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Human m6A demethylase FTO modulates the flowering time of tomato plants under low-temperature stress.

N6-methyladenosine (m6A) RNA modification plays an important role in plant development and environmental stress responses. However, whether m6A demethylation modulates flowering under low-temperature (LT) stress in tomatoes remains unclear. Here, we investigated whether ectopic expression of FTO, a well-characterized human m6A demethylase, influences flowering and post-transcriptional behaviour in tomato (Solanum lycopersicum) under LT conditions. Flowering of transgenic tomato plants expressing FTO was analyzed under LT and normal conditions (NC), and the impacts of FTO on transcripts-specific m6A level, mRNA stability and splicing efficiency of flowering-related genes were evaluated using RT-qPCR, LC-MS/MS, m6A-IP-qPCR, and RNA decay and splicing analyses. FTO-expressing plants exhibited accelerated flowering specifically under LT, whereas no significant differences were observed under normal growth conditions. This phenotype was accompanied by increased expression of positive floral regulators (SlMC, SlFCA, and SlJ2) and decreased expression of negative regulators (SlSVP, SlSP, and SlTMF) under LT conditions. Notably, these expression changes were associated with altered mRNA stability, with positive regulators showing increased stability and negative regulators showing reduced stability under LT conditions. m6A-IP-qPCR analysis indicated reduced m6A enrichment in these selected transcripts in FTO-expressing plants. In addition to effects on mRNA stability, FTO expression was associated with changes in the splicing efficiency of SlMC transcripts. Collectively, our findings indicate that human FTO functions as an mRNA m6A demethylase in tomatoes and is associated with altered RNA regulatory processes under LT conditions. These findings suggest that m6A-mediated post-transcriptional regulation contributes to stress-induced flowering plasticity under LT conditions, rather than direct activation of canonical flowering pathways.

Abiotic stress

A cis-regulatory allele of ZmNPR1I spatially uncouples flowering from stalk-rot resistance in maize.

Pleiotropic effects of adaptive genes frequently constrain crop improvement by coupling beneficial traits with unfavorable trade-offs. In maize, ZmCCT10 confers strong stalk-rot resistance but causes delayed flowering under long-day conditions, limiting its deployment in temperate breeding. Here, we identify qPss3 as a cis-regulatory locus that governs its downstream target gene, ZmNPR1I. The ZmNPR1I protein represses ZmCCT10 transcription and, together with ZmNPR1-3, facilitates ZmCCT10 protein degradation. The favorable qPss3A5 allele reduces ZmNPR1I expression in leaves, relieving repression of ZmCCT10, ZmSPL32, and ID1, which collectively enhance ZCN8 expression to accelerate flowering. In roots, however, pathogen-induced activation of the resistant ZmCCT10H5 allele largely bypasses qPss3 regulation, thereby preserving stalk-rot resistance. Introgression of qPss3A5 into ZmCCT10H5-containing maize germplasm restores flowering adaptation without compromising disease resistance, improving yield stability under disease pressure. Our work reveals a tissue-specific qPss3A5-ZmNPR1I regulatory module that uncouples the developmental and immune functions of a pleiotropic adaptive gene, providing a general strategy for optimizing beneficial alleles in crop breeding.

flowering time

Plasticity in morphogenetic expression in plant.

This article discusses the plasticity of morphogenetic programmes in plants. It outlines the numerous ways in which the flowering programme can be accelerated or modified into partial flower formation and describes the differentiation of naked ovules on anthers in intact plants. The generation of embryos and flowers from cultured cells is then described and some of the properties of hypohaploids outlined. It is concluded that in vitro culture provides a useful facility for manipulating plant development and that the interaction between the nucleus and the cytoplasm may exemplify and explain the process of developmental plasticity.

Culture Techniques

Speed breeding: protocols, application and achievements.

