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<title>bioRxiv Subject Collection: Genetics Plant Biology</title>
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<description>
This feed contains articles for bioRxiv Subject Collection "Genetics Plant Biology"
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<title>bioRxiv</title>
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<link>https://www.biorxiv.org</link>
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<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.07.723566v1?rss=1">
<title>
<![CDATA[
A novel mouse model of rare neurodevelopmental disorder, TBCK Syndrome 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.07.723566v1?rss=1
</link>
<description><![CDATA[
TBCK Syndrome is a rare Mendelian disorder caused by variants in the TBCK gene. Although symptoms affect multiple organ systems, hallmark features include intellectual and developmental disability, craniofacial differences, hypotonia, and premature death. At the cellular level, TBCK has been implicated in mTOR signaling, autophagy, mitophagy, and mRNA trafficking; however, the mechanisms underlying disease onset and progression remain unclear. To address this gap, we characterized a mouse model of TBCK Syndrome. These mice lack exon 5 of the TBCK gene, resulting in a whole-body knockout of Tbck, modeling the most severe known variant. We performed a comprehensive battery of developmental assays, along with microcomputed tomography and histological analyses, which revealed systemic alterations consistent with those observed in affected individuals. Notably, phenotypic changes arising from Tbck loss emerge early and are detectable in the brain, indicating a primary neurodevelopmental origin of disease pathology. Rigorous characterization of this Tbck-deficient mouse establishes the first in vivo platform to investigate disease mechanisms and provides a foundation for preclinical evaluation of gene and targeted pharmacological therapy strategies.
]]></description>
<dc:creator><![CDATA[ Melendez-Perez, A. J., Durham, E. L., Layo-Carris, D. E., Gonzalez, E. M., Lubin, E. E., Smith, S. M., Worthington, K. E., Katsura, K. A., Angireddy, R., Wang, X.-M., Abdalla, K. J., Nair, D., Black, A., Diaz-Rosado, A., Ciesielski, B., O'Brien, W. T., Bhoj, E. ]]></dc:creator>
<dc:date>2026-05-11</dc:date>
<dc:identifier>doi:10.64898/2026.05.07.723566</dc:identifier>
<dc:title><![CDATA[A novel mouse model of rare neurodevelopmental disorder, TBCK Syndrome]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.06.722921v1?rss=1">
<title>
<![CDATA[
Frequency-domain identification of photosynthetic regulation under fluctuating light 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.06.722921v1?rss=1
</link>
<description><![CDATA[
Plant photosynthesis operates under naturally fluctuating light, yet its dynamic responses across timescales remain incompletely understood. Here, we apply sinusoidal light modulation as a controlled periodic input and analyze the response in the frequency domain, enabling quantitative system identification of photosynthetic dynamics. Using a minimal biochemical model of photosynthetic electron transport and regulation, we show that the system exhibits distinct dynamic regimes separated by a characteristic timescale of approximately 10 s. In the high-frequency domain, the response is governed by constitutive processes and reflects steady-state properties such as the plastoquinone redox state. In the low-frequency domain, regulatory feedback dominates, particularly non-photochemical quenching (NPQ), which modulates both the amplitude and phase of the response. For small-amplitude perturbations, the system behaves linearly and can be characterized using transfer functions and Bode plots. We show that key physiological parameters, including relaxation times and regulatory gains, can be extracted directly from frequency-response features such as phase maxima and gain transitions. In the nonlinear regime, large-amplitude oscillations generate higher-harmonic structure and alter time-averaged photosynthetic performance relative to constant illumination. We further introduce the concept of regulation fingerprints, defined as ratios of transfer functions between regulated and unregulated systems. These fingerprints reveal distinct spectral signatures of fast (PsbS-mediated) and slow (zeaxanthin-dependent) NPQ processes, enabling their quantitative separation. Together, these results establish frequency-domain analysis as a framework for probing and identifying the dynamic regulation of photosynthesis under fluctuating light, with direct applicability to non-invasive measurements in laboratory and field conditions.
]]></description>
<dc:creator><![CDATA[ Nedbal, L. ]]></dc:creator>
<dc:date>2026-05-11</dc:date>
<dc:identifier>doi:10.64898/2026.05.06.722921</dc:identifier>
<dc:title><![CDATA[Frequency-domain identification of photosynthetic regulation under fluctuating light]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.06.723361v1?rss=1">
<title>
<![CDATA[
Cp36 serine recombinase as a new tool for zebrafish transgenesis 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.06.723361v1?rss=1
</link>
<description><![CDATA[
One feature key to the versatility of zebrafish as an animal model for biomedical research is the breadth of genetic tools available, including for transgenesis. While the Tol2 transposase system remains the gold standard, its efficiency can be highly variable. Here, we explored the potential of a complementary transgenesis system, Cp36, a large serine recombinase identified from Clostridium perfringens previously found to efficiently integrate target cargo into the human genome without a preinstalled attB site. We generated Cp36-based plasmid constructs for zebrafish transgenesis and compared their performance to matched Tol2 plasmids across multiple experimental contexts, including transient expression, germline transmission, and multi-transgene expression. Cp36 integrates small ~3.5kb cargo into the zebrafish genome and transmits to the next generation as efficiently as Tol2, but Cp36 performance declines substantially for larger ~7.5kb constructs. Both Cp36 and Tol2 have comparable efficiency in transiently expressing a second construct regardless of the transposase/recombinase used to integrate the first construct, indicating compatibility with sequential transgenesis strategies. In summary, we demonstrate that Cp36 functions as a new alternative transgenesis method in zebrafish.
]]></description>
<dc:creator><![CDATA[ Thrikawala, S., Naples, B., Rosowski, E. ]]></dc:creator>
<dc:date>2026-05-11</dc:date>
<dc:identifier>doi:10.64898/2026.05.06.723361</dc:identifier>
<dc:title><![CDATA[Cp36 serine recombinase as a new tool for zebrafish transgenesis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.07.723424v1?rss=1">
<title>
<![CDATA[
Allele-resolved monosome and polysome sequencing identifies functional cis-acting variants affecting mRNA translation efficiency 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.07.723424v1?rss=1
</link>
<description><![CDATA[
To prioritize germline genetic variants affecting mRNA fate at the post-transcriptional and translational levels, we leveraged sucrose-gradient-based isolation of 80S monosomes and polysomes, followed by mRNA retrieval and paired-end sequencing. Total cytoplasmic RNA was also sequenced for comparison. Experiments were performed in the non-transformed cell line RPE-1, cultured under basal conditions or upon p53 activation by Nutlin. Differential gene expression analysis confirmed a canonical p53 response. Heterozygous SNPs and SNVs were identified from the RNA-seq data, and allelic fractions (AF) were calculated for total, monosomal, and polysomal mRNAs. Variants showing reproducible AF differences across fractions beyond experimental variability were defined as tranSNPs. Among nearly 7000 heterozygous variants analyzable in polysomal or total RNA and over 5000 in monosomal mRNA, 1155 displayed a significant imbalance. Reporter assays performed in both RPE-1 and HCT116 cells validated allelic or haplotype effects for 17 selected variants in UTRs and coding regions, confirming differences in 15 cases, with evidence of cell line-specific responses. Proteomic analysis further supported allelic imbalance for selected missense variants. Overall, tranSNPs were identified in a non-transformed cell line at frequencies comparable to those in cancer cells, thereby extending their implications in human physiology. Further, monosome profiling enabled improved detection sensitivity of tranSNPs without positional bias, suggesting that 80S profiling improves detection of allele-specific translational regulation in RPE-1 cells.
