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Illuminating the mechanisms of inflorescence architecture of grain crops that feed our world may strengthen the goal towards sustainable agriculture. Lateral spikelet development of barley (Hordeum vulgare L.) is such an example of a floral architectural trait regulated by VRS1 (Vulgare Row-type Spike 1 or Six-rowed Spike 1, syn. HvHOX1). Its lateral spikelet-specific expression and the quantitative nature of suppressing spikelet development were previously shown in barley. However, the mechanistic function of this gene and its paralog HvHOX2 on spikelet development is still fragmentary.Here, we show that these duplicated transcription factors (TFs) have contrasting nucleotide diversity in various barley genotypes and several Hordeum species. Despite this difference, both proteins retain their basic properties of the homeodomain leucine zipper class I family of TFs. During spikelet development, these genes exhibit similar spatiotemporal expression patterns yet with anticyclic expression levels. A gene co-expression network analysis suggested that both have an ancestral relationship but their functions appear antagonistic to each other, i.e., HvHOX1 suppresses whereas HvHOX2 rather promotes spikelet development. Our transgenic promoter-swap analysis showed that HvHOX2 can restore suppressed lateral spikelets when expression levels are increased; however, at its low endogenous expression level, HvHOX2 appears dispensable for spikelet development. Collectively, this study proposes that the dosage of the two antagonistic TFs, HvHOX1 and HvHOX2, influence spikelet development in barley.

High-throughput root phenotyping in the soil became an indispensable quantitative tool for the assessment of effects of climatic factors and molecular perturbation on plant root morphology, development and function. To efficiently analyse a large amount of structurally complex soil-root images advanced methods for automated image segmentation are required. Due to often unavoidable overlap between the intensity of fore- and background regions simple thresholding methods are, generally, not suitable for the segmentation of root regions. Higher-level cognitive models such as convolutional neural networks (CNN) provide capabilities for segmenting roots from heterogeneous and noisy background structures, however, they require a representative set of manually segmented (ground truth) images. Here, we present a GUI-based tool for fully automated quantitative analysis of root images using a pre-trained CNN model, which relies on an extension of the U-Net architecture. The developed CNN framework was designed to efficiently segment root structures of different size, shape and optical contrast using low budget hardware systems. The CNN model was trained on a set of 6465 masks derived from 182 manually segmented near-infrared (NIR) maize root images. Our experimental results show that the proposed approach achieves a Dice coefficient of 0.87 and outperforms existing tools (e.g., SegRoot) with Dice coefficient of 0.67 by application not only to NIR but also to other imaging modalities and plant species such as barley and arabidopsis soil-root images from LED-rhizotron and UV imaging systems, respectively. In summary, the developed software framework enables users to efficiently analyse soil-root images in an automated manner (i.e. without manual interaction with data and/or parameter tuning) providing quantitative plant scientists with a powerful analytical tool.

Background Barley (Hordeum vulgare) is an important crop cultivated across the world. Drought is a major abiotic factor compromising barley yield worldwide, therefore in modern spring barley cultivars superior seed and malting quality characteristics should be combined with reasonable level of drought tolerance. Previously we have identified a number of barley lines demonstrating the superior yield performance under drought conditions. The aim of this work was to perform a QTL analysis of malting quality traits in a doubled haploid (DH) mapping population of two elite barley lines that differ in their reaction pattern to drought stress. Results A population of DH lines was developed by crossing two drought-tolerant elite breeding lines, Victoriana and Sofiara, exploiting distinct mechanism of drought tolerance, sustaining assimilation vs remobilization. The mapping population was assayed under field conditions at four distinct locations that differed in precipitation rate. DH lines were genotyped with the Illumina 9 K iSelect assay, and linkage map including 1782 polymorphic markers and covering a total map length of 1140 cM was constructed. The result of quantitative trait loci (QTL) analysis showed that majority of the traits were affected by several main effect QTL and/or QTL x environment (QE) interactions. In total, 57, 41, and 5 QTL were associated with yield-related traits, malting quality traits and seed quality traits, respectively. 11 and 29 of mapped QTL explained more than 10 and 5% of phenotypic variation, respectively. In several chromosomal regions co-localization between QTL for various traits were observed. The largest clusters were detected on chromosomes 3H and 4H. Conclusions Our QTL mapping results revealed several novel consistent genomic regions controlling malting quality which could be exploited in marker assisted selection. In this context, the complex QTL region on chromosome 3H seems of particular interest, as it harbors several large effect QTL. Electronic supplementary material The online version of this article (10.1186/s12870-018-1323-4) contains supplementary material, which is available to authorized users.

Quantitative characterization of root system architecture and its development is important for the assessment of a complete plant phenotype. To enable high-throughput phenotyping of plant roots efficient solutions for automated image analysis are required. Since plants naturally grow in an opaque soil environment, automated analysis of optically heterogeneous and noisy soil-root images represents a challenging task. Here, we present a user-friendly GUI-based tool for semi-automated analysis of soil-root images which allows to perform an efficient image segmentation using a combination of adaptive thresholding and morphological filtering and to derive various quantitative descriptors of the root system architecture including total length, local width, projection area, volume, spatial distribution and orientation. The results of our semi-automated root image segmentation are in good conformity with the reference ground-truth data (mean dice coefficient = 0.82) compared to IJ_Rhizo and GiAroots. Root biomass values calculated with our tool within a few seconds show a high correlation (Pearson coefficient = 0.8) with the results obtained using conventional, pure manual segmentation approaches. Equipped with a number of adjustable parameters and optional correction tools our software is capable of significantly accelerating quantitative analysis and phenotyping of soil-, agar- and washed root images.

