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The GRAS proteins belong to a plant transcriptional regulator family that function in the regulation of plant growth and development. Despite their important roles, in sunflower only one GRAS gene ( HaDella1) with the DELLA domain has been reported. Here, we provide a functional characterisation of a GRAS-like gene from Helianthus annuus ( Ha- GRASL) lacking the DELLA motif. The Ha- GRASL gene contains an intronless open reading frame of 1,743 bp encoding 580 amino acids. Conserved motifs in the GRAS domain are detected, including VHIID, PFYRE, SAW and two LHR motifs. Within the VHII motif, the P-H-N-D-Q-L residues are entirely maintained. Phylogenetic analysis reveals that Ha- GRASL belongs to the SCARECROW LIKE4/7 ( SCL4/7) subfamily of the GRAS consensus tree. Accumulation of Ha- GRASL mRNA at the adaxial boundaries from P6/P7 leaf primordia suggests a role of Ha- GRASL in the initiation of median and basal axillary meristems ( AMs) of sunflower. When Ha- GRASL is over-expressed in Arabidopsis wild-type plants, the number of lateral bolts increases differently from untransformed plants. However, Ha- GRASL slightly affects the lateral suppressor ( las-4-) mutation. Therefore, we hypothesise that Ha- GRASL and LAS are not functionally equivalent. The over-expression of Ha- GRASL reduces metabolic flow of gibberellins ( GAs) in Arabidopsis and this modification could be relevant in AM development. Phylogenetic analysis includes LAS and SCL4/7 in the same major clade, suggesting a more recent separation of these genes with respect to other GRAS members. We propose that some features of their ancestor, as well as AM initiation and outgrowth, are partially retained in both LAS and SCL4/7. [ABSTRACT FROM AUTHOR]

Rice ( Oryza sativa L.) can successfully germinate and grow even when flooded. Rice varieties possessing the submergence 1A ( Sub1A) gene display a distinct flooding-tolerant phenotype, associated with lower carbohydrate consumption and restriction of the fast-elongation phenotype typical of flooding-intolerant rice varieties. Calcineurin B-like interacting protein kinase 15 (CIPK15) was recently indicated as a key regulator of α-amylases under oxygen deprivation, linked to both rice germination and flooding tolerance in adult plants. It is still unknown whether the Sub1A- and CIPK15-mediated pathways act as complementary processes for rice survival under O deprivation. In adult plants Sub1A and CIPK15 may perhaps play an antagonistic role in terms of carbohydrate consumption, with Sub1A acting as a starch degradation repressor and CIPK15 as an activator. In this study, we analysed sugar metabolism in the stem of rice plants under water submergence by selecting cultivars with different traits associated with flooding survival. The relation between the Sub1A and the CIPK15 pathways was investigated. The results show that under O deprivation, the CIPK15 pathway is repressed in the tolerant, Sub1A-containing, FR13A variety. CIPK15 is likely to play a role in the up-regulation of Ramy3D in flooding-intolerant rice varieties that display fast elongation under flooding and that do not possess Sub1A. [ABSTRACT FROM AUTHOR]

Plant survival is greatly impaired when oxygen levels are limiting, such as during flooding or when anatomical constraints limit oxygen diffusion. Oxygen sensing in Arabidopsis thaliana is mediated by Ethylene Responsive Factor (ERF)-VII transcription factors, which control a core set of hypoxia- and anoxia-responsive genes responsible for metabolic acclimation to low-oxygen conditions. Anoxic conditions also induce genes related to reactive oxygen species (ROS). Whether the oxygen-sensing machinery coordinates ROS production under anoxia has remained unclear. Here we show that a low-oxygen-responsive universal stress protein (USP), Hypoxia Responsive Universal Stress Protein 1 (HRU1), is induced by RAP2.12 (Related to Apetala 2.12), an ERF-VII protein, and modulates ROS production in Arabidopsis. We found that HRU1 is strongly induced by submergence, but that this induction is abolished in plants lacking RAP2.12. Mutation of HRU1 through transfer DNA (T-DNA) insertion alters hydrogen peroxide production, and reduces tolerance to submergence and anoxia. Yeast two-hybrid and bimolecular fluorescence complementation (BiFC) analyses reveal that HRU1 interacts with proteins that induce ROS production, the GTPase ROP2 and the NADPH oxidase RbohD, pointing to the existence of a low-oxygen-specific mechanism for the modulation of ROS levels. We propose that HRU1 coordinates oxygen sensing with ROS signalling under anoxic conditions.

