Symplastic communication between the central cell and the egg apparatus cells in the embryo sac of Torenia fournieri Lind. before and during fertilization

 Various membrane-impermeable, water-soluble fluorescent tracers with different molecular weights were microinjected into the central cell of the embryo sac of Torenia fournieri Lind. before and during fertilization. Before anthesis, there was high symplastic permeability between the central cell and the egg apparatus cells. In this stage, fluorescent tracers up to 10 kDa could pass from the central cell into the egg apparatus cells, whereas those with larger molecular weight remained in the central cell. As the embryo sac matured, symplastic permeability decreased such that 2 d after anthesis only tracers less than 3 kDa could spread from the central cell into the egg cell. There appeared to be no symplastic permeability between the primary endosperm and zygote after fertilization, since tracers as small as 521 Da could not pass into the zygote in about half of the microinjected embryo sacs. This is the first report of a change in cell-to-cell communication among the cells of the female germ unit before and after fertilization. Received: 16 December 1999 / Accepted: 4 February 2000     

Questioning the relevance of shifting cultivation to Neolithic farming in the loess belt of Europe: evidence from the Hambach Forest experiment

Despite widespread criticism, the shifting cultivation model continues to inform discussion of Neolithic farming in Europe, beginning with early Neolithic (Linearbandkeramik or LBK) communities concentrated in the loess belt of western-central Europe. Hundreds of LBK and later Neolithic sites have been excavated in this region and many of them sampled for charred plant remains. Archaeobotanical data on the weed floras harvested with crops provide the most direct archaeological evidence of crop husbandry practices, including the permanence of crop fields, but have played a limited role in the debate over shifting cultivation. The Hambach Forest experiment, conducted in the 1970s-80s near Cologne, Germany, provides valuable comparative data on the weed floras growing in newly cleared cultivation plots in an area of longlived mixed oak woodland on loess-based soil. Correspondence analysis of the Hambach weed survey data suggests that weed floras of fields managed under a shifting cultivation regime would be rich in perennial species, including woodland perennials. Comparison of these results with Neolithic weed assemblages from the loess belt of western-central Europe strongly suggests that Neolithic crop fields were not recently cleared of woodland vegetation but were long-established. Received September 5, 2001 / Accepted February 26, 2002     

Archaeobotanical evidence for early agriculture at Ljubjansko barje (Ljubljana Moor), central Slovenia

Archaeobotanical evidence is presented for early agriculture at southwestern Ljubljansko barje (Ljubljana Moor), Slovenia. Archaeobotanical finds from the Eneolithic site at Hočevarica, and pollen records from an archaeological profile and from a nearby core were analysed. Numerous charred grains of cultivated cereals together with fossil seeds of Chenopodium sp. demonstrate that during the occupation of the settlement at Hočevarica, agriculture was well established. The majority of identified grains were of Hordeum vulgare (cultivated barley) and the rest were Triticum monococcum and T. turgidum ssp. dicoccum (cultivated wheats). Large amounts of cereal pollen and pollen of Chenopodiaceae also suggest strong human impact on the surrounding vegetation and landscape. Pollen and archaeobotanical data from Hočevarica show a large consistency in timing of the appearance of agriculture. In the pollen record from the core at Hočevarica a significant increase (up to 40%) in cereal pollen was detected at 4881 ± 50 B.P. (3770-3630 cal B.C.). Charred cereal grains were dated to 4800 ± 40 B.P. (3650-3520 cal B.C.). The grains of cultivated cereals from Hočevarica represent the oldest archaeobotanical evidence for agriculture in central Slovenia. Received February 18, 2002 / Accepted October 21, 2002     

The long history of Cannabis and its cultivation by the Romans in central Italy, shown by pollen records from Lago Albano and Lago di Nemi