One of the limiting factors in breeding and genetic research is the time required to develop pure lines. This is due, on the one hand, to the prolonged vegetative period of a single generation and, on the other hand, to the specifics of inbreeding, which typically requires 4-6 consecutive generations of self-pollination in plant material. Researchers have always sought approaches that enable the rapid development of homozygous plant lines. Consequently, methods such as greenhouse cultivation during the autumn-winter period, single-seed descent, shuttle breeding, embryo culture, and doubled haploid technology have been introduced into practice. All these methods have both advantages and limitations. One of the latest approaches facilitating a significant reduction in the vegetative period of plants is speed breeding (SB). This method is based on the application of factors that shorten the time from sowing to flowering, as well as techniques that accelerate the generative phase of development and overcome postharvest dormancy. This review provides a comprehensive list and characterization of all factors that influence the efficiency of speed breeding to varying degrees. Among the factors discussed that reduce the sowing-to-flowering period are photoperiod, light sources, spectral composition and light intensity, temperature, carbon dioxide levels, vernalization, mineral nutrition, substrate volume, mechanical shoot removal, and the use of plant growth regulators. To shorten the generative phase, the review summarizes the application of embryo culture and forced desiccation of immature seeds, along with methods to overcome postharvest dormancy. Additionally, applications of genetic approaches and genetic engineering for shortening generation time in speed breeding are described. The review also consolidates detailed protocols for approximately thirty crops. The high efficiency of speed breeding in reducing both the vegetative period per generation and the time required to develop pure lines has led to its increasing adoption in various research fields. This review highlights the application of speed breeding for hybridization and pure line development, introgression of target alleles, and genomic selection. A list of phenotypic traits exhibiting high correlation between controlled-environment and field conditions is provided.

accelerated flowering

A quantitative comparison of a position measurement system and accelerometry.

An electro-mechanical system to characterize the dynamic performance of a position measurement system was constructed. The system produced pure sine wave kinematics over the frequency range of 1-11 Hz. Synchronous measurements of the position (using infra-red light emitting diodes) and the acceleration were taken at discrete frequencies. The position signal was filtered and differentiated twice to obtain an estimated acceleration. The acceleration estimate was compared to the acceleration reading from the accelerometer, and both were compared to the theoretical acceleration. The comparison was based on extracting average features of the signal like amplitude, offset and noise. The results show that the accelerometer measurements matched the theoretical amplitude to within 1-3% over most of the range but showed significant offset drift. The acceleration estimates based on the position measurements were highly dependent on the filtering scheme, showed no significant offset but had higher levels of noise. The experimental measurements and the mathematical analysis quantitatively compared the dynamic performance of the position measurement system and the accelerometer. Such a system could be used to optimize the performance of position measurement devices, by comparing different filtering and/or differentiating schemes.

Acceleration

PtoeIF5A1: A Pleiotropic Regulator of Development, PCD, and Salt Tolerance in Populus tomentosa.

Eukaryotic translation initiation factor 5A (eIF5A) is a highly conserved protein family unique to eukaryotes, yet its functional characterization in woody plants remains limited. In this study, we identified four eIF5A genes (PtoeIF5A1-PtoeIF5A4) from the genome of Populus tomentosa, a fast-growing tree species indigenous to China, and characterized their expression patterns and functional roles through bioinformatics analysis, quantitative real-time PCR, stable overexpression in Arabidopsis thaliana, and transient expression in Nicotiana benthamiana leaves. Our results demonstrated that all PtoeIF5A proteins contain a conserved OB-fold domain and multiple phosphorylation sites, with PtoeIF5A1 showing predominant expression in roots and secondary xylem. Functional assays revealed that PtoeIF5A1 overexpression accelerated inflorescence stem elongation and early flowering in Arabidopsis, induced visible chlorosis and programmed cell death (PCD) in tobacco leaves, and significantly enhanced salt tolerance under NaCl treatment. Collectively, these findings establish PtoeIF5A1 in poplar as a pleiotropic regulator integrating developmental cues, programmed cell death, and stress responses; and as a valuable genetic resource for breeding stress-resilient woody plants.

Populus tomentosa

The role of prostaglandins in parturition, with special reference to the rat.

The evidence which suggests a role for prostaglandins in parturition is reviewed, with special reference to experimental observations on the rat. The pregnant uterus can both synthesize and metabolize prostaglandins. The biosynthetic capacity of the rat uterus increases as pregnancy proceeds. This increase, which is especially marked during the last few days of pregnancy, may be oestrogen-controlled. During pregnancy the tissue levels of the major prostaglandin-metabolizing enzyme, 15-hydroxyprostaglandin dehydrogenase (EC 1.1.1.141) is greatly increased. This may prevent aberrant production of prostaglandins from terminating the pregnancy prematurely. The capacity for prostaglandin metabolism begins to fall (perhaps under hormonal control) as the expected day of delivery approaches; this, coupled with an augmented biosynthesis, produces a net increase in uterine prostaglandins which contribute to the process of parturition by causing a decrease in progesterone concentration, increasing the muscular tone of the uterus, and altering uterine haemodynamics. Agents which block either prostaglandin synthesis or metabolism delay or accelerate the parturition process in the rat.