Plant adaptation to hypoxic conditions is mediated by the transcriptional activation of genes involved in the metabolic reprogramming of plant cells to cope with reduced oxygen availability. Recent studies indicated that members of the group VII of the Ethylene Responsive Transcription Factor (ERFs) family act as positive regulators of this molecular response. In the current study, the five ERF-VII transcription factors of Arabidopsis thaliana were compared to infer a hierarchy in their role with respect to the anaerobic response. When the activity of each transcription factor was tested on a set of hypoxia-responsive promoters, RAP2.2, RAP2.3 and RAP2.12 appeared to be the most powerful activators. RAP2.12 was further dissected in transactivation assays in Arabidopsis protoplasts to identify responsible regions for transcriptional activation. An ultimate C-terminal motif was identified as sufficient to drive gene transcription. Finally, using realtime RT-PCR in single and double mutants for the corresponding genes, we confirmed that RAP2.2 and RAP2.12 exert major control upon the anaerobic response., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

Plants often experience low oxygen conditions as the consequence of reduced oxygen availability in their environment or due to a high activity of respiratory metabolism. Recently, an oxygen sensing pathway was described in Arabidopsis thaliana which involves the migration of an ERF transcription factor (RAP2.12) from the plasma membrane to the nucleus upon hypoxia. Moreover, RAP2.12 protein level is controlled through an oxygen-dependent branch of the N-end rule pathway for proteasomal degradation. Inside the nucleus, RAP2.12 induces the expression of genes involved in the adaptation to reduced oxygen availability. In the present study, we describe the oxygen concentration and time-resolved characterization of RAP2.12 activity. A reduction of the oxygen availability to half the concentration in normal air is sufficient to trigger RAP2.12 relocalization into the nucleus, while nuclear accumulation correlates with the first induction of the molecular response to hypoxia. Nuclear presence of RAP2.12 may not only depend on relocalization of existing protein, but involves de novo synthesis of the transcription factor as well. After re-oxygenation of the tissue, degradation of RAP2.12 in the nucleus was observed within 3 h, concomitant with reduction in hypoxia responsive gene transcripts to normoxic levels., (© 2014 John Wiley & Sons Ltd.)

In plant and animal cells, amino-terminal cysteine oxidation controls selective proteolysis via an oxygen-dependent branch of the N-end rule pathway. It remains unknown how the N-terminal cysteine is specifically oxidized. Here we identify plant cysteine oxidase (PCO) enzymes that oxidize the penultimate cysteine of ERF-VII transcription factors by using oxygen as a co-substrate, thereby controlling the lifetime of these proteins. Consequently, ERF-VII proteins are stabilized under hypoxia and activate the molecular response to low oxygen while the expression of anaerobic genes is repressed in air. Members of the PCO family are themselves targets of ERF-VII transcription factors, generating a feedback loop that adapts the stress response according to the extent of the hypoxic condition. Our results reveal that PCOs act as sensor proteins for oxygen in plants and provide an example of how proactive regulation of the N-end rule pathway balances stress response to optimal growth and development in plants.

A plant's eventual size depends on the integration of its genetic program with environmental cues, which vary on a daily basis. Both efficient carbon metabolism and the plant hormone gibberellin are required to guarantee optimal plant growth. Yet, little is known about the interplay between carbon metabolism and gibberellins that modulates plant growth. Here, we show that sugar starvation in Arabidopsis thaliana arising from inefficient starch metabolism at night strongly reduces the expression of ent-kaurene synthase, a key regulatory enzyme for gibberellin synthesis, the following day. Our results demonstrate that plants integrate the efficiency of photosynthesis over a period of days, which is transduced into a daily rate of gibberellin biosynthesis. This enables a plant to grow to a size that is compatible with its environment.

