Morphological definition of Eurotestudo (Testudinidae,Chelonii): First part

This work details the morphological analysis upon which the creation of the testudinid genus Eurotestudo was based. The study is presented in two parts. The present first part includes an introduction to the subject, and the morphological data with illustrations, which formed the basis for the analysis. The results and lists of cited taxa appear in the second part (Ann. Pal., 92, 4). The analysis rests on a limited number of selected characters, principally those derived from shell osteology, which are preserved in fossils as well as in living specimens. Within the palaearctic genus Testudo s.l., Eurotestudo was separated from two other inclusive genera, Agrionemys and Testudo s.s. In a cladistic analysis, Eurotestudo is distinguished by the autapomorphy of the narrowing vertebrals. Several more external characters (not derivable from fossils and thus omitted from the present cladistic analysis) also support the existence of the genus. Eurotestudo shares homoplastic characters with the two other lineages, the frequency and the evolutionary degree of which vary according to species in the two other genera. Further osteological characters and characters known only from living specimens are added to the osteological study. The characters are examined on a series of taxa chosen among the better known fossil forms from the Oligocene to the Present, Quaternary forms of the Eu. hermanni group included, and representative extant forms. Possible members of the three stems are examined. The autapomorphies of each stem are evidenced, the polarization of characters is tested and the homoplasies are detected. The hypothesis of a common origin for Testudo and Eurotestudo has best support in the present analysis.     

Influx of double labelled glutamine into mistletoes (Viscum album) from the xylem sap of its host (Abies alba)

The flux of glutamine into the mistletoe Viscum album from the xylem sap of a coniferous host was analyzed. For this purpose, a perfusion system was used in which the xylem sap of the host was replaced by an artificial perfusion solution. With this system, flux rates into the mistletoe were determined in feeding experiments with the organic nitrogen source U(13)C/(15)N-Gln. At the end of the experiments the delta values of C and N were significantly depleted in the outflow compared to the percolation solution. Since this depletion was higher for C than for N, a combination of Gln uptake and simultaneous uploading of organic compounds in the host xylem can be assumed. Gln was strongly metabolized during its allocation in the mistletoe. As a consequence, the C skeleton of Gln was equally distributed between leaf and stem tissue, whereas N of Gln preferentially accumulated in the stem. Apparently, the C atoms of the Gln taken up are transported faster in the mistletoe to the sink tissues than the N atoms. It is concluded that C liberated from Gln is transported rapidly to different sink tissues, whereas N in the oversupplied mistletoes is transported slowly to sinks in the leaves.     

Different effects of chilling on respiration in leaves and roots of cucumber (Cucumis sativus)

The effects of chilling on respiration (SHAM-resistant, cytochrome pathway and KCN-resistant, alternative pathway), temperature sensitivity, relative electrolyte conductivity, and degrees of oxidative stress (H(2)O(2) and malonaldehyde (MDA) contents) were separately examined in leaves and roots of cucumber (Cucumis sativus L.). After chilling at 8 degrees C for 4 days, both total respiration and KCN-resistant respiration in roots increased at different measurement temperatures. In contrast, SHAM-resistant respiration remained unchanged. In comparison, chilling significantly decreased the total respiration in leaves and this decrease was mostly due to a decrease in SHAM-resistant respiration. Chilling apparently decreased the sensitivity of KCN-resistant respiration to changes of temperature. The reduction levels of ubiquinone pool (UQr/UQt) increased both in chilled leaves and roots whilst pyruvate content increased only in chilled roots, but not in chilled leaves. Furthermore increases of H(2)O(2) and MDA contents were much greater in leaves than in roots. The same trend was also observed for ion leakage from tissues. Taken together, the results suggested that the higher chilling tolerance of roots was associated with their high total respiration and KCN-resistant respiration.     

Spacing behaviour of juvenile corn mice, Calomys musculinus, at the beginning of the breeding period, in absence of adult males

This research was carried out to examine the hypothesis that the absence of fathers promotes a different spacing behaviour in juveniles Calomys musculinus at the beginning of the breeding period. The study was carried out in four 0.25-ha enclosures (two control and two experimental), in a natural pasture, between November 2003 and February 2004. In this study the fathers were removed from the experimental enclosures after juveniles were born. Home-range size depended on sex of juveniles and treatment (father removal). In control and experimental enclosures, female home-range sizes were always smaller than male home-ranges. Male home-ranges were always larger in experimental enclosures than in control enclosures. Treatment and overlap type (intra- and inter-sexual) were not independent. The overlap proportions of male home-ranges were greatest in experimental enclosures than in control enclosures, in both the overlap types (male/male, males/females). The intra- (females/females) and inter-sexual (females/males) overlap proportions of female home-ranges were independent of treatment. In C. musculinus, at the beginning of the breeding period and in absence of adult males, juvenile males increase their home-range size and therefore the degree of inter- and intra-sexual home-range overlap as a mechanism for enlarging the number of receptive females that they encounter.     