Animals

Rapid evolution of the plastid translational apparatus in a nonphotosynthetic plant: loss or accelerated sequence evolution of tRNA and ribosomal protein genes.

The vestigial plastid genome of Epifagus virginiana (beechdrops), a nonphotosynthetic parasitic flowering plant, is functional but lacks six ribosomal protein and 13 tRNA genes found in the chloroplast DNAs of photosynthetic flowering plants. Import of nuclear gene products is hypothesized to compensate for many of these losses. Codon usage and amino acid usage patterns in Epifagus plastic genes have not been affected by the tRNA gene losses, though a small shift in the base composition of the whole genome (toward A+T-richness) is apparent. The ribosomal protein and tRNA genes that remain have had a high rate of molecular evolution, perhaps due to relaxation of constraints on the translational apparatus. Despite the compactness and extensive gene loss, one translational gene (infA, encoding initiation factor 1) that is a pseudogene in tobacco has been maintained intact in Epifagus.

Amino Acid Sequence

Pan-genome-based resequencing of 2,320 accessions reveals structural variations and accelerates breeding advances in cultivated peanut.

The cultivated peanut is a crucial global legume crop that is essential for food security and nutrition, particularly in developing regions. However, its limited genetic variation hampers breeding progress and yield improvement. Here we constructed a graph-based pan-genome for peanut, incorporating 14 genomes that represent all 6 peanut varieties. Using this pan-genome, we genotyped 2,320 accessions, covering 88.03% of ICRISAT and 59.21% of USDA core germplasm, enriching valuable resources for genomic studies and breeding. We cataloged genomic structural variations and investigated the role of homoeologous exchanges in population divergence. Through our pan-genome approach, we overcame the challenges of genotyping posed by homoeologous exchanges and identified key genes associated with flowering and dwarfism in peanut. By integrating superior haplotypes and germplasm resources guided by the pan-genome, we further developed high-yield dwarf lines. This work provides essential genomic resources to accelerate functional gene discovery and modern peanut breeding.

Journal Article

Development and validation of a high-density 'Amahysnp' genotyping array in grain amaranth (Amaranthus hypochondriacus).

BACKGROUND: Grain amaranth has recently gained global attention as a promising crop alternative to traditional cereals due to its nutritional value and adaptability to various growing conditions. Although gene banks conserve extensive collections of amaranth germplasm, the genomic and phenotypic characterization of these resources is limited, which hinders their full utilization in breeding programs. A major challenge is the lack of high-throughput genotyping assays essential for comprehensive genomic characterization and trait mapping. High-density SNP arrays have become standard tools for genome-wide analysis across multiple loci, enabling molecular breeding across a range of crop species. RESULTS: In this study, we developed a 64 K high-throughput SNP genotyping array named "AmahySNP", using Affymetrix® Axiom® technology. The array contains 64,069 high-density SNPs distributed across both genic (55.17%) and non-genic (44.83%) regions of the Amaranthus hypochondriacus genome. The genic region includes 8,879 genes, which consist of 4,830 single-copy genes and 4,049 multi-copy genes distributed across 16 scaffolds. These genes cover various functional regions, including exons (10.5%), introns (40.1%), 5'UTRs (1.6%), and 3'UTRs (2.9%), respectively. The AmahySNP array was effectively utilized for population structure analysis, genetic diversity studies, core development, and genome wide association studies (GWAS) in amaranth germplasm. A representative core set of 112 accessions was identified, which includes two released varieties (Annapurna and Suvarna) and 100 diverse accessions from 12 different regions, representing 12% of the total 917 accessions evaluated. Phylogenetic analysis revealed three major genetic clusters, independent of their geographical origins. GWAS conducted using 22,763 polymorphic SNPs from 540 genotypes identified 13 novel loci associated days to flowering (DTF) trait, seven of which were located within annotated genes. CONCLUSIONS: The AmahySNP 64 K SNP chip a valuable genomic tool for amaranth research and breeding with a strong potential to accelerate its genetic improvement. It enables high-throughput genotyping for a wide range of applications, including GWAS and other genomic studies, and will significantly advance the exploration of natural genetic variations. Ultimately, this resource will empower amaranth breeders to develop improved amaranth cultivars with enhanced crop yield, resilience, and nutritional quality, contributing to global food security and sustainable agriculture.

