare di varietà di frumento tenero (Triticum aestivum L.), con diversi scopi congiunti: - effettuare la raccolta, la conservazione e lo scambio delle sementi; - conservare varietà autoctone che possono essere valorizzate attraverso la produzione di prodotti tipici, a tutela delle tradizioni locali; - effettuare descrizioni morfometriche corredate da immagini digitali; - applicare protocolli per l’analisi metabolica delle sementi. Ad ogni accessione della banca del germoplasma è assegnato un codice interno e quindi, dopo una prima fase di pulizia, selezione, misura di volume e peso del campione e stima del numero di cariossidi per campione, si procede all’essicazione e allo stoccaggio. Le cariossidi, confezionate in buste termosaldate, sono stoccate per la conservazione a medio (4 °C) e a lungo termine (-20 °C). Parte del materiale viene conservato in contenitori plastici in modo tale da avere sementi fruibili per l’analisi morfometrica, basata su descrittori standard e corredata da immagini digitali, e per le analisi biochimiche, i cui protocolli, in corso di perfezionamento, permetteranno di effettuare una caratterizzazione quali-quantitativa dei metaboliti secondari e del profilo aminoacidico. Sulla base di una convenzione con l’Istituto Alcide Cervi di Gattatico (RE), è inoltre disponibile un appezzamento di terreno presso l’annesso Parco Agroambientale, sul quale sono in atto sperimentazioni di campo e il rinnovo delle collezioni. Tutte le attività sono illustrate tramite adeguata cartellonistica per fornire informazioni sul materiale in coltivazione e per valorizzare il progetto anche a fini divulgativi e didattici

Micromorphological analysis in flowers and leaves of grapevine (Vitis vinifera ssp. sativa)

MICROMORPHOLOGICAL ANALYSIS IN FLOWERS AND LEAVES OF GRAPEVINE (Vitis vinifera ssp. sativa) SEM analysis represents an important tool to recognize some plant traits, morpho-physiological characters useful in studying reproductive fitness and plant-environmental relationships. Some cultivars of grapevine cultivated in Emilia, the Western part of Emilia-Romagna Region, have been studied to obtain a morpho-functional characterization of flowers. Malbo gentile, Trebbianina, Lambrusco di Sorbara, L. Montericco and L. Barghi showed hermaphrodite flowers with a normally developed gynoecium but back-reflexed stamens. Their flowers produced anomalous pollen, without furrows and pores-acolporated pollen-unable to let pollen tube growth. In Termarina and Lambrusco salamino, differently, hermaphrodite flowers had erect stamens at the anthesis and produced normal tricolporate pollen. These results have been related with those reported from OIV and IPGRI grapevine descriptors (1), relatively to the occurrence of morphologically hermaphrodite but functionally female flowers. Grapevine leaf traits are frequently related with susceptibility or tolerance to biotic and abiotic stress (2,3,4). Moreover, seventeen Vitis genotypes, from different Italian Regions, have been analysed at SEM to characterize leaves traits. Results showed that the genotypes differ for stomatal density and size: mean stomata density ranged between 100 and 200 stomata/mm2. Highest values of stomata density were recorded in the cultivar Nero d’Avola from Sicily and in V. Berlandieri, an American species from which several currently-used rootstocks have been obtained, while the lowest value was in wild grapevine (Vitis vinifera ssp. sylvestris). Evaluating density, position and types of trichomes, a wide variation - mainly with respect of density - resulted within the observed genotypes. These characters are usually taken in consideration as ampelographic descriptors (1) but they play also an important role in pathogen resistance and may be related with plant resistance to stress. References 1. IPGRI, UPOV, OIV. 1997. International Union for the Protection of New Varieties of Plants, Geneva, Switzerland/Office International de la Vigne et du Vin, Paris, France/International Plant Genetic Resources Institute, Rome, Italy. 2. Boso S., Alonso–Villaverde V., Santiago J.L., et al. (2010). Vitis, 49: 43-50. 3. Winkel T., Rambal S. (1993). Aust. J. Plant Physiol., 20: 143–157. 4. Boso S., Gago P., Alonso–Villaverde V., et al., (2011). Sci. Hort., 128: 228-238

Leaf traits: a micro-morphological characterization of grapevine (Vitis sp. pl.) genotypes

