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John goes for a ride in the sky on Ace Dragon's back and finds him not so limited after all.

Plants deploy several ammonium transporter (AMT) and nitrate transporter (NRT) genes to acquire NH 4 + and NO 3 − from the soil into the roots and then transport them to other plant organs. Coding sequences of wheat genes obtained from ENSEMBL were aligned to known AMT and NRT sequences of Arabidopsis , barley, maize, rice, and wheat to retrieve homologous genes. Bayesian phylogenetic relationships among these genes showed distinct classification of sequences with significant homology to NRT 1, NRT2 , and NRT3 ( NAR2 ). Inter-species gene duplication analysis showed that eight AMT and 77 NRT genes were orthologous to the AMT and NRT genes of aforementioned plant species. Expression patterns of these genes were studied via whole transcriptome sequencing of 21-day old seedlings of five spring wheat lines. Eight AMT and 52 NRT genes were differentially expressed between root and shoot; and 131 genes did not express neither in root nor in shoot of 21-day old seedlings. Homeologous genes in the A, B, and D genomes, characterized by high sequence homology, revealed that their counterparts exhibited different expression patterns. This complement and evolutionary relationship of wheat AMT and NRT genes is expected to help in development of wheat germplasm with increased efficiency in nitrogen uptake and usage.

\"Baffin Island: Sam and Remi Fargo are on a climate-control expedition in the Arctic, when to their astonishment they discover a Viking ship in the ice, perfectly preserved--and filled with pre-Columbian artifacts from Mexico. How can that be? As they investigate, tantalizing clues about a link between the Vikings and the legendary Toltec feathered-serpent god Quetzalcoatl--and a fabled object known as the Eye of Heaven--begin to emerge. But so do many dangerous people. Soon the Fargos find themselves on the run through jungles, temples, and secret tombs, caught among treasure-hunters, crime cartels, and those with a far more personal motivation for stopping them. The solution to a thousand-year-old mystery awaits them at the end of the chase--if they manage to survive it.\"--Amazon.com.

Wheat growing regions and seasons are diverse, mandating different varietal adaptation and management practices. Grain yield is the primary target for soft-red winter (SRW) wheat, due to lower protein content requirements. The growing season for SRW wheat in the eastern United States takes up to 9 months under variable environments, highlighting the importance of variety and management. In this study, we present the results of a 2-year field-based investigation of yield response of 30 wheat lines to different nitrogen treatments by dissecting yield to its components. For 5 out of the 30 lines, we performed in-tissue nitrogen analysis. Spring nitrogen (N) treatments were two levels of 0 kg N ha (low N) and 112 kg N ha (high N). On average, application of 112 kg N in the spring, in addition to fall N fertilizer, increased phytomass by 22% at maturity, enhanced fertile tiller numbers by 16%, and increased grain yield by 18% that coincided with a 26% increase in grain number per unit area. N in the grains, or the nitrogen harvest index, was lower (36% of total) in high N than in low N (40% of total) treatment, which indicated plants did not increase the in-grain utilization of N. The 18% higher grain yield with 112 kg N treatment occurred without considerable change in grain N content. However, lines with greater biomass produced greater yields in low N. Therefore, increasing tiller numbers and grain numbers for SRW wheat are the targeted traits for improving grain yield under N management, with less emphasis on the utilization of N in grains because N content is not critically influential for the marketability of soft wheat grains.

There are many rumors about the bay off Guadalcanal in the Solomon Islands. Some say it was the site of the lost empire of the Solomon king and that great treasure lies beneath the waters. Others say terrible things happened here, atrocities and disappearances at the hands of cannibal giants, and those who venture there do not return. It is cursed. Which is exactly what attracts the attention of husband-and-wife treasure-hunting team Sam and Remi Fargo. How could they resist? Clues and whispers lead them on a hunt from the Solomons to Australia to Japan, and what they find at the end of the trail is both wonderful and monstrous--and like nothing they have ever seen before.

