Aleurone Cells Research Articles

Hydrogen sulfide delays GA-triggered programmed cell death in wheat aleurone layers by the modulation of glutathione homeostasis and heme oxygenase-1 expression

Hydrogen sulfide (H2S) is considered as a cellular signaling intermediate in higher plants, but corresponding molecular mechanisms and signal transduction pathways in plant biology are still limited. In the present study, a combination of pharmacological and biochemical approaches was used to study the effect of H2S on the alleviation of GA-induced programmed cell death (PCD) in wheat aleurone cells. The results showed that in contrast with the responses of ABA, GA brought about a gradual decrease of l-cysteine desulfhydrase (LCD) activity and H2S production, and thereafter PCD occurred. Exogenous H2S donor sodium hydrosulfide (NaHS) not only effectively blocked the decrease of endogenous H2S release, but also alleviated GA-triggered PCD in wheat aleurone cells. These responses were sensitive to hypotaurine (HT), a H2S scavenger, suggesting that this effect of NaHS was in an H2S-dependent fashion. Further experiment confirmed that H2S, rather than other sodium- or sulphur-containing compounds derived from the decomposing of NaHS, was attributed to the rescuing response. Importantly, the reversing effect was associated with glutathione (GSH) because the NaHS triggered increases of endogenous GSH content and the ratio of GSH/oxidized GSH (GSSG) in GA-treated layers, and the NaHS-mediated alleviation of PCD was markedly eliminated by l-buthionine-sulfoximine (BSO, a selective inhibitor of GSH biosynthesis). The inducible effect of NaHS was also ascribed to the modulation of heme oxygenase-1 (HO-1), because the specific inhibitor of HO-1 zinc protoporphyrin IX (ZnPP) significantly suppressed the NaHS-related responses. By contrast, the above inhibitory effects were reversed partially when carbon monoxide (CO) aqueous solution or bilirubin (BR), two of the by-products of HO-1, was added, respectively. NaHS-triggered HO-1 gene expression in GA-treated layers was also confirmed. Together, the above results clearly suggested that the H2S-delayed PCD in GA-treated wheat aleurone cells was associated with the modulation of GSH homeostasis and HO-1 gene expression.

RNA-sequencing reveals previously unannotated protein- and microRNA-coding genes expressed in aleurone cells of rice seeds

The rice genome annotation has been greatly improved in recent years, largely due to the availability of full length cDNA sequences derived from many tissues. Among those yet to be studied is the aleurone layer, which produces hydrolases for mobilization of seed storage reserves during seed germination and post germination growth. Herein, we report transcriptomes of aleurone cells treated with the hormones abscisic acid, gibberellic acid, or both. Using a comprehensive approach, we identified hundreds of novel genes. To minimize the number of false positives, only transcripts that did not overlap with existing annotations, had a high level of expression, and showed a high level of uniqueness within the rice genome were considered to be novel genes. This approach led to the identification of 553 novel genes that encode proteins and/or microRNAs. The transcriptome data reported here will help to further improve the annotation of the rice genome.

Structural development of aleurone and its function in common wheat

The wheat aleurone is formed from surface endosperm cells, and its developmental status reflects its biogenesis, structural characteristics, and physiological functions. In this report, wheat caryopses at different development stages were embedded in Spurr's low-viscosity embedding medium for observation of the development of aleurone cells (ACs) by light microscopy, scanning electron microscopy, and fluorescence microscopy, respectively. According to their structures and physiological characterization, the ACs development process was divided into five stages: endosperm cellulization, spherosome formation, aleurone grain formation, filling material proliferation, and maturation. Furthermore, ACs in different parts of the caryopsis formed differently. ACs near the vascular bundle developed earlier and formed transfer cells, but other ACs formed slowly and did not form transfer cells. ACs on the caryopsis backside were a regular square shape; however, ACs in the caryopsis abdomen were mainly irregular. There were also differences in development between wheat varieties. ACs were rectangular in hard wheat but square in soft wheat. ACs were larger and showed a greater degree of filling in hard compared to soft wheat. The storage materials in ACs were different compared to inner endosperm cells (IECs). The concentrations of minerals such as sodium, magnesium, silicon, phosphorus and potassium were higher in ACs than in IECs. ACs contained many aleurone grains and spherosomes, which store lipids and mineral nutrients, respectively. The cell nucleus did not disappear and the cells were still alive during aleurone maturation. However, IECs were dead and mainly contained amyloplast and protein bodies, which store starch and protein, respectively. Overall, the above results characterized major structural features of aleurone and revealed that the wheat aleurone has mainly four functions.

