Genome-wide characterization of DELLA gene family in blueberry (Vaccinium darrowii) and their expression profiles in development and response to abiotic stress.

BACKGROUND: The DELLA proteins, a class of GA signaling repressors, belong to the GRAS family of plant-specific nuclear proteins. Members of DELLA gene family encode transcriptional regulators with diverse functions in plant development and abiotic stress responses. To date, DELLAs have been identified in various plant species, such as Arabidopsis thaliana, Malus domestica, Populus trichocarpa, and other land plants. Most information of DELLA family genes was obtained from A. thaliana, whereas little is known about the DELLA gene family in blueberry. RESULTS: In this study, we identified three DELLA genes in blueberry (Vaccinium darrowii, VdDELLA) and provided a complete overview of VdDELLA gene family, describing chromosome localization, protein properties, conserved domain, motif organization, and phylogenetic analysis. Three VdDELLA members, containing two highly conserved DELLA domain and GRAS domain, were distributed across three chromosomes. Additionally, cis-acting elements analysis indicated that VdDELLA genes might play a critical role in blueberry developmental processes, hormone, and stress responses. Expression analysis using quantitative real-time PCR (qRT-PCR) revealed that all of three VdDELLA genes were differentially expressed across various tissues. VdDELLA2 was the most highly expressed VdDELLA in all denoted tissues, with a highest expression in mature fruits. In addition, all of the three VdDELLA genes actively responded to diverse abiotic stresses. Based on qRT-PCR analysis, VdDELLA2 might act as a key regulator in V. darrowii in response to salt stress, whereas VdDELLA1 and VdDELLA2 might play an essential role in cold stress response. Under drought stress, all of three VdDELLA genes were involved in mediating drought response. Furthermore, their transiently co-localization with nuclear markers in A. thaliana protoplasts demonstrated their transcriptional regulator roles. CONCLUSIONS: In this study, three VdDELLA genes were identified in V. darrowii genome. Three VdDELLA genes were closely related to the C. moschata DELLA genes, S. lycopersicum DELLA genes, and M. domestica DELLA genes, respectively, indicating their similar biological functions. Expression analysis indicated that VdDELLA genes were highly efficient in blueberry fruit development. Expression patterns under different stress conditions revealed the differentially expressed VdDELLA genes responding to salt, drought, and cold stress. Overall, these results enrich our understanding of evolutionary relationship and potential functions of VdDELLA genes, which provide valuable information for further studies on genetic improvement of the plant yield and plant resistance.

Identification of Apple Flower Development-Related Gene Families and Analysis of Transcriptional Regulation.

Apple (Malus domestica Borkh.) stands out as a globally significant fruit tree with considerable economic importance. Nonetheless, the orchard production of 'Fuji' apples faces significant challenges, including delayed flowering in young trees and inconsistent annual yields in mature trees, ultimately resulting in suboptimal fruit yield due to insufficient flower bud formation. Flower development represents a pivotal process influencing plant adaptation to environmental conditions and is a crucial determinant of successful plant reproduction. The three gene or transcription factor (TF) families, C2H2, DELLA, and FKF1, have emerged as key regulators in plant flowering regulation; however, understanding their roles during apple flowering remains limited. Consequently, this study identified 24 MdC2H2, 6 MdDELLA, and 6 MdFKF1 genes in the apple genome with high confidence. Through phylogenetic analyses, the genes within each family were categorized into three distinct subgroups, with all facets of protein physicochemical properties and conserved motifs contingent upon subgroup classification. Repetitive events between these three gene families within the apple genome were elucidated via collinearity analysis. qRT-PCR analysis was conducted and revealed significant expression differences among MdC2H2-18, MdDELLA1, and MdFKF1-4 during apple bud development. Furthermore, yeast two-hybrid analysis unveiled an interaction between MdC2H2-18 and MdDELLA1. The genome-wide identification of the C2H2, DELLA, and FKF1 gene families in apples has shed light on the molecular mechanisms underlying apple flower bud development.

FveDREB1B improves cold tolerance of woodland strawberry by positively regulating FveSCL23 and FveCHS.