A trade-off between the amount and distance of pollen dispersal triggered by the mixed foraging behaviour of Sephanoides sephaniodes (Trochilidae) on Lapageria rosea (Philesiaceae)

Nectar thieves may increase or decrease pollinator-mediated pollen flow and thus may have positive or negative effects on plant reproductive success. In temperate rainforests of South America, the hummingbird Sephanoides sephaniodes acts as both a pollinator and non-destructive nectar thief on Lapageria rosea. Although pollinators that also act as nectar thieves have the potential to significantly modify plant reproductive success, no previous study has addressed this. To determine how the mixed behaviour of S. sephanoides affects pollen flow, we experimentally exposed some flowers to nectar theft and excluded nectar thieves from other flowers. We then assessed pollen dispersal into the floral neighbourhood. Thieved flowers exported less pollen, but the pollen exported was transferred farther into the neighbourhood. Our findings indicate a trade-off between distance and amount of pollen flow.     

Effect of trypsin inhibitor from Crotalaria pallida seeds on Callosobruchus maculatus (cowpea weevil) and Ceratitis capitata (fruit fly)

A proteinaceous trypsin inhibitor was purified from Crotalaria pallida seeds by ammonium sulfate precipitation, affinity chromatography on immobilized trypsin-Sepharose and TCA precipitation. The trypsin inhibitor, named CpaTI, had M(r) of 32.5 kDa as determined by SDS-PAGE and was composed of two subunits with 27.7 and 5.6 kDa linked by disulfide bridges. CpaTI was stable at 50 degrees C and lost 40% of activity at 100 degrees C. CpaTI was also stable from pH 2 to 12 at 37 degrees C. CpaTI weakly inhibited chymotrypsin and elastase and its inhibition of papain, a cysteine proteinase, were indicative of its bi-functionality. CpaTI inhibited, in different degrees, digestive enzymes from Spodoptera frugiperda, Alabama argillacea, Plodiainterpunctella, Anthonomus grandis and Zabrotes subfasciatus guts. In vitro and in vivo susceptibility of Callosobruchus maculatus and Ceratitis capitata to CpaTI was evaluated. C. maculatus and C. capitata enzymes were strongly susceptible, 74.4+/-15.8% and 100.0+/-7.3%, respectively, to CpaTI. When CpaTI was added to artificial diets and offered to both insect larvae, the results showed that C. maculatus was more susceptible to CpaTI with an LD(50) of 3.0 and ED(50) of 2.17%. C. capitata larvae were more resistant to CpaTI, in disagreement with the in vitro effects. The larvae were more affected at lower concentrations, causing 27% mortality and 44.4% mass decrease. The action was constant at 2-4% (w/w) with 15% mortality and 38% mass decrease.     

Shrubs as a source of spatial heterogeneity—the case of Retama sphaerocarpa in Mediterranean pastures of central Spain

The processes underlying the positive effect exerted by woody species on other plant species can be grouped into a physical effect due to the canopy itself (mainly amelioration of temperature and light extremes), and an edaphic effect (nutrient enrichment). Both groups of effects are present in the savanna-like systems originated by Retama sphaerocarpa in the Iberian Peninsula. The consequences of these shrubs for the spatial heterogeneity induced in the herbaceous community are not well known. The herbaceous community was sampled by means of radial transects around 20 adult shrubs, from the canopy centre to the open areas among the shrubs, both northwards and southwards. Floristic composition, diversity and pasture mean height change both along the inside–outside gradient, and from north to south. As a nutrient enriched environment, the centre of the understorey showed the lowest species richness and evenness, and the highest standing crop. In contrast, the environmentally stressful open areas showed the highest number of species and evenness, and the lowest standing crop. This general effect is only partially due to nutrient availability, and is not homogeneous around the shrub, as open area conditions penetrate into the understorey driving the herbaceous community towards that of open areas. An individual shrub effect is also suggested by our results. Different conditions are thus provided by R. sphaerocarpa understoreys, favouring different subsets of species from the general pool, and therefore defining shrub canopies as sources of spatial heterogeneity in the whole savanna-like system.     

