“Proteotranscriptomic analysis of advanced colorectal cancer patient derived organoids for drug sensitivity prediction”

BACKGROUND: Patient-derived organoids (PDOs) from advanced colorectal cancer (CRC) patients could be a key platform to predict drug response and discover new biomarkers. We aimed to integrate PDO drug response with multi-omics characterization beyond genomics. METHODS: We generated 29 PDO lines from 22 advanced CRC patients and provided a morphologic, genomic, and transcriptomic characterization. We performed drug sensitivity assays with a panel of both standard and non-standard agents in five long-term cultures, and integrated drug response with a baseline proteomic and transcriptomic characterization by SWATH-MS and RNA-seq analysis, respectively. RESULTS: PDOs were successfully generated from heavily pre-treated patients, including a paired model of advanced MSI high CRC deriving from pre- and post-chemotherapy liver metastasis. Our PDOs faithfully reproduced genomic and phenotypic features of original tissue. Drug panel testing identified differential response among PDOs, particularly to oxaliplatin and palbociclib. Proteotranscriptomic analyses revealed that oxaliplatin non-responder PDOs present enrichment of the t-RNA aminoacylation process and showed a shift towards oxidative phosphorylation pathway dependence, while an exceptional response to palbociclib was detected in a PDO with activation of MYC and enrichment of chaperonin T-complex protein Ring Complex (TRiC), involved in proteome integrity. Proteotranscriptomic data fusion confirmed these results within a highly integrated network of functional processes involved in differential response to drugs. CONCLUSIONS: Our strategy of integrating PDOs drug sensitivity with SWATH-mass spectrometry and RNA-seq allowed us to identify different baseline proteins and gene expression profiles with the potential to predict treatment response/resistance and to help in the development of effective and personalized cancer therapeutics

Immobilization of the white-rot fungus Anthracophyllum discolor to degrade the herbicide atrazine

Herbicides cause environmental concerns because they are toxic and accumulate in the environment, food products and water supplies. There is a need to develop safe, efficient and economical methods to remove them from the environment, often by biodegradation. Atrazine is such herbicide. White-rot fungi have the ability to degrade herbicides of potential utility. This study formulated a novel pelletized support to immobilize the white-rot fungus Anthracophyllum discolor to improve its capability to degrade the atrazine using a biopurification system (BS). Different proportions of sawdust, starch, corn meal and flaxseed were used to generate three pelletized supports (F1, F2 and F3). In addition, immobilization with coated and uncoated pelletized supports (CPS and UPS, respectively) was assessed. UPS-F1 was determined as the most effective system as it provided high level of manganese peroxidase activity and fungal viability. The half-life (t1/2) of atrazine decreased from 14 to 6 days for the control and inoculated samples respectively. Inoculation with immobilized A. discolor produced an increase in the fungal taxa assessed by DGGE and on phenoloxidase activity determined. The treatment improves atrazine degradation and reduces migration to surface and groundwater.Grant CONICYT/FONDAP/15130015Grant FONDECYT 112096

Transforming growth factor beta signaling: The master sculptor of fingers

Transforming growth factor beta (TGF?) constitutes a large and evolutionarily conserved superfamily of secreted factors that play essential roles in embryonic development, cancer, tissue regeneration, and human degenerative pathology. Studies of this signaling cascade in the regulation of cellular and tissue changes in the three-dimensional context of a developing embryo have notably advanced in the understanding of the action mechanism of these growth factors. In this review, we address the role of TGF? signaling in the developing limb, focusing on its essential function in the morphogenesis of the autopod. As we discuss in this work, modern mouse genetic experiments together with more classical embryological approaches in chick embryos, provided very valuable information concerning the role of TGF? and Activin family members in the morphogenesis of the digits of tetrapods, including the formation of phalanxes, digital tendons, and interphalangeal joints. We emphasize the importance of the Activin and TGF? proteins as digit inducing factors and their critical interaction with the BMP signaling to sculpt the hand and foot morphology

Archaeology, Architecture, and the Postcolonial Critique

Peer reviewe

Transcript levels of Toll-Like receptors 5, 8 and 9 correlate with inflammatory activity in Ulcerative Colitis