Localisation of Storage Reserves in Developing Seeds of Pongamia pinnata (L.) Pierre, a Potential Agroforestry Tree

AbstractPongamia pinnata is an important oil yielding perennial tree species. The aim of the present study was to document the histological and ultrastructural change that is occurring during pongamia seed development. The seeds were sampled at five stages of development at intervals of 3 weeks starting from 30 weeks after flowering up to 42 weeks. The seed development was followed microscopically using toluidine blue staining. The seed coat was made up of an external layer of palisade cells, an internal layer of hourglass cells followed by a parenchymatous cell layer and aleurone cell layer. The seed reserve compounds such as polysaccharides, proteins and starch showed distinct histochemical characterisation. Lignin was mainly found in the seed coat cell layers, while polysaccharides, proteins and starch granules in the cotyledon cells. The ultrastructural studies showed marked cellular changes during the seed development. The cell size varied from 9.4 to 78 μm during the seed development. The number of oil bodies per cell ranged from 200 to 300 at 42 weeks after flowering. Protein storage vacuoles were observed during the later stages of seed development. The plastids contained electron‐dense starch granules. The seeds harvested after 42 weeks after flowering had maximum physiological maturity with high oil content and other seed reserve materials. This basic knowledge on pongamia seed development could invariably be used for further understanding of biochemical changes that might be involved in the biosynthetic pathway of oil.

Visualization of the Anticaging Effect of Ronozyme WX Xylanase on Wheat Substrates

ABSTRACTA fluorescence microscope was used for visualization of the anticaging effect of a commercial xylanase on milled wheat, microtome cuts of wheat grains, and digesta samples obtained from piglets 1 or 4 h after feeding a wheat‐based diet (wheat 490 g/kg of diet, barley 100 g/kg of diet, and oats 100 g/kg of diet). Both starchy endosperm and aleurone cell walls were shown to be broken down by Ronozyme WX commercial xylanase. Data obtained by fluorescence microscopy was supplemented with measurements of starch and xylose released as a result of degradation of nonstarch polysaccharides by the xylanase. The results visualize and provide evidence that Ronozyme WX commercial xylanase can overcome the so‐called cage effect. This release of nutrients from their encapsulation in cereal cell wall structures has positive impact on nutrient digestibility and partially explains the positive effect of xylanase supplementation on livestock performance.

Dissociation of aleurone cell cluster from wheat bran by centrifugal impact milling

Wheat aleurone which contains most of the nutritionally important substances of wheat bran has been widely investigated, and electrostatic separation based on the distinct electrical properties of bran tissues has been proven to be effective for the enrichment of aleurone layer. In order to obtain a suitable starting material containing single-layer bran strips for the electrostatic separation, centrifugal impact milling was utilized to dissociate wheat bran constitutive tissues. During centrifugal impact milling, the outer pericarp was detached when the particle size of the fragments decreased to approximate 460μm, whereas the aleurone layer consisted of 20–30 intact cells in single-layer was completely dissociated until the particle size decreased to about 200μm. Moreover, the detached behaviors of bran tissues were well explained by the relationship of their elastoplasticities, the rupture-energy and peeling force generated by the rotating blade tip. After centrifugal impact milling and sieving, the bran fraction with the median particle diameter of 144μm contained 61% single-layer aleurone cell clusters, indicating that it could be a superior starting material for the electrostatic separation of wheat aleurone.