]]></description>
<dc:creator><![CDATA[ Alunno, L., Massignani, I., Hamadou, M. H., Mazza, F., Peroni, D., Belli, R., Dassi, E., Romanel, A., Inga, A. ]]></dc:creator>
<dc:date>2026-05-11</dc:date>
<dc:identifier>doi:10.64898/2026.05.07.723424</dc:identifier>
<dc:title><![CDATA[Allele-resolved monosome and polysome sequencing identifies functional cis-acting variants affecting mRNA translation efficiency]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.06.722817v1?rss=1">
<title>
<![CDATA[
AI platform for CRISPR functional mapping and function-based drug design 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.06.722817v1?rss=1
</link>
<description><![CDATA[
Conventional structure-based drug design has high clinical failure rates due to the disconnect where binding affinity does not guarantee safe functional modulation. To bridge this gap, we present CRISPRtile, a cloud-based platform for function-based drug design. By deriving library coverage optimization equations and leveraging AI to correct CRISPR guide biases, we generated toxicity and functional landscapes with over threefold error reduction compared to conventional methods. These maps bypass error and orders of magnitude higher computational cost in structure-based pipelines by enabling AI prediction of drug interaction directly from nontoxic functional sequences, while predicting brain penetration with benchmark leading performance. We demonstrate CRISPRtile by mapping the NLRP3 inflammasome and identifying FDA approved drugs with previously unrecognized ability to modulate it, revealing strategies to amplify or inhibit our immune response to homeostatic perturbations. These advances establish a generalizable strategy for the systematic discovery of safe functional modulators.
]]></description>
<dc:creator><![CDATA[ Ngo, J. C., Schoonenberg, V. A. C., Nandakumar, R., Wu, X., Sher, F. ]]></dc:creator>
<dc:date>2026-05-11</dc:date>
<dc:identifier>doi:10.64898/2026.05.06.722817</dc:identifier>
<dc:title><![CDATA[AI platform for CRISPR functional mapping and function-based drug design]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.06.723318v1?rss=1">
<title>
<![CDATA[
decepentaplegic directs the wiring of female-differentiated fruitless sex peptide response-inducing neurons 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.06.723318v1?rss=1
</link>
<description><![CDATA[
Female reproductive success requires coordinated behavioural responses following mating. In Drosophila melanogaster, these responses include reduced receptivity to further mating and increased oviposition, which are triggered by the male-derived sex-peptide (SP). Although the neural pathways controlling these behaviours have been partially characterised, the developmental mechanisms that establish and maintain these circuits remain poorly understood. Using a genetic approach, we identified an EMS-induced mutant that retains eggs and fails to reduce receptivity following SP exposure. We mapped this mutation to the dpp locus. Dpp acts within SP response inducing neurons (SPRINz) fru11/12 {cap} dsx and fru11/12 {cap} FD6, as downregulating dpp in subsets of sex-peptide target neurons disrupts receptivity. Here, dpp is required for correct neuronal wiring in dppHB3 mutants. Moreover, female sexual differentiation through the sex determination gene tra is required in SPRINz to display post-mating behaviours. Together, these findings suggest a new role for dpp in specifying neuronal connectivity in the context of sexual differentiation by the Drosophila canonical sex determination pathway to implement neuronal wiring for post-mating behaviours.
]]></description>
<dc:creator><![CDATA[ Singh, D. N. D., Soller, M. ]]></dc:creator>
<dc:date>2026-05-11</dc:date>
<dc:identifier>doi:10.64898/2026.05.06.723318</dc:identifier>
<dc:title><![CDATA[decepentaplegic directs the wiring of female-differentiated fruitless sex peptide response-inducing neurons]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.09.723998v1?rss=1">
<title>
<![CDATA[
A multi-species toolkit of TOP2 hypercleavage mutants for studying topoisomerase II-mediated DNA damage 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.09.723998v1?rss=1
</link>
<description><![CDATA[
DNA topoisomerase II (TOP2) generates transient DNA double-strand breaks that are trapped as TOP2-DNA covalent complexes (TOP2cc) by antibiotic and chemotherapy drugs. Here, we characterize tools for study of cellular responses to TOP2cc, exploiting a Saccharomyces cerevisiae TOP2 mutant (TOP2-F1025Y,R1128G) that generates spontaneous and inhibitor-induced covalent complexes at elevated frequencies. This Top2-hc (for "hypercleavage") mutant protein inhibits yeast cell growth when expressed alone or with endogenous Top2, and growth defects are exacerbated in DNA-repair-deficient genetic backgrounds and/or in the presence of low doses of the Top2 poison mAMSA. We generated analogous mutations in human and mouse TOP2A and TOP2B that gave increased TOP2cc, hypersensitization to topoisomerase poisons, increased DNA damage, and decreased cell survival in cultured cells. We further established knock-in mouse models with inducible, tissue-specific expression of each TOP2-hc isoform, demonstrating overt organismal toxicity and cellular markers of DNA damage responses. To illustrate the potential of these genetic tools, we carried out proof-of-principle screens in yeast and cultured human cells for sensitivity to TOP2-hc. The yeast screen revealed strong requirements for homologous recombination, moderate roles for sister chromatid cohesion and kinetochore function, and dependencies on vesicle and vacuolar functions. The pilot shRNA screen in human cells revealed shared requirements for resistance to expression of either TOP2A-hc or TOP2B-hc as well as examples of isoform specificity. These findings establish hypercleavage mutant proteins as effective tools for studying topoisomerase isoform-specific DNA damage and offer a foundation for exploring TOP2cc toxicity and tolerance in vivo.
]]></description>
<dc:creator><![CDATA[ Ontoso, D., Mehta, M., Shabro, A., Dittmar, J., Reid, R. J. D., Rothstein, R., Nitiss, J. L., Keeney, S. ]]></dc:creator>
<dc:date>2026-05-11</dc:date>
<dc:identifier>doi:10.64898/2026.05.09.723998</dc:identifier>
<dc:title><![CDATA[A multi-species toolkit of TOP2 hypercleavage mutants for studying topoisomerase II-mediated DNA damage]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.08.723766v1?rss=1">
<title>
<![CDATA[
Haplotype-based models improve sweep detection in ancient populations with complex demography 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.08.723766v1?rss=1
</link>
<description><![CDATA[
Identifying signatures of positive selection in humans is complicated by demographic processes such as bottlenecks, migration and admixture, all of which can distort or obscure the genomic patterns produced by selective sweeps. Ancient DNA offers a direct window into past allele and haplotype frequencies, yet most sweep scans in ancient populations rely on allele-frequency or site frequency spectrum (SFS) summaries, with limited use of haplotype-based approaches. Here, we evaluate the performance of haplotype and SFS-based methods for detecting selective sweeps under demographic scenarios that reflect the complex history of ancient and modern Europeans. We extend the haplotype-based likelihood framework saltiLASSI to accommodate pseudohaploid ancient genomes, enabling the use of truncated haplotype frequency spectra and their spatial decay to detect sweeps without requiring phased data. Using forward-in-time simulations, we examine sweeps of varying ages, two pulses of admixture with different source proportions, and cases where selection continues or ceases after admixture. We compare saltiLASSI to a widely used SFS-based approach (SweepFinder2). Our results show that haplotype-based likelihood models retain higher power than SFS methods in admixed populations, particularly when sweep haplotypes are introduced through migration or when selection has not had sufficient time to regenerate a clear SFS signature after admixture. These findings highlight the promise of haplotype-based inference for ancient DNA and demonstrate how model-based approaches can improve the detection of historical selective sweeps in populations with complex demographic histories.