Cold stress has seriously inhibited the growth and development of strawberry during production. CBF/DREB1 is a key central transcription factor regulating plant cold tolerance, but its regulatory mechanisms are varied in different plants. Especially in strawberry, the molecular mechanism of CBF/DREB1 regulating cold tolerance is still unclear. In this study, we found that FveDREB1B was most significantly induced by cold stress in CBF/DREB1 family of diploid woodland strawberry. FveDREB1B was localized to the nucleus, and DREB1B sequences were highly conserved in diploid and octoploid strawberry, and even similar in Rosaceae. And FveDREB1B overexpressed strawberry plants showed delayed flowering and increased cold tolerance, while FveDREB1B silenced plants showed early flowering and decreased cold tolerance. Under cold stress, FveDREB1B activated FveSCL23 expression by directly binding to its promoter. Meanwhile, FveDREB1B and FveSCL23 interacted with FveDELLA, respectively. In addition, we also found that FveDREB1B promoted anthocyanin accumulation in strawberry leaves by directly activating FveCHS expression after cold treatment and recovery to 25°C. DREB1B genes were also detected to be highly expressed in cold-tolerant strawberry resources 'Fragaria mandschurica' and 'Fragaria nipponica'. In conclusion, our study reveals the molecular mechanism of FveDREB1B-FveSCL23-FveDELLA module and FveDREB1B-FveCHS module to enhance the cold tolerance of woodland strawberry. It provides a new idea for improving the cold tolerance of cultivated strawberry and evaluating the cold tolerance of strawberry germplasm resources.

Gibberellin Signaling through RGA Suppresses GCN5 Effects on Arabidopsis Developmental Stages.

Histone acetyltransferases (HATs) modify the amino-terminal tails of the core histone proteins via acetylation, regulating chromatin structure and transcription. GENERAL CONTROL NON-DEREPRESSIBLE 5 (GCN5) is a HAT that specifically acetylates H3K14 residues. GCN5 has been associated with cell division and differentiation, meristem function, root, stem, foliar, and floral development, and plant environmental response. The flowers of gcn5 plants display a reduced stamen length and exhibit male sterility relative to the wild-type plants. We show that these effects may arise from gibberellin (GA)-signaling defects. The signaling pathway of bioactive GAs depends on the proteolysis of their repressors, DELLA proteins. The repressor GA (RGA) DELLA protein represses plant growth, inflorescence, and flower and seed development. Our molecular data indicate that GCN5 is required for the activation and H3K14 acetylation of genes involved in the late stages of GA biosynthesis and catabolism. We studied the genetic interaction of the RGA and GCN5; the RGA can partially suppress GCN5 action during the whole plant life cycle. The reduced elongation of the stamen filament of gcn5-6 mutants is reversed in the rga-t2;gcn5-6 double mutants. RGAs suppress the GCN5 effect on the gene expression and histone acetylation of GA catabolism and GA signaling. Interestingly, the RGA and RGL2 do not suppress ADA2b function, suggesting that ADA2b acts downstream of GA signaling and is distinct from GCN5 activity. In conclusion, we propose that the action of GCN5 on stamen elongation is partially mediated by RGA and GA signaling.

The MdNAC72-MdABI5 module acts as an interface integrating jasmonic acid and gibberellin signals and undergoes ubiquitination-dependent degradation regulated by MdSINA2 in apple.

Jasmonic acid (JA) and gibberellin (GA) coordinately regulate plant developmental programs and environmental cue responses. However, the fine regulatory network of the cross-interaction between JA and GA remains largely elusive. In this study, we demonstrate that MdNAC72 together with MdABI5 positively regulates anthocyanin biosynthesis through an exquisite MdNAC72-MdABI5-MdbHLH3 transcriptional cascade in apple. MdNAC72 interacts with MdABI5 to promote the transcriptional activation of MdABI5 on its target gene MdbHLH3 and directly activates the transcription of MdABI5. The MdNAC72-MdABI5 module regulates the integration of JA and GA signals in anthocyanin biosynthesis by combining with JA repressor MdJAZ2 and GA repressor MdRGL2a. MdJAZ2 disrupts the MdNAC72-MdABI5 interaction and attenuates the transcriptional activation of MdABI5 by MdNAC72. MdRGL2a sequesters MdJAZ2 from the MdJAZ2-MdNAC72 protein complex, leading to the release of MdNAC72. The E3 ubiquitin ligase MdSINA2 is responsive to JA and GA signals and promotes ubiquitination-dependent degradation of MdNAC72. The MdNAC72-MdABI5 interface fine-regulates the integration of JA and GA signals at the transcriptional and posttranslational levels by combining MdJAZ2, MdRGL2a, and MdSINA2. In summary, our findings elucidate the fine regulatory network connecting JA and GA signals with MdNAC72-MdABI5 as the core in apple.

Petunia PHYTOCHROME INTERACTING FACTOR 4/5 transcriptionally activates key regulators of floral scent.