The cores from the Albano and Nemi lakes, near Rome, were studied within the European Union funded PALICLAS project and provided high resolution records of the Late-glacial and Holocene. Pollen evidence of increasing human influence on vegetation was recorded in the Holocene parts of both diagrams, and the Cannabis (hemp) curve was one of the major signs. In this paper we present unambiguous pollen evidence from the Cannabaceae records for the cultivation of hemp in central Italy by the Romans. The oldest records of Cannabis and Humulus (hop) date from to the Late-glacial. Hop pollen values rise during the mid Holocene, while hemp pollen becomes more abundant from ca. 3000 cal B.P. onwards. The highest earliest hemp peak (21%) is dated to the 1st century A.D. This ‘Cannabis phase’, with the abrupt rise of hemp pollen soon after the rise of cultivated trees (Castanea, Juglans and Olea) is associated with the increase in cereals and ruderal plants. This unambiguous proof of cultivation by Romans around 2000 B.P. occurs as well as a long lasting pre-Roman presence of hemp in the area, which is natural and possibly also anthropogenic. Subsequent clear episodes of cultivation in the medieval period were found. Received February 4, 2002 / Accepted September 13, 2002 Correspondence to: Anna Maria Mercuri, e-mail: mercuri.annamaria@unimo.it     

Literature on archaeological remains of cultivated plants (1999/2000)

Publications on archaeological remains of cultivated plants have been collected, mainly from 1999, with some earlier and some from 2000. A list is given of the finds according to taxon, country, site, and age. Received November 22, 2000 / Accepted November 27, 2000     

Towards the archaeobotanical identification of intensive cereal cultivation: present-day ecological investigation in the mountains of Asturias, northwest Spain

The region of Asturias, northwest Spain, is highly unusual in that a cereal crop (spelt wheat) is cultivated on a garden scale using horticultural methods. A floristic survey was made of the weeds in 65 spelt plots in this region. The ecological attributes of the weed species were then measured and compared to an earlier study of the functional characteristics of weeds associated with pulse crops on the Greek island of Evvia. In this earlier study, it was possible to distinguish between plots cultivated intensively on a garden-scale and plots cultivated extensively in fields, on the basis of a suite of functional attributes of the weed species. The cereal plots from Asturias were correctly identified as gardens on the basis of the same suite of attributes. The Asturias plots were also compared to weed associations from autumn- and spring-sown crops in Germany, using a different suite of attributes, and were classified either as autumn-sown or ambiguously. This is consistent with the sowing time in Asturias, which is spread over late autumn to winter. These results demonstrate that the suites of functional attributes identified to distinguish intensive and extensive cultivation, and to recognise sowing time, can be applied in another geographical area and to another crop type. This paves the way for the application of these attributes to the identification of past agricultural practices from archaeological weed assemblages. Received September 12, 2001 / Accepted December 12, 2002     

Multi-disciplinary evidence of the Holocene history of a cultivated floodplain area in the wetlands of northern Colombia

An environmental reconstruction of the last 10,000 ¹⁴C years of a frequently flooded wetland ecosystem in the lower Magdalena valley in northern Colombia is presented, on the basis of a multi-disciplinary study of the sediments of the upper 15 m of the core from Boquillas (74°33'E, 9°7'N; 20 m a. s. l.). We used the following studies: pollen, lithology, organic structures, clay mineralogy, soil and sediment geochemistry, and δ¹³C values. The chronology is based on 13 AMS radiocarbon dates; the humic acid fractions were used in the case of seven samples. Pollen from local origin (swamps, open grass-rich vegetation, and gallery forest) show the development of the wetland area. River-transported pollen from a greater distance (dry forest, montane forest, Alnus) show changes in river activity and reflect large-scale changes of climatic conditions in the Momposina basin. From c. 10,010 to 9370 uncal B. P. (zone BQS-Ia) the river system was of high energy, as inferred by the lithological changes. The landscape was dominated by open grass-rich vegetation with gallery forest along the streams. A marked representation of Alnus and montane forest taxa indicate significant water transport and river dynamics. Climatic conditions were dry. From c. 9370-8430 uncal B. P. (zone BQS-Ib) wetlands were isolated from the main river system, and clayey sediments with kaolinite, smectite and illite as the main minerals accumulated in a lower-energy environment. Climatic conditions were dry and changes in the seasonal precipitation favoured the expansion of the gallery forest. From c. 8430 to 8040 uncal B. P. (zone BQS-Ic) low values of river-transported pollen indicate dry climatic conditions and open vegetation became more abundant. The flooding frequency of the Boquillas site diminished. From 8040 to 4900 uncal B. P. (zone BQS-Id) the Boquillas site was dominated by open vegetation with patches of gallery forest along the streams. Supply of river-transported allochthonous pollen (from many sources) was minimal. Clay minerals from the sediments suggest variable temperature and precipitation. From c. 4900 to 1550 uncal B. P. )zone BQS-II) the site was within the reach of the main river system as is the case today. Frequent flooding, coinciding with peaks of river-transported grains of Alnus and high sediment supply, point to high precipitation in the composite catchment area of the Magdalena, Cauca, San Jorge, and Cesár rivers. High values of phosphorous in the upper part of the core point to the presence of a pre-Hispanic civilization, approximately from 2000 uncal B. P. onward. Construction of an extensive drainage system allowed irrigation as well as drainage depending the annual cycle of precipitation. The landscape was significantly modified and allowed an extensive crop production on a system of raised fields. Received May 18, 2001 / Accepted June 15, 2001     