Changes in bran structure by bioprocessing with enzymes and yeast modifies the in vitro digestibility and fermentability of bran protein and dietary fibre complex

Bran is a good source of dietary fibre, phytochemicals, and also protein, but highly insoluble and recalcitrant structure of bran hinders accessibility of these components for gastrointestinal digestion. In the present work, influence of bioprocessing on the microstructure and chemical properties of rye bran and wheat bread fortified with the rye bran were studied. In vitro protein digestibility, and release of short chain fatty acids (SCFA) and ferulic acid in a gut model were studied. Bioprocessing of rye bran was performed with subsequent treatments with cell-wall hydrolysing enzymes (40 °C, 4 h) and yeast fermentation (20 °C, 20 h). Bioprocessing of rye bran resulted in reduced total dietary fibre content, caused mainly by degradation of fructan and β-glucan, and increased soluble fibre content, caused mainly by solubilisation of arabinoxylans. Microscopic analysis revealed degradation of aleurone cell wall structure of the bioprocessed rye bran. Bioprocessing caused release of protein from aleurone cells, assessed as a larger content of soluble protein in bran and a higher hydrolysis rate in vitro. Bioprocessed bran had also faster SCFA formation and ferulic acid release in the colon fermentation in vitro as compared to native bran.

Soil-Weed Seed Communication Systems

The nature of weeds is a complex adaptive, soil-seed communication system. The nature of weedy Setaria life history is an adaptable, changeable system in which complex behaviors emerge when self-similar plant components self-organize into functional traits possessing biological information about spatial structure and temporal behavior. Setaria life history behavior is a Markov chain of irreversible and reversible processes regulated by morpho-physiological traits acting through environment-plant communication systems (environment-plant-seed, soil-seed). Heritable functional traits are the physical reservoirs of information guiding life history development, emergent behavior. The consequence of structural self-similarity and behavioral self-organization has been the evolution of a complex adaptive seed-soil communication system. Weedy Setaria life history is represented in algorithmic form as FoxPatch, a model to forecast seed behavior. Weedy Setaria seed life history behaviors are controlled by environmental information (signals) flowing from the soil to the seed embryo. The specific signal to which Setaria is tuned affecting seed behavior in the soil is the amount of oxygen and heat (T, thermal) in soil water over time, oxy-hydro-thermal time (O2-H2O-T-Time). The Shannon environmental-biological communication system between the soil and the Setaria seed contains the five elements (E) and components:E1, information source, soil; E2, transmitter, soil particle contact with seed surface water films; E3, channel, continuous soil particle-seed surface water films; E4, receiver, living seed interior from the transfer aleurone cell layer (TACL) membrane to aleurone layer to embryo; E5, destination, embryo. The signal is soil O2-H2O-T-Time; the message is O2-H2O-T stimulating embryo respiration. The Setaria soil-seed communication system seed behavior can be also expressed as operations (processes) computed by seed algorithms. Information is physical: memory resides in several locations in the Setaria seed. Memory is expressed in the long-term by responsiveness to O2-H2O-heat messages as determined by the morpho-physiological soil-seed communication system (hull, TACL membrane, scavenger protein). The message is remembered: plants pass on a range of heteromorphic seeds appropriate to continuing, successful local adaptation.

