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At least 37 records · Page 2

The putative transporter MtUMAMIT14 participates in nodule formation in Medicago truncatula

Transport systems are crucial in many plant processes, including plant–microbe interactions. Nodule formation and function in legumes involve the expression and regulation of multiple transport proteins, and many are still uncharacterized, particularly for nitrogen transport. Amino acids originating from the nitrogen-fixing process are an essential form of nitrogen for legumes. This work evaluates the role of MtN21 (henceforth MtUMAMIT14), a putative transport system from the MtN21/EamA-like/UMAMIT family, in nodule formation and nitrogen fixation in Medicago truncatula. To dissect this transporter’s role, we assessed the expression of MtUMAMIT14 using GUS staining, localized the corresponding protein in M. truncatula root and tobacco leaf cells, and investigated two independent MtUMAMIT14 mutant lines. Our results indicate that MtUMAMIT14 is localized in endosomal structures and is expressed in both the infection zone and interzone of nodules. Comparison of mutant and wild-type M. truncatula indicates MtUMAMIT14, the expression of which is dependent on the presence of NIN, DNF1, and DNF2, plays a role in nodule formation and nitrogen-fixation. While the function of the transporter is still unclear, our results connect root nodule nitrogen fixation in legumes with the UMAMIT family.

58 GEOSCIENCES↗

Emplacement of Xenolith Nodules in the Kaupulehu Lava Flow, Hualalai Volcano, Hawaii

The basaltic Kaupulehu 1800-1801 lava flow of Hualalai Volcano, Hawaii contains abundant ultramafic xenoliths. Many of these xenoliths occur as bedded layers of semi-rounded nodules, each thinly coated with a veneer (typically 1 mm thick) of lava. The nodule beds are analogous to cobble deposits of fluvial sedimentary systems. Although several mechanisms have been proposed for the formation of the nodule beds, it was found that, at more than one locality, the nodule beds are overbank levee deposits. The geological occurrence of the nodules, certain diagnostic aspects of the flow morphology and consideration of the inferred emplacement process indicate that the Kaupulehu flow had an exceptionally low viscosity on eruption and that the flow of the lava stream was extremely rapid, with flow velocities of at least 10 m/s (more than 40 km/h. This flow is the youngest on Hualalai Volcano and future eruptions of a similar type would pose considerable hazard to life as well as property.

Guest, J. E.↗

In situ synchrotron investigation of degenerate graphite nodule evolution in ductile cast iron

Ductile cast irons (DCIs) are of increasing importance in the renewable energy and transportation sectors. The distribution and morphology of the graphite nodules, in particular the formation of degenerate features during solidification, dictate the mechanical performance of DCIs. In situ high-speed synchrotron X-ray tomography was used to capture the evolution of graphite nodules during solidification of DCI, including degenerate features and the effect of the carbon concentration field. The degeneration of nodules is observed to increase with re-melting cycles, which is attributed to Mg-loss. The dendritic primary austenite and carbon concentration gradients in the surrounding liquid phase were found to control nodule morphology by locally restricting and promoting growth. A coupled diffusion-mechanical model was developed, confirming the experimentally informed hypothesis that protrusions form through liquation cracking of the austenite shell and subsequent localised growth. In conclusion, these results provide valuable insights into the solidification kinetics of cast irons, supporting the design of advanced alloys.

36 MATERIALS SCIENCE↗

Convergent evolution of NFP -facilitated root nodule symbiosis

The origin and phylogenetic distribution of symbiotic associations between nodulating angiosperms and nitrogen-fixing bacteria have long intrigued biologists. Recent comparative evolutionary analyses have yielded alternative hypotheses: a multistep pathway of independent gains and losses of root nodule symbiosis vs. a single gain followed by numerous losses. A detailed reconstruction of the history of genes involved in signaling between nitrogen-fixing bacteria and potential hosts, particularly lipo-chitooligosaccharide (LCO) signaling, is needed to distinguish between these hypotheses. LCO recognition by plants involves the Nod Factor Perception ( NFP ) gene family; in the legume model Medicago truncatula (Fabales), MtNFP is essential for establishing rhizobial symbiosis. Here, we document convergent evolution of NFP , indicating multiple origins of LCO-driven symbiosis. In contrast to previous models that explain the recruitment of NFP via a single duplication in the ancestor of the nitrogen-fixing clade, our phylogenomic and synteny results suggest this duplication does not span the entire clade. Tandem duplication in a common ancestor of Cucurbitales and Rosales resulted in the NFP1 and NFP2 groups. In contrast, the phylogenetically closest paralog of MtNFP is MtLYR1 , located on a different chromosome within a large syntenic block. All available data indicate that a large-scale duplication resulted in MtNFP and MtLYR1 , likely corresponding to a whole-genome duplication in an ancestor of subfamily Papilionoideae of Fabaceae. We show that MtNFP and the NFP2 -like group are not orthologous, indicating multiple independent gains of NFP -based LCO signaling. This molecular convergence provides a possible mechanism for multiple gains of root nodule symbiosis across the nitrogen-fixing clade.