Transcriptome analysis reflects the status quo of transcribed genetic code present in the form of mRNA, which helps to infer biological processes and unravel metabolic status. Despite the increasing adoption of RNA-Seq technique in recent years, transcriptome analysis using the microarray platform remains the gold standard technique, which offers a simpler, more cost-effective, and efficient method for high-throughput gene expression profiling. In this chapter, we described a streamlined transcriptomic analyses pipeline employed to study developing rice grains that can also be applied to other tissue samples and species. We described a novel RNA extraction method that obviates the problem introduced by high-starch content during rice grain development that usually leads to reduction in RNA yield and quality. The detailed procedure of microarray analysis involved in cDNA synthesis, cRNA labeling, microarray hybridization, slide scanning, feature extraction to QC validation has been described. The description of a newly developed Indica- and Japonica-specific microarray slides developed from the genome information of subpopulation to study gene expression of 60,000 genes has been highlighted. The downstream bioinformatics analyses including expression QTL mapping and gene regulatory network analyses were mentioned.

DNA methylation is an important biomarker for gene activity. It contributes to gene silencing and is involved in regulating various seed developmental processes in plants. Many of these processes are involved in important traits associated with aspects of grain quality. A reliable, fast, and cheap method is the estimation of DNA methylation utilizing methylation sensitive restriction enzymes (MSRE) and quantitative real-time PCR (qPCR) for selected candidate regions. The presented method can be used to confirm an effect of RNAi constructs on their target genes or trans-activity. Analysis of promoter regions can contribute to estimation of gene activity and related traits.

DNA methylation is an important biomarker for gene activity. It contributes to gene silencing and is involved in regulating various seed developmental processes in plants. Many of these processes are involved in important traits associated with aspects of grain quality. A reliable, fast, and cheap method is the estimation of DNA methylation utilizing methylation sensitive restriction enzymes (MSRE) and quantitative real-time PCR (qPCR) for selected candidate regions. The presented method can be used to confirm an effect of RNAi constructs on their target genes or trans-activity. Analysis of promoter regions can contribute to estimation of gene activity and related traits.

Transcriptome analysis reflects the status quo of transcribed genetic code present in the form of mRNA, which helps to infer biological processes and unravel metabolic status. Despite the increasing adoption of RNA-Seq technique in recent years, transcriptome analysis using the microarray platform remains the gold standard technique, which offers a simpler, more cost-effective, and efficient method for high-throughput gene expression profiling. In this chapter, we described a streamlined transcriptomic analyses pipeline employed to study developing rice grains that can also be applied to other tissue samples and species. We described a novel RNA extraction method that obviates the problem introduced by high-starch content during rice grain development that usually leads to reduction in RNA yield and quality. The detailed procedure of microarray analysis involved in cDNA synthesis, cRNA labeling, microarray hybridization, slide scanning, feature extraction to QC validation has been described. The description of a newly developed Indica- and Japonica-specific microarray slides developed from the genome information of subpopulation to study gene expression of 60,000 genes has been highlighted. The downstream bioinformatics analyses including expression QTL mapping and gene regulatory network analyses were mentioned.

Strigolactones (SLs) represent a class of plant hormones that are involved in inhibiting shoot branching and in promoting abiotic stress responses. There is evidence that the biosynthetic pathways of SLs and abscisic acid (ABA) are functionally connected. However, little is known about the mechanisms underlying the interaction of SLs and ABA, and the relevance of this interaction for shoot architecture. Based on sequence homology, four genes (HvD27, HvMAX1, HvCCD7, and HvCCD8) involved in SL biosynthesis were identified in barley and functionally verified by complementation of Arabidopsis mutants or by virus-induced gene silencing. To investigate the influence of ABA on SLs, two transgenic lines accumulating ABA as a result of RNAi-mediated down-regulation of HvABA 8’-hydroxylase 1 and 3 were employed. LC-MS/MS analysis confirmed higher ABA levels in root and stem base tissues in these transgenic lines. Both lines showed enhanced tiller formation and lower concentrations of 5-deoxystrigol in root exudates, which was detected for the first time as a naturally occurring SL in barley. Lower expression levels of HvD27, HvMAX1, HvCCD7, and HvCCD8 indicated that ABA suppresses SL biosynthesis, leading to enhanced tiller formation in barley.

Terminal drought stress during grain filling is the major abiotic factor that limits crop yield in barley. The mother plant acclimates to the environment and perceives signals that result in a change of the physiological state within the grain and therefore affect the seed development and germination of the next generation. Small regulatory RNAs have been described to be involved in plant drought stress response by suppressing the respective target genes. Based on their origin and function, these small RNAs are classified as micro RNAs (miRNA), short interfering RNAs (siRNA) or heterochromatic small interfering RNA (hc-siRNA). In addition, 24mer sized hc-siRNAs are associated with RNA directed DNA methylation (RdDM) and transcriptional gene silencing (TGS). The analysis of hc-siRNA by small RNA sequencing in barley caryopses after imposition of terminal drought stress allowed the identification of stress specific 24mers. Based on the sequence homology of the siRNAs to the promoter region of CYTOKININ-OXIDASE 2.1 (HvCKX2.1), this putative target gene was selected for further investigation. Terminal drought stress leads to an increased level of DNA methylation at the HvCKX2.1 promoter and the seeds derived from drought stressed plants showed faster shoot emergence. Accumulation of cytokinin ribosides, which are the known substrates of cytokinin-oxidase, can explain the observed phenotype of faster shoot emergence from seeds of drought stressed mother plants. Analysis of transgenic plants with modulated levels of abscisic acid (ABA) in the grain confirmed the ABA/drought stress responsive ProHvCKX2.1 methylation and correlation with shoot emergence speed.