<p>Abstract</p> <p>Background</p> <p>Dysregulation of innate immune response by Toll-Like Receptors (TLRs) is a key feature in Ulcerative Colitis (UC). Most studies have focused on <it>TLR2, TLR3</it>, and <it>TLR4 </it>participation in UC. However, few studies have explored other TLRs. Therefore, the aim of this study was to evaluate the mRNA profiles of <it>TLR1 to 9 </it>in colonic mucosa of UC patients, according to disease activity.</p> <p>Methods</p> <p>Colonic biopsies were taken from colon during colonoscopy in 51 patients with Ulcerative Colitis and 36 healthy controls. mRNA levels of <it>TLR1 to 9, Tollip</it>, inflammatory cytokines <it>IL6 </it>and <it>TNF </it>were assessed by RT-qPCR with hydrolysis probes. Characterization of <it>TLR9 </it>protein expression was performed by Immunohistochemistry.</p> <p>Results</p> <p>Toll-like receptors <it>TLR8, TLR9</it>, and <it>IL6 </it>mRNA levels were significantly higher in the colonic mucosa from UC patients (both quiescent and active) as compared to healthy individuals (p < 0.04). In the UC patients group the <it>TLR2, TLR4, TLR8 </it>and <it>TLR9 </it>mRNA levels were found to be significantly lower in patients with quiescent disease, as compared to those with active disease (p < 0.05), whereas <it>TLR5 </it>showed a trend (p = 0.06). <it>IL6 </it>and <it>TNF </it>mRNA levels were significantly higher in the presence of active disease and help to discriminate between quiescent and active disease (p < 0.05). Also, <it>IL6 </it>and <it>TNF </it>mRNA positively correlate with TLRs mRNA with the exception for <it>TLR3</it>, with stronger correlations for <it>TLR5, TLR8</it>, and <it>TLR9 </it>(p < 0.0001). <it>TLR9 </it>protein expression was mainly in the lamina propria infiltrate.</p> <p>Conclusions</p> <p>This study demonstrates that <it>TLR2, TLR4, TLR8</it>, and <it>TLR9 </it>expression increases in active UC patients, and that the mRNA levels positively correlate with the severity of intestinal inflammation as well as with inflammatory cytokines.</p

Mast cell lineage diversion of T lineage precursors by the essential T cell transcription factor GATA-3

GATA-3 is essential for T cell development from the earliest stages. However, abundant GATA-3 can drive T lineage precursors to a non–T cell fate, depending on Notch signaling and developmental stage. Here, overexpression of GATA-3 blocked the survival of pro–T cells when Notch-Delta signals were present but enhanced viability in their absence. In fetal thymocytes at the double-negative 1 (DN1) stage and DN2 stage but not those at the DN3 stage, overexpression of GATA-3 rapidly induced respecification to the mast cell lineage with high frequency by direct transcriptional 'reprogramming'. Normal DN2 thymocytes also showed mast cell potential when interleukin 3 and stem cell factor were added in the absence of Notch signaling. Our results suggest a close relationship between the pro–T cell and mast cell programs and a previously unknown function for Notch in T lineage fidelity

AFLP analysis reveals high genetic diversity but low population structure in Coccidioides posadasiiisolates from Mexico and Argentina

BACKGROUND: Coccidioides immitis and C. posadasii cause coccidioidomycosis, a disease that is endemic to North and South America, but for Central America, the incidence of coccidioidomycosis has not been clearly established. Several studies suggest genetic variability in these fungi; however, little definitive information has been discovered about the variability of Coccidioides fungi in Mexico (MX) and Argentina (AR). Thus, the goals for this work were to study 32 Coccidioides spp. isolates from MX and AR, identify the species of these Coccidioides spp. isolates, analyse their phenotypic variability, examine their genetic variability and investigate the Coccidioides reproductive system and its level of genetic differentiation. METHODS: Coccidioides spp. isolates from MX and AR were taxonomically identified by phylogenetic inference analysis using partial sequences of the Ag2/PRA gene and their phenotypic characteristics analysed. The genetic variability, reproductive system and level of differentiation were estimated using AFLP markers. The level of genetic variability was assessed measuring the percentage of polymorphic loci, number of effective allele, expected heterocygosity and Index of Association (I(A)). The degree of genetic differentiation was determined by AMOVA. Genetic similarities among isolates were estimated using Jaccard index. The UPGMA was used to contsruct the corresponding dendrogram. Finally, a network of haplotypes was built to evaluate the genealogical relationships among AFLP haplotypes. RESULTS: All isolates of Coccidioides spp. from MX and AR were identified as C. posadasii. No phenotypic variability was observed among the C. posadasii isolates from MX and AR. Analyses of genetic diversity and population structure were conducted using AFLP markers. Different estimators of genetic variability indicated that the C. posadasii isolates from MX and AR had high genetic variability. Furthermore, AMOVA, dendrogram and haplotype network showed a small genetic differentiation among the C. posadasii populations analysed from MX and AR. Additionally, the I(A) calculated for the isolates suggested that the species has a recombinant reproductive system. CONCLUSIONS: No phenotypic variability was observed among the C. posadasii isolates from MX and AR. The high genetic variability observed in the isolates from MX and AR and the small genetic differentiation observed among the C. posadasii isolates analysed, suggest that this species could be distributed as a single genetic population in Latin America

Association of sleep, screen time and physical activity with overweight and obesity in Mexico

