New Tracer to Estimate Community Predation Rates of Phagotrophic Protists

Predation of eukaryotic microbes on prokaryotes is one of the most important trophic interactions on Earth, representing a major mortality term and shaping morphology and composition of prokaryotic communities. Here we introduce and validate a new tracer to determine predation rates on prokaryotes. Minicells of Escherichia coli marked with a bright green fluorescent protein (GFP) vector have many operational advantages over previously used prey analogs such as fluorescently labeled bacteria. GFP-minicells are similar in size to naturally occurring bacteria from a variety of environments including the oligotrophic open ocean and the deep sea. They are relatively stable against microbial and light degradation, are easy to grow and process, and can be produced inexpensively in large numbers. No chemical alteration of the particle surface due to heat killing and staining is involved. Grazing coefficients were compared between GFP-minicells and other GFP-modified bacteria, as well as 5-(4,6-dichlorotriazinyl) aminofluorescein (DTAF)stained cells. The grazing coefficients obtained from the removal of GFP-minicells compared favorably with estimates from tracer-independent estimates of grazing in the same experiments. Experiments with GFP-minicells resulted in community grazing coefficients similar to those reported for many different marine environments and those derived using various methods

Eukaryotic Microbes, Principally Fungi and Labyrinthulomycetes, Dominate Biomass on Bathypelagic Marine Snow

In the bathypelagic realm of the ocean, the role of marine snow as a carbon and energy source for the deep-sea biota and as a potential hotspot of microbial diversity and activity has not received adequate attention. Here, we collected bathypelagic marine snow by gentle gravity filtration of sea water onto μm filters from similar to 1000 to 3900 m to investigate the relative distribution of eukaryotic microbes. Compared with sediment traps that select for fast-sinking particles, this method collects particles unbiased by settling velocity. While prokaryotes numerically exceeded eukaryotes on marine snow, eukaryotic microbes belonging to two very distant branches of the eukaryote tree, the fungi and the labyrinthulomycetes, dominated overall biomass. Being tolerant to cold temperature and high hydrostatic pressure, these saprotrophic organisms have the potential to significantly contribute to the degradation of organic matter in the deep sea. Our results demonstrate that the community composition on bathypelagic marine snow differs greatly from that in the ambient water leading to wide ecological niche separation between the two environments

Examining Experience, Role, and LGBTQ Identity in Department Chairs

LGBTQ rights have progressed tremendously in recent times, not long ago LGBTQ individuals could be arrested simply for being themselves. Though many rights have been won, the fight for equity continues. This is especially true in the field of education, many think of higher education as a pathway to equity, but in reality it can serve to solidify societal inequities. Campus climate studies of LGBTQ faculty members in higher education show that climate is most impactful at the departmental level (Nichols & Scott, 2005), others highlight the importance of department chairs in fostering climate within their departments (Bystydzienski et al., 2017). Literature reveals a gap in examining the experiences of LGBTQ department chairs. Understanding these experiences and how LGBTQ identity impacts their various roles could provide insight to department chairs on how to improve their departmental climate for all faculty members, especially those within the LGTBQ community. This study utilizes Educational Criticism to gain a better understanding of how LGBTQ department chairs experience and work within their roles as faculty and departmental leaders. Two LGBTQ department chairs, Dani and Alex, highlight their experiences, how roles intersect with their queer identity, and examine how they challenge the norms of what it means to be a departmental leader. Their experiences are framed by not only their queer identity, but also in this case their gender presentation. Dani and Alex’s queer identity is present in many roles, but it is negotiated differently in each. Participants bring an outsider perspective to the department chair position, this perspective is influenced by their experiences and fuels their fight for equity in their departments and at their institutions

Sequential Transphosphorylation of the BRI1/BAK1 Receptor Kinase Complex Impacts Early Events in Brassinosteroid Signaling

SummaryBrassinosteroids (BRs) regulate plant development through a signal transduction pathway involving the BRI1 and BAK1 transmembrane receptor kinases. The detailed molecular mechanisms of phosphorylation, kinase activation, and oligomerization of the BRI1/BAK1 complex in response to BRs are uncertain. We demonstrate that BR-dependent activation of BRI1 precedes association with BAK1 in planta, and that BRI1 positively regulates BAK1 phosphorylation levels in vivo. BRI1 transphosphorylates BAK1 in vitro on specific kinase-domain residues critical for BAK1 function. BAK1 also transphosphorylates BRI1, thereby quantitatively increasing BRI1 kinase activity toward a specific substrate. We propose a sequential transphosphorylation model in which BRI1 controls signaling specificity by direct BR binding followed by substrate phosphorylation. The coreceptor BAK1 is then activated by BRI1-dependent transphosphorylation and subsequently enhances signaling output through reciprocal BRI1 transphosphorylation. This model suggests both conservation and distinct differences between the molecular mechanisms regulating phosphorylation-dependent kinase activation in plant and animal receptor kinases