Exposure or release of ferulic acid from wheat aleurone: Impact on its antioxidant capacity

The relationship between the aleurone cell integrity and the exposure or release of bioavailable ferulic acid (FA) with the antioxidant capacity of aleurone in in vitro and under simulated gastric conditions was explored. The antioxidant capacity of aleurone was increased by around 2-fold when its median particle size was reduced to under 50 μm. The opening of aleurone cells increased the physical exposure of FA bound to the insoluble polysaccharides, which seemed to be responsible of the increased antioxidant capacity. Synergistic combination of xylanase and feruloyl esterase was found to be the most efficient enzymatic treatment releasing up to 86% of total FA in bioaccessible forms. This enzymatic treatment significantly enhanced the radical scavenging activity of aleurone by up to 4-fold, which overlapped the overall antioxidant potential estimated from the total content of FA in aleurone. The improvement in the antioxidant capacity of aleurone was also observed in the simulated gastric digestion by inhibition of lipid oxidation.

Iron and zinc complexation in wild-type and ferritin-expressing wheat grain: implications for mineral transport into developing grain

We have used synchrotron-based X-ray fluorescence and absorption techniques to establish both metal distribution and complexation in mature wheat grains. In planta, extended X-ray absorption fine structure (EXAFS) spectroscopy reveals iron phytate and zinc phytate structures in aleurone cells and in modified aleurone cells in the transfer region of the grain: iron is coordinated octahedrally by six oxygen atoms and fewer than two phosphorous atoms. Zinc is coordinated tetrahedrally by four oxygen atoms and approximately 1.5 phosphorus atoms in an asymmetric coordination shell. We also present evidence of modified complexation of both metals in transgenic grain overexpressing wheat ferritin. For zinc, there is a consistent doubling of the number of complexing phosphorus atoms. Although there is some EXAFS evidence for iron phytate in ferritin-expressing grain, there is also evidence of a structure lacking phosphorus. This change may lead to an excess of phosphorus within the storage regions of grain, and in turn to the demonstrated increased association of phosphorus with zinc in ferritin-expressing grains. Derivative X-ray absorption spectra also suggest that mineral complexation in the transfer region of ferritin-expressing grains is quite different from that in wild-type grain. This may explain why the raised levels of minerals transported to the developing grain accumulate within the crease region of the transgenic grain.

Functional analysis of TaABF1 during abscisic acid and gibberellin signalling in aleurone cells of cereal grains

The wheat transcription factor TaABF1 physically interacts with the protein kinase PKABA1 and mediates both abscisic acid (ABA)-induced and ABA-suppressed gene expression. In bombarded aleurone cells of imbibing grains, the effect of TaABF1 in down-regulating the gibberellin (GA)-induced Amy32b promoter was stronger in the presence of exogenous ABA. As these grains contained low levels of endogenous ABA, the effect of TaABF1 may also be mediated by ABA-induced activation even in the absence of exogenous ABA. Levels of TaABF1 protein decreased slightly during imbibition of afterripened grains. However, TaABF1 levels (especially in aleurone layers) were not substantially affected by exogenous ABA or GA, indicating that changes in TaABF1 protein level are not an important part of regulating its role in hormone signalling. We found that TaABF1 was phosphorylated in vivo in aleurone cells, suggesting a role for post-translational modification in regulating TaABF1 activity. Induction of Amy32b by overexpression of the transcription factor GAMyb could not be prevented by TaABF1, indicating that TaABF1 acts upstream of GAMyb transcription in the signalling pathway. Supporting this view, knockdown of TaABF1 by RNA interference resulted in increased expression from the GAMyb promoter. These results are consistent with a model in which TaABF1 is constitutively present in aleurone cells, while its ability to down-regulate GAMyb is regulated in response to ABA.

Endosperm and aleurone cell structure in barley and wheat as studied by optical and Raman microscopy

Grain ultrastructure is of utmost importance when designing grain processing procedures in the food industry. In this study, wheat and barley grain components were localised using optical and Raman microscopy. The optical microscopic analyses were performed using several selective stains to localise β-glucan, protein and starch or autofluorescence to image the ferulic acid and other fluorescing substances. Alternatively, Raman microscopy was applied to localise the grain components without any need for preceding staining or other sample pretreatment. Both methods provided consistent information on the grain structures, illustrating the distribution of polysaccharides, aromatics and protein in endosperm and aleurone layers. In aleurone layers of both barley and wheat, a distinct difference between the anticlinal and periclinal cell walls was observed. The anticlinal cell walls were enriched with aromatic substances which were present in remarkably lower concentrations in the inner periclinal cell walls but for barley, an even higher concentration in the outer periclinal cell wall was observed. In addition, Raman spectroscopy illustrated the detailed distribution of substances across the aleurone cell walls: β-glucan was adjacent to proteins and it was deficient in the middle lamella whereas arabinoxylan was enriched in the outer cell wall layers and middle lamella.