PURPOSE: Approximately 70% of adults in Mexico are overweight or obese. Unhealthy lifestyle behaviors are also prevalent. We examined the association of three lifestyle behaviors with body mass index (BMI) categories in adults from Mexico. METHODS: We used publicly available data from the ENSANUT 2016 survey (n = 6419). BMI was used to categorize participants. Differences in sleep duration, suffering from symptoms of insomnia, TV watching time, time in front of any screen, vigorous physical activity (yes vs no), moderate physical activity (> 30 min/day-yes vs. no) and walking (> 60 min/day-yes vs. no) were compared across BMI groups using adjusted linear and logistic regression analyses. RESULTS: Thirty-nine percent of participants were overweight and 37% obese. Time in front of TV, in front of any screen, sleep duration and physical activity were significantly associated with overweight and obesity. Compared to normal weight participants, participants in the obese II category spend on average 0.60 h/day (95% CI 0.36-0.84, p = 0.001) and participants in the obese III category 0.54 h/day (95% CI 0.19-0.89, p < 0.001) more in front of any screen; participants in the obese II category reported 0.55 h/day less sleep (95% CI - 0.67 to - 0.43, p < 0.001); participants in the obese III category were less likely to engage in vigorous activity (OR = 0.60, 95% CI 0.43-0.84, p ≤ 0.003), or walking (OR = 0.65, 95% CI 0.49-0.88, p = 0.005). CONCLUSION: Screen time, sleeping hours, and physical activity were associated with overweight and obesity. However, these associations were not consistent across all BMI categories. Assuming established causal connections, overweight individuals and individuals with obesity would benefit from reduced screen time and engaging in moderate/vigorous physical activity. LEVEL OF EVIDENCE: Level III: observational case-control analytic study

Oxidative Stress Mediates Physiological Costs of Begging in Magpie (Pica pica) Nestlings

[Background] Theoretical models predict that a cost is necessary to guarantee honesty in begging displays given by offspring to solicit food from their parents. There is evidence for begging costs in the form of a reduced growth rate and immunocompetence. Moreover, begging implies vigorous physical activity and attentiveness, which should increase metabolism and thus the releasing of pro-oxidant substances. Consequently, we predict that soliciting offspring incur a cost in terms of oxidative stress, and growth rate and immune response (processes that generate pro-oxidants substances) are reduced in order to maintain oxidative balance. [Methodology/Principal Findings] We test whether magpie (Pica pica) nestlings incur a cost in terms of oxidative stress when experimentally forced to beg intensively, and whether oxidative balance is maintained by reducing growth rate and immune response. Our results show that begging provokes oxidative stress, and that nestlings begging for longer bouts reduce growth and immune response, thereby maintaining their oxidative status. [Conclusions/Significance] These findings help explaining the physiological link between begging and its associated growth and immunocompetence costs, which seems to be mediated by oxidative stress. Our study is a unique example of the complex relationships between the intensity of a communicative display (begging), oxidative stress, and life-history traits directly linked to viability.GM-R was supported by the Spanish Government (Ministerio de Ciencia y Tecnología, “Juan de la Cierva” program), and TR was supported by the Consejo Superior de Investigaciones Científicas (CSIC; Proyectos Intramurales Especiales)

The bHLH transcription factor SPATULA enables cytokinin signaling, and both activate auxin biosynthesis and transport genes at the medial domain of the gynoecium