New evidence for the use of root foods in pre-agrarian subsistence recovered from the late Mesolithic site at Halsskov, Denmark

Roots and tubers, together with other plant storage organs such as rhizomes, bulbs, corms, etc. are known to be a source of human food. Until very recently, however, remains of root foods were rarely identified from archaeological contexts in temperate Europe. New evidence for the use of root foods has been recently recovered from the early Erteb?lle settlement at Halsskov in Denmark. Remains included charred bulbs of Allium cf. ursinum and tubers of Conopodium majus. They were accompanied by charred remains of parenchymatous tissues derived from underground parts of other plants. All parenchymatous remains were recovered from features that could be interpreted as pit-cooking depressions. A minimum of two periods of root food gathering can be assumed, spring to early summer and autumn. The presence of charred seeds of Nuphar pumilum suggests that either the seeds or rhizomes (or both) were use as food. Received September 4, 2001 / Accepted February 27, 2002     

Vegetation in the mountains of northwest Iberia during the last glacial-interglacial transition

A new core taken from a site in the Sierra de Courel range of mountains in the northwest Iberian peninsula, Spain, enables a palaeoenvironmental reconstruction to be made of the biological events resulting from climatic changes and human activities during the last 17,000 years in the area. The sequence begins with a phase characterised by the dominance of Gramineae (Poaceae) and Pinus. A markedly arid period with prevalence of Artemisia pollen occurred between 17,5000 and 15,500 uncal. B.P. Subsequently, a succession of woodlands with Betula, Pinus and other mesophilous and thermophilous trees was recognised during the period 15,500-13,500 uncal. B.P. Between 13,500 and 10,300 uncal. B.P. herbaceous vegetation formations indicating moister conditions dominated the pollen record, although a maximum of Gramineae has been dated during the period 11,300-10,300 uncal. B.P. Comparison with other pollen data from neighbouring mountains allows an interpretation of the vegetation changes during the glacial/interglacial transition in the mountains of northwest Iberia. The Holocene pollen record from the site does not differ markedly from other records in the area, tree expansion occurring before 10,000 uncal. B.P. and high values of deciduous Quercus mixed with other trees and shrubs persisting until 3500 uncal. B.P., when increased human activities are shown by a fall of the tree pollen percentages. Received July 7, 1999 / Accepted May 22, 2000     

Purification and structural analysis of an abundant thaumatin-like protein from ripe banana fruit

 The pulp of ripe bananas (Musa acuminata) contains an abundant thaumatin-like protein (TLP). Characterization of the protein and molecular cloning of the corresponding gene from banana demonstrated that the native protein consists of a single polypeptide chain of 200 amino acid residues. Molecular modelling further revealed that the banana thaumatin-like protein (Ban-TLP) adopts an overall fold similar to that of thaumatin and thaumatin-like PR-5 proteins. Although the banana protein exhibits an electrostatically polarized surface, which is believed to be essential for the antifungal properties of TLPs, it is apparently devoid of antifungal activity towards pathogenic fungi. It exhibits a low but detectable in vitro endo-β-1,3-glucanase (EC 3.2.1.x) activity. As well as being present in fruits, Ban-TLP also occurs in root tips where its accumulation is enhanced by methyl jasmonate treatment of plants. Pulp of plantains (Musa acuminata) also contains a very similar TLP, which is even more abundant than its banana homologue. Our results demonstrate for the first time that fruit-specific (abundant) TLPs are not confined to dicots but occur also in fruits of monocot species. The possible role of the apparent widespread accumulation of fruit-specific TLPs is discussed. Received: 7 January 2000 / Accepted: 26 April 2000     