Floral scent emission of petunia flowers is regulated by light conditions, circadian rhythms, ambient temperature and the phytohormones GA and ethylene, but the mechanisms underlying sensitivity to these factors remain obscure. PHYTOCHROME INTERACTING FACTORs (PIFs) have been well studied as components of the regulatory machinery for numerous physiological processes. Acting redundantly, they serve as transmitters of light, circadian, metabolic, thermal and hormonal signals. Here we identified and characterized the phylogenetics of petunia PIF family members (PhPIFs). PhPIF4/5 was revealed as a positive regulator of floral scent: TRV-based transient suppression of PhPIF4/5 in petunia petals reduced emission of volatiles, whereas transient overexpression increased scent emission. The mechanism of PhPIF4/5-mediated regulation of volatile production includes activation of the expression of genes encoding biosynthetic enzymes and a key positive regulator of the pathway, EMISSION OF BENZENOIDS II (EOBII). The PIF-binding motif on the EOBII promoter (G-box) was shown to be needed for this activation. As PhPIF4/5 homologues are sensors of dawn and expression of EOBII also peaks at dawn, the prior is proposed to be part of the diurnal control of the volatile biosynthetic machinery. PhPIF4/5 was also found to transcriptionally activate PhDELLAs; a similar positive effect of PIFs on DELLA expression was further confirmed in Arabidopsis seedlings. The PhPIF4/5-PhDELLAs feedback is proposed to fine-tune GA signaling for regulation of floral scent production.

Genome-Wide Identification, Expression Analysis and Functional Study of DELLA Genes in Chinese Cabbage (Brassica rapa L. ssp. pekinensis).

BACKGROUND: DELLA protein is a crucial factor which played pivotal roles in regulating numerous intriguing biological processes in plant development and abiotic stress responses. However, little is known about the function and information of DELLA protein in Chinese cabbage. METHODS: Using 5 DELLA gene sequences in Arabidopsis Thaliana as probes, 5 DELLA genes in Chinese cabbage were identified by Blast search in Chinese cabbage database (Brassica database (BRAD)). The National Center for Biotechnology Information (NCBI), ExPaSy, SWISS-MODEL, DNAMAN, MEGA 11, PlantCARE were used to identify and analyze the DELLA gene family of Chinese cabbage. Gene expression was analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). The function of BraA10gRGL3 was verified by overexpression and phenotypic analysis of BraA10gRGL3 and yeast hybrid. RESULTS: In this study, 5 BraDELLAs homologous to Arabidopsis thaliana were identified and cloned based on the Brassica database, namely, BraA02gRGL1, BraA05gRGL2, BraA10gRGL3, BraA06gRGA and BraA09gRGA. All BraDELLAs contain the DELLA, TVHYNP, and GRAS conserved domains. Cis-element analysis revealed that the promoter regions of these 5 DELLA genes all contain light-responsive elements, TCT motif, I-box, G-box, and box 4, which are associated with GA signaling. Transcriptome analysis results proved that the expression of BraA02gRGL1, BraA05gRGL2, and BraA10gRGL3 in Y2 at different growth stages were lower than them in Y7, which is consistent with the phenotype that Y7 exhibited stronger stress tolerance than Y2. It is worth emphasizing that even through the overexpression of BraA10gRGL3-Y7 in Arabidopsis resulted in smaller leaf size and lower fresh weight compared to the wild type (WT) Arabidopsis: Columbia, a stronger response to abiotic stresses was observed in BraA10gRGL3-Y7. It indicated that BraA10gRGL3-Y7 can improve the stress resistance of plants by inhibiting their growth. Moreover, the yeast two-hybrid experiment confirmed that BraA10gRGL3-Y7 can interacted with BraA05gGID1a-Y7, BraA04gGID1b1, BraA09gGID1b3-Y2, and BraA06gGID1c, whereas BraA10gRGL3-Y2 cannot interact with any BraGID1. CONCLUSIONS: Collectively, BraDELLAs play important role in plant development and response to abiotic stress. The differences in amino acid sequences between BraA10gRGL3-Y2 and BraA10gRGL3-Y7 may result in variations in their protein binding sites, thus affecting their interaction with the BraGID1 family proteins. This systematic analysis lays the foundation for further study of the functional characteristics of DELLA genes of Chinese cabbage.

The Human Body is the Collective Unconscious: Archetypal Images as Innate Embodied Metaphors.

