Identification of Arabidopsis rat mutants

Limited knowledge currently exists regarding the roles of plant genes and proteins in the Agrobacterium tumefaciens-mediated transformation process. To understand the host contribution to transformation, we carried out root-based transformation assays to identify Arabidopsis mutants that are resistant to Agrobacterium transformation (rat mutants). To date, we have identified 126 rat mutants by screening libraries of T-DNA insertion mutants and by using various “reverse genetic” approaches. These mutants disrupt expression of genes of numerous categories, including chromatin structural and remodeling genes, and genes encoding proteins implicated in nuclear targeting, cell wall structure and metabolism, cytoskeleton structure and function, and signal transduction. Here, we present an update on the identification and characterization of these rat mutants.     

Reproductive measures and chick provisioning of Cory's Shearwater Calonectris diomedea borealis in the Azores

Capsule Timing of breeding influenced wing-length at fledging, and egg size may be an indicator of fledging weight and the amount of food received by chicks.

Aims To investigate chick growth, temporal patterns of chick food provisioning and the importance of indices of parental condition or quality, egg size and hatching date, to predict nestling body mass and wing-length at fledging, and compare breeding and chick feeding characteristics between colonies in the northeast Atlantic.

Methods A survey of Cory's Shearwater nests was carried out at Vila islet. A sample of 52 chicks, ringed and weighed at hatching, was selected to study chick growth and food provisioning.

Results Hatching success (51%) was much lower than fledging success (87%). Both hatching date and egg size contributed to explain wing-length at fledging, but hatching date, which was negatively correlated with wing-length at fledging, had the most important contribution (22%). There was some indication that egg size may explain variation in fledging weight and the amount of food received by chicks. Food delivery and feeding frequency of chicks varied throughout the chick development stage and three phases were distinguished: (1) 0–29 days, the highest feeding frequency values and a linear increase in food delivery; (2) 30–69 days, an oscillation in food delivery and medium feeding frequencies; (3) 70–90+ days, a sharp decrease in both food delivery and feeding frequency.

Conclusion Variation in food availability did not seem sufficient to override the overall importance of indices of parental quality in determining reproductive measures and chick provisioning. Breeding and feeding characteristics were similar between colonies in the northeast Atlantic, with variability in chick provisioning higher further south.     

Phytoplankton and chemico-physical parameters of the Scardovari Lagoon (Po Delta,North Adriatic Sea) during 1991 and 1992

Abstract

During research directed towards the employment of the biological resources of the North Adriatic lagoons, from January 1991 to December 1992, in both tidal phases, a survey was carried out on the phytoplankton and the chemico-physical parameters of the Scardovari lagoon. Data analyses allowed two different areas to be distinguished: one inner, which was generally characterized by high phytoplanktonic densities (St. 3–4), the other outer, which was more influced by marine load (St. 1–2). Photosynthetic picoplankton was dominant in terms of cell number in most samples. Phytoplanktonic fraction >2 μm was represented mainly by diatoms, which showed the higher species number. During the sample observation, some dinoflagellates, considered toxic or potentially toxic, were found. Their presence was important because of their effect on the bivalve mollusc cultures.     

Yolk hydrolases in the eggs of Anticarsia gemmatalis hubner (Lepidoptera: Noctuidae): A role for inorganic polyphosphate towards yolk mobilization

Despite being the main insect pest on soybean crops in the Americas, very few studies have approached the general biology of the lepidopteran Anticarsia gemmatalis and there is a paucity of studies with embryo formation and yolk mobilization in this species. In the present work, we identified an acid phosphatase activity in the eggs of A. gemmatalis (agAP) that we further characterized by means of biochemistry and cell biology experiments. By testing several candidate substrates, this enzyme proved chiefly active with phosphotyrosine; in vitro assays suggested a link between agAP activity and dephosphorylation of egg yolk phosphotyrosine. We also detected strong activity with endogenous and exogenous short chain polyphosphates (PolyP), which are polymers of phosphate residues involved in a number of physiological processes. Both agAP activity and PolyP were shown to initially concentrate in small vesicles clearly distinct from typically larger yolk granules, suggesting subcellular compartmentalization. As PolyP has been implicated in inhibition of yolk proteases, we performed in vitro enzymatic assays with a cysteine protease to test whether it would be inhibited by PolyP. This cysteine protease is prominent in Anticarsia egg homogenates. Accordingly, short chain PolyP was a potent inhibitor of cysteine protease. We thereby suggest that PolyP hydrolysis by agAP is a triggering mechanism of yolk mobilization in A. gemmatalis.     

