STM Expression Research Articles

Genetic and Epigenetic Evidence for Nonestrogenic Disruption of Otolith Development by Bisphenol A in Zebrafish.

Bisphenol A (BPA) is a well-known endocrine-disrupting chemical (EDC) that has estrogenic activities. In addition to disrupting reproductive development and function via estrogenic signaling pathways, BPA can also interfere with nonreproductive functions through nonestrogenic pathways; however, the mechanisms underlying such nonestrogenic activities are not well understood. In this study, we demonstrated that BPA could disrupt otolith formation during the early development of zebrafish with long-lasting ethological effects. Using multiple mutants of estrogen receptors, we provided strong genetic evidence that the BPA-induced otolith malformation was independent of estrogen signaling. Transcriptome analysis revealed that two genes related to otolith development, otopetrin 1 (otop1) and starmaker (stm), decreased their expression significantly after BPA exposure. Knockout of both otop1 and stm genes could phenocopy the BPA-induced otolith malformation, while microinjection of their mRNAs could rescue the BPA-induced abnormalities of otolith formation. Further experiments showed that BPA inhibited the expression of otop1 and stm by activating the MEK/ERK-EZH2-H3K27me3 signaling pathway. Taken together, our study provided comprehensive genetic and molecular evidence that BPA induced the otolith malformation through nonestrogenic pathway during zebrafish early development and its activities involved epigenetic control of key genes (e.g., otop1 and stm) participating in otolith formation.

ERF5 enhances protocorm-like body regeneration via enhancement of STM expression in Dendrobium orchid.

Orchid plants develop protocorms upon germination and produce protocorm-like structures called protocorm-like bodies (PLBs) from protocorms and somatic cells via tissue culture. Protocorm-like bodies have broad technical application potential in the orchid industry and their regeneration is a distinct developmental process in the plant kingdom. However, little is known about this unparalleled developmental program. In this study, we identified a PLB-abundant gene, ethylene response factor (ERF), and a transcription factor named DoERF5, and determined its important role in PLB regeneration in Dendrobium orchid. Overexpression of DoERF5 in Dendrobium greatly enhanced the PLB regeneration from PLB and stem explants, and upregulated the expression of WOUND-INDUCED DEDIFFERENTIATION (DoWIND) homologs and SHOOT MERISTEMLESS (DoSTM), as well as the genes involved in cytokinin biosynthesis (DoIPT) and the cytokinin response factors (DoARRs). However, silencing DoERF5 reduced the regeneration rate of PLBs, and downregulated the expression of DoWIND homologs, DoSTM and DoARRs. We demonstrated that DoERF5 is directly bound to the DoSTM promoter and regulates its expression. In addition, overexpression of DoSTM in Dendrobium orchid resulted in favorable regeneration of PLBs. Our results clarify that DoERF5 regulates the regeneration of PLB by enhancing DoSTM expression. Our findings provide new insights into how DoERF5 mediates PLB regeneration and offers technical potential in improving clonal propagation, preservation, and the bioengineering of orchids.

Cultivated strawberry HANABA TARANU: a tale of transcription factors, raging hormones, and asexual reproduction

Cultivated strawberry <scp><i>HANABA TARANU</i></scp>: a tale of transcription factors, raging hormones, and asexual reproduction

Early Low-Fluence Red Light or Darkness Modulates the Shoot Regeneration Capacity of Excised Arabidopsis Roots.

In plants, light is an important environmental signal that induces meristem development and interacts with endogenous signals, including hormones. We found that treatment with 24 h of low-fluence red light (24 h R) or 24 h of darkness (24 h D) following root excision greatly increased the frequency of shoot generation, while continuous low-fluence red light in callus and shoot induction stages blocked the explants’ ability to generate shoots. Shoot generation ability was closely associated with WUS expression and distribution pattern. 1-N-naphthylphtalamic acid (NPA) disrupted the dynamic distribution of the WUS signal induced by early 24 h R treatment, and NPA plus 24 R treatment increased the average shoot number compared with early 24 h R alone. Transcriptome analysis revealed that differentially expressed genes involved in meristem development and hormone signal pathways were significantly enriched during 24 R or 24 D induced shoot regeneration, where early 24 h R or 24 h D treatment upregulated expression of WOX5, LBD16, LBD18 and PLT3 to promote callus initiation and formation of root primordia, and also activated WUS, STM, CUC1 and CUC2 expression, leading to initiation of the shoot apical meristem (SAM). This finding demonstrates that early exposure of explants to transient low-fluence red light or darkness modulates the expression of marker genes related with callus development and shoot regeneration, and dynamic distribution of WUS, leading to an increased ability to generate shoots.