For a significant part of its history, archetype theory has been undermined by criticisms containing unexamined Cartesian assumptions. Such assumptions treat all cognition as disembodied, consisting of mere manipulation of abstract, inherently meaningless signs mimicked from verbal instruction or cultural learning. Since the 1980s, due to the results of many independent disciplines, however, this view is being replaced with one of embodied cognition. This shift has important consequences for archetype theory, allowing us to provide a non-reductive biological anchor that explains many characteristics of the archetypal image.

Pendant une partie importante de son histoire, la théorie des archétypes a été discréditée par des critiques contenant des hypothèses cartésiennes qui n'étaient pas remises en question. De telles hypothèses considèrent que toute capacité cognitive est désincarnée et consiste en une simple manipulation de signes abstraits et intrinsèquement dépourvus de sens, imités à partir d'instructions verbales ou d'apprentissage culturel. Néanmoins depuis les années 1980, du fait de résultats provenant de plusieurs disciplines indépendantes, cette façon de voir est remplacée par une autre: celle de la capacité cognitive incarnée. Ce déplacement a des conséquences importantes pour la théorie des archétypes, nous permettant de fournir un ancrage biologique non­réducteur qui explique un grand nombre de caractéristiques de l'image archétypale.

Durante una parte significativa de su historia, la teoría de los arquetipos se ha visto afectada por críticas que contenían supuestos cartesianos no examinados. Dichos supuestos dan cuenta de toda cognición como incorpórea, consistiendo en la mera manipulación de signos abstractos e intrínsecamente carentes de significado, imitados a partir de la instrucción verbal o el aprendizaje cultural. Sin embargo, desde la década de 1980, gracias a los resultados de muchas disciplinas independientes, este punto de vista se está sustituyendo por el de la cognición encarnada. Este cambio tiene importantes consecuencias para la teoría de los arquetipos, ya que nos permite ofrecer un anclaje biológico no reductivo que explica muchas características de la imagen arquetípica.

Renaissance-Europe-16th Century continued.

This chapter is limited to the text of Della Cruce, which contains the most comprehensive account of the instruments used in cranial surgery at the time. Of particular importance is Della Cruce's attitude to what he called non-perforating straight trepans, which in general he disliked. It may be noted that his text was the last to describe this sort of instrument. In the succeeding centuries, changes to penetrating instruments were all variations on the shape of different kinds of crown trepan. Like Berengario, Della Cruce described brace and bit trepans with interchangeable bits. Various methods were employed to prevent them penetrating too deeply.

DELLA proteins recruit the Mediator complex subunit MED15 to coactivate transcription in land plants.

DELLA proteins are negative regulators of the gibberellin response pathway in angiosperms, acting as central hubs that interact with hundreds of transcription factors (TFs) and regulators to modulate their activities. While the mechanism of TF sequestration by DELLAs to prevent DNA binding to downstream targets has been extensively documented, the mechanism that allows them to act as coactivators remains to be understood. Here, we demonstrate that DELLAs directly recruit the Mediator complex to specific loci in Arabidopsis, facilitating transcription. This recruitment involves DELLA amino-terminal domain and the conserved MED15 KIX domain. Accordingly, partial loss of MED15 function mainly disrupted processes known to rely on DELLA coactivation capacity, including cytokinin-dependent regulation of meristem function and skotomorphogenic response, gibberellin metabolism feedback, and flavonol production. We have also found that the single DELLA protein in the liverwort Marchantia polymorpha is capable of recruiting MpMED15 subunits, contributing to transcriptional coactivation. The conservation of Mediator-dependent transcriptional coactivation by DELLA between Arabidopsis and Marchantia implies that this mechanism is intrinsic to the emergence of DELLA in the last common ancestor of land plants.

C-TERMINAL DOMAIN PHOSPHATASE-LIKE 3 contributes to GA-mediated growth and flowering by interaction with DELLA proteins.

Gibberellic acid (GA) plays a central role in many plant developmental processes and is crucial for crop improvement. DELLA proteins, the core suppressors in the GA signaling pathway, are degraded by GA via the 26S proteasomal pathway to release the GA response. However, little is known about the phosphorylation-mediated regulation of DELLA proteins. In this study, we combined GA response assays with protein-protein interaction analysis to infer the connection between Arabidopsis thaliana DELLAs and the C-TERMINAL DOMAIN PHOSPHATASE-LIKE 3 (CPL3), a phosphatase involved in the dephosphorylation of RNA polymerase II. We show that CPL3 directly interacts with DELLA proteins and promotes DELLA protein stability by inhibiting its degradation by the 26S proteasome. Consequently, CPL3 negatively modulates multiple GA-mediated processes of plant development, including hypocotyl elongation, flowering time, and anthocyanin accumulation. Taken together, our findings demonstrate that CPL3 serves as a novel regulator that could improve DELLA stability and thereby participate in GA signaling transduction.