Expression of DNA methyltransferases is involved in Quercus suber cork quality

Cork oak (Quercus suber) is an important Portuguese species, mainly due to the economic value of the cork it produces. Cork results from phellogen, a meristematic tissue, which can locally produce lenticels or have discontinuities, originating “defects”: pores and nail inclusions that are detrimental to cork industrial use. Epigenetic processes control plant development and its deregulation can lead to altered phenotypes; therefore, the study of epigenetic players in the phellogen is important to understand the emergence of cork's defects. DNA methyltransferases (DNMTs) and one protein associated to MET1 (DMAP1) were characterized in Q. suber, and their gene expression was analyzed in phellogen and contiguous differentiating cell layers of trees producing high and low quality cork, after the evaluation of their defects by physical and image analysis methods. All classes of DNMTs (MET, DRM, and CMT) with the respective canonical motifs were identified in Q. suber. The expression analyses of these genes showed that QsDRM2 was the most active methyltransferases in the cells analyzed, and that all the genes were differentially expressed in trees with distinct cork quality, with a tendency for higher expression levels in low quality producers. Interestingly, the global methylation level was higher in cells with low expression of DNA methyltransferases. A positive and significant correlation was obtained between QsDMAP1 gene expression and the percentage of cork defects. This work provides the first evidence that cork quality in Q. suber is likely influenced by epigenetic mechanisms.     

Simulation of physiological loading in total hip replacements

The determination of biomechanical force systems of implanted femurs to obtain adequate strain measurements has been neglected in many published studies. Due to geometric alterations induced by surgery and those inherent to the design of the prosthesis, the loading system changes because the lever arms are modified. This paper discusses the determination of adequate loading of the implanted femur based on the intact femur-loading configuration. Four reconstructions with Lubinus SPII, Charnley Roundback, Muller Straight and Stanmore prostheses were used in the study. Pseudophysiologic and nonphysiologic implanted system forces were generated and assessed with finite element analysis. Using an equilibrium system of forces composed by the Fx (medially direction) component of the hip contact force and the bending moments Mx (median plane) and My (coronal plane) allowed adequate, pseudo-physiological loading of the implanted femur. We suggest that at least the bending moment at the coronal plane must be restored in the implanted femur-loading configuration.     

Comparison of Penicillium echinulatum and Trichoderma reesei cellulases in relation to their activity against various cellulosic substrates

Penicillium echinulatum has been identified as a potential cellulase producer for bioconversion processes but its cellulase system has never been investigated in detail. In this work, the volumetric activities of P. echinulatum cellulases were determined against filter paper (0.27 U/mL), carboxymethylcellulose (1.53 U/mL), hydroxyethylcellulose (4.68 U/mL), birchwood xylan (3.16 U/mL), oat spelt xylan (3.29 U/mL), Sigmacell type 50 (0.10 U/mL), cellobiose (0.19 U/mL), and p-nitrophenyl-glucopiranoside (0.31 U/mL). These values were then expressed in relation to the amount of protein and compared those of Trichoderma reesei cellulases (Celluclast 1.5L FG, Novozymes). Both enzyme complexes were shown to have similar total cellulase and xylanase activities. Analysis of substrate hydrolysates demonstrated that P. echinulatum enzymes have higher beta-glucosidase activity than Celluclast 1.5L FG, while the latter appears to have greater cellobiohydrolase activity. Unlike Celluclast 1.5L FG, P. echinulatum cellulases had enough beta-glucosidase activity to remove most of the cellobiose produced in hydrolysis experiments. However, Celluclast 1.5L FG became more powerful than P. echinulatum cellulases when supplemented with exogenous beta-glucosidase activity (Novozym 188). Both cellulase complexes displayed the same influence over the degree of polymerization of cellulose, revealing that hydrolyzes were carried out under the typical endo-exo synergism of fungal enzymes.     

Pollination in Brazilian Syngonanthus (Eriocaulaceae) species: evidence for entomophily instead of anemophily

BACKGROUND AND AIMS: The reproductive biology of Syngonanthus mucugensis and S. curralensis (Eriocaulaceae) was studied in areas of 'campo rupestre' vegetation in the Chapada Diamantina, north-eastern Brazil. These species are herbaceous and the individuals have a grouped distribution. Their leaves are united in a rosette, and their inflorescence is monoecious, of the capitulum type. The staminate and pistillate rings mature in a centripetal manner on the capitulum. METHODS: A field study was conducted, including observations concerning the morphology and biology of the flowers, fruit development, insect visits and anemophily, in both S. mucugensis and S. curralensis. Experimental pollinations were also carried out to study the mating systems of S. mucugensis. KEY RESULTS: Both species flower from June to August. The staminate cycle lasts approx. 7 d, and the pistillate cycle from 3 to 4 d, with no temporal overlap between them on the same capitulum. The pollen viability of S. mucugensis was 88.6%, and 92.5% for S. curralensis. The inflorescences of both species demonstrated ultraviolet absorbance, and a sweet odour was detected during both the staminate and pistillate phases. No nectar production was ever noted, although nectaries are present. Both species were visited by numerous groups of insects, with the Diptera being the principal pollinators, especially the species of Syrphidae and Bombyliidae. There were secondary pollinators among species of Coleoptera and Hymenoptera. There was no evidence of wind pollination. Syngonanthus mucugensis is a self-compatible species, and forms fruits by agamospermy at low frequencies. CONCLUSIONS: This is apparently the first report for pollination biology and mating systems of Eriocaulaceae. Conversely to that stated by some authors, entomophily, mainly effected by species of Diptera but also by species of Coleoptera and Hymenoptera, is probably the only pollination system in these species. In spite of the monoecious inflorescences without overlap of the staminate and pistillate phases, geitonogamy may occur in S. mucugensis, as the species is self-compatible and different capitula in the same plant at different phases is common.