Root systems architecture (RSA) and size properties are essential determinants of plant performance and need to be assessed in high-throughput plant phenotyping platforms. Thus, we tested a concept that involves near-infrared (NIR) imaging of roots growing along surfaces of transparent culture vessels using special long pass filters to block their exposure to visible light. Two setups were used to monitor growth of Arabidopsis, rapeseed, barley and maize roots upon exposure to white light, filter-transmitted radiation or darkness: root growth direction was analysed (1) through short-term cultivation on agar plates, and (2) using soil-filled transparent pots to monitor long-term responses. White light-triggered phototropic responses were detected for Arabidopsis in setup 1, and for rapeseed, barley and maize roots in setups 1 and 2, whereas light effects could be avoided by use of the NIR filter thus confirming its suitability to mimic darkness. NIR image-derived ‘root volume’ values correlated well with root dry weight. The root system fractions visible at the different pot sides and in different zones revealed species- and genotype-dependent variation of spatial root distribution and other RSA traits. Following this validated concept, root imaging setups may be integrated into shoot phenotyping facilities in order to enable root system analysis in the context of whole-plant performance investigations.

Rice lines with slower starch digestibility provide opportunities in mitigating the global rise in type II diabetes and related non-communicable diseases. However, screening for low glycemic index (GI) in rice breeding programs is not possible due to time and cost constraints. This study evaluated the feasibility of using in vitro cooked grain amylolysis, starch mobilization patterns during seed germination, and variation in starch structure and composition in the mature seed to differentiate patterns of starch digestibility. Mobilization patterns of total starch, resistant starch, amylose and amylopectin chains, and free sugars during seed germination revealed that the process is analogous to digestion in the human gastrointestinal tract. The combination of these biochemical markers can be used as an alternative measure to predict GI. Additionally, transcriptome analysis of stored mRNA transcripts in high and low GI lines detected differences in starch metabolism and confirmed the importance of seed storage pathways in influencing digestibility. Pathway analyses supported by metabolomics data revealed that resistant starch, cell wall non-starch polysaccharides and flavonoids potentially contribute to slower digestibility. These new insights can guide precision breeding programs to produce low GI rice with acceptable cooking quality to help mitigate the burden of diet-associated lifestyle diseases.

Eltern von Kindern mit Muskelhypotonie stellen sich die folgenden Fragen: Wir haben die Fruhbehandlung verpasst, ist unserem Kind noch zu helfen? Wird unser Kind immer motorische Schwierigkeiten haben? Kann sich die verzogerte Entwicklung auf den Verstand auswirken? Kommt es infolge der motorischen Probleme auch zu einem geistigen Stillstand, zum Zuruckbleiben in der geistigen Entwicklung? Wie wird unser Sprossling in einer Gruppe mit anderen Kindern zurechtkommen? Wirkt sich das langsame Vorankommen auf das schulische Lernen aus? Bremst es etwa die Schulkarriere? Erwarten unser Kind auch im spateren Leben Probleme mit der Alltagsbewaltigung?

Kinder mit Muskelhypotonie, Entwicklungsverzogerung und Behinderung halten oft viele Jahre lang eine enge Symbiose mit ihren Eltern aufrecht. Wenn das ausere und innere Gleichgewicht fehlen und jede Veranderung der Korperhaltung Unsicherheit hervorruft, reagieren die meisten Kinder intuitiv mit anklammerndem Verhalten. Die Angst zu fallen sitzt tief. Sie ist aus kindlicher Sicht berechtigt, denn infolge der motorischen Instabilitat ist auch die Wahrnehmung fur den eigenen Korper eingeschrankt. Auf dem Arm gehalten, im kuscheligen Korperkontakt fuhlt sich jedes Kind wohl. Mit zunehmender Mobilitat lockern die meisten Kleinkinder die naturliche Symbiose selbst. Bei Entwicklungsverzogerung verspatet sich die Eroberung der Umwelt, der Aktionsradius bleibt klein, das Kind will die enge Symbiose nicht losen.

Gut, dass Sie mir anfangs nicht gesagt haben, dass Sie am Therapieerfolg meiner Tochter zweifelten! Ich hatte grose Hoffnung in den neuen Therapieansatz nach Castillo Morales gesetzt, nachdem es mit der herkommlichen neurophysiologischen Behandlung keine nennenswerten Fortschritte gab. Schonungslos hatte ein Arzt bei der Fruhdiagnostik mir gegenuber die Vermutung geausert: „Ihr Kind wird nie laufen, sondern im Rollstuhl sitzen.“ Vielleicht hatte ich ohne Aussicht auf Verbesserung den Glauben an Angels Forderung aufgegeben. Fachlich fundierte Elternberatung ist vonnoten. Jedoch fuhlen sich viele Eltern nach einer arztlich gut gemeinten diagnostischen Aufklarung „im Regen stehend“. Der grundliche Facharzt nennt nicht selten alle prognostischen Eventualitaten, die das kleine Baby treffen konnten. Ausspruche wie „Ihr Kind wird nie …“ zerstoren Hoffnung und Lebensfreude. Wer weis denn sicher am Anfang des Lebens, wie viel oder wie wenig sich durch kontinuierliches Uben erreichen lasst oder sich von selbst entwickelt?