]]></description>
<dc:creator><![CDATA[ Sequeira, A. N., Szpiech, Z. A., Huber, C. D. ]]></dc:creator>
<dc:date>2026-05-11</dc:date>
<dc:identifier>doi:10.64898/2026.05.08.723766</dc:identifier>
<dc:title><![CDATA[Haplotype-based models improve sweep detection in ancient populations with complex demography]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-11</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.06.721854v1?rss=1">
<title>
<![CDATA[
The nonsense-mediated decay RNA-surveillance pathway facilitates the hypoxia response by C. elegans 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.06.721854v1?rss=1
</link>
<description><![CDATA[
Hypoxia (low O2) induces physiological, metabolic and behavioral changes. The major hypoxia response pathway is conserved from nematodes to mammals and is driven by activation of the HIF (hypoxia inducible factor) family of transcription factors. Despite a fundamental role of the HIF pathway in regulating cellular homeostasis in response to hypoxic stress, interactions between the HIF pathway and cellular quality-control pathways are largely unknown. Using an unbiased genetic screen, we discovered that in C. elegans nonsense-mediated decay (NMD), an evolutionarily conserved mechanism of RNA surveillance, acts in parallel to HIF-1 to selectively enhance specific aspects of the response to HIF-1 activation, possibly through co-regulation of a distinct subset of HIF-1-upregulated genes. Our findings reveal a functional integration between oxygen sensing and RNA surveillance and establish NMD as a key regulator of specific aspects of the HIF-1-driven transcriptional program and physiological response.
]]></description>
<dc:creator><![CDATA[ Diehl, C., Horvitz, H. R. ]]></dc:creator>
<dc:date>2026-05-10</dc:date>
<dc:identifier>doi:10.64898/2026.05.06.721854</dc:identifier>
<dc:title><![CDATA[The nonsense-mediated decay RNA-surveillance pathway facilitates the hypoxia response by C. elegans]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.09.724051v1?rss=1">
<title>
<![CDATA[
The variable wheat stripe rust effector AvrYr7 evades Yr7 recognition through sequence and expression polymorphisms 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.09.724051v1?rss=1
</link>
<description><![CDATA[
Wheat diseases, including wheat stripe (yellow) rust, significantly impact wheat production and lead to several billions of dollars in economic losses each year. Wheat stripe rust is caused by the fungal pathogen Puccinia striiformis f. sp. tritici (Pst) which is best controlled via integrated disease management including growing disease resistant wheat cultivars. To-date, there are over 80 catalogued and more than 10 cloned stripe rust resistance genes (Yr genes). Yet our knowledge of corresponding avirulence (Avr) genes lags far behind which contrasts with other important wheat pathogens. The absence of cloned Pst's Avrs reflects long-standing challenges associated with Pst complex, highly heterozygous genome and the lack of robust transformation and map-based cloning systems. Recent advances in high-quality genome assemblies and wheat defense reporter assays have now made Avr gene identification more feasible. Building on these step-change advancements, we identify and characterize AvrYr7 which is specifically recognized by Yr7. We further identify six additional alleles of avrYr7 that escape recognition due to non-synonymous genetic variations, transposable element activity, missense mutation, and expression polymorphism. These findings provide critical insights into virulence evolution in one of the world's most important wheat pathogens.
]]></description>
<dc:creator><![CDATA[ Baig, D. I., Moeller, M., Tam, R., Pereira, E. C., Rodriguez-Algaba, J., Mojerlou, S., Hovmoller, M. S., Fejer Justesen, A., Nha Ho, T., Zhang, J., Ding, Y., Li, J., Wu, J., Periyannan, S., Zhang, X., Rathjen, J., Schwessinger, B. ]]></dc:creator>
<dc:date>2026-05-10</dc:date>
<dc:identifier>doi:10.64898/2026.05.09.724051</dc:identifier>
<dc:title><![CDATA[The variable wheat stripe rust effector AvrYr7 evades Yr7 recognition through sequence and expression polymorphisms]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.08.723708v1?rss=1">
<title>
<![CDATA[
Evolutionary diversification of the SymRK receptor family in land plants 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.08.723708v1?rss=1
</link>
<description><![CDATA[
Plant receptor-like kinases (RLKs) are involved in diverse processes, ranging from growth and reproduction to interactions with microbes. Variation in the extracellular domains delineates several RLKs subfamilies, including the malectin-like domain leucine-rich repeat receptor-like kinases (MLD-LRR-RLKs). Symbiosis Receptor-like Kinase (SymRK) is the prototypical member of MLD-LRR-RLKs and is required for microbial accommodation in host roots during root endosymbiosis. Yet, comparative phylogenetic analysis of SymRK orthologs in the broader context of MLD-LRR-RLK subfamily evolution remains limited. In this study, we examined the inventory, phylogeny and clade-specific evolutionary and transcriptional characteristics of this receptor group. SymRK and its closest homologs are present in most land plant lineages and group into four major clades and six additional species-specific clades. These clades can be distinguished by their evolutionary characteristics as either conserved with reduced gene copy number changes (including SymRK) or expanded and diversified, as observed in clade IV. Clade IV dynamics are largely driven by tandem gene duplications, which often arise within gene clusters. We further analysed the evolutionary characteristics of MLD-LRR-RLKs at the population level in Arabidopsis thaliana accessions. We found that some genes are conserved across accessions and are therefore likely to be functionally important, whereas a subset of genes, often located within tandem clusters, are highly diverse and likely contribute to accession-specific adaptations. Finally, most MLD-LRR-RLKs in the A. thaliana Col-0 accession are expressed in roots and respond broadly to biotic stimuli at the transcriptional level. Notably, clustered genes frequently exhibited divergent expression profiles, suggesting transcriptional diversification. Together, we revealed two contrasting evolutionary characteristics among members of the MLD-LRR-RLK subfamily, potentially associated with their functions in plants.
]]></description>
<dc:creator><![CDATA[ Fougner-Okland, T., Rodrgiuez-Arevalo, I., Makris, A., Lian, Q., Kamal, N., Schneeberger, K., Parniske, M., Ried-Lasi, M. K., Parys, K. ]]></dc:creator>
<dc:date>2026-05-10</dc:date>
<dc:identifier>doi:10.64898/2026.05.08.723708</dc:identifier>
<dc:title><![CDATA[Evolutionary diversification of the SymRK receptor family in land plants]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.07.723432v1?rss=1">
<title>
<![CDATA[
LIME: a fully automated pipeline for high-throughput quantification of leaf lesions 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.07.723432v1?rss=1
</link>
<description><![CDATA[
Accurate quantification of leaf lesion severity is essential for plant disease research and phenotyping but is often limited by subjective visual scoring and time-intensive manual image analysis. We present LIME, a fully automated, open-source image analysis pipeline for high-throughput quantification of leaf lesions from disease assay images. LIME integrates zero-shot leaf segmentation using the Segment Anything Model with a convolutional neural network for lesion area estimation. Applied to Arabidopsis thaliana leaves infected with Sclerotinia sclerotiorum, the proposed approach achieved a mean absolute percentage error of 12.9%, comparable to observed intrarater variability in manual scoring. Stratified evaluation across lesion-size groups demonstrated consistent prediction accuracy for small, intermediate, and large lesions, and comparative analysis showed that the deep learning-based model substantially outperformed color-based baseline methods. Under GPU-accelerated execution, LIME processed complete assays containing approximately 200 leaves in 15 minutes, representing an approximate 13-fold reduction in processing time relative to manual annotation. Together, these results indicate that LIME enables objective, reproducible, and scalable quantification of leaf lesion severity in standardized plant pathology assays. The pipeline is released as an open-source tool to support quantitative phenotyping studies.