Complex control of seed germination timing by ERF50 involves RGL2 antagonism and negative feedback regulation of DOG1.

Seed dormancy governs germination timing, with both evolutionary and applied consequences. Despite extensive studies on the hormonal and genetic control of these processes, molecular mechanisms directly linking dormancy and germination remain poorly understood. By screening a collection of lines overexpressing Arabidopsis transcription factors, we identified ERF50 as a key gene to control dormancy and germination. To study its regulation, we measured seed-related physiological parameters in loss-of-function mutants and carried out transactivation, protein interaction and ChIP-PCR analyses. We found direct ERF50-mediated repression of DOG1 and activation of EXPA2 transcription, which results in enhanced seed germination. Although ERF50 expression is increased by DOG1 in dormant seeds, ERF50 germination-promoting activity is blocked by RGL2. The physiological, genetic and molecular evidence gathered here supports that ERF50 controls germination timing by regulating DOG1 levels to leverage its role as enhancer of seed germination, via RGL2 antagonism on EXPA2 expression. Our results highlight the central role of ERF50 as a feedback regulator to couple and fine-tune seed dormancy and germination.

Scapulohumeral subluxation and luxation in five alpacas.

INTRODUCTION: The patient histories, findings from clinical examinations, diagnostic imaging techniques, the surgical procedures, complications, necropsy findings and the outcomes from five male or castrated male alpacas with scapulohumeral sub-/luxation are presented. These alpacas each had a history of severe forelimb lameness for one week (n: 1), four weeks (n: 2) and for two-to-three months (n: 2). Two of the five alpacas were euthanized due to severe osteoarthritic changes that developed during the two-to-three months of scapulohumeral luxation. Three alpacas were treated with open reduction and internal stabilisation by placing tension band sutures between one 4,5 mm cortical screw anchored in the scapular neck and two 4,5 mm cortical screws anchored in the greater humeral tubercle, all of them provided with washers. Post-surgery a carpal flexion sling was applied to avoid postoperative weight-bearing. An exercise programme was started after removal of the carpal sling and continued for 12 weeks. In one of the three alpacas an additional non-displaced fracture of the acromion occurred two weeks after surgery. In conclusion, all three treated alpacas had good-to-excellent long-term outcomes and are still alive 123, 15 and 12 months after surgical repair of the scapulohumeral sub-/luxation. As four weeks, or even up to three months elapsed in four of these five alpacas until a definitive diagnosis was made, more education should be provided to alpaca owners that severely lame animals should be presented to a veterinarian with adequate diagnostic possibilities and expertise as soon as possible in order not to compromise treatment success.

INTRODUCTION: Dans cette étude rétrospective, on présente les antécédents des patients, les résultats des examens cliniques, les techniques d'imagerie diagnostique, les procédures chirurgicales, les complications, les résultats de l'autopsie et les résultats finaux de cinq alpagas mâles ou mâles castrés souffrant d'une subluxation ou luxation scapulo-humérale. Ces alpagas présentaient chacun des antécédents de boiterie sévère des membres antérieurs depuis une semaine (n: 1), quatre semaines (n: 2) et deux à trois mois (n: 2). Deux des cinq alpagas ont été euthanasiés en raison des changements arthrosiques graves qui s'étaient développés au cours des deux à trois mois de luxation scapulo-humérale. Trois alpagas ont été traités par réduction ouverte et stabilisation interne en plaçant des sutures à bande de tension entre une vis corticale de 4,5 mm ancrée dans le col de l'omoplate et deux vis corticales de 4,5 mm ancrées dans le tubercule huméral supérieur, toutes munies de rondelles. Après l'opération, une écharpe de flexion du carpe a été mise en place pour éviter la mise en charge postopératoire. Un programme d'exercices a été mis en place après le retrait de l'écharpe carpienne et a été poursuivi pendant 12 semaines. Chez l'un des trois alpagas, une fracture non déplacée de l'acromion s'est produite deux semaines après l'opération. En conclusion, les trois alpagas traités ont eu des résultats à long terme bons à excellents et sont toujours en vie 123, 15 et 12 mois après la réparation chirurgicale de la subluxation scapulo-humérale. Dans la mesure où il s'est écoulé quatre semaines voire jusqu'à trois mois chez quatre de ces cinq alpagas avant qu'un diagnostic définitif ne soit posé, il convient d'informer davantage les propriétaires d'alpagas que les animaux gravement boiteux doivent être présentés à un vétérinaire disposant des possibilités de diagnostic et de l'expertise adéquates le plus rapidement possible afin de ne pas compromettre le succès du traitement.