Accumulation of tocopherols and tocotrienols during seed development of grape (Vitis vinifera L. cv. Albert Lavallée)

Tocopherols and tocotrienols are present in mature seeds. Yet, little is known about the physiological role and the metabolism of these compounds during seed development. Here we present data on tocopherol and tocotrienol accumulation during seed development in Vitis vinifera L. cv. Albert Lavallée (Royal). This species was chosen for its ability to synthesize both tocopherols and tocotrienols. It is shown here for the first time that during seed development there are significant differences in localization and accumulation kinetics of tocopherols and tocotrienols. Tocopherols are found homogeneously dispersed throughout all tissues of the seed, in concentrations ranging from 20 to 100 microg tocopherol per g dry weight. Tocopherol levels decrease gradually during seed development. In contrast, tocotrienols are only found in the endosperm of the seeds, accumulating in a sigmoid fashion during the maturation period of seed development. Tocotrienol levels were found to be (54+/-7.4) microg/g dry seed in 90-day-old seeds of V. vinifera L. Furthermore, tocotrienol biosynthesis is demonstrated in these seeds during tocotrienol accumulation and in an endosperm fraction isolated at 75 days after flowering.     

The subfossil occurrence and paleoecological implications of Macrotarsomys petteri (Rodentia: Nesomyidae) in extreme southeastern Madagascar

Remains of a large-bodied species of endemic nesomyid rodent, Macrotarsomys petteri Goodman and Soarimalala, 2005, were identified from subfossil deposits recovered from Andrahomana Cave in extreme southeastern Madagascar. This recently described extant species was previously only known from a single specimen collected at a site about 450 km northwest of Andrahomana and with distinctly different forest habitat than currently found in the vicinity of the cave. Radiocarbon dating of remains of M. petteri from the cave site indicates that it persisted in the region subsequent to human settlement. Previous reports of a large member of Macrotarsomys from other Quaternary sites in southern Madagascar are almost certainly of M. petteri. It is proposed that this species once had a broad distribution across the southern portion of the island during a more mesic period and subsequent aridification of the region has led to its local extirpation across most of its former range. To cite this article: S.M. Goodman et al., C. R. Palevol 5 (2006).     

Possible role of jasmonic acid in the regulation of floral induction, evocation and floral differentiation in Lemna minor L.

Jasmonic acid (JA) is implicated in a wide variety of developmental and physiological processes in plants. Here, we studied the effects of JA and the combination of JA and ethylenediamine-dio-hydroxyphenyl-acetic acid (EDDHA) on flowering in Lemna minor in axenical cultures. JA (0.475-47.5 nmol l(-1)) enhanced floral induction in L. minor under long-day (LD) conditions. Under the same conditions, at a concentration of 237.5 nmol l(-1), JA inhibited floral induction, and at a concentration of 475 nmol l(-1) it prevented floral induction. Under LD conditions with LD preculture, a combination of EDDHA (20,500 nmol l(-1)) and JA (47.5 nmol l(-1)) had a synergistic effect on the promotion of floral induction. Floral induction was enhanced to the greatest extent in experiments with LD precultures. Microscopic examination of microphotographs of histological sections showed that JA and, to an even greater extent, JA+EDDHA at optimal concentrations promote apical floral induction (evocation). Furthermore, JA, and to an even greater extent JA in combination with EDDHA in an optimal concentration, also promote flower differentiation, especially the development of stamens, as is evident from the microphotographs. The experimental results show that JA promotes floral induction in other species of Lemnaceae from various groups according to their photoperiodic response. The results support our hypothesis that, in addition to previously ascribed functions, JA may regulate floral induction, evocation and floral differentiation. Our hypothesis is supported also by the results obtained by quantitative determination of endogenous JA levels in L. minor at three growth stages. The levels of endogenous JA decreased from 389 ng JA g(-1) (fresh weight) of L. minor during the vegetative stage to 217 ng JA g(-1) during the evocation stage, and to 37.5 ng JA g(-1) during the flowering stage, which proves that JA is used for flowering.     

The influence of abiotic stress and phenotypic plasticity on the distribution of invasive Alternanthera philoxeroides along a riparian zone