The microgeometry of the cellular microenvironment profoundly impacts cellular behaviors, yet the link between it and the ubiquitously expressed mechanosensitive ion channel PIEZO1 remains unclear. Herein, we describe a fluorescent micropipette aspiration assay that allows for simultaneous visualization of intracellular calcium dynamics and cytoskeletal architecture in real-time, under varied micropipette geometries. By integrating elastic shell finite element analysis with fluorescent lifetime imaging microscopy and employing PIEZO1-specific transgenic red blood cells and HEK cell lines, we demonstrate a direct correlation between the microscale geometry of aspiration and PIEZO1-mediated calcium signaling. We reveal that increased micropipette tip angles and physical constrictions lead to a significant reorganization of F-actin, accumulation at the aspirated cell neck, and subsequently amplify the tension stress at the dome of the cell to induce more PIEZO1’s activity. Disruption of the F-actin network or inhibition of its mobility leads to a notable decline in PIEZO1 mediated calcium influx, underscoring its critical role in cellular mechanosensing amidst geometrical constraints. Cells sense mechanical force through PIEZO1 channels. Here, authors show that both microgeometry and aspiration force mediate the PIEZO1 activity, and also alter F-actin organization, further amplifying PIEZO1 activity along membrane.

Annually, over 100 million tons of nitrogen fertilizer are applied in wheat fields to ensure maximum productivity. This amount is often more than needed for optimal yield and can potentially have negative economic and environmental consequences. Monitoring crop nitrogen levels can inform managers of input requirements and potentially avoid excessive fertilization. Standard methods assessing plant nitrogen content, however, are time-consuming, destructive, and expensive. Therefore, the development of approaches estimating leaf nitrogen content in vivo and in situ could benefit fertilization management programs as well as breeding programs for nitrogen use efficiency (NUE). This study examined the ability of hyperspectral data to estimate leaf nitrogen concentrations and nitrogen uptake efficiency (NUpE) at the leaf and canopy levels in multiple winter wheat lines across two seasons. We collected spectral profiles of wheat foliage and canopies using full-range (350–2500 nm) spectroradiometers in combination with leaf tissue collection for standard analytical determination of nitrogen. We then applied partial least-squares regression, using spectral and reference nitrogen measurements, to build predictive models of leaf and canopy nitrogen concentrations. External validation of data from a multi-year model demonstrated effective nitrogen estimation at leaf and canopy level (R2 = 0.72, 0.67; root-mean-square error (RMSE) = 0.42, 0.46; normalized RMSE = 12, 13; bias = −0.06, 0.04, respectively). While NUpE was not directly well predicted using spectral data, NUpE values calculated from predicted leaf and canopy nitrogen levels were well correlated with NUpE determined using traditional methods, suggesting the potential of the approach in possibly replacing standard determination of plant nitrogen in assessing NUE. The results of our research reinforce the ability of hyperspectral data for the retrieval of nitrogen status and expand the utility of hyperspectral data in winter wheat lines to the application of nitrogen management practices and breeding programs.

Cloning from historically cryopreserved cells offers a potential means to restore lost genetic variation or increase the representation of particular lineages within bottlenecked species, provided such biobanked materials are archived for such genetic rescue applications. One species for which cloning can provide genetic management benefits is Przewalski’s horse, Equus przewalskii. All ~1800 living Przewalski’s horses, distributed across ex situ breeding facilities and in situ reintroduction sites, are descended from one or more of the five founder lineages established by the 12 horses captured from the wild between 1898 and 1947. Since the 1970s, the San Diego Zoo Wildlife Alliance Biodiversity Bank’s Frozen Zoo® (Escondido, CA, USA) has biobanked cells or tissues of 575 individuals spanning many generations. A pedigree analysis of a subset of deceased individuals represented in the San Diego Zoo Wildlife Alliance Biodiversity Bank’s Frozen Zoo® revealed an underrepresented male that lived from 1975 to 1998, Studbook Number 615 (also known as Kuporovitch), who would be of high value for breeding if cloned. Here, we report that two healthy clones were produced from this cell line using cross-species somatic cell nuclear transfer from 2020 to 2023. Their identification as clones was verified by a standard horse-pedigree genotyping panel, and, for one clone, a whole genome sequencing comparison to the original donor was performed. This is the first time that multiple healthy clones surviving the perinatal period have been produced for an endangered species.