Amaranthus

Analysis of deep-resequencing data of 984 soybean accessions reveals structural variations underlying agronomic traits.

Genomic structural variants (SVs) are major sources of genetic variation and have profound impacts on phenotypic traits. However, their functional effects remain largely unexplored in soybean. Here, we resequence 940 soybean accessions. Together with 44 publicly available datasets, we identify 602,281 SVs. Using a graph-based genome, we detect an additional 58,760 presence/absence variations (PAVs) that broadly affect gene expression. Population genomic analyses reveal that SVs serve as a core driving force for soybean domestication and improvement. Integrating SVs with QTLs for oil and protein content, and performing GWAS on 27 traits, we identify key functional SVs. These include transposable element insertions altering seed coat color, multiple insertions within a cytochrome P450 gene modifying flower and hypocotyl color, and a GmMATE1 deletion enhancing seed size. Together, our study establishes a comprehensive SV map of soybean, offering a valuable resource for dissecting the genetic basis of complex traits to accelerate molecular breeding.

Glycine max

Accidental exposure of veterinary students to rabies.

Accidental exposure to rabies occurred in more than 200 veterinary students at Texas A&M University from 1970 to 1977. Few of the animals to which the students were exposed had typical signs of rabies prior to the exposures. An accelerated preexposure rabies prophylaxis program coupled with retention of suspect tissues suitable for fluorescent antibody procedures has reduced the number of postexposure prophylaxis series.

Animals

Heparin metabolism and heparin-release lipase activity during long-term estrogen-progestin treatment.

Short-term oral contraceptive therapy has been reported to decrease postheparin lipolytic activity (PHLA). Resistance to heparin has been held responsible for this effect. To test several alternative explanations, we studied both PHLA and heparin concentrations in nine control women and nine women receiving long-term estrogen-progestin therapy after they were given heparin intravenously (10 units/kg). There were no significant differences in the concentration of heparin, its rate of disappearance, or calculated space of distribution between control and treated groups. PHLA was depressed (p less than 0.05) by approximately 50% during long-term estrongen-progestin therapy. PHLA disappearance was similar in both groups. Thus, reduced PHLA in women receiving long-term oral contraceptive therapy cannot be related to altered heparin metabolism or to accelerated enzyme disappearance from plasma. Long-term estrogen-progestin administration may decrease the heparin-facilitated release of individual triglyceride hydrolase activities or enhance the affinity of enzyme binding to tissues.

Contraceptives, Oral

Stable lyophilized reagents for the serum ferritin assay.

Two monoclonal antibodies to human ferritin, including one that was coupled to horseradish peroxidase, were lyophilized. These reagents show little loss of activity on reconstitution and demonstrate acceptable stability in the accelerated degradation test. When applied in a simple ELISA for the assay of serum ferritin along with the WHO standard for serum ferritin (80/602) they provide a robust assay with standardized reagents which is potentially suitable for use as a reference assay.

Antibodies, Monoclonal

Haplotype Blocks Are Associated With Rapid Local Adaptation to Environmental Shifts in Wild Barley.

Genomic mechanisms of local adaptation must be highly responsive in geographic regions where climate is changing rapidly. The Levant region is a critical biodiversity hotspot and the distribution edge for many species, including the wild ancestor of domesticated barley. This region is under an accelerated desertification process, thus enforcing a rapid genomic response to the projected environmental changes. To elucidate the genomic basis of rapid local adaptation, we studied wild barley populations using an ecological-genetic sampling design that decouples environmental variation from demographic background. We collected and sequenced 300 wild barley individuals and evaluated the phenotypes of 3600 progeny plants over 3 years. Our genomic analyses revealed that local adaptation is associated with clusters of candidate genes forming haplotype blocks. These clusters are enriched with environment and stress responsive genes, including flowering time regulators, drought and heat responsive genes. We identified six candidate adaptive haplotype blocks which span 1-8 Mbp and are distributed across chromosomes 1H, 2H, 4H and 5H, each segregating as two major haplotypes. Additionally, we integrated over 2600 occurrence records into ecological and evolutionary modelling to assess the genomic vulnerability of populations to projected future climates. Our study identifies candidate genomic regions and environmental drivers of local adaptation in wild barley and highlights the advantage of haplotype blocks architecture in orchestrating an efficient response to rapid environmental change. We highlight the ecological factors most strongly associated with the observed evolutionary responses and provide insights and guidelines for biodiversity conservation and implementation of crop wild relatives in breeding.