The analysis of leaf structure provides important elements to ecological studies and useful information to evaluate the susceptibility or tolerance to biotic and abiotic stress. A number of histological studies have been carried out on grapevine cultivars, relating leaf traits to resistance against fungal pathogens (1), water stress response (2) and other environmental conditions (3). In the present work the leaves of 17 different grapevine genotypes, Vitis Berlandieri, Vitis vinifera ssp. sylvestris and cultivars of Vitis vinifera ssp. sativa from six different Italian Regions, have been observed at the Scanning Electron Microscope (SEM). The leaves of all genotypes showed stomata only on the abaxial surface and scattered in the epidermis, but with a great variability, in terms of stomatal density and size, occurring among species and cultivars: mean stomata density in fact ranged between 100 and 200 stomata/mm2. High values of stomata density (more of 200 stomata/mm2) were recorded in Nero d’Avola and V. Berlandieri. The lowest values occurred in wild grapevine and in Sangiovese but, concerning the size of stomata, these last genotypes showed significantly larger and wider stomata when compared to the others. Stomata density appears to be a very variable parameter, related to genotype (4), but also to many environmental conditions, i.e. water stress and light irradiance (5) and CO2 concentration (6). Furthermore, raised and sunken stomata were visible in the same leaf with different ratios. In Vitis Berlandieri more than 99% of the stomata was protruding from the surface of the epidermis while Nero d’Avola exhibited about 89% of sunken stomata. Density, position and types of trichomes differently characterized the leaves of studied Vitis accessions; prostrate and erect hairs were mainly observed on abaxial side but in some genotypes they were observed also on upper epidermis. Leaf hairs are morphological characters that are taken in consideration as ampelographic descriptors of Vitis species and cultivars but they play also an important role in pathogen resistance (7). In this study the analysis of leaf traits allowed to obtain a micro-morphological characterization of grapevine genotypes; some characters may be related with plant resistance to stress. References 1) Boso S., Alonso–Villaverde V., Santiago J.L., et al. (2010). Macro– and microscopic leaf characteristics of six grapevine genotypes (Vitis spp.) with different susceptibilities to grapevine downy mildew. Vitis, 49: 43-50. 2) Winkel T., Rambal S. (1993). Influence of water stress on grapevine growing in the field, from leaf to whole plant response. Aust J Plant Physiol, 20: 143–157. 3) Rogiers S.Y., Hardie W.J., Smith J.P. (2011). Stomatal density of grapevine leaves (Vitis vinifera L.) responds to soil temperature and atmospheric carbon dioxide. Aust J Grape Wine R, 17:147-152. 4) Boso S., Gago P., Alonso–Villaverde V.,et al. (2011). Variability at the electron microscopic level in leaves of members of the genus Vitis. Sci Hort, 128: 228-238. 5) Palliotti A., Cartechini A., Ferranti F. (2000). Morpho–anatomical and physiological characteristics of primary and lateral shoot leaves of Cabernet Franc and Trebbiano toscano grapevines under two irradiance regimes. Am J Enol Viticult, 51: 122–130. 6) Moutinho-Pereira J.M., Gonçalves B., Bacelar E.A.,et al. (2009). Effects of elevated CO2 on grapevine (Vitis vinifera L.): physiological and yield attributes. Vitis, 48:159–165. 7) Levin D.A. (1973). The role of trichomes in plant defence. Q Rev Biol, 48: 3-15

Caratterizzazione micromorfologica di foglie in genotipi di Vitis

The leaf micromorphological features of 17 different genotypes of grapevine have been analyzed at Environmental Scanning Electron Microscope in order to achieve useful information for a better understanding of the relationship with the sensibility/tolerance to biotic and abiotic stress. The analysis showed leaves with stomata only on the abaxial side, scattered in the epidermis, with a great variability of position among accessions. Furthermore, raised and sunken stomata were visible in the same leaf with different ratios. In Vitis Berlandieri more than 99% of the stomata was protruding from the surface of the epidermis while the cv Nero d\u2019Avola exhibited about 89% of sunken stomata. Mean stomatal density always ranged between 100 and 200 stomata/mm2, with significant differences amongst the accessions. The lowest values occurred in the wild grapevine and in Sangiovese but, concerning the size of stomata, these genotypes showed significantly larger and wider stomata when compared to the others. Density and types of trichomes differently characterize the leaves of studied Vitis accessions. Prostrate and erect hairs were mainly observed on abaxial side but in some genotypes they occurred also on upper epidermis. Key words: grapevine, electronic microscopy, stomata, hairs, abiotic stres

POLLEN, MACROREMAINS AND FRUITS’ aDNA FROM THE TAKARKORI ROCKSHELTER: AN INTEGRATED ARCHAEOBOTANICAL RESEARCH IN CENTRAL SAHARA