LCO signaling↗

An LCO-responsive homolog of NODULE INCEPTION positively regulates lateral root formation in Populus sp.

Abstract The transcription factor NODULE INCEPTION (NIN) has been studied extensively for its multiple roles in root nodule symbiosis within plants of the nitrogen-fixing clade (NFC) that associate with soil bacteria, such as rhizobia and Frankia. However, NIN homologs are present in plants outside the NFC, suggesting a role in other developmental processes. Here, we show that the biofuel crop Populus sp., which is not part of the NFC, contains eight copies of NIN with diversified protein sequence and expression patterns. Lipo-chitooligosaccharides (LCOs) are produced by rhizobia and a wide range of fungi, including mycorrhizal ones, and act as symbiotic signals that promote lateral root formation. RNAseq analysis of Populus sp. treated with purified LCO showed induction of the PtNIN2 subfamily. Moreover, the expression of PtNIN2b correlated with the formation of lateral roots and was suppressed by cytokinin treatment. Constitutive expression of PtNIN2b overcame the inhibition of lateral root development by cytokinin under high nitrate conditions. Lateral root induction in response to LCOs likely represents an ancestral function of NIN retained and repurposed in nodulating plants, as we demonstrate that the role of NIN in LCO-induced root branching is conserved in both Populus sp. and legumes. We further established a visual marker of LCO perception in Populus sp. roots, the putative sulfotransferase PtSS1 that can be used to study symbiotic interactions with the bacterial and fungal symbionts of Populus sp.

Irving, Thomas B. (ORCID:0000000330404543)↗

Isolation of potential plant growth-promoting bacteria from nodules of legumes grown in arid Botswana soil

As the world population increases, improvements in crop growth and yield will be needed to meet rising food demands, especially in countries that have not developed agricultural practices optimized for their own soils and crops. In many African countries, farmers improve agricultural productivity by applying synthetic fertilizers and pesticides to crops, but their continued use over the years has had serious environmental consequences including air and water pollution as well as loss of soil fertility. To reduce the overuse of synthetic amendments, we are developing inocula for crops that are based on indigenous soil microbes, especially those that enhance plant growth and improve agricultural productivity in a sustainable manner. We first isolated environmental DNA from soil samples collected from an agricultural region to study the composition of the soil microbiomes and then used Vigna unguiculata (cowpea), an important legume crop in Botswana and other legumes as “trap” plants using the collected soil to induce nitrogen-fixing nodule formation. We have identified drought-tolerant bacteria from Botswana soils that stimulate plant growth; many are species of Bacillus and Paenibacillus . In contrast, the cowpea nodule microbiomes from plants grown in these soils house mainly rhizobia particularly Bradyrhizobium , but also Methylobacterium spp. Hence, the nodule microbiome is much more limited in non-rhizobial diversity compared to the soil microbiome, but also contains a number of potential pathogenic bacteria.

Kosty, Melissa↗

Final Scientific/Technical Report: Real Time Particle Size and Settling Velocity for In-Situ Monitoring of Deep-Sea Polymetallic Nodule Mining