[Five case reports on granulosa cell tumors in cattle with practical information on diagnosis and possible progression]. / Fünf Fallberichte zum Granulosazelltumor beim Rind mit praktischen Hinweisen zur Diagnostik und zum möglichen Verlauf.

INTRODUCTION: Five cases of ovarian tumors (granulosa cell tumors) in cattle are presented from the patient load of the Vetsuisse University of Zurich and Bern. The aim of this work was to demonstrate the variable development of the illness and to indicate diagnostic and therapeutic possibilities to the practicing veterinarians. Case 1 shows bilateral appearance and the development of malignancy and metastases. The main symptoms in case 2 were the development of the mammary gland in a juvenile animal and the behavior modification due to a hormonal imbalance. The cases 3, 4 and 5 underwent surgery, case 4 restarted reproductive activity resulting in five subsequent pregnancies. The initial presumption is a result of a gynecological including ultrasonographic examination and can be verified by the analysis of Müllerian Inhibiting Hormone in serum. The decision to perform surgery should be done rapidly, as normal fertility can be achieved if the tumor is located unilaterally. Tumor growth and potential malignancy can provoke fatal health issues and also make it impossible to use meat of these animals for consumption.

INTRODUCTION: Cinq cas de tumeurs ovariennes (tumeurs des cellules de la granulosa) chez les bovins sont présentés à partir de la patientèle de l'Université Vetsuisse de Zurich et de Berne. Le but de ce travail était de montrer l'évolution variable de la maladie et d'indiquer les possibilités diagnostiques et thérapeutiques aux vétérinaires praticiens. Le cas 1 montre la possibilité d'une apparition bilatérale avec développement d'une tumeur maligne et de métastases. Les principaux symptômes du cas 2 étaient le développement de la glande mammaire chez un animal juvénile et la modification du comportement due à un déséquilibre hormonal. Les cas 3, 4 et 5 ont subi une intervention chirurgicale, le cas 4 a repris une activité de reproduction avec cinq gestations ultérieures. La présomption initiale résulte d'un examen gynécologique et peut être vérifiée par l'analyse de l'hormone anti-müllérienne dans le sérum. La décision d'opérer doit être prise rapidement, car une intervention chirurgicale pour enlever l'ovaire atteint peut, dans l'idéal, permettre une reprise de l'activité de reproduction. La croissance de la tumeur et la malignité potentielle peuvent provoquer des problèmes de santé fatals et rendre la viande de ces animaux impropre à la consommation.

GRAS gene family in rye (Secale cereale L.): genome-wide identification, phylogeny, evolutionary expansion and expression analyses.

BACKGROUND: The GRAS transcription factor family plays a crucial role in various biological processes in different plants, such as tissue development, fruit maturation, and environmental stress. However, the GRAS family in rye has not been systematically analyzed yet. RESULTS: In this study, 67 GRAS genes in S. cereale were identified and named based on the chromosomal location. The gene structures, conserved motifs, cis-acting elements, gene replications, and expression patterns were further analyzed. These 67 ScGRAS members are divided into 13 subfamilies. All members include the LHR I, VHIID, LHR II, PFYRE, and SAW domains, and some nonpolar hydrophobic amino acid residues may undergo cross-substitution in the VHIID region. Interested, tandem duplications may have a more important contribution, which distinguishes them from other monocotyledonous plants. To further investigate the evolutionary relationship of the GRAS family, we constructed six comparative genomic maps of homologous genes between rye and different representative monocotyledonous and dicotyledonous plants. The response characteristics of 19 ScGRAS members from different subfamilies to different tissues, grains at filling stages, and different abiotic stresses of rye were systematically analyzed. Paclobutrazol, a triazole-based plant growth regulator, controls plant tissue and grain development by inhibiting gibberellic acid (GA) biosynthesis through the regulation of DELLA proteins. Exogenous spraying of paclobutrazol significantly reduced the plant height but was beneficial for increasing the weight of 1000 grains of rye. Treatment with paclobutrazol, significantly reduced gibberellin levels in grain in the filling period, caused significant alteration in the expression of the DELLA subfamily gene members. Furthermore, our findings with respect to genes, ScGRAS46 and ScGRAS60, suggest that these two family members could be further used for functional characterization studies in basic research and in breeding programmes for crop improvement. CONCLUSIONS: We identified 67 ScGRAS genes in rye and further analysed the evolution and expression patterns of the encoded proteins. This study will be helpful for further analysing the functional characteristics of ScGRAS genes.