A sustained CYCLINB1;1 and STM expression in the neoplastic tissues induced by Rhodococcus fascians on Arabidopsis underlies the persistence of the leafy gall structure

ABSTRACT Rhodococcus fascians is a gram-positive phytopathogen that infects a wide range of plant species. The actinomycete induces the formation of neoplastic growths, termed leafy galls, that consist of a gall body covered by small shoots of which the outgrowth is arrested due to an extreme form of apical dominance. In our previous work, we demonstrated that in the developing gall, auxin drives the transdifferentiation of parenchyma cells into vascular elements. In this work, with the use of transgenic Arabidopsis thaliana plants carrying molecular reporters for cell division (pCYCB1;1:GUS) and meristematic activity (pSTM:GUS), we analyzed the fate of cells within the leafy gall. Our results indicate that the size of the gall body is determined by ongoing mitotic cell divisions as illustrated by strong CYCB1;1 expression combined with the de novo formation of new meristematic areas triggered by STM expression. The shoot meristems that develop in the peripheral parts of the gall are originating from high ectopic STM expression. Altogether the presented data provide further insight into the cellular events that accompany the development of leafy galls in response to R. fascians infection.

Establishment of the Embryonic Shoot Meristem Involves Activation of Two Classes of Genes with Opposing Functions for Meristem Activities.

The shoot meristem, a stem-cell-containing tissue initiated during plant embryogenesis, is responsible for continuous shoot organ production in postembryonic development. Although key regulatory factors including KNOX genes are responsible for stem cell maintenance in the shoot meristem, how the onset of such factors is regulated during embryogenesis is elusive. Here, we present evidence that the two KNOX genes STM and KNAT6 together with the two other regulatory genes BLR and LAS are functionally important downstream genes of CUC1 and CUC2, which are a redundant pair of genes that specify the embryonic shoot organ boundary. Combined expression of STM with any of KNAT6, BLR, and LAS can efficiently rescue the defects of shoot meristem formation and/or separation of cotyledons in cuc1 cuc2 double mutants. In addition, CUC1 and CUC2 are also required for the activation of KLU, a cytochrome P450-encoding gene known to restrict organ production, and KLU counteracts STM in the promotion of meristem activity, providing a possible balancing mechanism for shoot meristem maintenance. Together, these results establish the roles for CUC1 and CUC2 in coordinating the activation of two classes of genes with opposite effects on shoot meristem activity.

Overexpression of TCP transcription factor OsPCF7 improves agronomic trait in rice

Plant morphogenetic regulation is an important part of plant development biology. An ideal plant architecture can be created to allow plant to fully capture and use solar energy and maximize rice yields per unit area. The TCP (THEOSINTE BRANCHCHED1-CYCLOIDEA-PROLIFERATING CELL FACTOR1) family transcription factors participate in plant developmental processes associated with cell proliferation and growth. In this study, OsPCF7 gene was isolated from Oryza sativa L., which encodes a class II TCP protein with 270 amino acids. The qPCR analysis showed that OsPCF7 expressed in all tissues and organs at all times. The expressions of OsPCF7 in the tiller stage were dramatically increased, and OsPCF7 protein was predominately targeted to the nucleus. The overexpression of OsPCF7 increased shoot height, root length, and total root number of transgenic rice seedlings and promoted the tiller and heading of rice plant, causing increase of panicle numbers and filled grain numbers per plant, finally leading to increase of grain yield per plant. In addition, OsPCF7 could positively regulate the expression of STM, KNAT2, KNAT6, LOX2, AS1, and IAA3 in overexpressing OsPCF7 plant. By contrast, RNAi of OsPCF7 decreased the tiller, panicle numbers, filled grain numbers, and grain yield per plant. RNAi of OsPCF7 also led to a significant decrease in the expression of STM, KNAT2, KNAT6, LOX2, AS1, and IAA3. Taken together, OsPCF7 played multiple roles in rice plant architecture by regulating different biological processes during rice growth and development, which made it a potential gene in genetic improvement for agronomic trait of crops.