In the coming decade, it is anticipated that deep-sea mining activities will commence throughout the deep ocean, at depths around 4000-6000m where vast deposits of baseball-sized rocks called polymetallic nodules lay on the seabed. These nodules contain resources such as cobalt, copper and nickel which are needed to produce batteries for electric vehicles. Collector machines driving on the seabed and gathering these nodules will stir up clouds of sediment, and there is concern about how the resulting concentration of sediment in the near-bottom ocean water will affect deep and mid-ocean biology. There is currently no technology capable of comprehensively monitoring the concentrations and properties of stirred up sediment in the deep. This project addressed this critical gap by developing a deep-sea particle measurement system (Real-Time Size and Settling Velocity, RTSSV) that uses a multi-camera high-resolution video imaging system, with accompanying image processing, to measure the concentration, size distribution and settling speed of sediment. Importantly, these measurements are made in-situ, so that delicate sediment aggregates will not be disturbed, as is the case when collecting samples at depth to be measured in the lab. The instrument can be mounted on a variety of established deep-sea platforms such as moorings, AUVs and ROVs, providing data in near real time, where the key measurements need to be made—from the source of the sediment disturbance and throughout the resulting plume—vital for determining if deep-sea mining can be done in an environmentally responsible manner. The developed system (RTSSV) will possess the only such capability worldwide, giving a US-based group a substantial competitive advantage as this newly evolving global industry develops. As added value, the technology will also have widespread application to the offshore oil and gas industry, dredging industry, and to sediment science research.

47 OTHER INSTRUMENTATION↗

Spatial mapping of plant N-glycosylation cellular heterogeneity inside soybean root nodules provided insights into legume-rhizobia symbiosis

Although ubiquitously present, information on the function of complex N-glycan posttranslational modification in plants is very limited and it is often neglected. In this work, we adopted an enzyme-assisted matrix-assisted laser desorption/ionization mass spectrometry imaging strategy to visualize the distribution and identity of N-glycans in soybean root nodules at a cellular resolution. We additionally performed proteomics analysis to probe the potential correlation to proteome changes during symbiotic rhizobia-legume interactions. Our ion images reveal that intense N-glycosylation occurs in the sclerenchyma layer, and inside the infected cells within the infection zone, while morphological structures such as the cortex, uninfected cells, and cells that form the attachment with the root are less N-glycosylated. Notably, we observed different N-glycan profiles between soybean root nodules infected with wild type rhizobia and those infected with mutant rhizobia incapable of efficiently fixing atmospheric nitrogen. The majority of complex N-glycan structures, particularly those with characteristic Lewis-a epitopes, are overexpressed in the mutant nodules. Our proteomic results revealed that these glycans likely originated from proteins that maintain the redox balance crucial for proper nitrogen fixation, but also from enzymes involved in N-glycan and phenylpropanoid biosynthesis. These findings indicate possible involvement of Lewis-a glycans in these critical pathways during legume-rhizobia symbiosis.

59 BASIC BIOLOGICAL SCIENCES↗

Discovery top-down proteomics in symbiotic soybean root nodules

Proteomic methods have been widely used to study proteins in complex biological samples to understand biological molecular mechanisms. Most well-established methods (known as bottom-up proteomics, BUP) employ an enzymatic digestion step to cleave intact proteins into smaller peptides for liquid chromatography (LC) mass spectrometry (MS) detection. In contrast, top-down proteomics (TDP) directly characterizes intact proteins including all possible post-translational modifications (PTMs), thus offering unique insights into proteoform biology where combinations of individual PTMs may play important roles. We performed TDP on soybean root nodules infected by the symbiotic Bradyrhizobium japonicum in both the wildtype bacterium and a nifH- mutant, which lacks the ability to fix nitrogen in the soybean root nodule. TDP captured 1648 proteoforms derived from 313 bacterial genes and 178 soybean genes. Leghemoglobin, the most abundant protein in the sample, existed in many truncated proteoforms. Interestingly, these truncated proteoforms were considerably more abundant in the wildtype relative to the nifH- mutant, implicating protease activity as an important factor in nitrogen fixation. Proteoforms with various PTMs and combinations thereof were identified using an unrestricted open modification search. This included less common PTMs such as myristoylation, palmitoylation, cyanylation, and sulfation. In parallel, we collected high resolution MS imaging (MSI) data of intact proteins and biopolymers (<20 kDa due to current technical limitations) from sections of the soybean root nodules using matrix-assisted laser desorption/ionization (MALDI) coupled to high resolution Orbitrap. Several detected proteoforms exhibited unique spatial distributions inside the infection zone and cortex, suggesting functional compartmentalization in these regions. A subset of peaks from the MALDI-MSI were assigned to proteoforms detected in TDP LCMS data based on matching accurate masses. Many of the proteins detected in both LCMS and MALDI-MSI are currently uncharacterized in UniProt: the PTM and spatial information presented here will be valuable in understanding their biological functions. Taken together, our study demonstrates how untargeted TDP approach can provide unique insights into plant proteoform biology. On-going technology developments are expected to further improve TDP coverage for more comprehensive high-throughput analysis of proteoforms.

