Iron deficiency prolongs seed dormancy in Arabidopsis plants

The understanding of seed dormancy, germination and longevity are important goals in plant biology, with relevant applications for agriculture, food industry and also human nutrition. Reactive Oxygen Species (ROS) are key molecules involved in the release of dormancy, when their concentrations fall within the so called 'oxidative window'. The mechanisms of ROS distribution and sensing in seeds, from dormant to germinating ones, still need elucidation. Also, the impact of iron (Fe) deficiency on seed dormancy is still unexplored; this is surprising, given the known pro-oxidant role of Fe when in a free form. We provide evidence of a link between plant Fe nutrition and dormancy of progeny seeds by using different Arabidopsis ecotypes and mutants with different dormancy strengths grown in control soil or under severe Fe deficiency. The latter condition extends the dormancy in several genotypes. The focus on the mechanisms involved in the Fe deficiency-dependent alteration of dormancy and longevity promises to be a key issue in seed (redox) biology

Microtubule depolymerization affects endocytosis and exocytosis in the tip and influences endosome movement in tobacco pollen tubes

Polarized organization of the cytoplasm of growing pollen tubes is maintained by coordinated function of actin filaments (AFs) and microtubules (MTs). AFs convey post-Golgi secretory vesicles to the tip where some fuse with specific domains of the plasma membrane (PM). Secretory activity is balanced by PM retrieval that maintains cell membrane economy and regulates the polarized composition of the PM, by dividing lipids/proteins between the shank and the tip. Although AFs play a key role in PM internalization in the shank, the role of MTs in exoendocytosis needs to be characterized. The present results show that integrity of the MT cytoskeleton is necessary to control exoendocytosis events in the tip. MT polymerization plays a role in promoting PM invagination in the apex of tobacco pollen tubes since Nocodazole affected PM internalization in the tip and subsequent migration of endocytic vesicles from the apex for degradation. MT depolymerization in the apex and shank was associated with misallocation of a significantly greater amount of internalized PM to the Golgi apparatus and its early recycling to the secretory pathway. FRAP experiments also showed that MT depolymerization in the tip region influenced the rate of exocytosis in the central domain of the apical PM

Transitions in morphologies, fluid regimes, and feeding mechanisms during development of the medusa Lychnorhiza lucerna

Author Posting. © The Author(s), 2016. This is the author's version of the work. It is posted here by permission of Inter-Research for personal use, not for redistribution. The definitive version was published in Marine Ecology Progress Series 557 (2016): 145-159, doi:10.3354/meps11855.The early ontogeny of scyphomedusae involves morphological and functional transitions in body plans that affect the predators’ propulsive and feeding strategies. We applied high-speed videography, digital particle image velocimetry (DPIV) and dye visualization techniques to evaluate alterations in swimming and feeding mechanisms during ontogeny of the rhizostome medusa Lychnorhiza lucerna Haeckel, 1880 (Scyphozoa, Rhizostomeae). During early ontogeny, the ephyral mouth lips develop into complex filtering structures along the oral arms. The viscous environments (Reynolds number <100) experienced by ephyrae constrain the feeding mechanisms that transport fluid during ephyral bell pulsations. In contrast, adult medusan fluid flows are dominated by inertial forces and bell pulsations effectively transport fluids and entrained prey toward the oral arms. The oral arm surfaces are covered by motile epidermal cilia that drive these entrained flows through filtering gaps in the oral arms where food particles are retained. In addition to this process within the oral arms, vortices generated during bell pulsation flow downstream along the outside of the medusae and continuously transport prey toward the exterior oral arm surfaces. Although calanoid copepods are capable of escape velocities that greatly exceed L. lucerna’s feeding current speeds, copepods often fail to detect the predator’s feeding currents or inadvertently jump into medusan capture surfaces during failed escape attempts. Consequently, the comparatively weak predator feeding currents successfully capture a portion of the copepods encountered by swimming medusae. These results clarify the processes that enable rhizostome medusae to play key roles as consumers in tropical and subtropical coastal environments.The study was partially funded by grants 2011/00436-8, 2013/19478-8, and 2014/00824-6 São Paulo Research Foundation (FAPESP), and CAPES PROEX2017-09-2

Restricted O2 consumption in pea roots induced by hexanoic acid is linked to depletion of Krebs cycle substrates

Plant roots are exposed to hypoxia in waterlogged soils, and they are further challenged by specific phytotoxins produced by microorganisms in such conditions. One such toxin is hexanoic acid (HxA), which, at toxic levels, causes a strong decline in root O2 consumption. However, the mechanism underlying this process is still unknown. We treated pea (Pisum sativum L.) roots with 20 mM HxA at pH 5.0 and 6.0 for a short time (1 h) and measured leakage of key electrolytes such as metal cations, malate, citrate and nonstructural carbohydrates (NSC). After treatment, mitochondria were isolated to assess their functionality evaluated as electrical potential and O2 consumption rate. HxA treatment resulted in root tissue extrusion of K+, malate, citrate and NSC, but only the leakage of the organic acids and NSC increased at pH 5.0, concomitantly with the inhibition of O2 consumption. The activity of mitochondria isolated from treated roots was almost unaffected, showing just a slight decrease in oxygen consumption after treatment at pH 5.0. Similar results were obtained by treating the pea roots with another organic acid with a short carbon chain, that is, butyric acid. Based on these results, we propose a model in which HxA, in its undissociated form prevalent at acidic pH, stimulates the efflux of citrate, malate and NSC, which would, in turn, cause starvation of mitochondrial respiratory substrates of the Krebs cycle and a consequent decline in O2 consumption. Cation extrusion would be a compensatory mechanism in order to restore plasma membrane potential