Low oxygen stress often occurs during the life of green organisms, mostly due to the environmental conditions affecting oxygen availability. Both plants and algae respond to low oxygen by resetting their metabolism. The shift from mitochondrial respiration to fermentation is the hallmark of anaerobic metabolism in most organisms. This involves a modified carbohydrate metabolism coupled with glycolysis and fermentation. For a coordinated response to low oxygen, plants exploit various molecular mechanisms to sense when oxygen is either absent or in limited amounts. In Arabidopsis thaliana, a direct oxygen sensing system has recently been discovered, where a conserved N-terminal motif on some ethylene responsive factors (ERFs), targets the fate of the protein under normoxia/hypoxia. In Oryza sativa, this same group of ERFs drives physiological and anatomical modifications that vary in relation to the genotype studied. The microalga Chlamydomonas reinhardtii responses to low oxygen seem to have evolved independently of higher plants, posing questions on how the fermentative metabolism is modulated. In this review, we summarize the most recent findings related to these topics, highlighting promising developments for the future.

The crucial role of carbohydrate in plant growth and morphogenesis is widely recognized. In this study, we describe the characterization of nana, a dwarf Arabidopsis (Arabidopsis thaliana) mutant impaired in carbohydrate metabolism. We show that the nana dwarf phenotype was accompanied by altered leaf morphology and a delayed flowering time. Our genetic and molecular data indicate that the mutation in nana is due to a transfer DNA insertion in the promoter region of a gene encoding a chloroplast-located aspartyl protease that alters its pattern of expression. Overexpression of the gene (oxNANA) phenocopies the mutation. Both nana and oxNANA display alterations in carbohydrate content, and the extent of these changes varies depending on growth light intensity. In particular, in low light, soluble sugar levels are lower and do not show the daily fluctuations observed in wild-type plants. Moreover, nana and oxNANA are defective in the expression of some genes implicated in sugar metabolism and photosynthetic light harvesting. Interestingly, some chloroplast-encoded genes as well as genes whose products seem to be involved in retrograde signaling appear to be down-regulated. These findings suggest that the NANA aspartic protease has an important regulatory function in chloroplasts that not only influences photosynthetic carbon metabolism but also plastid and nuclear gene expression.

Reactive oxygen species (ROS) play an important role as triggers of gene expression during biotic and abiotic stresses, among which is low oxygen (O(2)). Previous studies have shown that ROS regulation under low O(2) is driven by a RHO-like GTPase that allows tight control of hydrogen peroxide (H(2)O(2)) production. H(2)O(2) is thought to regulate the expression of heat shock proteins, in a mechanism that is common to both O(2) deprivation and to heat stress. In this work, we used publicly available Arabidopsis (Arabidopsis thaliana) microarray datasets related to ROS and O(2) deprivation to define transcriptome convergence pattern. Our results show that although Arabidopsis response to anoxic and hypoxic treatments share a common core of genes related to the anaerobic metabolism, they differ in terms of ROS-related gene response. We propose that H(2)O(2) production under O(2) deprivation is a trait present in a very early phase of anoxia, and that ROS are needed for the regulation of a set of genes belonging to the heat shock protein and ROS-mediated groups. This mechanism, likely not regulated via the N-end rule pathway for O(2) sensing, is probably mediated by a NADPH oxidase and it is involved in plant tolerance to the stress.

Background: Calprotectin, a major cytosolic protein of neutrophils, is increased in inflammatory bowel disease (IBD) and may be considered a suitable marker of intestinal inflammation. Abdominal MRI is becoming more frequently used for the evaluation of IBD patients. Aim of this study was to investigate the role of MRI in IBD for the assessment of disease activity in comparison with faecal calprotectin levels., Patients and Methods: Twenty-four consecutive hospitalized pts (12 F, 12 M, median age: 56; range: 22-77) with a proven diagnosis of CD were studied. At the time of the MRI examination, pts provided a single stool sample for calprotectin measurement. Calprotectin was measured by ELISA (Calprest(R)). Pathological values were considered more than 50 microg/g. All pts underwent MRI, performed at 1.5 T, with HASTE T2w with and without fat-saturation, FLASH T1w fat-saturated sequences pre and post iv injection of 0.1 ml/kg of Gadolinium. Presence, degree and length of wall inflammation were evaluated. The MRI degree of wall inflammation was graded with a 0-3 scoring system (0=absent 1=light 2=moderate 3=severe) by considering findings observed on T1 post Gd and T2 fat-suppressed images, as well as the degree of wall thickness. The length of extension was considered as less than 15 cm, between 15 cm and 30 cm, or more than 30 cm. Spearman's correlation coefficient was used to evaluated differences in calprotectin levels among the groups obtained by MRI findings., Results: Grade 0 MRI was found in 1 pt with a faecal calprotectin measurement of 206.25 microg/g; Grade 1 MRI was found in 4 pts with a median faecal calprotectin of 100 microg/g (5-325); Grade 2 MRI was found in 10 pts with a median faecal calprotectin of 243.75 microg/g (7.5-606.25); Grade 3 MRI was found in 9 pts with a median faecal calprotectin of 1012.5 microg/g (30-1268.8). A trend of positive correlation was therefore found between MRI scores of activity and calprotectin levels (p less than 0.0001) and between MRI scores of thickening of intestinal involvement and calprotectin levels (p = 0.005). No apparent correlation was observed between faecal calprotectin concentration and length., Conclusions: Data presenting show that faecal calprotectin levels well correlate with the degree of mucosal inflammation are in agreement with previous studies. Considering the correlation obtained between calprotectin level and MRI findings, we believe that MRI is helpful in assessing and monitoring the degree of disease in Crohn's disease.