Differences in phosphatidic acid signalling and metabolism between ABA and GA treatments of barley aleurone cells

Phosphatidic acid (PA) is the common lipid product in abscisic acid (ABA) and gibberellic acid (GA) response. In this work we investigated the lipid metabolism in response to both hormones. We could detect an invivo phospholipase D activity (PLD, EC 3.1.4.4). This PLD produced [(32)P]PA (phosphatidic acid) rapidly (minutes) in the presence of ABA, confirming PA involvement in signal transduction, and transiently, indicating rapid PA removal after generation. The presence of PA removal by phosphatidate phosphatase 1 and 2 isoforms (E.C. 3.1.3.4) was verified in isolated aleurone membranes invitro, the former but not the latter being specifically responsive to the presence of GA or ABA. The invitro DGPP phosphatase activity was not modified by short time incubation with GA or ABA while the invitro PA kinase - that allows the production of 18:2-DGPP from 18:2-PA - is stimulated by ABA. The long term effects (24h) of ABA or GA on lipid and fatty acid composition of aleurone layer cells were then investigated. An increase in PC and, to a lesser extent, in PE levels is the consequence of both hormone treatments. ABA, in aleurone layer cells, specifically activates a PLD whose product, PA, could be the substrate of PAP1 and/or PAK activities. Neither PLD nor PAK activation can be monitored by GA treatment. The increase in PAP1 activity monitored after ABA or GA treatment might participate in the increase in PC level observed after 24h hormone incubation.

Whole grains, type 2 diabetes, coronary heart disease, and hypertension: Links to the aleurone preferred over indigestible fiber

Higher whole grain cereal intakes are associated with substantially lower risks of type 2 diabetes, coronary heart disease, and hypertension. These reduced risks have been established in large prospective studies that now include millions of person-years of follow-up. We analyze the results of 11 major prospective studies to provide recommendations about whole grain consumption. The following review establishes the amount of whole grains that should ideally be consumed based on prospective evidence; defines the nature of whole grains; identifies that the whole grain evidence is robust and not due to confounding; and provides a detailed assessment of several potential mechanisms for the effect of whole grains on health. We draw the following conclusions. Firstly, to maintain health, 40 grams or more of whole grains should be consumed daily. This is about a bowl of whole grain breakfast cereal daily, but 80% of the population does not achieve this. Secondly, aleurone in bran is a critical grain component generally overlooked in favor of indigestible fiber. Live aleurone cells constitute 50% of millers' bran. They store minerals, protein, and the antioxidant ferulic acid, and are clearly more than just indigestible fiber. Finally, we suggest potential roles for magnesium, zinc, and ferulic acid in the development of chronic disease. If the results of prospective studies were applied to the life-style practices of modern societies there exists the potential for enormous personal health and public financial benefits.

Barley HvHMA1 is a heavy metal pump involved in mobilizing organellar Zn and Cu and plays a role in metal loading into grains.