The combination of two-photon calcium imaging and targeted two-photon optogenetic stimulation, termed all-optical interrogation, provides spatial and temporal precision when recording and manipulating neural circuit activity . All-optical experiments often use red-shifted opsins in combination with green fluorescent reporters of neuronal activity. However, their excitation spectra still partially overlap, meaning that the imaging laser can excite the opsin. Although some care has been taken in the past to understand the effects of this spectral overlap; further work is required to understand its impact on the findings of all-optical studies. We aimed to investigate whether two-photon imaging of the green fluorescent calcium reporter GCaMP6s at 920 nm increase the rate of response desensitization in neurons targeted for two-photon stimulation at 1035 nm expressing the red-shifted opsin C1V1. We systematically varied either the inter-stimulus interval or the duration of two-photon calcium imaging during targeted two-photon optogenetic stimulation of mouse layer 2/3 barrel cortex or visual cortex neurons. We found that two-photon imaging at 920 nm decreases trial-by-trial photostimulation responses in targeted C1V1-expressing neurons-an effect that is exacerbated at shorter inter-stimulus intervals. This is consistent with the imaging laser increasing the rate of opsin desensitization. Reduced photostimulation responses are not limited to targeted cells and are found across the field of view. Such network effects are less pronounced at shorter imaging doses. Our results provide methodological optimizations that enable trial-by-trial decreases in photostimulation response to be mitigated in all-optical experiments. This will reduce an external source of trial-by-trial variability in future all-optical experiments.

Wheat is a major source of calories and protein for humans worldwide. Wheat is the most widely grown crop, with cultivation areas and production systems on every continent. The cultivated land area is vast because of its importance and adaptability to various environmental conditions. Global wheat production has not kept up with the growing population, provoking the need to develop new methods and techniques to increase genetic gains. The first research chapter of this Ph.D. dissertation involves performing genome-wide association studies (GWAS) to identify and examine transferability of marker-trait associations (MTAs) across environments. I evaluated yield and yield components traits among 270 soft red winter (SRW) wheat varieties. The population consists of experimental breeding lines adapted to the Midwestern and eastern United States and developed by public university breeding programs. Phenotypic data from a two-year field study and a 45K-SNP marker dataset were analyzed by FarmCPU model to identify MTAs for yield related traits. Grain yield was positively correlated with thousand kernel weight, biomass, and grain weight per spike while negatively correlated with days to heading and maturity. Sixty-one independent loci were identified for agronomic traits, including a region that with –logP of 16.35, which explained 18% of the variation in grain yield. Using 12 existing datasets from other states and seasons, in addition to my own data, I examined the transferability of significant MTAs for grain yield and days to heading across homogenous environments. For grain yield and days to heading, I only observed 6 out of 28 MTAs to hold up across homogenous environments. I concluded that not all marker-trait associations can be detected in other environments.In the second research chapter of this Ph.D. dissertation, I dissected yield component traits under contrasting nitrogen environments by using field-based low-throughput phenotyping. I characterized grain yield formation and quality attributes in soft red winter wheat. Using a splitblock design, I studied responses of 30 experimental lines, as sub-plot, to high nitrogen and low nitrogen environment, as main-plot, for two years. Differential N environments were imposed by the application, or lack thereof, of spring nitrogen application in a field, following a previous corn harvest. In this study, I measured agronomic traits, in-tissue nitrogen concentrations, nitrogen use efficiency, nitrogen harvest index and end-use quality traits on either all or subset of the germplasm. My data showed that biomass, number of spikes and total grain numbers per unit area were most sensitive to low nitrogen while kernel weight remained stable across environments. Significant genotype x N-environment interaction allowed me to select N-efficient germplasm, that can be used as founding parents for a potential breeding population specifically for low-N environments. I did this selection on the basis of superior agronomic traits and the presence of the desirable gluten quality alleles such as Glu-A1b (2*) and Glu-D1d (5+10).