Relatively few studies have compared invasibility and species invasiveness among microhabitats within communities, synchronously. We surveyed the abundance and performance of non-native Alternanthera philoxeroides (Mart.) Griseb. (alligator weed), its co-occurring native congener, Alternanthera sessilis (L.) DC. (sessile joyweed), and other species in a wetland community along a riparian zone in southeast China to test the hypotheses that: i) degree of invasion differs between different types of microhabitats within the community; and ii) microhabitat types that differ in invasibility also differ in soil resource availability or in sediment characteristics likely to affect resource availability; iii) phenotypic plasticity of A. philoxeroides may play a key role in its adaptation to diverse habitats as can be concluded from its extremely low genetic diversity in China. The study riparian zone comprises different types of microhabitats including wet abandoned field, swamp, marsh dunes and gravel dunes. Consistent with these hypotheses, cover of A. philoxeroides was high in abandoned fields (73 ± 2.9%) and swamps (94 ± 1.3%), which had high soil nutrients and water availability. On the contrary, cover of native A. sessilis was relatively high in marsh dunes and grave dunes, which had coarse gravel surfaces, low soil nutrients and low water availability. A. philoxeroides showed greater morphological plasticity in response to habitat variation. In abiotically harsh habitats, stems had limited growth, and were prostrate with weak adventitious roots at nodes, forming thin, scattered patches. In the two richer habitats, the highly branched plants spread over the water or soil surface, supporting dense stronger leaf-bearing stems which grew vertically. The growth pattern of A. sessilis among microhabitats did not exhibit significant variations. These results suggest that morphological plasticity and microhabitat types with high soil resources may facilitate invasions of A. philoxeroides.     

Cloning and functional expression of an acyl-ACP thioesterase FatB type from Diploknema (Madhuca) butyracea seeds in Escherichia coli

A cDNA of fatty acyl-acyl carrier protein (ACP) thioesterase (Fat) from developing seed of Madhuca butyracea has been cloned. The deduced amino acid sequence of the cDNA corresponding to the mature polypeptide showed 30-40% and 60-75% identity to the reported FatA and FatB class of plant thioesterases, respectively. This gene, MbFatB, is present as a single copy in M. butyracea genome and the MbFatB protein was detected clearly in seed tissues of this plant but not in that of Indian mustard (Brassica juncea). Heterologous expression of the MbFatB gene driven by different promoters in E. coli wild type and fatty acid beta-oxidation mutant (fadD88) strains resulted production of the recombinant protein with various fusion tags either as biologically inactive (insoluble) or functionally active forms. Expression of functionally active recombinant MbFatB in E. coli affected bacterial growth and cell morphology as well as changed the fatty acid profiles of the membrane lipid and the culture supernatant. Alteration of the fatty acid composition was directed predominantly towards palmitate and to a lesser extent myristate and oleate due to acyl chain termination activity of plant thioesterase in bacteria. Thus, this new MbFatB gene isolated from a non-traditional oil-seed tree can be used in future for transgenic development of oil-seed Brassica, a widely cultivated crop that expresses predominantly oleoyl-ACP thioesterase (FatA) in its seed tissue and has high amount of unwanted erucic acid in edible oil in order to alter the fatty acid profile in a desirable way.     

Gene silencing of cinnamyl alcohol dehydrogenase in Pinus radiata callus cultures

Xylem-derived Pinus radiata cell cultures, which can be induced to differentiate tracheary elements (TEs), were transformed with an RNAi construct designed to silence cinnamyl alcohol dehydrogenase (CAD), an enzyme involved in the biosynthesis of monolignols. Quantitative enzymatic CAD measurements revealed reduced CAD activity levels in most transclones generated. TEs from transclones with approximately 20% residual CAD activity did not release elevated levels of vanillin, which was derived from coniferyl-aldehyde through a mild alkali treatment. However, the activation of the phenylpropanoid pathway in transclones with approximately 20% residual CAD activity through the application of non-physiological concentrations of sucrose and l-phenylalanine produced phenotypic changes. The accumulation of metabolites such as dihydroconiferyl-alcohol (DHCA), which also accumulates in the P. taeda CAD mutant cad-n1, was observed. These results indicate that a substantial reduction in CAD activity is necessary for this enzyme to become a rate-limiting step in lignin biosynthesis in conifers such as P. radiata and confirm that transformable P. radiata callus cultures can be useful to investigate the function of xylogenesis-related genes in conifers.     

Biochemical and molecular characterization of flavonoid 7-sulfotransferase from Arabidopsis thaliana

Flavonoid compounds play important roles as flower pigments, stress metabolites formed in response to UV, during pollen germination and for polar auxin transport (Trends Plant Sci. 1 (1996) 377). Flavonoid sulfate esters are common in plants, especially the Asteraceae; however, due to the lack of information regarding the factors that regulate their accumulation, their exact role remains to be elucidated. The biosynthesis of flavonol sulfate esters is catalyzed by a number of position specific flavonol sulfotransferases (STs). An Arabidopsis thaliana database search has allowed us to identify and classify 18 putative ST coding sequences. We report here the cloning and characterization of the AtST3a member of this family that is expressed at early stages of seedling development and in the inflorescence stem and siliques of mature plants. The recombinant AtST3a protein exhibits strict specificity for position 7 of flavonoids. In contrast to previously characterized flavonol 7-ST from Flaveria bidentis that sulfonates only flavonol disulfates, AtST3a was found to accept as substrates a number of flavonols and flavone aglycones, as well as their monosulfate esters. The discovery of a flavonol ST from A. thaliana suggests that flavonol sulfates are more widely distributed than originally believed and this model plant could be used to study their biological significance.     