Heavy metal transporters belonging to the P1B-ATPase subfamily of P-type ATPases are key players in cellular heavy metal homeostasis. Heavy metal transporters belonging to the P1B-ATPase subfamily of P-type ATPases are key players in cellular heavy metal homeostasis. In this study we investigated the properties of HvHMA1, which is a barley orthologue of Arabidopsis thaliana AtHMA1 localized to the chloroplast envelope. HvHMA1 was localized to the periphery of chloroplast of leaves and in intracellular compartments of grain aleurone cells. HvHMA1 expression was significantly higher in grains compared to leaves. In leaves, HvHMA1 expression was moderately induced by Zn deficiency, but reduced by toxic levels of Zn, Cu and Cd. Isolated barley chloroplasts exported Zn and Cu when supplied with Mg-ATP and this transport was inhibited by the AtHMA1 inhibitor thapsigargin. Down-regulation of HvHMA1 by RNA interference did not have an effect on foliar Zn and Cu contents but resulted in a significant increase in grain Zn and Cu content. Heterologous expression of HvHMA1 in heavy metal-sensitive yeast strains increased their sensitivity to Zn, but also to Cu, Co, Cd, Ca, Mn, and Fe. Based on these results, we suggest that HvHMA1 is a broad-specificity exporter of metals from chloroplasts and serve as a scavenging mechanism for mobilizing plastid Zn and Cu when cells become deficient in these elements. In grains, HvHMA1 might be involved in mobilizing Zn and Cu from the aleurone cells during grain filling and germination.

Ultra-fine grinding increases the antioxidant capacity of wheat bran

In order to study the influence of wheat bran particle size on its antioxidant capacity, the wheat bran was ground under normal and cryogenic conditions with variable intensity to produce ten fractions with different physical structures. The high energy grinding increased 3-fold the specific surface of the bran fractions and also the proportion of particles smaller than 50 μm assimilated to the proportion of disrupted aleurone cells. All the ground bran fractions presented similar total ferulic acid concentration and chemical form (free, conjugated, linked). A positive effect of the grinding on the antioxidant capacity of the bran fractions was noticed. The antioxidant capacity increased from 30 to 45 mmol TEAC/kg when the specific surface increased from 0.09 to 0.26–0.30 m2/g. The antioxidant capacity of the bran fractions was linearly correlated with the specific surface, with the D50 values and with the proportion of particles smaller than 50 μm. In in vitro gastric conditions, the finely ground bran inhibited the accumulation of conjugated dienes more efficiently than coarse bran. In conclusion, the bran structure affects its antioxidant capacity. This effect remained in gastric conditions showing that grinding can be used to produce wheat bran fractions with higher nutritional value.

TamiR159 Directed Wheat TaGAMYB Cleavage and Its Involvement in Anther Development and Heat Response

In Arabidopsis and rice, miR159-regulated GAMYB-like family transcription factors function in flower development and gibberellin (GA) signaling in cereal aleurone cells. In this study, the involvement of miR159 in the regulation of its putative target TaGAMYB and its relationship to wheat development were investigated. First, we demonstrated that cleavage of TaGAMYB1 and TaGAMYB2 was directed by miR159 using 5′-RACE and a transient expression system. Second, we overexpressed TamiR159, TaGAMYB1 and mTaGAMYB1 (impaired in the miR159 binding site) in transgenic rice, revealing that the accumulation in rice of mature miR159 derived from the precursor of wheat resulted in delayed heading time and male sterility. In addition, the number of tillers and primary branches in rice overexpressing mTaGAMYB1 increased relative to the wild type. Our previous study reported that TamiR159 was downregulated after two hours of heat stress treatment in wheat (Triticum aestivum L.). Most notably, the TamiR159 overexpression rice lines were more sensitive to heat stress relative to the wild type, indicating that the downregulation of TamiR159 in wheat after heat stress might participate in a heat stress-related signaling pathway, in turn contributing to heat stress tolerance.