The peptide molecular links between the central nervous and the immune systems

Summary. The central nervous system (CNS) and the immune system were for many years considered as two autonomous systems. Now, the reciprocal connections between them are generally recognized and very well documented. The links are realized mainly by various immuno- and neuropeptides. In the review the influence of the following immunopeptides on CNS is presented: tuftsin, thymulin, thymopoietin and thymopentin, thymosins, and thymic humoral factor. On the other side, the activity in the immune system of such neuropeptides as substance P, neurotensin, some neurokinins, enkephalins, and endorphins is discussed.     

Archaeobotanical analysis of some early Neolithic settlements in the Kujawy region, central Poland, with potential plant gathering activities emphasised

Neolithic settlements in the Kujawy region of central Poland are represented by seven archaeological sites which have botanical material archaeologically dated to the Linear Pottery culture (LBK) (ca. 5400-5000 cal. B.C.) and the Lengyel culture (ca. 4400-4000 cal. B.C.). The composition of plant remains suggests that Stipa pennata s.l. played a certain role in the economy of the Neolithic settlers. The presence of this xerothermic grass is best explained by local gathering rather than distant transport or coming into the sediment by chance. The finding of Hierochlo? cf. australis grains represents the first identification of this plant in archaeobotanical material from Poland. This, now rare, plant contains coumarin and for this reason could have been useful in prehistory. Other plants such as Bromus spp., Chenopodium album type, Fallopia convolvulus and Galium spp. were found in large quantities, and although common weeds now, they could also have been collected by the Neolithic settlers. Plants such as Corylus avellana and Vaccinium vitis-idaea that are typically considered to be collected as foods are present in the studied material but in very small quantities. Received September 17, 2001 / Accepted March 13, 2002     

Continuity and changes in plant resources during the Neolithic period in western Switzerland

The Neolithic and Bronze Age (4400-1570 B.C.) pile dwellings of Concise-sous-Colachoz on the shore of Lake Neuchatel (Canton of Vaud, western Switzerland), known as an archaeological site for more than 140 years, were recently re-investigated due to rescue excavations. Plant macrofossil analyses were done for several Neolithic occupation phases (3868-2440 B.C. the Cortaillod classique, Cortaillod moyen, Cortaillod tardif and Auvernier periods) with a focus on cereal remains, and additionally, archaeological deposits dating to the Cortaillod moyen culture (3710-3677 B.C.) were studied in detail. The preliminary study of cereal macrofossil remains from all the mentioned Neolithic phases show that the most important cereals were Triticum aestivum/durum/turgidum (naked wheat), T. monococcum (einkorn) and Hordeum vulgare (barley). The preferences for specific crops did obviously not alter significantly through time, but, extraordinarily, significant amounts of einkorn continued to be grown for at least 1400 years during the Neolithic period. Other cultivated plants were Pisum sativum (pea), Linum usitatissimum (flax), and Papaver somniferum (opium poppy). Additionally to the seeds, capsule fragments of opium poppy were found in the Cortaillod moyen deposits. These represent the first finds of uncharred capsule fragments in Europe. Compared with other central European sites, opium poppy was very common during the 38th and 37th cent. B.C. and obviously less appreciated towards the end of the Neolithic in the western part of Switzerland. In central Switzerland the trends seem different: there opium poppy was mainly used during the Late Neolithic period. This may be due to cultural differences within contemporaneous human societies. Wild fruits which were collected as plant resources during the Cortaillod moyen period included Prunus spinosa (sloe), Cornus sanguinea (dogwood), Malus sylvestris (apple), Rubus idaeus/caesius/fruticosus (raspberry/dewberry/blackberry), Fragaria vesca (wild strawberry), Rosa sp. (hip), Quercus sp. (acorn), Corylus avellana (hazelnut), and Fagus sylvatica (beechnut), among others. Compared with other Neolithic sites in westem and central Switzerland the local population of Concise-sous-Colachoz used few sloes, while dogwood fruits were in use throughout the Neolithic period at Lake Neuchatel. Received September 4, 2001 / Accepted May 13, 2002     

