Lipoxygenase6-Dependent Oxylipin Synthesis in Roots Is Required for Abiotic and Biotic Stress Resistance of Arabidopsis

Jasmonates are oxylipin signals that play important roles in the development of fertile flowers and in defense against pathogens and herbivores in leaves. The aim of this work was to understand the synthesis and function of jasmonates in roots. Grafting experiments with a jasmonate-deficient mutant demonstrated that roots produce jasmonates independently of leaves, despite low expression of biosynthetic enzymes. Levels of 12-oxo-phytodienoic acid, jasmonic acid, and its isoleucine derivative increased in roots upon osmotic and drought stress. Wounding resulted in a decrease of preformed 12-oxo-phytodienoic acid concomitant with an increase of jasmonic acid and jasmonoyl-isoleucine. 13-Lipoxygenases catalyze the first step of lipid oxidation leading to jasmonate production. Analysis of 13-lipoxygenase-deficient mutant lines showed that only one of the four 13-lipoxygenases, LOX6, is responsible and essential for stress-induced jasmonate accumulation in roots. In addition, LOX6 was required for production of basal 12-oxo-phytodienoic acid in leaves and roots. Loss-of-function mutants of LOX6 were more attractive to a detritivorous crustacean and more sensitive to drought, indicating that LOX6-derived oxylipins are important for the responses to abiotic and biotic factors.Oxylipins are ubiquitous signaling molecules that are derived from polyunsaturated fatty acids by enzymatic and nonenzymatic processes. In plants, the biosynthesis and function of oxylipins of the jasmonate family in aboveground tissues has been investigated in detail. Jasmonates comprise 12-oxo-phytodienoic acid (OPDA), jasmonic acid (JA), and derivatives of JA. In leaves, jasmonates accumulate in response to abiotic factors such as wounding, drought, osmotic stress, darkness, and ozone and during interactions with organisms such as herbivores, pathogens, and mutualistic organisms (Wasternack, 2007). The relevance of jasmonates in wound response, ozone tolerance, and the defense against herbivores and necrotrophic pathogens in leaves has been well investigated using mutants in JA biosynthesis and signaling (Browse, 2009a). In addition, jasmonates play an important role in flower development, and Arabidopsis (Arabidopsis thaliana) mutants in the JA pathway are male sterile (Browse, 2009b). The first step in jasmonate biosynthesis is catalyzed by 13-lipoxygenases (LOXs). The resulting 13(S)-hydroperoxyoctadecatrienoic acid (13-HPOTE) is converted by allene oxide synthase (AOS) and allene oxide cyclase to OPDA (Wasternack, 2007). These enzymatic steps are located in plastids. OPDA is transported to peroxisomes and converted to JA. JA can be further metabolized to different derivatives that take place mainly in the cytosol. The conjugation of JA with Ile is an important step because jasmonoyl-Ile (JA-Ile) has been identified as a biologically active jasmonate (Staswick and Tiryaki, 2004). OPDA is also biologically active without conversion to JA derivatives. In contrast to all other jasmonates, the OPDA structure contains an electrophilic α,β-unsaturated carbonyl group that renders OPDA more reactive than JA. Therefore, OPDA is classified as a reactive electrophile species with unique signaling properties different from other jasmonates (Farmer and Davoine, 2007).Of the six lipoxygenase genes present in Arabidopsis, four genes encode 13-LOX. For the respective enzymes LOX2, LOX3, LOX4, and LOX6, it was shown that linolenic acid is the preferred substrate and that 13-HPOTE is formed in vitro (Bannenberg et al., 2009). All four enzymes are proposed to be located in plastids. LOX2 is highly expressed in leaves; expression is up-regulated by jasmonates and stress treatments such as wounding and osmotic stress (Bell and Mullet, 1993; Seltmann et al., 2010a). LOX2 was shown to contribute the majority of jasmonate synthesis upon wounding and osmotic stress and during senescence in leaves (Bell et al., 1995; Glauser et al., 2009). LOX2 is also responsible for the accumulation of arabidopsides (Glauser et al., 2009), which are galactolipids containing esterified OPDA in plastids by direct oxidation of galactolipids (Zoeller et al., 2012). LOX3 and LOX4 are required for the development of fertile flowers (Caldelari et al., 2011). LOX6 shows overall low expression (Bannenberg et al., 2009). Recently, it was reported that LOX6 contributes to the fast accumulation of JA and JA-Ile in wounded leaves and is required for the fast increase of JA and JA-Ile in distal leaves after wounding (Chauvin et al., 2013).In contrast to leaves and flowers, little is known on jasmonate biosynthesis and function in roots. Expression of the plastid-localized enzymes of jasmonate synthesis LOX2, AOS, and allene oxide cyclase2 is very low in roots (Zimmermann et al., 2004). By contrast, enzymes such as 9-LOX and α-dioxygenase1 are strongly expressed in roots. These enzymes are involved in the biosynthesis of oxylipins different from jasmonates, and 9-LOX products have been shown to regulate lateral root development because mutants in LOX1 and LOX5 produce more lateral roots (Vellosillo et al., 2007). However, jasmonate function in roots is still obscure. Here, we analyzed jasmonate accumulation in roots upon different stress treatments and show that mutants defective in LOX6 are impaired in stress-induced jasmonate synthesis and are more susceptible to drought and detritivore feeding.     