]]></description>
<dc:creator><![CDATA[ Tan, D. ]]></dc:creator>
<dc:date>2026-05-10</dc:date>
<dc:identifier>doi:10.64898/2026.05.07.723432</dc:identifier>
<dc:title><![CDATA[LIME: a fully automated pipeline for high-throughput quantification of leaf lesions]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-10</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.08.723809v1?rss=1">
<title>
<![CDATA[
A massively parallel reporter assay of MECP2 cis-regulatory elements reveals genetic candidates for male-biased autism 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.08.723809v1?rss=1
</link>
<description><![CDATA[
Autism affects males four times more often than females, yet the basis of this sex bias remains unclear. One hypothesis is that hypomorphic variants in X-linked genes--genes where loss-of-function alleles cause syndromic neurodevelopmental disorders (NDDs) predominantly in females--produce milder, non-syndromic phenotypes in hemizygous males. We tested this by investigating cis-regulatory elements (CREs) of MECP2, a dosage-sensitive X-linked gene. Using a massively parallel reporter assay in human neurons, we mapped transcription factor binding sites within MECP2 CREs and tested autism-associated variants for functional impact. We identified two noncoding variants that change CRE activity, each with a male-biased phenotype. One of these, a promoter variant, disrupts NFY binding and reduces MECP2 expression by ~30%, a magnitude that produces autism-like phenotypes in mice. These findings suggest noncoding MECP2 variants can cause non-syndromic, male-biased autism, and provide a framework for uncovering regulatory variants in other X-linked NDD genes that may contribute to autism's missing heritability.
]]></description>
<dc:creator><![CDATA[ Meyer-Schuman, R., Cherry, F., Sui, Y., Papastathopoulos-Katsaros, A., Zhong, Y., Li, Y., Wang, T., Hennick, K., Karunakaran, D., Berk-Rauch, H., Liu, Z., Chakravarti, A., Nowakowski, T. J., Eichler, E., Zoghbi, H. Y. ]]></dc:creator>
<dc:date>2026-05-09</dc:date>
<dc:identifier>doi:10.64898/2026.05.08.723809</dc:identifier>
<dc:title><![CDATA[A massively parallel reporter assay of MECP2 cis-regulatory elements reveals genetic candidates for male-biased autism]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.06.723280v1?rss=1">
<title>
<![CDATA[
IRES-TrAPPr reveals novel insights into viral and cellular mRNA translation 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.06.723280v1?rss=1
</link>
<description><![CDATA[
Ribosome recruitment to human mRNAs is thought to occur primarily via cap-dependent initiation (CDI). This process is suppressed during a variety of cellular stresses, including viral infection, suggesting stress-response genes and viral mRNAs use alternative mechanisms to initiate translation. Indeed, many viruses recruit ribosomes directly via Internal Ribosome Entry Sites (IRESes). Hundreds of human mRNAs have been reported to also contain IRESes due to their ability to enhance expression in bicistronic and backspliced circRNA plasmid reporters. These DNA-based screens also reported hundreds of novel IRESes from more than fifty human viruses. However, such assays are prone to false-positives due to promoter and splicing activity, do not compare CDI and IRES translation, and lack the temporal resolution necessary for stress-response studies. To address these issues, we developed IRES-Translating Affinity Protein Profiling (IRES-TrAPPr), a massively parallel reporter assay that simultaneously quantifies CDI and IRES activity from thousands of co-transfected mRNAs. We validated this new method using luciferase assays and structure-function analyses of established viral IRESes, demonstrating exquisite sensitivity and specificity. Using IRES-TrAPPr, we quantified the activities of IRES elements from hundreds of viruses from a diversity of hosts. Our results provide evidence that viral IRESes from warm-blooded hosts have adapted higher structural stability to maintain folding at higher temperatures. Finally, we find hundreds of candidate human and viral IRESes from DNA-based screens have negligible IRES activity. Altogether, our results show that IRES-TrAPPr provides a novel, accurate platform for IRES research.
]]></description>
<dc:creator><![CDATA[ May, G. E., McManus, J. ]]></dc:creator>
<dc:date>2026-05-09</dc:date>
<dc:identifier>doi:10.64898/2026.05.06.723280</dc:identifier>
<dc:title><![CDATA[IRES-TrAPPr reveals novel insights into viral and cellular mRNA translation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.08.723725v1?rss=1">
<title>
<![CDATA[
Dim Green Light Enables Day-and-Night Monitoring of Leaf Movements 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.08.723725v1?rss=1
</link>
<description><![CDATA[
Understanding plant growth dynamics requires imaging across day-and-night cycles to quantify growth, movement and development in the aerial plant body and to capture the rhythmic nature of these processes. This requires imaging in light during the day and in darkness at night without perturbing plant physiology. Nighttime imaging has typically depended on infrared (IR) illumination, producing monochrome datasets that require specialised hardware and separate analysis pipelines when combined with daytime RGB imaging. Here, we evaluated very low-intensity green (dimG) illumination from standard LEDs as a practical alternative for colour-consistent nighttime imaging and assessed its physiological impact in Arabidopsis thaliana and Lactuca sativa (lettuce). We show that high resolution colour images can be obtained under dimG using low- cost cameras, with sufficient consistency between full-spectrum and dimG images to allow direct comparison and unified image analysis. We show that very low-fluence green light (<0.5 mol m-2 s-1) does not sustain circadian oscillations of gene activity under continuous exposure and does not perturb rhythms when applied during the dark phase of diel cycles. DimG imaging enabled accurate detection of diel leaf movement profiles in Arabidopsis circadian mutants, revealing genotype-specific phase differences under varying photoperiods. In lettuce, dimG pulses and continuous dimG enabled accurate quantification of diel leaf movement without affecting growth, stomatal opening, electron transport rate or chlorophyll content. Motion profiles under continuous dimG mirrored those under darkness. Our findings establish dim green illumination as a cost-effective solution for night-time imaging, simplifying phenotyping workflows with minimal impact on physiology.
]]></description>
<dc:creator><![CDATA[ Herrero, E., Gill, A. R., Wijeweera, S., Ginzburg, D., Stamford, J. D., Antoniades, A., Bromley, J. R., Mortimer, J., Gilliham, M., Millar, H., Webb, A. A. ]]></dc:creator>
<dc:date>2026-05-09</dc:date>
<dc:identifier>doi:10.64898/2026.05.08.723725</dc:identifier>
<dc:title><![CDATA[Dim Green Light Enables Day-and-Night Monitoring of Leaf Movements]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.08.723783v1?rss=1">
<title>
<![CDATA[
Turnip mosaic virus co-opts host RNA methylation to orchestrate plant infection 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.08.723783v1?rss=1
</link>
<description><![CDATA[
N6-methyladenosine (m6A) is a key RNA modification that regulates transcript stability and translation. However, its function in plant viruses remains largely unclear. Here, we show that the positive-sense single-stranded +ssRNA Turnip mosaic virus (TuMV) relies on the host m6A machinery to support efficient infection. Our findings uncover a previously unrecognized nuclear phase in the TuMV life cycle, during which viral RNA undergoes extensive methylation by host enzymes. We identify a complex and non-canonical methylation landscape on the TuMV genome, where canonical DRACH motifs are embedded within clusters of additional virus-specific non-canonical m6A sites. Notably, we also detect the presence of another RNA modification, m5C (5-methylcytosine), in close proximity to m6A-marked regions. This coordinated methylation landscape appears to be critical for efficient viral polyprotein synthesis. In its absence, the virus displays aberrant methylation and reduced infectivity as observed in m6A writer and reader mutants. Based on this, we propose a "seeding" model in which initial m6A deposition at canonical sites nucleates the formation of a broader network of m6A and m5C marks, likely guided by RNA structure rather than sequence motifs alone.