Blood amino acids concentration during insulin induced hypoglycemia in rats: the role of alanine and glutamine in glucose recovery

Summary. Our purpose was to determine the blood amino acid concentration during insulin induced hypoglycemia (IIH) and examine if the administration of alanine or glutamine could help glycemia recovery in fasted rats. IIH was obtained by an intraperitoneal injection of regular insulin (1.0 U/kg). The blood levels of the majority of amino acids, including alanine and glutamine were decreased (P < 0.05) during IIH and this change correlates well with the duration than the intensity of hypoglycemia. On the other hand, the oral and intraperitoneal administration of alanine (100 mg/kg) or glutamine (100 mg/kg) accelerates glucose recovery. This effect was partly at least consequence of the increased capacity of the livers from IIH group to produce glucose from alanine and glutamine. It was concluded that the blood amino acids availability during IIH, particularly alanine and glutamine, play a pivotal role in recovery from hypoglycemia.     

Mangroves of Oman during the late Holocene; climatic implications and impact on human settlements

Pollen and micropalaeontological analyses carried out on mangrove swamp sediments of Suwayh, Oman (22°05.589'N, 50°40.033'E) reveal environmental changes linked both to climate (monsoon) and geomorphological (sea-level) variations during the Late Holocene. A Rhizophora mangrove developed at Suwayh around 6000 years B. P. under climate conditions marked by an increased tropical influence as compared to the modern situation, with dominant summer rains. The later extension of Prosopis cineraria at Suwayh provides evidence for a different rainfall pattern, with a winter rainy season. Pollen and micropalaeontological composition shows three episodes influenced by the sea water at Suwayh. The most important corresponded to the mangrove episode centred ca 6000 B. P. This was followed by two episodes of slight seawater incursion at ca. 5100 and 4500 B. P. responsible for the formation of a brackish laagon. Comparison based on ¹⁴C measurements on shell recovered from both the sedimentary sequence of Suwayh and the nearby archaeological sites demonstrates that close relations existed between man and mangroves during the Neolithic and the Early Bronze Age. Received October 31, 2001 / Accepted January 24, 2002 Correspondence to: Anne-Marie Lézine     

Observations on the relationship between the extracellular changes of taurine and glutamate during cortical spreading depression, during ischemia, and within the area surrounding a thrombotic infarct

Summary. Taurine and glutamate were monitored by microdialysis technique during various cerebral insults: a. Application of K⁺ triggered a cortical spreading depression (CSD). Taurine and glutamate increased concomitantly but recovery of glutamate was faster than that of taurine. b. Application of NMDA induced also CSD but only taurine increased. c. Induction of an infarct triggered repetitive CSDs. Taurine increased rapidly whereas glutamate rose slowly starting with some delay. d. After induction of ischemia, taurine and glutamate increased after onset of depolarisation. The increase of glutamate occurred late after a small, transient increase in parallel with the depolarisation. These data suggest a close functional relationship between the changes of both amino acids. Therefore, they should be monitored together especially in clinical settings: during excitation, only taurine will increase; during overexcitation, taurine will also increase but to a higher maximum followed by a moderate rise of glutamate; after energy failure, taurine will accumulate to its highest level followed by a continuous rise of glutamate. Received January 25, 2000/Accepted January 31, 2000     

Conversion of guaiacyl to syringyl moieties on the cinnamyl alcohol pathway during the biosynthesis of lignin in angiosperms

