An automatic device to record the force necessary to compensate plant movements

A device, which automatically and continuously keeps moving plants in fixed positions, has been constructed. The apparatus is based on optical detection of any movements of the plant organ under study. The plant organ is kept in the desired position by means of wires, and the force necessary to apply to the wires to achieve this is recorded. The force reflects any tendency of the plant organ to move. The system is controlled by an Apple II computer. The device can work in one or two dimensions and record compensation forces in a wide range. In the experiments mentioned below, the magnitude of the forces necessary to keep the plants in a fixed position was of the order of 10^-4N.
Circadian leaf movements of Oxalis regnellii Mig. were studied in the device. The leaf rhythm continued, although the leaflets were clamped and the light input on the leaf therefore constant. Circumnutation of hypocotyls of Helianthus annus L. cv. Californicus were drastically reduced in amplitude when the hypocotyls were kept in vertical position by the wires. Since the gravitropic input signal to this system was zeroed by the equipment, the results demonstrated that in the absence of gravitropic inputs the circumnutation reactions drastically diminish. This confirmed that circumnutations of these hypocotyls are influenced by gravity. Finally, the apparatus was used to study phototropic reactions: By clamping phototropically stimulated coleoptiles of Avena sativa L. cv. Seger no gravitational stimulations were involved and the phototropic reaction without interference from gravity could be studied.     

Differences in chemical composition of plants grown at constant relative growth rates with stable mineral nutrition

Summary Leaf chemistry of a willow clone (Salix aquatica Smith) differed significantly when grown at constant relative growth rates depending upon the relative availability of nutrients and light. Concentration of amino acids and nitrate were high in plants grown with a relative surplus of nutrients. Concentrations of starch, tannin, and lignin, on the other hand, were high in plants grown with a relative surplus of carbon. Photosynthetic rates, expressed per unit leaf area, were similar when plants were grown under high light conditions, regardless of nutrient availability. Dark respiration was much higher in plants supplied with abundant nutrients than in those with a more limited supply, reflecting differences in nitrogen concentration of the tissue. The experimental approach allows plants to be grown to a standard size with differing, but highly uniform chemistry. Plants grown in such a manner may provide good experimental material to evaluate interactions between herbivores or pathogens and their hosts.     

Low degree of ubiquitination of histone 2A in the dipteran Chironomus tentans

A polyclonal antibody to ubiquitin has been prepared and shown to react with both ubiquitin and ubiquitinated histone 2A (uH2A). Applying this antibody in Western blotting experiments, we have observed that the salivary glands of Chironomus tentans contain an unusually low amount of uH2A (1% of histone 2A), while the amount of free ubiquitin is as abundant as in other animal cells, e.g. HeLa cells. The same low content of uH2A was also found in diploid epidermal cells of Chironomus origin suggesting that the low amount is not a characteristic of the polytene state of chromatin in salivary gland cells but rather a property of C. tentans as a species. The significance of the low degree of ubiquitination is discussed in relation to the information available on the organization of Chironomus chromatin into unusually large chromomeric entities.     

Analysis of Replication Intermediates Indicates That Drosophila melanogaster Mitochondrial DNA Replicates by a Strand-Coupled Theta Mechanism

Mitochondrial DNA synthesis is necessary for the normal function of the organelle and for the eukaryotic organism as a whole. Here we demonstrate, using two-dimensional agarose gel electrophoresis to analyse replication intermediates, that unidirectional, strand-coupled DNA synthesis is the prevalent mode of mtDNA replication in Drosophila melanogaster. Commencing within the single, extended non-coding region (NCR), replication proceeds around the circular genome, manifesting an irregular rate of elongation, and pausing frequently in specific regions. Evidence for a limited contribution of strand-asynchronous DNA synthesis was found in a subset of mtDNA molecules, but confined to the ribosomal RNA gene region, just downstream of the NCR. Our findings imply that strand-coupled replication is widespread amongst metazoans, and should inform future research on mtDNA metabolism in D. melanogaster.     

Structural and functional insights into the modulation of the activity of a flax cytokinin oxidase by flax rust effector AvrL567-A

During infection, plant pathogens secrete effector proteins to facilitate colonization. In comparison with our knowledge of bacterial effectors, the current understanding of how fungal effectors function is limited. In this study, we show that the effector AvrL567-A from the flax rust fungus Melampsora lini interacts with a flax cytosolic cytokinin oxidase, LuCKX1.1, using both yeast two-hybrid and in planta bimolecular fluorescence assays. Purified LuCKX1.1 protein shows catalytic activity against both N6-(Δ2-isopentenyl)-adenine (2iP) and trans-zeatin (tZ) substrates. Incubation of LuCKX1.1 with AvrL567-A results in increased catalytic activity against both substrates. The crystal structure of LuCKX1.1 and docking studies with AvrL567-A indicate that the AvrL567 binding site involves a flexible surface-exposed region that surrounds the cytokinin substrate access site, which may explain its effect in modulating LuCKX1.1 activity. Expression of AvrL567-A in transgenic flax plants gave rise to an epinastic leaf phenotype consistent with hormonal effects, although no difference in overall cytokinin levels was observed. We propose that, during infection, plant pathogens may differentially modify the levels of extracellular and intracellular cytokinins.     

Making teeth to order: conserved genes reveal an ancient molecular pattern in paddlefish (Actinopterygii)

Ray-finned fishes (Actinopterygii) are the dominant vertebrate group today (+30 000 species, predominantly teleosts), with great morphological diversity, including their dentitions. How dental morphological variation evolved is best addressed by considering a range of taxa across actinopterygian phylogeny; here we examine the dentition of Polyodon spathula (American paddlefish), assigned to the basal group Acipenseriformes. Although teeth are present and functional in young individuals of Polyodon, they are completely absent in adults. Our current understanding of developmental genes operating in the dentition is primarily restricted to teleosts; we show that shh and bmp4, as highly conserved epithelial and mesenchymal genes for gnathostome tooth development, are similarly expressed at Polyodon tooth loci, thus extending this conserved developmental pattern within the Actinopterygii. These genes map spatio-temporal tooth initiation in Polyodon larvae and provide new data in both oral and pharyngeal tooth sites. Variation in cellular intensity of shh maps timing of tooth morphogenesis, revealing a second odontogenic wave as alternate sites within tooth rows, a dental pattern also present in more derived actinopterygians. Developmental timing for each tooth field in Polyodon follows a gradient, from rostral to caudal and ventral to dorsal, repeated during subsequent loss of teeth. The transitory Polyodon dentition is modified by cessation of tooth addition and loss. As such, Polyodon represents a basal actinopterygian model for the evolution of developmental novelty: initial conservation, followed by tooth loss, accommodating the adult trophic modification to filter-feeding.