Leaf growth - complex regulation of a seemingly simple process.

Understanding the underlying mechanisms of plant development is crucial to successfully steer or manipulate plant growth in a targeted manner. Leaves, the primary sites of photosynthesis, are vital organs for many plant species, and leaf growth is controlled by a tight temporal and spatial regulatory network. In this review, we focus on the genetic networks governing leaf cell proliferation, one major contributor to final leaf size. First, we provide an overview of six regulator families of leaf growth in Arabidopsis: DA1, PEAPODs, KLU, GRFs, the SWI/SNF complexes, and DELLAs, together with their surrounding genetic networks. Next, we discuss their evolutionary conservation to highlight similarities and differences among species, because knowledge transfer between species remains a big challenge. Finally, we focus on the increase in knowledge of the interconnectedness between these genetic pathways, the function of the cell cycle machinery as their central convergence point, and other internal and environmental cues.

A DELAY OF GERMINATION 1 (DOG1)-like protein regulates spore germination in the moss Physcomitrium patens.

DELAY OF GERMINATION 1 is a key regulator of dormancy in flowering plants before seed germination. Bryophytes develop haploid spores with an analogous function to seeds. Here, we investigate whether DOG1 function during germination is conserved between bryophytes and flowering plants and analyse the underlying mechanism of DOG1 action in the moss Physcomitrium patens. Phylogenetic and in silico expression analyses were performed to identify and characterise DOG1 domain-containing genes in P. patens. Germination assays were performed to characterise a Ppdog1-like1 mutant, and replacement with AtDOG1 was carried out. Yeast two-hybrid assays were used to test the interaction of the PpDOG1-like protein with DELLA proteins from P. patens and A. thaliana. P. patens possesses nine DOG1 domain-containing genes. The DOG1-like protein PpDOG1-L1 (Pp3c3_9650) interacts with PpDELLAa and PpDELLAb and the A. thaliana DELLA protein AtRGA in yeast. Protein truncations revealed the DOG1 domain as necessary and sufficient for interaction with PpDELLA proteins. Spores of Ppdog1-l1 mutant germinate faster than wild type, but replacement with AtDOG1 reverses this effect. Our data demonstrate a role for the PpDOG1-LIKE1 protein in moss spore germination, possibly alongside PpDELLAs. This suggests a conserved DOG1 domain function in germination, albeit with differential adaptation of regulatory networks in seed and spore germination.

The 2022 community-based integrated care reform in Italy: From desiderata to implementation.

The Italian National Recovery and Resilience Plan allocated € 7 Bn for community care. In May 2022, the Italian government issued a Decree to define the strategy for the development of community-based integrated care. The reform aims to create uniformly a network of services close to where patients live, thus overcoming geographical disparities between regions. The strategy is based on a strong role of the central government in community care, but still leaves autonomy to regions. Levelling availability of services across territories, setting uniform targets with a short period horizon and disregarding starting points may create important implementation problems. Financial constraints will also hamper the implementation of the reform. Ultimately the development of Italian community care will depend on the institutional and managerial capabilities of regions and local health authorities. Firstly, they should shape the actual implementation of community care services by defining organizational arrangements, priority targets and models of care delivery. Secondly, they should develop strategies to face the lack of financial resources and the shortage of healthcare workforce. This contribution informs international readers about a major policy in a European country and its implementation challenges. It offers insights into inter-government relations in NHS-type healthcare systems (Nordic countries and Spain), showcasing the complexity of policymaking involving multiple political actors and resulting indeterminacy of policies and their implementation.

The tomato EAR-motif repressor, SlERF36, accelerates growth transitions and reduces plant life cycle by regulating GA levels and responses.