]]></description>
<dc:creator><![CDATA[ Hirt, H., Almeida-Trapp, M., Nawaz, K., Secco, N., Sheikh, A. ]]></dc:creator>
<dc:date>2026-05-09</dc:date>
<dc:identifier>doi:10.64898/2026.05.08.723783</dc:identifier>
<dc:title><![CDATA[Turnip mosaic virus co-opts host RNA methylation to orchestrate plant infection]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.08.723321v1?rss=1">
<title>
<![CDATA[
The common symbiosis pathway controls plant root microbiomes in a host-specific manner 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.08.723321v1?rss=1
</link>
<description><![CDATA[
Crop nutrition depends on plant-microbe interactions, yet it remains unclear whether conserved genetic pathways impose universal rules on root microbiome assembly across plant hosts. Here, we show that the Common Symbiosis Signalling Pathway (CSSP), a conserved genetic module controlling endosymbiosis with arbuscular mycorrhizal fungi and nitrogen-fixing bacteria, regulates root microbiome assembly in a host-specific manner across contrasting fertilisation regimes. Using Lotus japonicus and Hordeum vulgare, we demonstrate that mutations in orthologous CSSP genes remodel root bacterial communities in both species, but with distinct taxonomic outcomes. In Lotus, CSSP disruption reduces rhizobial colonisation and promotes niche replacement by commensal taxa, whereas in Hordeum, the same mutations broadly restructure bacterial lineages without converging on Lotus-like responses. Root exudate profiling reveals host-specific metabolic differences, particularly in phenylpropanoid (flavonoids and coumarins) and gibberellin pathways, linking CSSP activity to chemically distinct rhizosphere environments that correlate with divergent microbiome assembly patterns across hosts. Moreover, root bacterial community composition accurately predicts plant nutritional status, highlighting tight coupling between host physiology and microbiome composition. Together, our results show that conserved symbiosis signalling regulates root microbiome assembly, while host-specific metabolic environments determine taxonomic outcomes. This extends CSSP function beyond canonical endosymbioses and positions symbiosis signalling as a general determinant of plant-microbiome interactions with implications for crop nutrition.
]]></description>
<dc:creator><![CDATA[ Martyn, A., Thorsgaard Jensen, I., Lind Salomonsen, C., Blahovska, Z., Tao, K., Dings, H., Nott Bjoergvinsdottir, T., Tolstrup Christensen, B., Oldroyd, G., Waagepetersen, R., Glasius, M., Radutoiu, S. ]]></dc:creator>
<dc:date>2026-05-09</dc:date>
<dc:identifier>doi:10.64898/2026.05.08.723321</dc:identifier>
<dc:title><![CDATA[The common symbiosis pathway controls plant root microbiomes in a host-specific manner]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.07.723636v1?rss=1">
<title>
<![CDATA[
The Case for Kinases:A Phosphorylation Driven Model for Circadian Temperature Compensation 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.07.723636v1?rss=1
</link>
<description><![CDATA[
Circadian rhythms, ~24-hour biological cycles, enable organisms to anticipate rhythmic environmental cycles so they can assign proper day and night functions that align with those cycles. Circadian rhythms are defined by their ability to be reset by external cues, their capacity to continue to oscillate in the absence of those cues, and their capacity to maintain the rate of the clock across a range of ambient temperatures, a property known as temperature compensation. In the Neurospora clock, the White Collar Complex (WCC) drives expression of FRQ which nucleates a complex including FRH and CK1a that phosphorylates and thereby represses WCC activity. Work to date has suggested that kinases may be involved in temperature compensation and that in Neurospora the primary target of these is FRQ. Here we investigate the genetic relationship between two clock kinases, Casein Kinase I (ck-1a) and Casein Kinase II (cka), in their regulation of temperature compensation using novel alleles, ck-1aD135G and Delta cka. We find that that the clock relies on Casein Kinase I more at cold temperature, but this changes as temperature increases, and the clock relies more on Casein Kinase II at warm temperatures. Using quantitative proteomics on FRQ across temperatures, we find that the FRQ phosphorylation landscape is dependent on temperature and is altered in temperature compensation mutants. This leads to the development of a phosphorylation driven model for temperature compensation, where key temperature compensation specific domains on FRQ are phosphorylated to regulate period length in response to temperature, including by Casein Kinase I and Casein Kinase II.
]]></description>
<dc:creator><![CDATA[ Stevenson, E.-L., Kelliher, C. M., Kettenbach, A. N., Loros, J. J., Dunlap, J. C. ]]></dc:creator>
<dc:date>2026-05-09</dc:date>
<dc:identifier>doi:10.64898/2026.05.07.723636</dc:identifier>
<dc:title><![CDATA[The Case for Kinases:A Phosphorylation Driven Model for Circadian Temperature Compensation]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-09</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.05.722758v1?rss=1">
<title>
<![CDATA[
Reaction Norm Modeling of High-Dimensional Genomic and Environmental Data Improves Prediction Accuracy in Winter Wheat 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.05.722758v1?rss=1
</link>
<description><![CDATA[
Genomic prediction models that account genotype-by-environment (G*E) have the potential to accelerate the rate of genetic gain for yield and agronomic performance, yet relatively few studies have applied G*E prediction in public soft red winter wheat (Triticum aestivum) breeding programs. In this study, we extended a reaction norm-based genomic prediction framework by integrating weather-based environmental covariates to more effectively capture genotype-environment interactions. Key agronomic traits, including seed yield, plant height, test weight, and heading date, were evaluated across 33 environments (location-year) using over 3,200 breeding lines from the North Carolina State University small grains breeding program. Multiple genomic prediction models were compared using several cross-validation (CV) schemes representing common breeding scenarios. Across traits, the reaction norm M5 model, which incorporates both G*E and genotype-by-environmental covariate interactions (G*O), achieved the highest prediction accuracy (PA) in CV2 (predicting incomplete field trials) and CV1 for yield and test weight (predicting new lines). The highest PA was observed for test weight under CV2 (0.54) and for yield under CV1 (0.41). Under CV0 (predicting new environments), the M3 model incorporating G*E produced highest PA across traits, with the greatest accuracy for plant height (0.45), although differences among M2, M3, and M4 were small. Prediction under CV00 (predicting new lines in new environments) remained more challenging, with PA values 0.10-0.20 across traits. Overall, our results demonstrate that integrating environmental covariates into genomic prediction models can improve predictive performance across diverse wheat-growing environments in North Carolina, supporting their utility for applied breeding efforts.
]]></description>
<dc:creator><![CDATA[ Acharya, S. R., Garcia-Abadillo, J., Lyerly, J., Brown-Guedira, G., Jarquin, D., Bandillo, N. ]]></dc:creator>
<dc:date>2026-05-08</dc:date>
<dc:identifier>doi:10.64898/2026.05.05.722758</dc:identifier>
<dc:title><![CDATA[Reaction Norm Modeling of High-Dimensional Genomic and Environmental Data Improves Prediction Accuracy in Winter Wheat]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.05.722931v1?rss=1">
<title>
<![CDATA[
Genomic and Transcriptomic Basis of Salinity Tolerance in Dry Pea 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.05.722931v1?rss=1
</link>
<description><![CDATA[
Salinity is a major crop production constraint in dry pea (Pisum sativum L.), making the development of salt-tolerant varieties essential to improve crop productivity and land-use efficiency. The genetic mechanisms of salt tolerance in dry pea is largely unknown, and research on salt-tolerant genes is limited. In this study, we established comprehensive genomic and transcriptomic resources, along with a robust screening protocol, to dissect the genetic basis of salinity tolerance using two germplasm sets: the USDA pea diversity panel, consisting of approximately 200 globally sourced accessions, and a set of 300 modern elite lines from the NDSU Pulse Crops Breeding Program. Genetic variation for the salinity response was assessed based on ten phenotypic traits, with root dry weight, shoot dry weight, and specific root length identified as key indicators based on their heritability. Genome-wide association mapping uncovered significant genomic regions and several candidate genes linked to salt stress, with the strongest association found on chromosome 6. Overlapping QTL signals across traits suggest a shared genetic architecture underlying salinity tolerance. Field-based transcriptomic analysis further identified five putative genes involved in salinity response conserved across multiple crop species. Notably, Psat5g000800, encoding a glycosyl hydrolase gene, was markedly upregulated under salinity stress. These findings highlight the complex, multi-gene regulatory nature of salinity tolerance in dry pea and underscore the importance of functional validation of candidate genes. This study provides key insights and practical tools to support breeding efforts aimed at improving salt tolerance in dry pea.