Hydatid cyst disease still is a serious public-health problem in endemic areas. It is a tissue infestation caused by the larval stage of a parasite, Echinococcus granulosus. Although liver and lung are the most commonly involved organs, hydatid disease can occur in all viscera and soft tissues. In 10% of cases, hydatid disease arises in the viscera: mainly in the spleen (0.9-8%), but also in kidney, bone, heart and peritoneal cavity (0.5-5%). Other rare locations such as muscles have been described in less than 1% of cases of hydatid disease. We report magnetic resonance imaging findings of a case of diffuse abdominal hydatidosis.

Electrophysiological findings in 14 patients with non-Friedreich early onset cerebellar ataxia are reported. Nerve conduction studies showed reduction of sensory action potential amplitudes in 7 cases associated in 3 with a decrease of sensory conduction velocities. Six subjects also exhibited a chronic neurogenic pattern to standard needle electromyography. Motor conduction velocities were normal in all cases; only two cases showed an increase in distal motor latencies. Short-latency somatosensory evoked potentials following median nerve stimulation revealed a prolonged central conduction time (N13-N20 interpeak latency) in 7 cases, compatible with supraspinal damage of the somatosensory pathways. These electrophysiological data are compared with those obtained in two reference groups of patients, respectively affected by Friedreich's ataxia and olivo-ponto-cerebellar atrophy.

The possibility was investigated that impulses in group Ib afferents from fore-arm flexors have access to cerebral cortex in man. A long-lasting increase in the threshold (Th) of group Ia afferent fibres from flexor carpi radialis muscle (Fcr) was obtained after prolonged (100 Hz for 20 min) tendon vibration at the wrist. It was assumed that under this condition a weak electrical stimulation of the median nerve at the elbow, insufficient to reactivate Ia fibres because of the rise in their threshold (as verified by the method of homonymous Ia facilitation of Fcr H reflex), engaged Ib fibres only. Peripheral volleys at Erb's point and cerebral cortical potentials to median nerve stimulation at the elbow were evoked before and after prolonged vibration of Fcr. During 10-30 min after the end of vibration, in which homonymous facilitation of Fcr H reflex was abolished, both the ongoing peripheral volley and the cortical responses were markedly reduced with respect to their control values. Recovery to pre-vibration control amplitudes coincided with recovery of Fcr H reflex homonymous facilitation. In order to verify if activity in afferents other than group Ib fibres might contribute to the cortical response after vibration, specially designed experiments were also performed. It is concluded that the cortical wave recorded after 20 min vibration represents the arrival of Ib impulses from flexor carpi radialis to the human cerebral cortex.

We report a case of classic phenylketonuria (PKU) treated by dietary restriction until the age of 5 years. At the age of 18 years, she developed anxious-depressive symptoms and deterioration of school performance. Neurophysiological investigations showed changes in VEPs and BAEPs and brain MRI showed white matter abnormalities. The return to a diet low in phenylalanine reduced psychiatric disturbances and reversed neurophysiological and MRI changes. Our data confirm previous observations of white matter involvement in PKU and the utility of lifelong dietary therapy to prevent neurological and/or psychiatric deterioration.