Eine ausgepragte muskulare Hypotonie vom Ausmas einer globalen Bewegungsstorung beginnt sich bei dem Kleinkind zu etablieren. Alle Korperbereiche sind betroffen – der Mund, die Zunge und die Mimik jedoch nicht. Elena kann mit den Augen sprechen! Sie beobachtet genau ihre Umgebung und wirkt nachdenklich, in sich versunken. Trotz ihrer Passivitat ist sie ein zufriedenes Kind, das nicht fordert, nicht schreit und stets zufrieden erscheint. Das kleine Madchen liebt zwei Korperhaltungen, die es selbst nicht verandert: auf dem Schos der Eltern kuscheln oder auf dem Rucken liegend in die wedelnden Hande klatschen. Stundenlang kann es liegend verweilen, den freundlichen Stimmen lauschen und die Gerausche des Haushalts aufnehmen. Es scheint nichts zu vermissen. Die liebevolle Zuwendung der Eltern ist ihm sicher. Es bleibt frohlich und genugsam auf dem Rucken liegen, ohne den Wunsch zu verspuren, sich aufzurichten. Diese Art der Antriebstorung blockiert die ohnehin schleppende Entwicklung.

Die willkurliche Muskulatur reagiert auf Wahrnehmung von Druck und Zug, Widerstand, Bewegung und Vibration mit erhohter Spannung oder Entspannung. Druck auf die Fussohle regt z. B. die Korperaufrichtung an. Bei Widerstand baut sich Kraft auf. Bei Zug kommt es als Gegenreaktion zu vermehrtem Zusammenziehen von Muskeln. Neurophysiologische Behandlung nutzt mechanische Stimulierung zur Tonusregulation. Bei der Vojta-Methode wird Druck angewendet. Das Castillo Morales®-Konzept nutzt Vibration zur Tonusregulation. Vibration starkt schwache Muskulatur bei Muskelhypotonie; sie kann auch spastische, hypertone Muskeln entspannen. Es kommt auf den Rhythmus und die Dosierung der manuellen Stimulierung an. Vibration wird von Sauglingen und Kindern gut akzeptiert. Das Ungeborene wurde von vibrierendem Pulsieren umgeben. Bei Fruhgeborenen sollten Eltern und Therapeuten bereits im Brutkasten mit vibratorischer Anregung beginnen.

Umsichtige Eltern schauen sich um, welche Einrichtung mit den Entwicklungsproblemen ihres Sprosslings umgehen kann. Sie entscheiden nicht nach optischen Kriterien wie der Ausstattung, sondern nach der personellen Besetzung und der Fachkenntnis der Erzieherinnen: Wird mein Kind in der Kita dort abgeholt, wo es in seiner Entwicklung steht? Gewahrt man ihm eine zeitlich ausreichende Ubergangsphase zur Eingewohnung, in der ein Elternteil anwesend ist? Hat das Kind ein bis zwei nicht wechselnde Bezugspersonen, an die es sich jederzeit wenden kann? Ist eine zusatzliche sonderpadagogisch geschulte Fachfrau vor Ort, die sich um mein Kind zumindest stundenweise kummert? Werden die beantragten Kosten fur eine Integrationshilfe oder die Inklusion rechtzeitig mit dem Kindergarteneintritt ubernommen? Welches padagogische Konzept und welche ethischen Erziehungsziele hat sich der Kindergarten auf die Fahne geschrieben? Sollen die Kinder selbsttatig mit freien Wahlmoglichkeiten Spiel- und Umgangsformen erlernen, oder werden sie in familiar organisierten Gruppen mit konstant anwesenden Kontakterzieherinnen angeleitet?

Wie gut, wenn Eltern entwicklungsverzogerter Kinder ihre Erfahrungen austauschen. Das Internet bietet nahezu unbegrenzte Moglichkeiten, sodass heute keine Familie in Isolation stecken bleiben muss. Einige Fragen bleiben jedoch ohne personlichen Kontakt und ohne besondere Hilfestellung in nebuloser Unsicherheit stecken: Gibt es Hilfen, die das alltagliche Handling eines entwicklungsverzogerten Kindes erleichtern? Welche Hilfsmittel wecken Bewegungsfreude bei einem antriebslosen Kind? Welche Fahrgerate sind fur Kinder mit Gleichgewichtsproblemen geeignet? Gibt es Alternativen zum obligatorischen Laufradfahren? Welche Angebote des Spielgeratemarktes sind sinnvoll, und welche sind weniger oder nicht empfehlenswert? Kann das hypotone Kleinkind aus einem Becher trinken lernen?

Wir werden seit Fabians drittem Lebensmonat im Castillo Morales Zentrum super betreut. Zwischendurch versuchten wir es mal mit einer Behandlung nach Bobath. Diese Therapie hat nach unserer Meinung nicht so gut gepasst. Das Team im Castillo Morales Zentrum ist super und gibt uns sehr hilfreiche Ubungen fur Zuhause mit. Allerdings glauben wir, dass noch mehr drin ist, und darum schreibe ich Ihnen. Was Sie schreiben, hat Hand und Fus, und wir sind uberzeugt, dass wir von Ihren Erfahrungen profitieren konnen. Das groste Manko ist nach unserer Meinung der schwache Schulterbereich, einschlieslich der Arme. Auch die Haltung des Rumpfes ist steif und unflexibel. Oft fehlt Fabian die Kraft, Handlungen, die er sich vornimmt, umzusetzen. Er wirkt mit seinen ruckartigen Bewegungen eher tollpatschig.