Phylogenetic signals in thermal traits remain stronger in the tropics if we can believe published physiological data. A reply to McKechnie et al., “Data quality problems undermine analyses of endotherm upper critical temperatures”

Due to concerns about data quality, McKechnie, Coe, Gerson, and Wolf ( 2016 ) questioned the conclusions of our study (Khaliq et al., 2015 ) published in this journal. Here, we argue that most of the questioned data points are in fact useful for macrophysiological analyses, mostly because the vast majority of data are explicitly reported in the peer‐reviewed physiological literature. Furthermore, we show that our conclusions remain largely robust irrespective of the data inclusion criterion. While we think that constructive debates about the adequate use of primary data in meta‐studies as well as more transparency in data inclusion criteria are indeed useful, we also emphasize that data suitability should be evaluated in the light of the scope and scale of the study in which they are used. We hope that this discussion will not discourage the exchange between disciplines such as biogeography and physiology, as this integration is needed to address some of the most urgent scientific challenges.     

The Diurnal Pattern of Nitrate Uptake and Reduction by Spinach (Spinacia oleracea L.)

Spinach plants were grown in bowls of aerated nutrient solutionin a controlled environment chamber for 24 h, and harvestedevery 3·5-5 h to record their growth, nitrate and wateruptake, and plant nitrate concentration. Twelve such experimentsare described, either with a 14/10 h dark/light regime, or continuouslight or darkness. The irradiance was either 110, 320, or 510µmol m^-2 s^-1 (PPFD). All these regimes began at the endof the light period of a 14/10 h dark/light regime (510 µmolm^-2 s^-1) lasting approximately 2 weeks. Nitrate uptake rate per g of dry weight of plant continued almostunabated at about 17 µmol h^-1 through the initial 14-hdark period, and then fell away sharply if the light was notrestored, but increased slightly when it was. With continuouslight at 510 µmol m^-2 s^-1, uptake rate rose steadily forthe first 24 h of light, and then fell sharply for about 6 h.Shoot nitrate concentration increased about three-fold in thedark phase, and declined in the light at a rate which was positivelyrelated to the irradiance. Root nitrate concentration was severaltimes higher than that of the shoot: its diurnal change wassmaller (relative to the mean) than that of the shoot. Nitratereduction occurred to a small extent in the dark, and increasedrapidly as soon as the lights came on, to remain at a roughlyconstant rate (related to the irradiance) throughout the lightphase. Dry matter increase in the light was related to irradiance,but with little increase above 320 µmol m^-2 s^-1. Respiratoryweight loss in the dark was not detectable. Rate of fresh weightincrease was approximately constant throughout light and darkperiods. The results compare quite well with the predictions of a simplesimulation model, based on the pump/leak principle.Copyright1994, 1999 Academic Press Spinacia oleracea, nitrate, uptake, reduction, influx, efflux, diurnal, regulation, model, simulation     