New insights into iron transport from maternal tissues to endosperm in mature wheat seed using synchrotron radiation

Iron deficiency is estimated to affect over half of the world's population (Zheng et al., 2010). Wheat flour is the main ingredient of bread, and is a staple food for many civilizations. In wheat grains, iron is located in the outer layer, called bran and is lost substantially during milling and processing (Zhang et al., 2010). Thus, the flour is almost devoid of iron. Wheat grain with iron-enriched endosperm is of great interest. It is desirable to understand the bottlenecks which prevent iron translocation from the outer bran layers into the endosperm in cereals. Synchrotron radiation can be useful in identifying the transport and state of metals in plant tissues. We generated element localization maps in grains of contrasting genotypes of wheat by using synchrotron powered beam line, VESPERS (very sensitive elemental and structural probe employing radiation), at Canadian Light Source, Saskatoon, Canada. The X-ray Fluorescence analysis was focused at crease, aleurone and endosperm of mature seeds from Triticum aestivum and Aegilops kotschyi. In high iron genotypes, iron was present at higher concentration in the vascular cells than the aleurone. In contrast, in the low iron genotypes, most of the Fe was mobilized from vascular cells to aleurone cells. To further validate this result and to get accurate quantification of Fe in different tissues, micro-PIXE (Particle induced x-ray emission) analysis of the same samples is in progress at the Biotechnical Faculty and Jozef Stefan Institute, Ljubljana, Slovenia. We recorded XANES (X-ray Absorption Near Edge Structure) spectra at different points of the crease and aleurone layer. The oxidation state of iron in all cases was Fe 3+. The pre-edge structures showed one bump, suggesting tetraedric or octaedric Fe-oxygen coordination. To know the chemical coordination environment around iron centre in the cells of different tissues (such as vascular cells, transfer cells, aleurone cells, and endosperm cells), EXAFS (extended X-ray absorption fine structure) is being run on the wheat grain samples.

Proteomic Profiling of the Aleurone Layer of Mature Arabidopsis thaliana Seed

The aleurone layer of mature Arabidopsis thaliana seed plays important roles in seed germination and dormancy. However, the proteomic profile of this cell layer is unknown partly because it is difficult to separate this thin cell layer from the mature seeds. In this study, we have used a simple technique to separate the aleurone layer along with the seed coat following germination of seeds and determined for the first time the putative protein composition of this cell layer. By subjecting the total proteins extracted from the seed coat to 2D gel electrophoresis followed by liquid chromatography/tandem mass spectrometry, we identified four AGI loci, AT4G28520, AT5G44120, AT1G03880, and AT1G03890; all of which belong to the seed storage family of proteins. Because in Arabidopsis the diploid aleurone cells of the seed coat perform protein storage functions similar to that of triploid endosperm of other plant species, it is assumed that the above AGI loci are associated with the aleurone layer of the seed coat.

Comparative phytohormone profiles, lipid kinase and lipid phosphatase activities in barley aleurone, coleoptile, and root tissues

We analyzed lipid kinase and lipid phosphatase activities and determined endogenous phytohormone levels by liquid chromatography-tandem mass spectrometry in root and coleoptile tissues following germination of barley (Hordeum vulgare) seeds. The enzymes showing highest activity in aleurone cells were diacylglycerol kinase (DAG-k, EC 2.7.1.107) and phosphatidate kinase (PA-k). The ratio of gibberellins (GAs) to abscisic acid (ABA) was 2-fold higher in aleurone than in coleoptile or root tissues. In coleoptiles, phosphatidylinositol 4-kinase (PI4-k, EC 2.7.1.67) showed the highest enzyme activity, and jasmonic acid (JA) level was higher than in aleurone. In roots, activities of PI4-k, DAG-k, and PA-k were similar, and salicylic acid (SA) showed the highest concentration. In the assays to evaluate the hydrolysis of DGPP (diacylglycerol pyrophosphate) and PA (phosphatidic acid) we observed that PA hydrolysis by LPPs (lipid phosphate phosphatases) was not modified; however, the diacylglycerol pyrophosphate phosphatase (DGPPase) was strikingly higher in coleoptile and root tissues than to aleurone. Relevance of these findings in terms of signaling responses and seedling growth is discussed.