]]></description>
<dc:creator><![CDATA[ Acharya, S. R., Bredu, E., Navasca, H., Worral, H., Piche, L., Saludares, R. A., Johnson, J. P., Coyne, C., Mcphee, K., Zhang, Q., Ostlie, M., Bandillo, N. ]]></dc:creator>
<dc:date>2026-05-08</dc:date>
<dc:identifier>doi:10.64898/2026.05.05.722931</dc:identifier>
<dc:title><![CDATA[Genomic and Transcriptomic Basis of Salinity Tolerance in Dry Pea]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.06.723131v1?rss=1">
<title>
<![CDATA[
The lack of peroxisomal Glycolate Oxidases 1 and 2 influences mitochondrial electron transport chain and its redox state under control and cadmium stress 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.06.723131v1?rss=1
</link>
<description><![CDATA[
The production of reactive oxygen species (ROS) in response to cadmium (Cd) has been extensively studied, demonstrating that they play a key role in the plants response to this heavy metal. While the role of enzymes like RBOHs has been thoroughly studied, the function of other ROS-producing enzymes, such as peroxisomal glycolate oxidase (GOX), remains largely overlooked. Peroxisomal GOX is a core metabolic enzyme of the photorespiratory pathway occurring in chloroplasts, mitochondria and peroxisomes. Using Arabidopsis (Arabidopsis thaliana) mutants lacking the main peroxisomal GOX genes, GOX1 (gox1-1) and GOX2 (gox2-1) we explored their function in plant response to Cd. Although photosynthetic capacity appears to be affected to the same extent in both mutants under control and Cd stress conditions, GOX2 seems to play a greater role in ROS production in response to the metal. Transcriptomic analyses on WT and gox2-1 pointed to the mitochondrial electron transport chain (mETC) as a target of Cd stress. We further investigated the individual GOX1 and GOX2 functions in mETC regulation and redox state. Although oxidative ratio of mitochondria was higher in both mutants, it was more pronounced in the absence of GOX1. Furthermore, the mETC is affected in both mutants but the regulation of its components differs in each mutant. These results point out the different functions of the two photorespiratory GOX isoforms in Arabidopsis, leading to a better understanding of the photorespiratory pathway.
]]></description>
<dc:creator><![CDATA[ Collado-Arenal, A. M., Rodriguez-Serrano, M., Pelaez-Vico, M. A., Terron-Camero, L. C., Perez-Gordillo, F. L., Ranea-Robles, P., Lopez, L. C., Sandalio, L., Romero-Puertas, M. C. ]]></dc:creator>
<dc:date>2026-05-08</dc:date>
<dc:identifier>doi:10.64898/2026.05.06.723131</dc:identifier>
<dc:title><![CDATA[The lack of peroxisomal Glycolate Oxidases 1 and 2 influences mitochondrial electron transport chain and its redox state under control and cadmium stress]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.07.723536v1?rss=1">
<title>
<![CDATA[
Zea Lip: An atlas of glycerolipid profiles across leaf development in maize 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.07.723536v1?rss=1
</link>
<description><![CDATA[
Lipids are the predominant building blocks of plant membranes and are essential for plant growth and development. They are crucial for survival during times of stress as lipids are involved in multiple signaling pathways, and their relative abundances can change in response to environmental factors. To better characterize the lipid composition of the vital food crop maize, we generated a comprehensive glycerolipid atlas using ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. We surveyed the lipid profiles of three different maize genotypes: B73, a temperate inbred; CML312, a subtropical inbred; and Palomero Toluqueno, an open-pollinated variety from the Mexican highlands. We collected leaf samples from 4 developmental stages and 6 leaves. From one growth stage, we also sampled along with three leaf zones: base, center, and tip. The genotype and leaf number were the major drivers of lipid differences. Phosphatidylcholine, lysophosphatidylcholine, and triacylglycerol genotypic differences were particularly high. We generated an eFP browser to be integrated into the maize genome browser, as well as a separate web interface to easily browse and compare lipid levels across tissues and genotypes, available at https://rrellan.shinyapps.io/Zea-Lip/.

SIGNIFICANCE STATEMENTThis work creates a spatial map of lipids in maize leaves across four growth stages for three genotypes: a lowland, a sub-tropical, and a highland. The resources generated here will directly benefit both the maize and lipid communities by creating a large dataset that can be used to generate new hypotheses in understanding lipid metabolism and environmental responses in maize.
]]></description>
<dc:creator><![CDATA[ Juarez Nunez, K. A., Lobet, G., Tandukar, N., Jadidzadeh, E., Pasha, A., Provart, N. J., Holland, J. B., Rellan-Alvarez, R., Barnes, A. C. ]]></dc:creator>
<dc:date>2026-05-08</dc:date>
<dc:identifier>doi:10.64898/2026.05.07.723536</dc:identifier>
<dc:title><![CDATA[Zea Lip: An atlas of glycerolipid profiles across leaf development in maize]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.07.723495v1?rss=1">
<title>
<![CDATA[
PAT: An Image Analysis Tool for Automated Scoring of Pollen in Alexander-Stained Anthers 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.07.723495v1?rss=1
</link>
<description><![CDATA[
Quantitative pollen viability analysis is a critical but labor-intensive step in plant reproductive biology. Existing deep-learning Segment Anything Models (SAM) fail to reliably segment viable pollen in Alexander-stained anthers. To address this, we fine-tuned an existing Cellpose-SAM model for pollen segmentation. We integrated it into PAT (Pollen Analysis Tool), a cross-platform desktop application. PAT features instance segmentation with interactive quality control, an in-app model retraining module, and publication-ready statistical outputs. We deployed PAT in an EMS suppressor screen of semi-sterile Arabidopsis smg7-6 mutants, enabling efficient candidate prioritization for whole genome sequencing and mapping candidate mutation. This screen led to the identification of a point mutation in CAP-D2 (capd2-2), a Condensin I subunit, that rescues the smg7-6 meiotic phenotype. Notably, mutation in a Condensin II subunits (CAP-D3 and CAP-H2) does not confer rescue. Further characterization suggests the capd2-2 allele is hypomorphic, showing no defects in vegetative growth, chromocenter compaction, or transposable element silencing. Collectively, we demonstrate that accessible AI tools have the potential to bridge gaps in plant phenotyping and accelerate the pace of biological discovery.

HighlightWe combined AI-powered image analysis with an easy-to-use desktop app to automate plant pollen counting, then used it to identify a new genetic suppressor of meiotic defects.