The archaeobotanical record from the Takarkori rockshelter (Fezzan, south-western Libya) was systematically collected in the 2003-2006 excavation seasons. The archaeological excavation was carried out over a surface of ca. 120 square metres and brought to light occupation layers radiocarbon dated between ca. 4500 and 8900 uncal. years bp. The rockshelter hosts a 1.6 m thick archaeological deposit including structures, fireplaces and burials of women and children interred near the shelter wall. An impressive amount of mainly desiccated and well preserved plant macroremains was brought to light from Late Acacus and Pastoral layers. Known amount of sediment (3 or 6 l) were dry sieved. Charcoals and seeds/fruits concentrations visible to naked eye were also collected. Pollen was sampled from on-site trenches including different amount of organic matter: the best results were obtained from a sequence of thirty samples. The main pollen zones were in agreement with main cultural changes in the archaeological record. Moreover, the exceptionally well-preserved fruits of Poaceae encouraged analyses on aDNA (done in the Centre for Ancient DNA Research of the University of Copenhagen). The extraction of ancient DNA was carried out on records from three samples of Poaceae fruits accumulation (spot n° 8136, 1423 and A2908). Two of the spot samples are dated to ca. 8000 uncal. years bp; one is dated to ca. 6000-5500 uncal. years bp. Notwithstanding the common doubts about the possibility of DNA preservation in arid and hot environments, the lab study was quite successful, with 5 out of 6 samples extracted and with 3 out of 5 correctly identified. DNA sequences (blind analyses) from Echinochloa, Panicum and Sorghum agree with the identifications made on the basis of morphological analysis. Further elaborations of the sequences obtained allowed to reach a more detailed identification of the genus Panicum, which compared with the sequences currently available in the online database seems to be P. laetum.The integrated archaeobotanical studies on macroremains and pollen, carried out within a multidisciplinary research framework, provided information on the past human-environment relationships and on the Holocene landscape evolution of the area

Ricostruzione ambientale e climatica dei siti archeologici olocenici del Tadrart Acacus (Sahara Centrale, Libia) basata sui reperti botanici

Central Sahara rock shelters and caves offer the possibility to work on a fairly detailed archaeobotanical reconstruction, giving a coherent picture of plant cover during the early and middle Holocene and helping the investigation of past landscape evolution towards the present desert condition. This paper summarises palynological research carried out within a multidisciplinary archaeological research on the Wadi Teshuinat area (Tadrart Acacus Mts., in South-Western Fezzan, Libya). The sites were occupied by hunter-gatherer and pastoralist cultures. On-site pollen data, treated as a single \u2018regional site\u2019, showed that different pollen stratigraphies and flora characterised the different past phases. Plant macro-remains also helped to understand local plant exploitation and landscape evolution. Pollen spectra showed that some climate oscillations had occurred in the Wadi Teshuinat area in the early and middle Holocene: wet and cool (approx. 8800-8250 cal.BC), dry and warm (approx. 7920-7520 cal. BC), wet (approx. 7550-7200 cal.BC), dry (6340-6210 cal.BC - geoarchaeological evidence), wet and warm (approx. 6250-4300 cal.BC, including a wetter and warmer phase at approx. 5500-4600 cal.BC); dry and warm (4250-2900 cal.BC); drier and warm (2900-1600 cal.BC, up to the present). Since the early Holocene, both climatic and anthropic factors have played an important and strictly interconnected role in transforming the environment. Thus, while subsistence strategies were adjusting to climatic and environmental changes, the plant landscape was also being slowly and continuously shaped by humans

Pollen as indicator of food consumption from the archaeological sites of the Wadi Teshuinat area (Tadrart Acacus, Libyan Sahara)

Palynology applied to archaeological sciences puts particular emphasis on the pollen produced by plants handled during human activities. In fact, the palynology of archaeological sites (on-site spectra) helps to recognize anthropogenic pollen, and to investigate the human role in exploiting or changing the plant cover near the site. As pollen is mainly transported into the site by humans, and in general only a minor part arrives by air or water, high percentages or concentrations of pollen are indicative of transport of organic matter, including both collections of plants and faeces containing pollen from eaten plants. This poster presents the list of pollen from archaeological sites, and related to food consumption by humans and their domestic animals. 13 archaeological sites were investigated, all of which are located in the Tadrart Acacus, a mountainous area of the Fezzan, in South-Western Libya, and date to the early and middle Holocene. A relationship between pollen accumulation and food plants is evident from the pollen spectra. Emphasis was made on the anthropogenic pollen indicators and grasses. Macroremains of wild cereals (mainly millets such Brachiaria, Urochloa, Panicum, etc.) are particularly abundant in the deposits, and were used as a parallel tool to better understand plant exploitation in this region. During the Holocene, the sites were occupied by pre-Pastoral (hunter-gatherers) and Pastoral (pastoralists) cultures. Different pollen stratigraphies and floras characterised the diverse sites and the relevant cultural phases. At the early Holocene, hunter-gatherers harvested plants for food and, less commonly, for fodder (mainly Panicum type, Artemisia and Typha); at the middle Holocene, pastoralists occupied the sites, and a large part of the deposits were filled with a few wild cereals and many domestic animal excrements (therefore, pollen of grasses, including wild cereals, and Echium were frequent in the spectra)