Manche Eltern beschreiben ihr Baby mit Muskelhypotonie in den ersten Lebensmonaten als pflegeleicht, wenn es sich ruhig und wenig anspruchsvoll verhalt. Aber sind die alltagliche Korperpflege, das Saugen und Trinken, das Tragen und Halten wirklich so leicht? Lassen Sie uns genauer hinschauen: Bewegungsanpassungen beim Anziehen bleiben aus. Das Kind hilft sozusagen nicht mit, streckt die Arme nicht vor, die Finger verfangen sich im Armel. Es hebt das Gesas beim Wickeln und Hoseanziehen nicht an. Es kann keine Brucke machen. Das Baby strampelt wenig. Die Beine liegen gestreckt wie auseinandergefallen auf der Matratze. Es verdreht die Fuse nach ausen. In Ruckenlage stemmt es die Fersen nicht gegen den Widerstand der Unterlage und beim Wickeln nicht gegen den Korper der Eltern. Beim Hochnehmen und Tragen wirkt die Haltung puppenartig mit schlenkernden, baumelnden Gliedmasen. Das Kleine fuhlt sich schwer an und droht zu entgleiten. Es halt sich gar nicht fest, sondern lehnt sich an den Korper des Erwachsenen an oder uberstreckt sich.

Abscisic acid (ABA) is a central player in plant responses to drought stress. How variable levels of ABA under short-term versus long-term drought stress impact assimilation and growth in crops is unclear. We addressed this through comparative analysis, using two elite breeding lines of barley (Hordeum vulgare) that show senescence or stay-green phenotype under terminal drought stress and by making use of transgenic barley lines that express Arabidopsis (Arabidopsis thaliana) 9-cis-epoxycarotenoid dioxygenase (AtNCED6) coding sequence or an RNA interference (RNAi) sequence of ABA 8'-hydroxylase under the control of a drought-inducible barley promoter. The high levels of ABA and its catabolites in the senescing breeding line under long-term stress were detrimental for assimilate productivity, whereas these levels were not perturbed in the stay-green type that performed better. In transgenic barley, drought-inducible AtNCED expression afforded temporal control in ABA levels such that the ABA levels rose sooner than in wild-type plants but also subsided, unlike as in the wild type , to near-basal levels upon prolonged stress treatment due to down-regulation of endogenous HvNCED genes. Suppressing of ABA catabolism with the RNA interference approach of ABA 8'-hydroxylase caused ABA flux during the entire period of stress. These transgenic plants performed better than the wild type under stress to maintain a favorable instantaneous water use efficiency and better assimilation. Gene expression analysis, protein structural modeling, and protein-protein interaction analyses of the members of the PYRABACTIN RESISTANCE1/PYRABACTIN RESISTANCE1-LIKE/REGULATORY COMPONENT OF ABA RECEPTORS, TYPE 2C PROTEIN PHOSPHATASE Sucrose non-fermenting1-related protein kinase2, and ABA-INSENSITIVE5/ABA-responsive element binding factor family identified specific members that could potentially impact ABA metabolism and stress adaptation in barley.

Inflorescence architecture of barley ( Hordeum vulgare L.) is common among the Triticeae species, which bear one to three single-flowered spikelets at each rachis internode. Triple spikelet meristem is one of the unique features of barley spikes, in which three spikelets (one central and two lateral spikelets) are produced at each rachis internode. Fertility of the lateral spikelets at triple spikelet meristem gives row-type identity to barley spikes. Six-rowed spikes show fertile lateral spikelets and produce increased grain yield per spike, compared with two-rowed spikes with sterile lateral spikelets. Thus, far, two loci governing the row-type phenotype were isolated in barley that include Six-rowed spike1 ( Vrs1 ) and Intermedium-C . In the present study, we isolated Six-rowed spike4 ( Vrs4 ), a barley ortholog of the maize ( Zea mays L.) inflorescence architecture gene RAMOSA2 ( RA2 ). Eighteen coding mutations in barley RA2 ( HvRA2 ) were specifically associated with lateral spikelet fertility and loss of spikelet determinacy. Expression analyses through mRNA in situ hybridization and microarray showed that Vrs4 ( HvRA2 ) controls the row-type pathway through Vrs1 ( HvHox1 ), a negative regulator of lateral spikelet fertility in barley. Moreover, Vrs4 may also regulate transcripts of barley SISTER OF RAMOSA3 ( HvSRA ), a putative trehalose-6-phosphate phosphatase involved in trehalose-6-phosphate homeostasis implicated to control spikelet determinacy. Our expression data illustrated that, although RA2 is conserved among different grass species, its down-stream target genes appear to be modified in barley and possibly other species of tribe Triticeae.

Drought is a major limiting factor for barley production, especially in the primary areas of its cultivation. Wild barley represents a major source of favourable alleles for increasing the genetic variation for multiple traits including resistance to both biotic and abiotic stresses. We used advanced backcross quantitative trait locus (AB-QTL) analysis of a BC3-doubled haploid population developed between the cultivated parent Brenda (Hordeum vulgare ssp. vulgare) and the wild accession HS584 (H. vulgare ssp. spontaneum) to study the contribution of wild barley in improving various agronomic and seed quality traits under post-anthesis drought. The experiment was carried out at two different locations (IPK, Gatersleben and Nordsaat, Bohnshausen) and terminal drought was imposed by withholding water or spraying with potassium iodide at 10 days after flowering under greenhouse or field conditions, respectively. QTL analysis indicated that wild barley contributed favourably to most of the traits studied under both control and drought conditions. A total of seven hot-spot QTL regions with co-localizing QTL for various traits harboured more than 80 % of the stable QTL detected in the present study. For yield and thousand-grain weight and their respective drought tolerance indices, most of the QTL were derived from Brenda. On the other hand, for traits like seed length and seed nitrogen content, all the QTL were contributed by HS584, the parent having higher trait value. A significantly reduced carbon/nitrogen (C/N) ratio in the selected contrasting inferior lines compared to superior ones suggests that C/N ratio could be a potential parameter for screening not just seed quality parameters but also grain weight performance under terminal drought.