]]></description>
<dc:creator><![CDATA[ Volkava, D., Raxwal, V. K., Riha, K. ]]></dc:creator>
<dc:date>2026-05-08</dc:date>
<dc:identifier>doi:10.64898/2026.05.07.723495</dc:identifier>
<dc:title><![CDATA[PAT: An Image Analysis Tool for Automated Scoring of Pollen in Alexander-Stained Anthers]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.07.722400v1?rss=1">
<title>
<![CDATA[
Cryptic variation alters gene dosage sensitivity to shape inflorescence architecture in tomato 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.07.722400v1?rss=1
</link>
<description><![CDATA[
Phenotypic diversity arises in large part from genetic variants at multiple interacting loci, many of which alter gene dosage rather than abolish gene function. Dosage-sensitive variants, which often produce nonlinear phenotypic outcomes, can be exploited to fine-tune quantitative traits for crop improvement using genome editing. However, the phenotypic outcomes of individual variants can differ substantially across genetic backgrounds, as segregating alleles may modulate allelic effects in unexpected ways. Yet, how genetic background shapes gene dosage effects remains underexplored. Here, we show that MADS-box gene dosage effects, which can be used to tune tomato inflorescence architecture for optimal fruit yield, differ profoundly between distinct genetic backgrounds. We mapped the genetic basis of this background dependency and identified the cryptic modifier locus suppressor of branching 2 (sb2), which contains the conserved floral identity gene ANANTHA. We show that natural variation at sb2 modulates how inflorescence architecture responds to MADS-box dosage effects from natural and engineered loss-of-function mutations. Our findings illustrate how cryptic genetic variants can reshape gene dosage relationships and underscore the importance of characterizing such hidden variation for predictive engineering of quantitative traits using genome editing.

Significance StatementAdvances in crop genome editing enable precise modifications of gene dosage to fine-tune quantitative traits in crop improvement, but the predictability of such strategies remains limited. We show that hidden genetic differences, known as cryptic variation, can alter how gene dosage changes influence plant growth and development. Using tomato inflorescence architecture as a model, we characterize a natural cryptic modifier locus, suppressor of branching 2 (sb2), that modifies the effects of natural and engineered mutations in dosage-sensitive MADS-box genes. Our findings demonstrate that gene dosage effects depend on genetic background and highlight an often-unrecognized constraint on precision breeding by genome editing. Accounting for similar cases of cryptic variation will be essential for predictable engineering of quantitative traits in crops.
]]></description>
<dc:creator><![CDATA[ Swinnen, G., Afonso, S., Lacchini, E., Stolz, S., Lize, E., Soyk, S. ]]></dc:creator>
<dc:date>2026-05-08</dc:date>
<dc:identifier>doi:10.64898/2026.05.07.722400</dc:identifier>
<dc:title><![CDATA[Cryptic variation alters gene dosage sensitivity to shape inflorescence architecture in tomato]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.07.723153v1?rss=1">
<title>
<![CDATA[
Sequence adaptations in the intracellular domain of Symbiosis receptor-like kinase (SymRK) promoted infection thread progression in root nodule primordia 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.07.723153v1?rss=1
</link>
<description><![CDATA[
The uptake of nitrogen-fixing bacteria into living plant cells and the intracellular accommodation of arbuscular mycorrhiza (AM) fungi requires the plasma membrane-localised Symbiosis Receptor-like Kinase (SymRK). AM is widespread across terrestrial vascular plant lineages, while the nitrogen-fixing root nodule symbiosis (RNS) is restricted to one clade within the eurosids. This distribution led to the concept that SymRK was adopted during evolution to mediate RNS. Comparative analyses revealed that SymRK orthologs from the eurosid clade support RNS while SymRK from the phylogenetically distant species Solanum lycopersicum (tomato) does not. To dissect the molecular basis for this different functionality, we carried out complementation analyses of the Lotus japonicus symrk-3 mutant which is unable to form AM or RNS. Domains swap chimera from the tomato and L. japonicus SymRK orthologs revealed that the intracellular domain of L. japonicus SymRK is necessary and for cortical infection thread (IT) and symbiosome development at 21 days post inoculation. Notably, this signalling specificity could be overcome by ectopic expression of tomato SymRK, pointing to altered protein dosage as a potential determinant of function. Consistent with this idea, SINA family E3 ubiquitin ligases interacted with and ubiquitinylated L. japonicus SymRK, but not tomato SymRK. In yeast two hybrid analysis, the interaction of SymRK with SINA2 and SINA4 depended on the C-terminal intrinsically disordered tail region of L. japonicus SymRK. We conclude that the SymRK intracellular domain evolved interaction capabilities with SINA E3 ligases which correlates with its ability to support RNS.
]]></description>
<dc:creator><![CDATA[ Spezzati, M., Seidler, A. I., Ried-Lasi, M. K., Jelen, J. V., Buchner, J., Abele, M., Fougner-Okland, T. H., Beckers, C., Klingl, A., Ludwig, C., Parys, K., Parniske, M. ]]></dc:creator>
<dc:date>2026-05-08</dc:date>
<dc:identifier>doi:10.64898/2026.05.07.723153</dc:identifier>
<dc:title><![CDATA[Sequence adaptations in the intracellular domain of Symbiosis receptor-like kinase (SymRK) promoted infection thread progression in root nodule primordia]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.06.723330v1?rss=1">
<title>
<![CDATA[
CSLB4-mediated cell wall remodeling decouples phloem access from aphid performance in Arabidopsis thaliana 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.06.723330v1?rss=1
</link>
<description><![CDATA[
The plant cell wall (CW) is a key determinant of plant defense; however, the extent to which natural variation in CW architecture contributes to resistance against phloem-feeding insects remains unclear. Here, we combined genome-wide association studies (GWAS) with functional analyses to identify genetic determinants of resistance against the specialist aphid Brevicoryne brassicae in Arabidopsis thaliana. GWAS conducted on 200 natural accessions identified a single locus on chromosome 2 associated with aphid performance. Integration of haplotype and epidermis-specific expression data prioritized CSLB4, a member of the cellulose synthase-like B family. Loss-of-function cslb4 mutants showed reduced aphid offspring, indicating enhanced resistance to B. brassicae, whereas performance of the generalist aphid Myzus persicae was unaffected. Electrical penetration graph analyses revealed earlier phloem access on cslb4 mutants despite reduced performance, indicating a decoupling between phloem access and aphid success. Biochemical and immunolocalization analyses showed that CSLB4 disruption altered CW architecture, including increased xyloglucan epitope accessibility in mesophyll cell walls and reduced callose deposition upon aphid infestation. In addition, CSLB4 localized to Golgi-associated compartments, and in silico analyses are consistent with a role in non-cellulosic polysaccharide biosynthesis. Together, these findings identified CSLB4 as a modulator of CW architecture that uncouples phloem access from aphid performance.

HighlightA GWAS identifies CSLB4 as a regulator of cell wall architecture that uncouples aphid-feeding from performance, revealing a new mechanism of plant resistance to specialist insects.
]]></description>
<dc:creator><![CDATA[ Moraga, F., Arias-G, D., Sanhueza, D., Delgado-Rioseco, J., Fuenzalida-Valdivia, I., Inostroza-Aguirre, C., Peppino-Margutti, M., Ramos, M., Zavala-Torres, D., Ormeno, F., Sepulveda-, R. V., Fusari, C. M., Herrera-Vasquez, A., Saez, S., Blanco-Herrera, F. ]]></dc:creator>
<dc:date>2026-05-08</dc:date>
<dc:identifier>doi:10.64898/2026.05.06.723330</dc:identifier>
<dc:title><![CDATA[CSLB4-mediated cell wall remodeling decouples phloem access from aphid performance in Arabidopsis thaliana]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.07.723502v1?rss=1">
<title>
<![CDATA[
A natural soil-derived microbiota reshapes nitrogen form and plant mineral nutrition 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.07.723502v1?rss=1
</link>
<description><![CDATA[
Nitrogen form in soil has a major influence on plant growth and mineral nutrition, yet whether and how nitrogen form is controlled by microbiota in natural soils remains poorly understood. Here we show that, in a high-organic-matter soil from Danish nature, the native soil microbiota determines nitrogen form and thereby controls plant mineral nutrition. Eliminating the microbiota by sterilization disrupted nitrification, causing ammonium accumulation and loss of nitrate, which resulted in impaired growth and a pronounced reduction in shoot Mg and Ca associated with chlorosis. Reintroduction of a simplified soil-derived microbiota restored nitrification and re-established a balanced NO{square}{square}/NH{square}{square} regime, which rescued Mg and Ca accumulation, alleviated chlorosis, and promoted plant growth. Metagenomic analyses of bulk soil, rhizosphere, and root-associated communities revealed enrichment of nitrogen-cycling functions, including nitrification-related genes, supporting the capacity of the restored microbiota to modulate nitrogen form in soil and the rhizosphere. Moreover, this microbiota alleviated mineral deficiency symptoms in an organic agricultural soil. Together, our findings reveal a natural microbiota-dependent mechanism by which soil microbes determine nitrogen form and thereby regulate plant mineral nutrition, particularly Mg and Ca homeostasis.