Developmental changes in antioxidant metabolites, enzymes, and pigments in fruit exocarp of four tomato (Lycopersicon esculentum Mill.) genotypes: β-carotene, high pigment-1, ripening inhibitor, and ‘Rutgers’

In surface cell layers of fleshy fruit, antioxidants must limit photooxidative reactions that generate reactive oxygen species (ROS) in high light. Our objective was to measure changes in the concentrations of antioxidant metabolites and pigments, and the activities of enzymes of the Mehler-peroxidase, ascorbate-glutathione cycle in fruit exocarp tissue under non-stress conditions of the following fruit-specific tomato (Lycopersicon esculentum Mill.=Solanum lycopersicum) mutants and their parent: (1) beta-carotene (B), (2) high pigment (hp-1), (3) ripening inhibitor (rin), and (4) the nearly isogenic wild-type 'Rutgers'. Developmental variables included days after anthesis (DAA) and fruit surface color. The highest total ascorbic acid (AsA) concentration was in the exocarp of immature green fruit of hp-1, being 32% higher than 'Rutgers'. The hp-1 mutant also had the highest chlorophyll and total carotenoid concentrations, comprised mostly of lycopene in red ripe fruit; whereas, beta-carotene comprised 90% of the carotenoids in B. Although enzyme activities varied within genotype, they generally increased with development, then decreased as fruit maturity was reached, being coupled with AsA and glutathione (GSH) concentrations. In all mutants, dark-green (DG) exocarp had more chlorophyll and protein, higher concentrations of reduced AsA and GSH, and usually lower enzyme activities than light-green (LG) exocarp taken from the same fruit.     

Patterning of impoundment impact on chironomid assemblages and their environment with use of the self-organizing map (SOM)

The paper assesses the impact of the Jeziorsko dam reservoir on chironomid assemblages and selected environmental factors in the Warta River, Poland, by means of patterns recognized with the self-organizing map (SOM, Kohonen unsupervised artificial neural network). Over 1988–1996, in four annual cycles, a total of 233 monthly samples were collected in a seven order section of the river at two sites: WAA (backwater) located about 2 km upstream from the Jeziorsko Reservoir, and WAB (tailwater) located about 1.5 km downstream from the reservoir's dam. At each site three habitats were selected: H₁, H₂ and H₃ at WAA, and H₁₁, H₁₂ and H₁₃ at WAB. H₁ and H₁₁ were located in the depositional area close to the banks, H₂ and H₁₂ about 6–7 m towards the mid-river and H₃ and H₁₃ in the mid-river. SOM effectively vertically separated H₁ and H₁₁ (bank habitats) from H₃ and H₁₃ (the mid-river zone of both sites) and H₂ (the transition zone of the upstream site). The H₁₂ samples were scattered all over SOM but still exhibited a slight temporal gradient. At the end of the study the water discharge, especially in summers, stabilized at WAB at a level lower than natural and as a result submerged macrophytes appeared at H₁₂ making the abundance of macroinvertebrates increase very quickly. Moreover, a weaker horizontal grouping of samples by season and by site of collection (upstream or downstream from the reservoir) was observed over SOM: 1) bank upstream habitat H₁, with hydrological regime resembling natural, was separated from the downstream H₁₁, which enlarged and contracted in response to dam operation, 2) deeper habitats were less dependent on water level and this is why they underwent seasonal fluctuations. To sum up, the deepest habitats were most resistant to water level fluctuations, while the formerly most productive habitat at the tailwater WAB site, H₁₁, became the most negatively impacted. Nevertheless, the reservoir has not negatively influenced chironomid density, because the latter increased closer the mid-river, at H₁₂, where large patches of macrophytes developed.     

Two new heterakid (Nematoda) species from Kinixys erosa (Schweigger, 1812), (Chelonii: Testudinidae) in the Democratic Republic of Congo

Two species of Africana Tavassos, 1920 (Africana kinixysae n. sp. and Africana congoensis n. sp.) are described from Kinixys erosa (Schweigger, 1812) in the Democratic Republic of Congo. Light microscopy and scanning electron microscopy studies revealed morphological differences in the structure of the male caudal end and cephalic end, which helped to differentiate these new species from the other species of the genus, and from each other. A key for determination of all species in the genus is provided.