Recent developments in defining the functional basis of abscisic acid in regulating growth, development and stress response have provided essential components for its actions. We are yet to envision the impact of how differential levels of ABA influence plant growth across life cycle. Here we reviewed the information arising from the recent unprecedented advancement made in the field of ABA signaling operative under calcium-dependent and calcium-independent pathways mediating the transcriptional reprogramming under short-term stress response. Advancement made in the field of ABA receptors and transporters has started to fill major gaps in our understanding of the ABA action. However, ABA just not only regulates guard cell movement but impacts other reproductive tissue development through massive transcriptional reprogramming events affecting various stages of the plant life cycle. Therefore many questions still remain unanswered. One such intriguing question is the contradictory role of ABA known to mediate two opposite faces of the coin: regulating abiotic stress tolerance and imparting growth retardation. In this review, we critically assessed the impact of substantial elevated levels of ABA on impairment of photosynthesis and growth alteration and its subsequent influence on seed yield formation. Excess biosynthesis of ABA under stress may deprive the same precursor pool necessary for chlorophyll biosynthesis pathway, thereby triggering growth retardation. Further, we emphasized the importance of ABA homeostasis for integrating stress cues towards coordinating sustainable plant growth. Also we provided a pertinent background on ABA biosynthesis and degradation pathway manipulation to highlight the genes and processes used in genetic engineering of plants for changed ABA content.

Recent studies suggest that abscisic acid (ABA) at its basal level plays an important role during seed set and grain filling events. Under drought stress ABA levels were found to be significantly enhanced in the developing seed. Until now we lacked an understanding of (1) ABA homeostasis in developing seeds under terminal drought and (2) the interactive role of ABA in regulating the starch biosynthesis pathway in developing grains under terminal drought. We have recently reported the possible regulation of ABA homeostasis in source (flag leaf) and sink (developing grains) tissues under post-anthesis drought stress in barley and concluded that significantly enhanced ABA levels in developing grains are due to strong activation of the ABA deconjugation pathway and fine regulation of the ABA biosynthesis-degradation pathway.1 Additionally, we provided evidence for the role of ABA in differential regulation of starch biosynthesis genes and a significant upregulation of starch degradation beta amylase genes under drought, i.e., ABA not only influences the rate of starch accumulation but also starch quality.

Repression of ABA degradation in transgenic barley alters the expression of genes involved in strigolactone biosynthesis, revealing cross-talk between ABA and strigolactone that affects the development of tillers., Strigolactones (SLs) represent a class of plant hormones that are involved in inhibiting shoot branching and in promoting abiotic stress responses. There is evidence that the biosynthetic pathways of SLs and abscisic acid (ABA) are functionally connected. However, little is known about the mechanisms underlying the interaction of SLs and ABA, and the relevance of this interaction for shoot architecture. Based on sequence homology, four genes (HvD27, HvMAX1, HvCCD7, and HvCCD8) involved in SL biosynthesis were identified in barley and functionally verified by complementation of Arabidopsis mutants or by virus-induced gene silencing. To investigate the influence of ABA on SLs, two transgenic lines accumulating ABA as a result of RNAi-mediated down-regulation of HvABA 8’-hydroxylase 1 and 3 were employed. LC-MS/MS analysis confirmed higher ABA levels in root and stem base tissues in these transgenic lines. Both lines showed enhanced tiller formation and lower concentrations of 5-deoxystrigol in root exudates, which was detected for the first time as a naturally occurring SL in barley. Lower expression levels of HvD27, HvMAX1, HvCCD7, and HvCCD8 indicated that ABA suppresses SL biosynthesis, leading to enhanced tiller formation in barley.

We herein report on a 23-year-old female patient who suffered from Crohn's disease and anorexia nervosa and died after long-term malnutrition and a perforated colitis. At autopsy, her pancreas displayed two peculiar findings. First, there was a constant and aberrant expression of CA 19-9 in the acinar cells. The expression of the carbohydrate antigen CA 19-9 is normally confined to duct-like epithelia, including centroacinar cells, while islet and acinar cells have repeatedly been reported to be immunohistochemically negative. Thus, to the best of our knowledge, our case is the first to show aberrant acinar CA 19-9 expression, and its potential meaning is discussed. Second, the pancreas showed a heterogeneity in acinar morphology, with peri-insular acini being considerably larger than tele-insular acini. The existence of enlarged peri-insular acini, mainly in animals, has occasionally been reported. Its origin, however, is still unclear. Some authors have proposed an influence of high insulin concentrations, exerted via the insulo-acinar capillary axis. We agree with the concept of an islet-derived mechanism. However, as we have observed a similar heterogeneity in streptozotocin- and autoimmune-diabetic rats, we presume that other islet hormones, in particular glucagon, might be more important for this phenomenon in the animals, as well as in the present case.