Heterogeneous perfusion is a consequence of uniform shear stress in optimized arterial tree models

Using optimized computer models of arterial trees we demonstrate that flow heterogeneity is a necessary consequence of a uniform shear stress distribution. Model trees are generated and optimized under different modes of boundary conditions. In one mode flow is delivered to the tissue as homogeneously as possible. Although this primary goal can be achieved, resulting shear stresses between blood and the vessel walls show very large spread. In a second mode, models are optimized under the condition of uniform shear stress in all segments which in turn renders flow distribution heterogeneous. Both homogeneous perfusion and uniform shear stress are desirable goals in real arterial trees but each of these goals can only be approached at the expense of the other. While the present paper refers only to optimized models, we assume that this dual relation between the heterogeneities in flow and shear stress may represent a more general principle of vascular systems.     

Isocitrate dehydrogenase 1 mutant R132H sensitizes glioma cells to BCNU-induced oxidative stress and cell death

Isocitrate dehydrogenase 1 (IDH1) decarboxylates isocitrate to α-ketoglutarate (α-KG) leading to generation of NADPH, which is required to regenerate reduced glutathione (GSH), the major cellular ROS scavenger. Mutation of R132 of IDH1 abrogates generation of α-KG and leads to conversion of α-KG to 2-hydroxyglutarate. We hypothesized that glioma cells expressing mutant IDH1 have a diminished antioxidative capacity and therefore may encounter an ensuing loss of cytoprotection under conditions of oxidative stress. Our study was performed with LN229 cells stably overexpressing IDH1 R132H and wild type IDH1 or with a lentiviral IDH1 knockdown. Quantification of GSH under basal conditions and following treatment with the glutathione reductase inhibitor BCNU revealed significantly lower GSH levels in IDH1 R132H expressing cells and IDH1 KD cells compared to their respective controls. FACS analysis of cell death and ROS production also demonstrated an increased sensitivity of IDH1-R132H-expressing cells and IDH1 KD cells to BCNU, but not to temozolomide. The sensitivity of IDH1-R132H-expressing cells and IDH1 KD cells to ROS induction and cell death was further enhanced with the transaminase inhibitor aminooxyacetic acid and under glutamine free conditions, indicating that these cells were more addicted to glutaminolysis. Increased sensitivity to BCNU-induced ROS production and cell death was confirmed in HEK293 cells inducibly expressing the IDH1 mutants R132H, R132C and R132L. Based on these findings we propose that in addition to its established pro-tumorigenic effects, mutant IDH1 may also limit the resistance of gliomas to specific death stimuli, therefore opening new perspectives for therapy.     

Myosin in onion (Allium cepa) bulb scale epidermal cells: involvement in dynamics of organelles and endoplasmic reticulum

Studied with the fluorochrome 3,3-dihexyloxacarbocyanine iodide [(DIOC₆(3)], the dynamic system of the endoplasmic reticulum (ER) in epidermal cells of onion bulb scales consists of long, tubular strands moving together with organelles in the deeper cytoplasm, and of a less mobile network composed of tubular and lamellar elements at the cell periphery. Treatment with the sulfhydryl-reagent N-ethylmaleimide (NEM) inhibited organelle and ER movement, and caused the fusion of ER-tubules into flat sheets. Fixed, long, tubular ER strands were formed by lowering the cytosolic pH of NEM-treated cells. Both these observations indicate the involvement of myosin in the dynamics of organelles and ER. Using a monoclonal antibody against murine skeletal muscle myosin (known to cross-react with plant myosin; Tang et al. 1989, J. Cell Sci. 92: 569–574), myosin was identified by immunofluorescence microscopy. Mapping the distribution of myosin, actin filaments, ER, and organelles in different phases of recovery after centrifugation of epidermal cells, co-localization of myosin with ER and organelles but not with actin filaments was observed, supporting the hypothesis that a membrane bound motor protein exists in onion epidermal cells, which translocates organelles and the endoplasmic reticulum along actin filaments.     