Aglycons derived from 4-O-β-D-glucosides of both caffeyl and 5-hydroxyconiferyl alcohols were incorporated into guaiacyl (G) and syringyl (S) units in the lignin of newly formed xylem of several angiosperms. It is likely that these aglycons enter the cinnamyl alcohol pathway as intermediates in the introduction of methoxyl groups onto aromatic rings, and serve as precursors for the biosynthesis of lignin. The S/G ratio in this pathway was coincident with the ratio in the cell wall lignin of each tree. Our results indicate that the cinnamyl alcohol pathway involves the same mechanisms as the cinnamic acid and cinnamyl CoA pathways and they suggest that this novel pathway might be part of a metabolic grid in the biosynthesis of lignin. Received: 8 September 1999 / Accepted: 4 October 1999     

Effects of glucocorticoids on polyamine metabolism in liver and spleen of guinea pig during sensitization

Summary. Glucocorticoids are potent anti-inflammatory and immunosuppressive agents. As endogenous inhibitors of cytokine synthesis, glucocorticoids suppress immune activation and uncontrolled overproduction of cytokines, preventing tissue injury. Also, polyamine spermine is endogenous inhibitor of cytokine production (inhibiting IL-1, IL-6 and TNF synthesis). The idea of our work was to examine dexamethasone effects on the metabolism of polyamines, spermine, spermidine and putrescine and polyamine oxidase activity in liver and spleen during sensitization of guinea pigs. Sensitization was done by application of bovine serum albumin with addition of complete Freund’s adjuvant. Our results indicate that polyamine amounts and polyamine oxidase activity increase during immunogenesis in liver and spleen. Dexamethasone application to sensitized and unsensitized guinea pigs causes depletion of polyamines in liver and spleen. Dexamethasone decreases polyamine oxidase activity in liver and spleen of sensitized guinea pigs, increasing at the same time PAO activity in tissues of unsensitized animals.     

Genetic study of glutathione accumulation during cold hardening in wheat

The effect of cold hardening on the accumulation of glutathione (GSH) and its precursors was studied in the shoots and roots of wheat (Triticum aestivum L.) cv. Cheyenne (Ch, frost-tolerant) and cv. Chinese Spring (CS, moderately frost-sensitive), in a T. spelta L. accession (Tsp, frost-sensitive) and in chro- mosome substitution lines CS (Ch 5A) and CS (Tsp 5A). The fast induction of total glutathione accumulation was detected during the first 3 d of hardening in the shoots, especially in the frost-tolerant Ch and CS (Ch 5A). This observation was corroborated by the study of de novo GSH synthesis using [³⁵S]sulfate. In Ch and CS (Ch 5A) the total cysteine, γ-glutamylcysteine (precursors of GSH), hydroxymethylglutathione and GSH contents were greater during the 51-d treatment than in the sensitive genotypes. After 35 d hardening, when the maximum frost tolerance was observed, greater ratios of reduced to oxidised hydroxymethylglutathione and glutathione were detected in Ch and CS (Ch 5A) compared to the sensitive genotypes. A correspondingly greater glutathione reductase (EC 1.6.4.2) activity was also found in Ch and CS (Ch 5A). It can be assumed that chromosome 5A of wheat has an influence on GSH accumulation and on the ratio of reduced to oxidised glutathione as part of a complex regulatory function during hardening. Consequently, GSH may contribute to the enhancement of frost tolerance in wheat. Received: 24 March 1999 / Accepted: 19 July 1999     

DNA damage during glycation of lysine by methylglyoxal: assessment of vitamins in preventing damage

Summary. Amino acids react with methylglyoxal to form advanced glycation end products. This reaction is known to produce free radicals. In this study, cleavage to plasmid DNA was induced by the glycation of lysine with methylglyoxal in the presence of iron(III). This system was found to produce superoxide as well as hydroxyl radicals. The abilities of various vitamins to prevent damage to plasmid DNA were evaluated. Pyridoxal-5-phosphate showed maximum protection, while pyridoxamine showed no protection. The protective abilities could be directly correlated to inhibition of production of hydroxyl and superoxide radicals. Pyridoxal-5-phosphate exhibited low radical scavenging ability as evaluated by its TEAC, but showed maximum protection probably by interfering in free radical production. Pyridoxamine did not inhibit free radical production. Thiamine and thiamine pyrophosphate, both showed protective effects albeit to different extents. Tetrahydrofolic acid showed better antioxidant activity than folic acid but was found to damage DNA by itself probably by superoxide generation.