59 BASIC BIOLOGICAL SCIENCES↗

Magnetite-sulfide chondrules and nodules in CK carbonaceous chondrites - Implications for the timing of CK oxidation

CK carbonaceous chondrites contain rare (about 0.1 vol pct) magnetite-sulfide chondrules that range from about 240 to 500 microns in apparent diameter and have ellipsoidal to spheroidal morphologies, granular textures, and concentric layering. They resemble the magnetite-sulfide nodules occurring inside mafic silicate chondrules in CK chondrites. It seems likely that the magnetite-sulfide chondrules constitute the subset of magnetite-sulfide nodules that escaped as immiscible droplets from their molten silicate chondrule hosts during chondrule formation. The intactness of the magnetite-sulfide chondrules and nodules implies that oxidation of CK metal occurred before agglomeration. Hence, the pervasive silicate darkening of CK chondrites was caused by the shock mobilization of magnetite and sulfide, not metallic Fe-Ni and sulfide as in shock-darkened ordinary chondrites.

Rubin, Alan E.↗

Crustal and subcrustal nodules in ejecta from Kilbourne Hole Maar, New Mexico

Nodules retrieved from the ejecta of volcanic craters serve as the source of two major items of information. The first is in providing details of the geochemistry and mineralogy of the Earth's interior by supplying samples of materials that cannot be obtained by existing drilling techniques. The other is in providing information regarding the process which led to their transport from the Earth's interior to the surface. The primary purpose of the present study was to examine the morphology of the nodules in an attempt to place some constraints on the process that brought them to the surface. This attempt is briefly discussed.

Whitford-Stark, J. L.↗

Compositions of large metal nodules in mesosiderites - Links to iron meteorite group IIIAB and the origin of mesosiderite subgroups

The compositions of large metal nodules in 12 mesosiderites, ranging from 3 mm to 4 cm in minimum dimensions and from 10 to 200 mg/g in silicate content, were analyzed by neutron activation; also analyzed were two additional samples (Pennyweight Point and Murchison Downs) which were originally designated iron meteorites but were later classified as Mes nodules. It was found that mesosiderites subgroups classified on the basis of metal composition are virtually the same as those based on silicate composition. The general covariation of the silicate and the metal compositional characteristics suggests that these features were established in the same process or in linked processes. A mesosiderite formation model is proposed.

Hassanzadeh, Jamshid↗

Thresholds of Detection and Identification of Halite Nodule Habitats in the Atacama Desert Using Remote Imaging

The guiding theme of Mars exploration is shifting from global and regional habitability assessment to biosignature detection. To locate features likely to contain biosignatures, it is useful to focus on the reliable identification of specific habitats with high biosignature preservation potential. Proposed chloride deposits on Mars may represent evaporitic environments conducive to the preservation of biosignatures. Analogous chloride- bearing, salt-encrusted playas (salars) are a habitat for life in the driest parts of the Atacama Desert, and are also environments with a taphonomic window. The specific geologic features that harbor and preserve microorganisms in Atacama salars are sub- meter to meter scale salt protuberances, or halite nodules. This study focuses on the ability to recognize and map halite nodules using images acquired from an unmanned aerial vehicle (UAV) at spatial resolutions ranging from mm/pixel to that of the highest resolution orbital images available for Mars.

Phillips, M. S.↗

Shifts in evolutionary lability underlie independent gains and losses of root-nodule symbiosis in a single clade of plants

Abstract Root nodule symbiosis (RNS) is a complex trait that enables plants to access atmospheric nitrogen converted into usable forms through a mutualistic relationship with soil bacteria. Pinpointing the evolutionary origins of RNS is critical for understanding its genetic basis, but building this evolutionary context is complicated by data limitations and the intermittent presence of RNS in a single clade of ca. 30,000 species of flowering plants, i.e., the nitrogen-fixing clade (NFC). We developed the most extensive de novo phylogeny for the NFC and an RNS trait database to reconstruct the evolution of RNS. Our analysis identifies evolutionary rate heterogeneity associated with a two-step process: An ancestral precursor state transitioned to a more labile state from which RNS was rapidly gained at multiple points in the NFC. We illustrate how a two-step process could explain multiple independent gains and losses of RNS, contrary to recent hypotheses suggesting one gain and numerous losses, and suggest a broader phylogenetic and genetic scope may be required for genome-phenome mapping.

59 BASIC BIOLOGICAL SCIENCES↗