Written expression in boys with ADHD: The mediating roles of working memory and oral expression

ABSTRACTThe written expression difficulties experienced by children with ADHD are widely recognized; however, scant empirical evidence exists concerning the cognitive mechanisms and processes underlying these deficiencies. The current study investigated the independent and potentially interactive contributions of two developmentally antecedent cognitive processes – viz., working memory (WM) and oral expression – hypothesized to influence written expression ability in boys. Thirty-three boys with ADHD-Combined Presentation and 27 neurotypical (NT) boys 8–12 years of age were administered standardized measures of oral and written expression, and multiple counterbalanced tasks to assess WM central executive (CE) processes, WM phonological short-term memory (PH STM), and WM visuospatial short-term memory (VS STM). Bias-corrected bootstrapped mediation analyses revealed a significant mediation effect, wherein the independent and interactive effects of PH STM and oral expression collectively explained 76% of the diagnostic status to written expression relation. The implications of the obtained results for clinical practice suggest that children with ADHD may benefit by incorporating a blended approach that simultaneously strengthens PH STM capacity and oral expression abilities as antecedents to engaging in writing-related activities.

Smoothelin as a marker of muscularis propria in biopsies of colorectal carcinomas.

e14076 Background: Preoperative staging is of eminent importance for the treatment of colorectal carcinomas (CRCs). Especially the distinction of carcinoma in situ (not permigrating the muscularis mucosae (MM)), the pT1 stage (limited to the submucosa) and pT2 and above, infiltrating the muscularis propria(MP) in endoscopic biopsies could change the therapeutic regimen. Smoothelin (SMT) is a robust marker of the MP in normal colon, which is not expressed in MM; therefore, we analysed its expression in biopsies and resections of CRCs, to test its predictive value in staging of CRCs. Methods: 50 CRCs, diagnosed in biopsy particles and staged in the surgical specimens were analysed by IHC for expression of smoothelin (MoAb R4A, DCS Hamburg), Desmin and smooth muscle antigen SMA (all DAKO ) in MP and MM in the vicinity of invading carcinoma cells and in distance to the tumor. Semiquantitative grading REMMELE. Results: Desmin and SMA were consistently expressed in the smooth muscle cells of MM and MP near and distant to carcinoma cells. SMT showed no or little expression in the muscle cells of MM. The penetration of the MM by tumorcells did not alter their reactivity. In the MP however, the invasion by carcinoma cells caused a total or subtotal loss of SMT in the cytoplasm of the smooth muscle cells. The diameter of the nucleus and the longitudinal axis in SMT-deprivated cells did not change, but the transversal diameters of the cells were decreased. Reducing the mechanical stress of smooth muscle cells diminishes also the expression of SMT. No predictive correlation between biopsy-estimated depth and real depth of tumorinfiltration was seen. Conclusions: Expression of SMT correlates to functional status of the cells. The reliable difference in the IHC-expression of SMT in MM and MP in normal colon is not seen in the colon near carcinomas: the muscle cells of MP also lose their STM expression. Thus, negativity for SMT in smooth muscle cells near tumorcells does not necessarely proof, that this will be MM; as well, it could be SMT-deprivated cells of the MP. To rely upon SMT in biopsies for the determination of the depth of invasion is highly dangerous.

Functions of Rhomboid Family Protease RHBDL2 and Thrombomodulin in Wound Healing

The expression of thrombomodulin (TM), a membrane glycoprotein, is upregulated in neoepidermis during cutaneous wound healing. Rhomboid-like-2 (RHBDL2), an intramembrane serine protease, specifically cleaves TM at the transmembrane domain and causes the release of soluble TM (sTM). However, the physiological functions of TM and RHBDL2 in wound healing remain unclear. We demonstrated that both TM and RHBDL2 are upregulated in HaCaT cells stimulated by scratch wounds; furthermore, increased sTM was found in culture medium. Conversely, inhibition of RHBDL2 by 3,4-dichloroisocoumarin (DCI) or short hairpin RNA significantly inhibited wound-induced TM ectodomain shedding and wound healing. Both conditioned media from multiple-scratch-wounded HaCaT and recombinant sTM accelerated wound healing in HaCaT cells; such effects were abrogated by anti-TM antibodies. The RNA released from injured cells is involved in the induction of TM and RHBDL2. RHBDL2 and sTM were upregulated in ex vivo tissue culture of the injured skin. Furthermore, DCI inhibited sTM production and wound healing; this was reversed by recombinant sTM in mice. Thus, RHBDL2 and TM have important roles in wound healing via the release of sTM from keratinocytes; this may function as an autocrine/paracrine signal promoting wound healing.