We present new estimates on the national road kill for nine large and medium-sized mammals in Sweden. Our estimates are based on 705 drivers' reports on the number of animals accidentally hit during individually chosen reference periods. During 1960–2000, a total of 881 animal-vehicle collisions were reported based on 243.6 million driven kilometres, representing 0.37% of the overall mileage driven in Sweden during 1992, the mean reference year of all replies. The collision frequencies ranged from 0.07 incidents per million kilometres for medium-sized mustelids, over 0.42 for badgers Meles meles to 1.11 for hares Lepus spp. Our data suggest that during 1992, 7,000–13,500 moose Alces alces, 43,500–59,000 roe deer Capreolus capreolus, 63,500–81,500 hares, 22,000–33,000 badgers and 6,500–12,500 red foxes Vulpes vulpes may have been killed on Swedish roads. Among these game species, the extrapolated nation-wide road kill ranged between 7 and 97% of the average annual harvest, and between 1 and 12% of the assessed total populations in 1992. Our results are in agreement with other independent road kill estimates for Sweden. The data suggested an overall increase in the frequency of road kills over the past 40 years, which partly can be attributed to changes in traffic volume and wildlife population sizes (game bags). In most species, the estimated levels of nation-wide road mortality are not alarmingly high, although local impacts may be significant. In badgers and hares, the ratio of the estimated road kill to the annual harvest increased two fold, suggesting a steady increase in the relative importance of road mortality. We conclude that interviews with drivers can provide a cheap and useful index of wildlife traffic mortality.

Tumour progression can be investigated in liposarcomas showing a transition from a low-grade well-differentiated (WD) to a high-grade dedifferentiated (DD) variant. As RB1 gene alterations are common defects in sarcomas, this study examined the frequency of RB1 loss of heterozygosity (LOH) in a group of 14 well-differentiated liposarcomas (WDLs) and 17 well-differentiated/dedifferentiated liposarcomas (WD/DDLs), using a microdissection approach (PALM laser pressure catapulting) that allows the two histological components to be separated for polymerase chain reaction (PCR) analysis. In addition, RB1 protein expression and the Mib1 proliferation index were determined by immunohistochemistry and interphase FISH was performed with an RB1 probe at 13q14. By the use of four intragenic polymorphic RB1 markers (introns 1, 17, 20, and 25) for PCR, allelic losses were found only in the DD parts, but never in the pure WDLs or in the WD components of the WD/DDLs investigated. Furthermore, DD areas characterized by a heterogeneous RB1 protein expression pattern (35–65% immunopositivity), as compared with 90–100% RB1 positivity in WD areas, showed a marked increase in Mib1 proliferation index (19.6% versus 1.8% in WD areas; p

Crop plants are regularly challenged by a range of environmental stresses which typically retard their growth and ultimately compromise economic yield. The stress response involves the reprogramming of approximately 4% of the transcriptome. Here, the behavior of AtRD22 and AtUSPL1, both members of the Arabidopsis thaliana BURP (BNM2, USP, RD22 and polygalacturonase isozyme) domain-containing gene family, has been characterized. Both genes are up-regulated as part of the abscisic acid (ABA) mediated moisture stress response. While AtRD22 transcript was largely restricted to the leaf, that of AtUSPL1 was more prevalent in the root. As the loss of function of either gene increased the plant's moisture stress tolerance, the implication was that their products act to suppress the drought stress response. In addition to the known involvement of AtUSPL1 in seed development, a further role in stress tolerance was demonstrated. Based on transcriptomic data and phenotype we concluded that the enhanced moisture stress tolerance of the two loss-of-function mutants is a consequence of an enhanced basal defense response.

In the changing environment, water is the major limiting factor for crop productivity throughout the world, and there is every need to generate climate-resilient crops. Since drought is a complex phenomenon, we need to dissect various mechanisms at the physiological, biochemical, and molecular levels in order to generate crop plants with better drought tolerance but without any yield penalties. Accumulated literature points out that improvement at both source and sink levels are needed to elevate final yields under water deficit conditions. Here, we summarize the current status of plant adaptation mechanisms and the strategies that we need to carve for generating drought stress-tolerant crops like barley.

Drought is one of the most severe environmental stress factors limiting crop yield especially when occurring during anthesis and seed filling. This terminal drought is characterized by an excess production of the phytohormone abscisic acid (ABA) which plays an important role during seed development and dormancy. All the genes putatively involved in ABA biosynthesis and inactivation in barley were identified and their expression studied during plant ontogeny under standard and drought-stress conditions to learn more about ABA homeostasis and the possible mode of cross-talk between source and sink tissues. Out of 41 genes related to ABA biosynthesis and inactivation 19 were found to be differentially regulated under drought stress in both flag leaves and developing seed during seed filling. Transcripts of plastid-located enzymes are regulated similarly in flag leaf and seed under terminal drought whereas transcripts of cytosolic enzymes are differentially regulated in the two tissues. Detailed information on the expression of defined gene family members is supplemented by measurements of ABA and its degradation and conjugation products, respectively. Under drought stress, flag leaves in particular contain high concentrations of both ABA and the ABA degradation products phaseic acid (PA) and diphaseic acid (DPA); whereas, in seeds, besides ABA, DPA was mainly found. The measurements also revealed a positive correlation between ABA level and starch content in developing seeds for the following reasons: (i) genes of the ABA controlled SnRK2.6 and RCAR/PP2C-mediated signal transduction pathway to the ABF transcription factor HvABI5 are activated in the developing grain under drought, (ii) novel ABA- and dehydration-responsive cis-elements have been found in the promoters of key genes of starch biosynthesis (HvSUS1, HvAGP-L1) and degradation (HvBAM1) and these transcripts/activity are prominently induced in developing seeds during 12 and 16 DAF, (iii) spraying of fluridone (an ABA biosynthesis inhibitor) to drought-stressed plants results in severely impaired starch content and thousand grain weight of mature seeds.