[EN] Fruits and seeds are the major food source on earth. Both derive from the gynoecium and, therefore, it is crucial to understand the mechanisms that guide the development of this organ of angiosperm species. In Arabidopsis, the gynoecium is composed of two congenitally fused carpels, where two domains: medial and lateral, can be distinguished. The medial domain includes the carpel margin meristem (CMM) that is key for the production of the internal tissues involved in fertilization, such as septum, ovules, and transmitting tract. Interestingly, the medial domain shows a high cytokinin signaling output, in contrast to the lateral domain, where it is hardly detected. While it is known that cytokinin provides meristematic properties, understanding on the mechanisms that underlie the cytokinin signaling pattern in the young gynoecium is lacking. Moreover, in other tissues, the cytokinin pathway is often connected to the auxin pathway, but we also lack knowledge about these connections in the young gynoecium. Our results reveal that cytokinin signaling, that can provide meristematic properties required for CMM activity and growth, is enabled by the transcription factor SPATULA (SPT) in the medial domain. Meanwhile, cytokinin signaling is confined to the medial domain by the cytokinin response repressor ARABIDOPSIS HISTIDINE PHOSPHOTRANSFERASE 6 (AHP6), and perhaps by ARR16 (a type-A ARR) as well, both present in the lateral domains (presumptive valves) of the developing gynoecia. Moreover, SPT and cytokinin, probably together, promote the expression of the auxin biosynthetic gene TRYPTOPHAN AMINOTRANSFERASE OF ARABIDOPSIS 1 (TAA1) and the gene encoding the auxin efflux transporter PIN-FORMED 3 (PIN3), likely creating auxin drainage important for gynoecium growth. This study provides novel insights in the spatiotemporal determination of the cytokinin signaling pattern and its connection to the auxin pathway in the young gynoecium.IRO, VMZM, HHU and PLS were supported by the Mexican National Council of Science and Technology (CONACyT) with a PhD fellowship (210085, 210100, 243380 and 219883, respectively). Work in the SDF laboratory was financed by the CONACyT grants CB-2012-177739, FC-2015-2/1061, and INFR-2015-253504, and NMM by the CONACyT grant CB-2011-165986. SDF, CF and LC acknowledge the support of the European Union FP7-PEOPLE-2009-IRSES project EVOCODE (grant no. 247587) and H2020-MSCARISE-2015 project ExpoSEED (grant no. 691109). SDF also acknowledges the Marine Biological Laboratory (MBL) in Woods Hole for a scholarship for the Gene Regulatory Networks for Development Course 2015 (GERN2015). IE acknowledges the International European Fellowship-METMADS project and the Universita degli Studi di Milano (RTD-A; 2016). Research in the laboratory of MFY was funded by NSF (grant IOS-1121055), NIH (grant 1R01GM112976-01A1) and the Paul D. Saltman Endowed Chair in Science Education (MFY). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Reyes Olalde, J.; Zuñiga, V.; Serwatowska, J.; Chávez Montes, R.; Lozano-Sotomayor, P.; Herrera-Ubaldo, H.; Gonzalez Aguilera, K.... (2017). The bHLH transcription factor SPATULA enables cytokinin signaling, and both activate auxin biosynthesis and transport genes at the medial domain of the gynoecium. PLoS Genetics. 13(4):1-31. https://doi.org/10.1371/journal.pgen.1006726S131134Reyes-Olalde, J. I., Zuñiga-Mayo, V. M., Chávez Montes, R. A., Marsch-Martínez, N., & de Folter, S. (2013). Inside the gynoecium: at the carpel margin. Trends in Plant Science, 18(11), 644-655. doi:10.1016/j.tplants.2013.08.002Alvarez-Buylla, E. R., Benítez, M., Corvera-Poiré, A., Chaos Cador, Á., de Folter, S., Gamboa de Buen, A., … Sánchez-Corrales, Y. E. (2010). Flower Development. The Arabidopsis Book, 8, e0127. doi:10.1199/tab.0127Bowman, J. L., Baum, S. F., Eshed, Y., Putterill, J., & Alvarez, J. (1999). 