Design and Experimental Characterization of a Niti-Based, High-Frequency, Centripetal Peristaltic Actuator

Development and experimental testing of a peristaltic device actuated by a single shape-memory NiTi wire are described. The actuator is designed to radially shrink a compliant silicone pipe, and must work on a sustained basis at an actuation frequency that is higher than those typical of NiTi actuators. Four rigid, aluminum-made circular sectors are sitting along the pipe circumference and provide the required NiTi wire housing. The aluminum assembly acts as geometrical amplifier of the wire contraction and as heat sink required to dissipate the thermal energy of the wire during the cooling phase. We present and discuss the full experimental investigation of the actuator performance, measured in terms of its ability to reduce the pipe diameter, at a sustained frequency of 1.5 Hz. Moreover, we investigate how the diameter contraction is affected by various design parameters as well as actuation frequencies up to 4 Hz. We manage to make the NiTi wire work at 3% in strain, cyclically providing the designed pipe wall displacement. The actuator performance is found to decay approximately linearly with actuation frequencies up to 4 Hz. Also, the interface between the wire and the aluminum parts is found to be essential in defining the functional performance of the actuator

Anti-oxidant potential and gap junction-mediated intercellular communication as early biological markers of mercuric chloride toxicity in MDCK cell line.

In this study, the early nephrotoxic potential of mercuric chloride (HgCl(2)) has been evaluated in vitro, by exposing a renal-derived cell system, the tubular epithelial Madin-Darby canine kidney (MDCK) cell line, to the presence of increasing HgCl(2) concentrations (0.1-100 microM) for different periods of time (from 4 to 72 h). As possible biological markers of the tubular-specific toxicity of HgCl(2) in exposed-MDCK cultures we analysed: (i) critical biochemical parameters related to oxidative stress conditions and (ii) gap-junctional function (GJIC). HgCl(2) cytotoxicity was evaluated by cell-density assay. The biochemical analysis of the pro-oxidant properties of the mercuric ion (Hg(2+)) was performed by evaluating the effect of the metal salt on the antioxidant status of the MDCK cells. The cell glutathione (GSH) content and the activity of glutathione peroxidase (Gpx) and catalase (Cat), two enzymes engaged in the H(2)O(2) degradation, were quantified. HgCl(2) influence on MDCK GJIC was analysed by the microinjection/dye-transfer assay. HgCl(2)-induced morphological changes in MDCK cells were also taken into account. Our results, proving that subcytotoxic (0.1-10 microM) HgCl(2) concentrations affect either the antioxidant defences of MDCK cells or their GJIC, indicate these critical functions as suitable biological targets of early mercury-induced tubular cell injury

Genomic organization and evolutionary conservation of plant D-type cyclins

Plants contain more genes encoding core cell cycle regulators than other organisms but it is unclear whether these represent distinct functions. D-type cyclins (CYCD) play key roles in the G1-to-S-phase transition, and Arabidopsis (Arabidopsis thaliana) contains 10 CYCD genes in seven defined subgroups, six of which are conserved in rice (Oryza sativa). Here, we identify 22 CYCD genes in the poplar (Populus trichocarpa) genome and confirm that these six CYCD subgroups are conserved across higher plants, suggesting subgroup-specific functions. Different subgroups show gene number increases, with CYCD3 having three members in Arabidopsis, six in poplar, and a single representative in rice. All three species contain a single CYCD7 gene. Despite low overall sequence homology, we find remarkable conservation of intron/exon boundaries, because in most CYCD genes of plants and mammals, the first exon ends in the conserved cyclin signature. Only CYCD3 genes contain the complete cyclin box in a single exon, and this structure is conserved across angiosperms, again suggesting an early origin for the subgroup. The single CYCD gene of moss has a gene structure closely related to those of higher plants, sharing an identical exon/intron structure with several higher plant subgroups. However, green algae have CYCD genes structurally unrelated to higher plants. Conservation is also observed in the location of potential cyclin-dependent kinase phosphorylation sites within CYCD proteins. Subgroup structure is supported by conserved regulatory elements, particularly in the eudicot species, including conserved E2F regulatory sites within CYCD3 promoters. Global expression correlation analysis further supports distinct expression patterns for CYCD subgroups

Methods for suspension culture, protoplast extraction, and transformation of high-biomass yielding perennial grass Arundo donax

Arundo donax L. is a promising biofuel feedstock in the Mediterranean region. Despite considerable interest in its genetic improvement, Arundo tissue culture and transformation remains arduous. The authors developed methodologies for cell- and tissue culture and genetic engineering in Arundo. A media screen was conducted, and a suspension culture was established using callus induced from stem axillary bud explants. DBAP medium, containing 9 \ub5M 2,4-D and 4.4 \ub5M BAP, was found to be the most effective medium among those tested for inducing cell suspension cultures, which resulted in a five-fold increase in tissue mass over 14 days. In contrast, CIM medium containing 13 \ub5M 2,4-D, resulted in just a 1.4-fold increase in mass over the same period. Optimized suspension cultures were superior to previously-described solidified medium-based callus culture methods for tissue mass increase. Suspension cultures proved to be very effective for subsequent protoplast isolation. Protoplast electroporation resulted in a 3.3 \ub1 1.5% transformation efficiency. A dual fluorescent reporter gene vector enabled the direct comparison of the CAMV 35S promoter with the switchgrass ubi2 promoter in single cells of Arundo. The switchgrass ubi2 promoter resulted in noticeably higher reporter gene expression compared with that conferred by the 35S promoter in Arundo