Leptomeningeal carcinomatosis is a clinically important and severe complication in patients with cancer. Leptomeningeal involvement as a secondary event in gastric carcinoma is rarely reported and usually occurs late in advanced disease. Herein, we report a case of leptomeningeal carcinomatosis as the initial manifestation of a previously asymptomatic gastric adenocarcinoma. The clinical features and the appropriate diagnostic procedures are discussed.

Tumour progression can be investigated in liposarcomas showing a transition from a low-grade well-differentiated (WD) to a high-grade dedifferentiated (DD) variant. As RB1 gene alterations are common defects in sarcomas, this study examined the frequency of RB1 loss of heterozygosity (LOH) in a group of 14 well-differentiated liposarcomas (WDLs) and 17 well-differentiated/dedifferentiated liposarcomas (WD/DDLs), using a microdissection approach (PALM laser pressure catapulting) that allows the two histological components to be separated for polymerase chain reaction (PCR) analysis. In addition, RB1 protein expression and the Mib1 proliferation index were determined by immunohistochemistry and interphase FISH was performed with an RB1 probe at 13q14. By the use of four intragenic polymorphic RB1 markers (introns 1, 17, 20, and 25) for PCR, allelic losses were found only in the DD parts, but never in the pure WDLs or in the WD components of the WD/DDLs investigated. Furthermore, DD areas characterized by a heterogeneous RB1 protein expression pattern (35-65% immunopositivity), as compared with 90-100% RB1 positivity in WD areas, showed a marked increase in Mib1 proliferation index (19.6% versus 1.8% in WD areas; p0.001). Interphase fluorescence in situ hybridization (FISH) detected a higher RB1-LOH rate in the DD components of WD/DDLs. Considering the different detection sensitivities of the three methodologies, it is concluded that loss of RB1 function already begins in the WDL, and that the tumour cell population with RB1-LOH starts prevailing in the tumour mass during progression of a WDL.

The rapid increase in heat shock proteins upon exposure to damaging stresses and during plant development related to desiccation events reveal their dual importance in plant development and stress tolerance. Genome-wide sequence survey identified 20 non-redundant small heat shock proteins (sHsp) and 22 heat shock factor (Hsf) genes in barley. While all three major classes (A, B, C) of Hsfs are localized in nucleus, the 20 sHsp gene family members are localized in different cell organelles like cytoplasm, mitochondria, plastid and peroxisomes. Hsf and sHsp members are differentially regulated during drought and at different seed developmental stages suggesting the importance of chaperone role under drought as well as seed development. In silico cis-regulatory motif analysis of Hsf promoters showed an enrichment with abscisic acid responsive cis-elements (ABRE), implying regulatory role of ABA in mediating transcriptional response of HvsHsf genes. Gene regulatory network analysis identified HvHsfB2c as potential central regulator of the seed-specific expression of several HvsHsps including 17.5CI sHsp. These results indicate that HvHsfB2c is co-expressed in the central hub of small Hsps and therefore it may be regulating the expression of several HvsHsp subclasses HvHsp16.88-CI, HvHsp17.5-CI and HvHsp17.7-CI. The in vivo relevance of binding specificity of HvHsfB2C transcription factor to HSE-element present in the promoter of HvSHP17.5-CI under heat stress exposure is confirmed by gel shift and LUC-reporter assays. Further, we isolated 477 bp cDNA from barley encoding a 17.5 sHsp polypeptide, which was predominantly upregulated under drought stress treatments and also preferentially expressed in developing seeds. Recombinant HvsHsp17.5-CI protein was expressed in E. coli and purified to homogeneity, which displayed in vitro chaperone activity. The predicted structural model of HvsHsp-17.5-CI protein suggests that the α-crystallin domain is evolutionarily highly conserved.

Background Sucrose phosphate synthase (SPS) is an important component of the plant sucrose biosynthesis pathway. In the monocotyledonous Poaceae, five SPS genes have been identified. Here we present a detailed analysis of the wheat SPSII family in wheat. A set of homoeologue-specific primers was developed in order to permit both the detection of sequence variation, and the dissection of the individual contribution of each homoeologue to the global expression of SPSII. Results The expression in bread wheat over the course of development of various sucrose biosynthesis genes monitored on an Affymetrix array showed that the SPS genes were regulated over time and space. SPSII homoeologue-specific assays were used to show that the three homoeologues contributed differentially to the global expression of SPSII. Genetic mapping placed the set of homoeoloci on the short arms of the homoeologous group 3 chromosomes. A resequencing of the A and B genome copies allowed the detection of four haplotypes at each locus. The 3B copy includes an unspliced intron. A comparison of the sequences of the wheat SPSII orthologues present in the diploid progenitors einkorn, goatgrass and Triticum speltoides, as well as in the more distantly related species barley, rice, sorghum and purple false brome demonstrated that intronic sequence was less well conserved than exonic. Comparative sequence and phylogenetic analysis of SPSII gene showed that false purple brome was more similar to Triticeae than to rice. Wheat - rice synteny was found to be perturbed at the SPS region. Conclusion The homoeologue-specific assays will be suitable to derive associations between SPS functionality and key phenotypic traits. The amplicon sequences derived from the homoeologue-specific primers are informative regarding the evolution of SPSII in a polyploid context.