Advancing the Research and Development of Assured Artificial Intelligence and Machine Learning Capabilities

Artificial intelligence (AI) and machine learning (ML) have become increasingly vital in the development of novel defense and intelligence capabilities across all domains of warfare. An adversarial AI (A2I) and adversarial ML (AML) attack seeks to deceive and manipulate AI/ML models. It is imperative that AI/ML models can defend against these attacks. A2I/AML defenses will help provide the necessary assurance of these advanced capabilities that use AI/ML models. The A2I Working Group (A2IWG) seeks to advance the research and development of assured AI/ML capabilities via new A2I/AML defenses by fostering a collaborative environment across the U.S. Department of Defense and U.S. Intelligence Community. The A2IWG aims to identify specific challenges that it can help solve or address more directly, with initial focus on three topics: AI Trusted Robustness, AI System Security, and AI/ML Architecture Vulnerabilities.Comment: Presented at AAAI FSS-20: Artificial Intelligence in Government and Public Sector, Washington, DC, US

The Arabidopsis leucine-rich repeat receptor kinase BIR3 negatively regulates BAK1 receptor complex formation and stabilizes BAK1

BAK1 is a co-receptor and positive regulator of multiple ligand-binding leucine-rich-repeat receptor kinases (LRR-RKs) and is involved in brassinosteroid (BR)-dependent growth and development, innate immunity and cell death control. The BAK1-interacting LRR-RKs BIR2 and BIR3 were previously identified by proteomics analyses of in vivo BAK1 complexes. Here we show that BAK1-related pathways such as innate immunity and cell death control are affected by BIR3 in Arabidopsis thaliana. BIR3 also has a strong negative impact on BR signaling. BIR3 directly interacts with the BR receptor BRI1 and other ligand-binding receptors and negatively regulates BR signaling by competitive inhibition of BRI1. BIR3 is released from BAK1 and BRI1 after ligand exposure and directly affects the formation of BAK1 complexes with BRI1 or FLAGELLIN SENSING2. Double mutants of bak1 and bir3 show spontaneous cell death and constitutive activation of defense responses. BAK1 and its closest homolog BKK1 interact with and are stabilized by BIR3, suggesting that bak1 bir3 double mutants mimic the spontaneous cell death phenotype observed in bak1 bkk1 mutants via destabilization of BIR3 target proteins. Our results provide evidence for a negative regulatory mechanism for BAK1 receptor complexes in which BIR3 interacts with BAK1 and inhibits ligand-binding receptors to prevent BAK1 receptor complex formation

The BRASSINOSTEROID INSENSITIVE1–LIKE3 Signalosome Complex Regulates Arabidopsis

Brassinosteroid (BR) hormones are primarily perceived at the cell surface by the leucine-rich repeat receptor-like kinase BRASSINOSTEROID INSENSITIVE1 (BRI1). In Arabidopsis thaliana, BRI1 has two close homologs, BRI1-LIKE1 (BRL1) and BRL3, respectively, which are expressed in the vascular tissues and regulate shoot vascular development. Here, we identify novel components of the BRL3 receptor complex in planta by immunoprecipitation and mass spectrometry analysis. Whereas BRI1 ASSOCIATED KINASE1 (BAK1) and several other known BRI1 interactors coimmunoprecipitated with BRL3, no evidence was found of a direct interaction between BRI1 and BRL3. In addition, we confirmed that BAK1 interacts with the BRL1 receptor by coimmunoprecipitation and fluorescence microscopy analysis. Importantly, genetic analysis of brl1 brl3 bak1-3 triple mutants revealed that BAK1, BRL1, and BRL3 signaling modulate root growth and development by contributing to the cellular activities of provascular and quiescent center cells. This provides functional relevance to the observed protein–protein interactions of the BRL3 signalosome. Overall, our study demonstrates that cell-specific BR receptor complexes can be assembled to perform different cellular activities during plant root growth, while highlighting that immunoprecipitation of leucine-rich repeat receptor kinases in plants is a powerful approach for unveiling signaling mechanisms with cellular resolution in plant development