4 Molecular Genetics of Gynoecium Development in Arabidopsis. Current Topics in Developmental Biology Volume 45, 155-205. doi:10.1016/s0070-2153(08)60316-6Chávez Montes, R. A., Herrera-Ubaldo, H., Serwatowska, J., & de Folter, S. (2015). Towards a comprehensive and dynamic gynoecium gene regulatory network. Current Plant Biology, 3-4, 3-12. doi:10.1016/j.cpb.2015.08.002Marsch-Martínez, N., & de Folter, S. (2016). Hormonal control of the development of the gynoecium. Current Opinion in Plant Biology, 29, 104-114. doi:10.1016/j.pbi.2015.12.006Marsch-Martínez, N., Ramos-Cruz, D., Irepan Reyes-Olalde, J., Lozano-Sotomayor, P., Zúñiga-Mayo, V. M., & de Folter, S. (2012). The role of cytokinin during Arabidopsis gynoecia and fruit morphogenesis and patterning. The Plant Journal, 72(2), 222-234. doi:10.1111/j.1365-313x.2012.05062.xZhao, Z., Andersen, S. U., Ljung, K., Dolezal, K., Miotk, A., Schultheiss, S. J., & Lohmann, J. U. (2010). Hormonal control of the shoot stem-cell niche. Nature, 465(7301), 1089-1092. doi:10.1038/nature09126Ashikari, M. (2005). Cytokinin Oxidase Regulates Rice Grain Production. Science, 309(5735), 741-745. doi:10.1126/science.1113373Bartrina, I., Otto, E., Strnad, M., Werner, T., & Schmülling, T. (2011). Cytokinin Regulates the Activity of Reproductive Meristems, Flower Organ Size, Ovule Formation, and Thus Seed Yield in Arabidopsis thaliana. The Plant Cell, 23(1), 69-80. doi:10.1105/tpc.110.079079Hwang, I., Sheen, J., & Müller, B. (2012). Cytokinin Signaling Networks. Annual Review of Plant Biology, 63(1), 353-380. doi:10.1146/annurev-arplant-042811-105503Schaller, G. E., Bishopp, A., & Kieber, J. J. (2015). The Yin-Yang of Hormones: Cytokinin and Auxin Interactions in Plant Development. The Plant Cell, 27(1), 44-63. doi:10.1105/tpc.114.133595Kieber, J. J., & Schaller, G. E. (2010). The Perception of Cytokinin: A Story 50 Years in the Making: Figure 1. Plant Physiology, 154(2), 487-492. doi:10.1104/pp.110.161596Long, J. A., Moan, E. I., Medford, J. I., & Barton, M. K. (1996). A member of the KNOTTED class of homeodomain proteins encoded by the STM gene of Arabidopsis. Nature, 379(6560), 66-69. doi:10.1038/379066a0Jasinski, S., Piazza, P., Craft, J., Hay, A., Woolley, L., Rieu, I., … Tsiantis, M. (2005). KNOX Action in Arabidopsis Is Mediated by Coordinate Regulation of Cytokinin and Gibberellin Activities. Current Biology, 15(17), 1560-1565. doi:10.1016/j.cub.2005.07.023Yanai, O., Shani, E., Dolezal, K., Tarkowski, P., Sablowski, R., Sandberg, G., … Ori, N. (2005). Arabidopsis KNOXI Proteins Activate Cytokinin Biosynthesis. Current Biology, 15(17), 1566-1571. doi:10.1016/j.cub.2005.07.060Scofield, S., Dewitte, W., Nieuwland, J., & Murray, J. A. H. (2013). The Arabidopsis homeobox gene SHOOT MERISTEMLESS has cellular and meristem-organisational roles with differential requirements for cytokinin and CYCD3 activity. The Plant Journal, 75(1), 53-66. doi:10.1111/tpj.12198Gordon, S. P., Chickarmane, V. S., Ohno, C., & Meyerowitz, E. M. (2009). Multiple feedback loops through cytokinin signaling control stem cell number within the Arabidopsis shoot meristem. Proceedings of the National Academy of Sciences, 106(38), 16529-16534. doi:10.1073/pnas.0908122106Chickarmane, V. S., Gordon, S. P., Tarr, P. T., Heisler, M. G., & Meyerowitz, E. M. (2012). Cytokinin signaling as a positional cue for patterning the apical-basal axis of the growing Arabidopsis shoot meristem. Proceedings of the National Academy of Sciences, 109(10), 4002-4007. doi:10.1073/pnas.1200636109Leibfried, A., To, J. P. C., Busch, W., Stehling, S., Kehle, A., Demar, M., … Lohmann, J. U. (2005). WUSCHEL controls meristem function by direct regulation of cytokinin-inducible response regulators. Nature, 438(7071), 1172-1175. doi:10.1038/nature04270Werner, T., Motyka, V., Laucou, V., Smets, R., Van Onckelen, H., & Schmülling, T. (2003). Cytokinin-Deficient Transgenic Arabidopsis Plants Show Multiple Developmental Alterations Indicating Opposite Functions of Cytokinins in the Regulation of Shoot and Root Meristem Activity. The Plant Cell, 15(11), 2532-2550. doi:10.1105/tpc.014928Larsson, E., Franks, R. G., & Sundberg, E. (2013). Auxin and the Arabidopsis thaliana gynoecium. Journal of Experimental Botany, 64(9), 2619-2627. doi:10.1093/jxb/ert099Weijers, D., & Wagner, D. (2016). Transcriptional Responses to the Auxin Hormone. Annual Review of Plant Biology, 67(1), 539-574. doi:10.1146/annurev-arplant-043015-112122Robert, H. S., Crhak Khaitova, L., Mroue, S., & Benková, E. (2015). The importance of localized auxin production for morphogenesis of reproductive organs and embryos inArabidopsis. Journal of Experimental Botany, 66(16), 5029-5042. doi:10.1093/jxb/erv256Kuusk, S., Sohlberg, J. J., Magnus Eklund, D., & Sundberg, E. (2006). Functionally