Impact of dry mouth conditions on oral health-related quality of life in older people

Objective: The aim of the present study was to evaluate the impact of dry mouth conditions on oral health‐related quality of life in frail old people, residents at community care centers. Further, reliability and validity of a visual analogue scale (VAS) for dry mouth symptoms were determined within the study cohort. Background: In old people functional, social and psychological impacts of oral conditions are associated with an overall sense of well being and general health. Subjective dry mouth and reduced saliva flow are common disorders in old people caused by disease and medication. Thus, dry mouth conditions may be determinants for compromised oral health‐related quality of life in old people. Method: In total, 50 old people living at service homes for the old people were asked to answer questionnaires on subjective dry mouth (VAS) and Oral Health Impact Profile (OHIP14) for oral health‐related quality of life. Saliva flow was estimated by absorbing saliva into a pre‐weighed cotton roll. Results: The final study cohort comprised 41 old people (aged 83–91 years). Significant associations were identified between both objective and subjective dry mouth and overall or specific aspects of oral health‐related quality of life. Conclusion: Dry mouth (objective and subjective) is significantly associated with oral health‐related quality of life strengthening the value of monitoring dry mouth conditions in the care of frail old people.     

Maturation and maintenance of cholinergic medial septum neurons require glucocorticoid receptor signaling

Glucocorticoids have been shown to influence trophic processes in the nervous system. In particular, they seem to be important for the development of cholinergic neurons in various brain regions. Here, we applied a genetic approach to investigate the role of the glucocorticoid receptor (GR) on the maturation and maintenance of cholinergic medial septal neurons between P15 and one year of age by using a mouse model carrying a CNS-specific conditional inactivation of the GR gene (GRNesCre). The number of choline acetyltransferase and p75NTR immuno-positive neurons in the medial septum (MS) was analyzed by stereology in controls versus mutants. In addition, cholinergic fiber density, acetylcholine release and cholinergic key enzyme activity of these neurons were determined in the hippocampus. We found that in GRNesCre animals the number of medial septal cholinergic neurons was significantly reduced during development. In addition, cholinergic cell number further decreased with aging in these mutants. The functional GR gene is therefore required for the proper maturation and maintenance of medial septal cholinergic neurons. However, the loss of cholinergic neurons in the medial septum is not accompanied by a loss of functional cholinergic parameters of these neurons in their target region, the hippocampus. This pinpoints to plasticity of the septo-hippocampal system, that seems to compensate for the septal cell loss by sprouting of the remaining neurons.     

The survivin/Aurora B complex: its role in coordinating tension and attachment

Proper chromosome segregation relies on the action of the spindle checkpoint. Recent data have shown that the chromosomal passenger proteins survivin and Aurora B play an important auxiliary role in spindle checkpoint surveillance. Knock-down experiments in human cells indicate that the function of the survivin/Aurora B complex is required to correct improper microtubule-kinetochore interactions. Combined data of four different groups show that the survivin/Aurora B complex is not an integral component of the spindle checkpoint, but it enables the cell to communicate lack of tension back to the attached microtubules. Moreover, they show that the affinity of BubR1 for kinetochores is directly influenced by the absence or presence of the survivin/Aurora B complex. These functions of the survivin/Aurora B complex are essential for chromosome biorientation, a prerequisite for proper chromosome segregation. As such, this complex plays an important role in the maintenance of a stable genome.