]]></description>
<dc:creator><![CDATA[ Dethier, L., Xie, J., Yang, X., Bai, Y., Xu, D. ]]></dc:creator>
<dc:date>2026-05-08</dc:date>
<dc:identifier>doi:10.64898/2026.05.07.723502</dc:identifier>
<dc:title><![CDATA[A natural soil-derived microbiota reshapes nitrogen form and plant mineral nutrition]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.07.723431v1?rss=1">
<title>
<![CDATA[
Herbivory-induced alterations in cytosolic proteins of pigeon pea (Cajanus cajan) leaves 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.07.723431v1?rss=1
</link>
<description><![CDATA[
Insect herbivory triggers cytosolic proteome reprogramming by activating defense pathways and modulating key metabolic processes. We found that simulated herbivory in pigeon pea (Cajanus cajan) induced reactive oxygen species (ROS) production and molecular alterations within 12 hours (h) of post treatment. We compared the leaf proteome profiles of two cultivated genotypes, ICPL 332 (moderately resistant) and ICPL 87 (susceptible), using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) coupled with mass spectrometry (MS). More than 220 protein spots were detected in ICPL 332 and over 200 in ICPL 87. Comparative analysis revealed 75 differentially accumulated proteins (DAPs), of which 40 were consistently reproducible across biological replicates. These included 11 unique to ICPL 87, 9 unique to ICPL 332, and 10 common to both genotypes. Among the shared DAPs, ICPL 332 showed five upregulated and five downregulated, whereas ICPL 87 exhibited only two upregulated and eight downregulated. Functional categorization grouped DAPs into primary metabolism, stress response, and growth and development. Proteins related to primary metabolism were largely downregulated in both genotypes, while stress-associated proteins exhibited substantial downregulation in ICPL 87 compared to ICPL 332. Overall, the results demonstrate proteomic adjustments underlying defense responses in pigeon pea genotypes.
]]></description>
<dc:creator><![CDATA[ S, A., Kalita, P. J., Meshram, S. K., Das, A., Patil, R. I., Das, S., Jaba, J., Das, D., Acharjee, S. ]]></dc:creator>
<dc:date>2026-05-08</dc:date>
<dc:identifier>doi:10.64898/2026.05.07.723431</dc:identifier>
<dc:title><![CDATA[Herbivory-induced alterations in cytosolic proteins of pigeon pea (Cajanus cajan) leaves]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-08</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.06.723179v1?rss=1">
<title>
<![CDATA[
A high-resolution atlas of cattle regulatory variants and their cross-species activity in matched human cells 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.06.723179v1?rss=1
</link>
<description><![CDATA[
Identifying causal noncoding variants underlying complex traits in cattle remains challenging because high-resolution functional maps of regulatory variation are lacking. Here we combine massively parallel reporter assays with graph genomics to measure autonomous transcriptional activity from >1.5 billion DNA fragments spanning both cattle subspecies. In primary bovine cells, we assay >15 million variants and identify >150,000 expression-modulating variants enriched at cattle eQTL and GWAS loci. This enables the refinement of broad association signals to small sets of candidate functional regulatory variants. Our haplotype-aware framework captures rare, multi-allelic and tightly linked variants poorly resolved by conventional eQTL studies, and quantifies the disproportionate impact of larger variants on transcription. Furthermore, we use these data to train a deep-learning model that successfully predicts bovine promoter activity directly from sequence. Profiling the same cattle DNA in matched primary human cells reveals widespread conservation of promoter and enhancer activity, allelic effects and regulatory grammar, supporting the transfer of annotations and models across species. However, species-dependent effects are enriched in evolutionarily young sequences and p53-family motifs, highlighting the limits to simple cross-species extrapolation. Together, these data provide a high-resolution atlas of cattle regulatory variation and a framework for prioritising causal noncoding variants for cattle trait improvement.
]]></description>
<dc:creator><![CDATA[ Zhao, R., Plenderleith, L., Debnath, T., Owen, R., Pagie, L., Bisht, V., Metheringham, C., Marr, M., Powell, J., Talenti, A., Zhu, C., Paxton, E., Jensen, K., van Arensbergen, J., Connelley, T., Morrison, L., Hassan, M., Prendergast, J. G. ]]></dc:creator>
<dc:date>2026-05-07</dc:date>
<dc:identifier>doi:10.64898/2026.05.06.723179</dc:identifier>
<dc:title><![CDATA[A high-resolution atlas of cattle regulatory variants and their cross-species activity in matched human cells]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
<item rdf:about="https://www.biorxiv.org/content/10.64898/2026.05.05.721418v1?rss=1">
<title>
<![CDATA[
Genome-wide CRISPR knockout cell screening platform for the disease vector tick species Ixodes scapularis 
]]>
</title>
<link>
https://www.biorxiv.org/content/10.64898/2026.05.05.721418v1?rss=1
</link>
<description><![CDATA[
The black legged tick, Ixodes scapularis, is a vector of the bacterium that causes Lyme disease and several other illnesses, including anaplasmosis, babesiosis, and tick-borne encephalitis. Although high-quality genome annotations are available for I. scapularis, functional understanding of I. scapularis genes is limited. To address this, we developed a platform for genome-wide CRISPR-Cas9 knockout screening in I. scapularis cells. To evaluate the platform, we performed a screen to identify genes associated with cellular fitness, and screens for resistance to treatment with copper chloride, Antimycin A, or Destruxin A (DA), a cyclic hexadepsipeptide produced by the pathogenic fungus Metarhizium anisopliae. In each case, the screens implicate specific sets of conserved and non-conserved I. scapularis genes in relevant cellular functions, providing the first experimental evidence of function for a large set of I. scapularis genes. Altogether, in this first-of-its-kind effort for the arthropod subclass Acari, we present an unbiased genome-wide CRISPR-Cas9 knockout cell screening platform, related resources, and datasets that will be broadly useful to efficiently uncover cellular functions of I. scapularis genes.
]]></description>
<dc:creator><![CDATA[ Butnaru, M., McKenna, W., Goswami, S., Wu-Chuang, A., Mameli, E., Wilcox, A., Quennesson, L., Kim, A.-R., Veal, A., Chen, W., Verzone, H., Lane, E. A., Laukaitis-Yousey, H. J., Araneo, C., Singh, N., Pedra, J., Hu, Y., Viswanatha, R., Perrimon, N., Mohr, S. E. ]]></dc:creator>
<dc:date>2026-05-07</dc:date>
<dc:identifier>doi:10.64898/2026.05.05.721418</dc:identifier>
<dc:title><![CDATA[Genome-wide CRISPR knockout cell screening platform for the disease vector tick species Ixodes scapularis]]></dc:title>
<dc:publisher>Cold Spring Harbor Laboratory</dc:publisher>
<prism:publicationDate>2026-05-07</prism:publicationDate>
<prism:section></prism:section>
</item>
</rdf:RDF>