redundantSHIfamily genes regulate Arabidopsis gynoecium development in a dose-dependent manner. The Plant Journal, 47(1), 99-111. doi:10.1111/j.1365-313x.2006.02774.xSohlberg, J. J., Myrenås, M., Kuusk, S., Lagercrantz, U., Kowalczyk, M., Sandberg, G., & Sundberg, E. (2006). STY1regulates auxin homeostasis and affects apical-basal patterning of the Arabidopsis gynoecium. The Plant Journal, 47(1), 112-123. doi:10.1111/j.1365-313x.2006.02775.xStåldal, V., Sohlberg, J. J., Eklund, D. M., Ljung, K., & Sundberg, E. (2008). Auxin can act independently ofCRC,LUG,SEU,SPTandSTY1in style development but not apical-basal patterning of theArabidopsisgynoecium. New Phytologist, 180(4), 798-808. doi:10.1111/j.1469-8137.2008.02625.xVan Gelderen, K., van Rongen, M., Liu, A., Otten, A., & Offringa, R. (2016). An INDEHISCENT-Controlled Auxin Response Specifies the Separation Layer in Early Arabidopsis Fruit. Molecular Plant, 9(6), 857-869. doi:10.1016/j.molp.2016.03.005José Ripoll, J., Bailey, L. J., Mai, Q.-A., Wu, S. L., Hon, C. T., Chapman, E. J., … Yanofsky, M. F. (2015). microRNA regulation of fruit growth. Nature Plants, 1(4). doi:10.1038/nplants.2015.36Larsson, E., Roberts, C. J., Claes, A. R., Franks, R. G., & Sundberg, E. (2014). Polar Auxin Transport Is Essential for Medial versus Lateral Tissue Specification and Vascular-Mediated Valve Outgrowth in Arabidopsis Gynoecia. Plant Physiology, 166(4), 1998-2012. doi:10.1104/pp.114.245951Nole-Wilson, S., Azhakanandam, S., & Franks, R. G. (2010). Polar auxin transport together with AINTEGUMENTA and REVOLUTA coordinate early Arabidopsis gynoecium development. Developmental Biology, 346(2), 181-195. doi:10.1016/j.ydbio.2010.07.016De Folter, S. (2016). Auxin Is Required for Valve Margin Patterning in Arabidopsis After All. Molecular Plant, 9(6), 768-770. doi:10.1016/j.molp.2016.05.005Moubayidin, L., & Østergaard, L. (2014). Dynamic Control of Auxin Distribution Imposes a Bilateral-to-Radial Symmetry Switch during Gynoecium Development. Current Biology, 24(22), 2743-2748. doi:10.1016/j.cub.2014.09.080Girin, T., Paicu, T., Stephenson, P., Fuentes, S., Körner, E., O’Brien, M., … Østergaard, L. (2011). INDEHISCENT and SPATULA Interact to Specify Carpel and Valve Margin Tissue and Thus Promote Seed Dispersal in Arabidopsis. The Plant Cell, 23(10), 3641-3653. doi:10.1105/tpc.111.090944Ioio, R. D., Nakamura, K., Moubayidin, L., Perilli, S., Taniguchi, M., Morita, M. T., … Sabatini, S. (2008). A Genetic Framework for the Control of Cell Division and Differentiation in the Root Meristem. Science, 322(5906), 1380-1384. doi:10.1126/science.1164147Bishopp, A., Help, H., El-Showk, S., Weijers, D., Scheres, B., Friml, J., … Helariutta, Y. (2011). A Mutually Inhibitory Interaction between Auxin and Cytokinin Specifies Vascular Pattern in Roots. Current Biology, 21(11), 917-926. doi:10.1016/j.cub.2011.04.017De Rybel, B., Adibi, M., Breda, A. S., Wendrich, J. R., Smit, M. E., Novák, O., … Weijers, D. (2014). Integration of growth and patterning during vascular tissue formation in Arabidopsis. Science, 345(6197), 1255215. doi:10.1126/science.1255215Pernisova, M., Klima, P., Horak, J., Valkova, M., Malbeck, J., Soucek, P., … Hejatko, J. (2009). Cytokinins modulate auxin-induced organogenesis in plants via regulation of the auxin efflux. Proceedings of the National Academy of Sciences, 106(9), 3609-3614. doi:10.1073/pnas.0811539106Cheng, Z. J., Wang, L., Sun, W., Zhang, Y., Zhou, C., Su, Y. H., … Zhang, X. S. (2012). Pattern of Auxin and Cytokinin Responses for Shoot Meristem Induction Results from the Regulation of Cytokinin Biosynthesis by AUXIN RESPONSE FACTOR3. Plant Physiology, 161(1), 240-251. doi:10.1104/pp.112.203166Alvarez, J., & Smyth, D. R. (2002). CRABS CLAWandSPATULAGenes Regulate Growth and Pattern Formation during Gynoecium Development inArabidopsis thaliana. International Journal of Plant Sciences, 163(1), 17-41. doi:10.1086/324178Groszmann, M., Bylstra, Y., Lampugnani, E. R., & Smyth, D. R. (2010). Regulation of tissue-specific expression of SPATULA, a bHLH gene involved in carpel development, seedling germination, and lateral organ growth in Arabidopsis. Journal of Experimental Botany, 61(5), 1495-1508. doi:10.1093/jxb/erq015Smyth, D. R., Bowman, J. L., & Meyerowitz, E. M. (1990). Early flower development in Arabidopsis. The Plant Cell, 2(8), 755-767. doi:10.1105/tpc.2.8.755Müller, B., & Sheen, J. (2008). Cytokinin and auxin interaction in root stem-cell specification during early embryogenesis. Nature, 453(7198), 1094-1097. doi:10.1038/nature06943Argyros, R. D., Mathews, D. E., Chiang, Y.