Faster vegetative growth and early maturity/harvest reduce plant life cycle time and are important agricultural traits facilitating early crop rotation. GA is a key hormone governing developmental transitions that determine growth speed in plants. An EAR-motif repressor, SlERF36 that regulates various growth transitions, partly through regulation of the GA pathway and GA levels, was identified in tomato. Suppression of SlERF36 delayed germination, slowed down organ growth and delayed the onset of flowering time, fruit harvest and whole-plant senescence by 10-15 days. Its over-expression promoted faster growth by accelerating all these transitions besides increasing organ expansion and plant height substantially. The plant life cycle and fruit harvest were completed 20-30 days earlier than control without affecting yield, in glasshouse as well as net-house conditions, across seasons and generations. These changes in life cycle were associated with reciprocal changes in expression of GA pathway genes and basal GA levels between suppression and over-expression lines. SlERF36 interacted with the promoters of two GA2 oxidase genes, SlGA2ox3 and SlGA2ox4, and the DELLA gene, SlDELLA, reducing their transcription and causing a 3-5-fold increase in basal GA3/GA4 levels. Its suppression increased SlGA2ox3/4 transcript levels and reduced GA3/GA4 levels by 30%-50%. SlERF36 is conserved across families making it an important candidate in agricultural and horticultural crops for manipulation of plant growth and developmental transitions to reduce life cycles for faster harvest.

[Pestiviruses in sheep and goats in Austria: Options for integration into the bovine viral diarrhea (BVDV) monitoring program]. / Pestiviren bei Schafen und Ziegen in ­Österreich: Möglichkeiten zur ­Einbindung in die BVDV-Überwachung.

INTRODUCTION: After the successful eradication of the bovine viral diarrhea virus (BVDV) in cattle in Austria, the risk of infections with the border disease virus (BDV) remains. Both viruses belong to the pestivirus genus. BDV infections lead to false-positive results in BVDV surveillance. This can be attributed to the contact to small ruminant populations. In particular, keeping cattle together with sheep or goats on a farm or alpine pasture are significant risk factors. Between 2015 and 2022, BDV type 3 was detected in 15 cattles in Austria. These animals were almost exclusively persistently infected calves. However, a positive antibody result for pestiviruses can lead to an extremely time-consuming and costly, and not always successful search for the source of the infection if no active virus excretor is found. This study documents how small ruminants can be integrated into pestivirus monitoring with a manageable amount of work and costs. 23 406 sheep and goat samples from two brucellosis surveillance programs in small ruminants were analyzed retrospectively. Blood samples were examined using pestivirus real-time pool RT-PCR (qPCR). Direct virus detection of BDV-3 was achieved in 40 sheep from five different federal states. Over the entire investigation period a further 37 detections of BDV-3 were found in cattle, sheep and goats outside of this study throughout Austria. This study accounts for 52 % of all border disease detections from 2015 to 2022. By including small ruminants in pestivirus monitoring, the disruptive factor BDV and the risk of its introduction into cattle herds can be significantly minimized in the future.

INTRODUCTION: Après l'éradication réussie du virus de la diarrhée virale bovine (BVDV) chez les bovins en Autriche, le risque d'infections par le virus de la Border Disease (BDV) demeure. Ces deux virus appartiennent au genre des pestivirus. Les infections par le BDV entraînent des résultats faussement positifs dans la surveillance du BVDV. Ce phénomène peut être attribué aux contacts avec les populations de petits ruminants. En particulier, la détention de bovins avec des moutons ou des chèvres sur une exploitation ainsi que les pâturages alpins sont des facteurs de risque importants pour les infections. Entre 2015 et 2022, le BDV de type 3 a été détecté chez 15 bovins en Autriche. Ces animaux étaient presque exclusivement des veaux infectés de manière persistante. Cependant, un résultat positif aux anticorps contre les pestivirus peut conduire à une recherche extrêmement longue et coûteuse et pas toujours fructueuse de la source de l'infection si aucun excréteur de virus actif n'est trouvé. Cette étude montre comment les petits ruminants peuvent être intégrés dans la surveillance des pestivirus avec une quantité de travail et des coûts gérables. À cette fin, 23 460 échantillons d'ovins et de caprins provenant de deux programmes de surveillance de la brucellose chez les petits ruminants ont été utilisés de façon rétrospective. Les échantillons de sang ont été examinés à l'aide de la RT-PCR en temps réel des pestivirus (qPCR). La détection directe du virus BDV-3 a été réalisée chez 40 moutons provenant de cinq länder différents. Sur l'ensemble de la période d'investigation (2015 ­ 2022), 37 autres détections de BDV-3 ont été effectuées chez des bovins, des ovins et des caprins en dehors de cette étude, dans toute l'Autriche. Cette étude représente 52 % de toutes les détections de Border Disease entre 2015 et 2022. En incluant les petits ruminants dans la surveillance des pestivirus, le facteur de perturbation qu'est le BDV et le risque de son introduction dans les troupeaux de bovins peuvent être considérablement minimisés à l'avenir.