Ectopic expression of the Arabidopsis MINI ZINC FINGER1 and MIF3 genes induces shoot meristems on leaf margins

A leaf undergoes determinate growth from a primordium on flank of the shoot apical meristem. Several intrinsic pathways restrict meristematic activity in the leaf of Arabidopsis; however, other factors remain to be defined. We report here that the overexpression of MINI ZINC FINGER1 (MIF1) or MIF3 disrupted the leaf determinate growth by inducing ectopic shoot meristems on leaf margins. These ectopic meristems occurred along margins of late rosette leaves at serration sinuses in an ERECTA-dependent manner. Expression of STM was activated in these ectopic meristems but not other leaf regions. The formation of ectopic meristems was independent of the BP gene but suppressed by exogenous gibberellic acid. In addition, reduced auxin response along leaf margins and subsequent response peak in the sinus were correlated with the occurrence of ectopic meristems. Our results suggest that the sinus of leaf serration is a quiescent domain possessing the potential for meristem formation. MIF1- or MIF3-overexpressing transgenic plants may provide a new genetic system for dissecting the molecular mechanism that maintains leaf determinate growth, and for understanding the interactions between hormone actions and meristematic activity.

Shoot apical meristem structure and STM expression in has mutant of Arabidopsis thaliana

Wild-type and the handshake (has) mutants in Arabidopsis thaliana were analyzed. Compared to the wild-type, has mutants display a number of morphological alterations, which can largely be traced back to altered meristem function. Analyses of apical meristem of mutant plants showed that mutation affected meristem structure and patterns of STM expression.

Gene TAENIATA Is a Novel Negative Regulator of the Arabidopsis thaliana Homeobox Genes KNAT1, KNAT2, KNAT6, and STM

Mutant Arabidopsis thaliana taeniata (tae) plants are characterized by an altered morphology of leaves and the inflorescence. At the beginning of flowering, the inflorescence produces fertile flowers morphologically intermediate between a shoot and a flower. The recessive mutation tae also causes the formation of ectopic meristems and shoot rosettes on leaves. The expressivity of the mutant characters depend on the temperature and photoperiod. Analysis of the activity of KNOX class I genes in the leaves of the tae mutant has demonstrated the expression of genes KNAT2 and STM and an increase in the expression of genes KNAT1 and KNAT6 compared to wild-type leaves. These data indicate that the TAE gene negatively regulates the KNAT1, KNAT2, KNAT6, and STM genes.

Investigation of cytokinin-deficient phenotypes in Arabidopsis by ectopic expression of orchid DSCKX1

The plant hormone cytokinin plays a major role in regulating plant growth and development. Here we generated cytokinin-reduction Arabidopsis plants by overexpressing a heterologous cytokinin oxidase gene DSCKX1 from Dendrobium orchid. These transgenic plants exhibited reduced biomass, rapid root growth, decreased ability to form roots in vitro, and reduced response to cytokinin in growing calli and roots. Furthermore, the expression of KNAT1, STM, and CycD3 genes was significantly reduced in the transgenic plants, suggesting that cytokinin may function to control the cell cycles and shoot/root development via regulation of these genes.

Axillary meristem development in Arabidopsis thaliana.

Axillary shoot apical meristems initiate post-embryonically in the axils of leaves. Their developmental fate is a main determinant of the final plant body plan. In Arabidopsis, usually a single axillary meristem initiates in the leaf axil even though there is developmental potential for formation of multiple branches. While the wild-type plants rarely form multiple branches in the leaf axil, tfl1-2 plants regularly develop two or more branches in the axils of the rosette leaves. Axillary meristem formation in Arabidopsis occurs in two waves: an acropetal wave forms during plant vegetative development, and a basipetal wave forms during plant reproductive development. We report here the morphological and anatomical changes, and the STM expression pattern associated with the formation of axillary and accessory meristems during Arabidopsis vegetative development.