-H., Palmer, C. M., Thibault, D. M., Etheridge, N., … Schaller, G. E. (2008). Type B Response Regulators of Arabidopsis Play Key Roles in Cytokinin Signaling and Plant Development. The Plant Cell, 20(8), 2102-2116. doi:10.1105/tpc.108.059584Mason, M. G., Mathews, D. E., Argyros, D. A., Maxwell, B. B., Kieber, J. J., Alonso, J. M., … Schaller, G. E. (2005). Multiple Type-B Response Regulators Mediate Cytokinin Signal Transduction in Arabidopsis. The Plant Cell, 17(11), 3007-3018. doi:10.1105/tpc.105.035451Ishida, K., Yamashino, T., Yokoyama, A., & Mizuno, T. (2008). Three Type-B Response Regulators, ARR1, ARR10 and ARR12, Play Essential but Redundant Roles in Cytokinin Signal Transduction Throughout the Life Cycle of Arabidopsis thaliana. Plant and Cell Physiology, 49(1), 47-57. doi:10.1093/pcp/pcm165Yokoyama, A., Yamashino, T., Amano, Y.-I., Tajima, Y., Imamura, A., Sakakibara, H., & Mizuno, T. (2006). Type-B ARR Transcription Factors, ARR10 and ARR12, are Implicated in Cytokinin-Mediated Regulation of Protoxylem Differentiation in Roots of Arabidopsis thaliana. Plant and Cell Physiology, 48(1), 84-96. doi:10.1093/pcp/pcl040Schuster, C., Gaillochet, C., & Lohmann, J. U. (2015). Arabidopsis HECATE genes function in phytohormone control during gynoecium development. Development, 142(19), 3343-3350. doi:10.1242/dev.120444Toledo-Ortiz, G., Huq, E., & Quail, P. H. (2003). The Arabidopsis Basic/Helix-Loop-Helix Transcription Factor Family. The Plant Cell, 15(8), 1749-1770. doi:10.1105/tpc.013839Reymond, M. C., Brunoud, G., Chauvet, A., Martínez-Garcia, J. F., Martin-Magniette, M.-L., Monéger, F., & Scutt, C. P. (2012). A Light-Regulated Genetic Module Was Recruited to Carpel Development in Arabidopsis following a Structural Change to SPATULA. The Plant Cell, 24(7), 2812-2825. doi:10.1105/tpc.112.097915Ballester, P., Navarrete-Gómez, M., Carbonero, P., Oñate-Sánchez, L., & Ferrándiz, C. (2015). Leaf expansion in Arabidopsis is controlled by a TCP-NGA regulatory module likely conserved in distantly related species. Physiologia Plantarum, 155(1), 21-32. doi:10.1111/ppl.12327Hellens, R., Allan, A., Friel, E., Bolitho, K., Grafton, K., Templeton, M., … Laing, W. (2005). Plant Methods, 1(1), 13. doi:10.1186/1746-4811-1-13Makkena, S., & Lamb, R. S. (2013). The bHLH transcription factor SPATULA regulates root growth by controlling the size of the root meristem. BMC Plant Biology, 13(1), 1. doi:10.1186/1471-2229-13-1Stepanova, A. N., Robertson-Hoyt, J., Yun, J., Benavente, L. M., Xie, D.-Y., Doležal, K., … Alonso, J. M. (2008). TAA1-Mediated Auxin Biosynthesis Is Essential for Hormone Crosstalk and Plant Development. Cell, 133(1), 177-191. doi:10.1016/j.cell.2008.01.047Bhargava, A., Clabaugh, I., To, J. P., Maxwell, B. B., Chiang, Y.-H., Schaller, G. E., … Kieber, J. J. (2013). Identification of Cytokinin-Responsive Genes Using Microarray Meta-Analysis and RNA-Seq in Arabidopsis. Plant Physiology, 162(1), 272-294. doi:10.1104/pp.113.217026Sakai, H., Aoyama, T., & Oka, A. (2000). Arabidopsis ARR1 and ARR2 response regulators operate as transcriptional activators. The Plant Journal, 24(6), 703-711. doi:10.1046/j.1365-313x.2000.00909.xSakai, H. (2001). ARR1, a Transcription Factor for Genes Immediately Responsive to Cytokinins. Science, 294(5546), 1519-1521. doi:10.1126/science.1065201Moubayidin, L., Di Mambro, R., Sozzani, R., Pacifici, E., Salvi, E., Terpstra, I., … Sabatini, S. (2013). Spatial Coordination between Stem Cell Activity and Cell Differentiation in the Root Meristem. Developmental Cell, 26(4), 405-415. doi:10.1016/j.devcel.2013.06.025Benková, E., Michniewicz, M., Sauer, M., Teichmann, T., Seifertová, D., Jürgens, G., & Friml, J. (2003). Local, Efflux-Dependent Auxin Gradients as a Common Module for Plant Organ Formation. Cell, 115(5), 591-602. doi:10.1016/s0092-8674(03)00924-3Okada, K., Ueda, J., Komaki, M. K., Bell, C. J., & Shimura, Y. (1991). Requirement of the Auxin Polar Transport System in Early Stages of Arabidopsis Floral Bud Formation. The Plant Cell, 677-684. doi:10.1105/tpc.3.7.677Blilou, I., Xu, J., Wildwater, M., Willemsen, V., Paponov, I., Friml, J., … Scheres, B. (2005). The PIN auxin efflux facilitator network controls growth and patterning in Arabidopsis roots. Nature, 433(7021), 39-44. doi:10.1038/nature03184Mahonen, A. P. (2006). Cytokinin Signaling and Its Inhibitor AHP6 Regulate Cell Fate During Vascular Development. Science, 311(5757), 94-98. doi:10.1126/science.1118875Besnard, F., Refahi, Y., Morin, V., Marteaux, B., Brunoud, G., Chambrier, P., … Vernoux, T. (2013). Cytokinin signalling inhibitory fields provide robustness to phyllotaxis. Nature, 505(7483), 417-421. doi:10.1038/nature12791Longabaugh, W. J. R., Davidson, E. H., & Bolouri, H. (2005). Computational representation of developmental genetic regulatory networks. Developmental Biology, 283(1), 1-16. doi:10.1016/j.ydbio.2005.04.023Faure, E., Peter, I. S., & Davidson, E. H. (2013). A New Software Package for Predictive Gene Regulatory Network Modeling and Redesign. Journal of Computational Biology, 20(6), 419-423. doi:10.1089/cmb.2012.0297Mangan, S., & Alon, U. (2003). Structure and function of the feed-forward loop network motif. Proceedings of the National Academy of Sciences, 100(21), 11980-11985. doi:10.1073/pnas.2133841100Chen, Q., Liu, Y., Maere, S., Lee, E., Van Isterdael, G., Xie, Z., … Vanneste, S. (2015). A coherent transcriptional feed-forward motif model for mediating auxin-sensitive PIN3 expression during lateral root development. Nature Communications, 6(1). doi:10.1038/ncomms9821Qiu, K., Li, Z., Yang, Z., Chen, J., Wu, S., Zhu, X., … Zhou, X. (2015). EIN3 and ORE1 Accelerate Degreening during Ethylene-Mediated Leaf Senescence by Directly Activating Chlorophyll Catabolic Genes in Arabidopsis. PLOS Genetics, 11(7), e1005399. doi:10.1371/journal.pgen.1005399Seaton, D. D., Smith, R. W., Song, Y. H., MacGregor, D. R., Stewart, K., Steel, G., … Halliday, K. J. (2015). Linked circadian outputs control elongation growth and flowering in response to photoperiod and temperature. Molecular Systems Biology, 11(1), 776. doi:10.15252/msb.20145766Roeder, A. H. K., & Yanofsky, M. F. (2006). Fruit Development in Arabidopsis. The Arabidopsis Book, 4, e0075. doi:10.1199/tab.0075Marsch-Martínez, N., Reyes-Olalde, J. I., Ramos-Cruz, D., Lozano-Sotomayor, P., Zúñiga-Mayo, V. M., & de Folter, S. (2012). Hormones talking. Plant Signaling & Behavior, 7(12), 1698-1701. doi:10.4161/psb.22422Balanza, V., Navarrete, M., Trigueros, M., & Ferrandiz, C. (2006). Patterning the female side of Arabidopsis: the importance of hormones. Journal of Experimental Botany, 57(13), 3457-3469. doi:10.1093/jxb/erl188Kamiuchi, Y., Yamamoto, K., Furutani, M., Tasaka, M., & Aida, M. (2014). The CUC1 and CUC2 genes promote carpel margin meristem formation during Arabidopsis gynoecium development. Frontiers in Plant Science, 5. doi:10.3389/fpls.2014.00165Scofield, S., Dewitte, W., & Murray, J. A. H. (2007). The KNOX gene SHOOT MERISTEMLESS is required for the development of reproductive meristematic tissues in Arabidopsis. The Plant Journal, 50(5), 767-781. doi:10.1111/j.1365-313x.2007.03095.xLi, K., Yu, R., Fan, L.-M., Wei, N., Chen, H., & Deng, X. W. (2016). DELLA-mediated PIF degradation contributes to coordination of light and gibberellin signalling in Arabidopsis. Nature Communications, 7(1). doi:10.1038/ncomms11868Oh, E., Zhu, J.-Y., & Wang, Z.-Y. (2012). Interaction between BZR1 and PIF4 integrates brassinosteroid and environmental responses. Nature Cell Biology, 14(8), 802-809. doi:10.1038/ncb2545Sharma, N., Xin, R., Kim, D.-H., Sung, S., Lange, T., & Huq, E. (2016). NO FLOWERING IN SHORT DAY (NFL) is a bHLH transcription factor that promotes flowering specifically under short-day conditions inArabidopsis. Development, 143(4), 682-690. doi:10.1242/dev.128595Varaud, E., Brioudes, F., Szécsi, J., Leroux, J., Brown, S., Perrot-Rechenmann, C., & Bendahmane, M. (2011). AUXIN RESPONSE FACTOR8 Regulates Arabidopsis Petal Growth by Interacting with the bHLH Transcription Factor BIGPETALp. The Plant Cell, 23(3), 973-983. doi:10.1105/tpc.110.081653Savaldi-Goldstein, S., & Chory, J. (2008). Growth coordination and the shoot epidermis. Current Opinion in Plant Biology, 11(1), 42-48. doi:10.1016/j.pbi.2007.10.009Schuster, C., Gaillochet, C., Medzihradszky, A., Busch, W., Daum, G., Krebs, M., … Lohmann, J. U. (2014). A Regulatory Framework for Shoot Stem Cell Co