Hordeum

Prophylactic tocainide or lidocaine in acute myocardial infarction.

Twenty-nine patients with acute myocardial infarction (AMI) were studied in a randomized double-blind trial of intravenous lidocaine and tocainide, followed by either oral tocainide or placebo without regard to previous therapy, for the prophylaxis of arrhythmias associated with acute infarction. No patient had symptomatic ventricular tachycardia or fibrillation, although 1 patient taking lidocaine was withdrawn from therapy because of breakthrough arrhythmias. One patient in each group died from mechanical complications of AMI. Tocainide was administered to 16 patients and lidocaine to 13. Seven of the 13 patients receiving lidocaine had ventricular tachycardia or accelerated idioventricular rhythm, compared with 2 of 16 receiving tocainide (p less than 0.05). Adverse effects were noted in 11 of the 13 patients receiving lidocaine and 6 of the 16 patients receiving tocainide. The infusions used provided therapeutic levels of lidocaine or tocainide and the transition to oral tocainide was accomplished safely with maintenance of therapeutic antiarrhythmic levels. Thus, tocainide appears to be at least as efficacious and may be safer than lidocaine for the prophylaxis of ventricular arrhythmias associated with AMI. The transition to oral tocainide is well tolerated and can be accomplished with minimal difficulty.

Administration, Oral

Antisense oligonucleotide-mediated MSH3 suppression reduces somatic CAG repeat expansion in Huntington's disease iPSC-derived striatal neurons.

Expanded CAG alleles in the huntingtin (HTT) gene that cause the neurodegenerative disorder Huntington's disease (HD) are genetically unstable and continue to expand somatically throughout life, driving HD onset and progression. MSH3, a DNA mismatch repair protein, modifies HD onset and progression by driving this somatic CAG repeat expansion process. MSH3 is relatively tolerant of loss-of-function variation in humans, making it a potential therapeutic target. Here, we show that an MSH3-targeting antisense oligonucleotide (ASO) effectively engaged with its RNA target in induced pluripotent stem cell (iPSC)-derived striatal neurons obtained from a patient with HD carrying 125 HTT CAG repeats (the 125 CAG iPSC line). ASO treatment led to a dose-dependent reduction of MSH3 and subsequent stalling of CAG repeat expansion in these striatal neurons. Bulk RNA sequencing revealed a safe profile for MSH3 reduction, even when reduced by >95%. Maximal knockdown of MSH3 also effectively slowed CAG repeat expansion in striatal neurons with an otherwise accelerated expansion rate, derived from the 125 CAG iPSC line where FAN1 was knocked out by CRISPR-Cas9 editing. Last, we created a knock-in mouse model expressing the human MSH3 gene and demonstrated effective in vivo reduction in human MSH3 after ASO treatment. Our study shows that ASO-mediated MSH3 reduction can prevent HTT CAG repeat expansion in HD 125 CAG iPSC-derived striatal neurons, highlighting the therapeutic potential of this approach.

Huntington Disease

Alteration in membrane protein band 3 associated with accelerated erythrocyte aging.

We report a human band 3 alteration that is associated with anemia as determined by a reticulocyte count of 20%. Erythrocyte defects included increased IgG binding, increased breakdown products of band 3, and altered anion- and glucose-transport activity in middle-aged cells. These changes were observed during normal erythrocyte aging in situ. Binding of ankyrin to band 3 was normal. Serum/cell crossover studies indicated that a neoantigen appears on the propositus' erythrocytes to which IgG from both propositus and control serum binds as measured with a protein A binding assay. IgG eluted from the propositus' erythrocytes appeared to have a specificity for senescent cell antigen as determined by a phagocytosis inhibition assay. Immunoelectron microscopy showed that antibodies to band 3, which do not normally bind to intact erythrocytes, bound to the propositus' erythrocytes. Antibody 980 binds to normal old cells but not young or middle-aged cells. It also binds to a distinct region of band 3 in immunoblots of membranes from the propositus' middle-aged cells. Cells from both of the propositus' parents exhibited increased IgG binding and altered anion and glucose transport. The results of these studies suggest that (i) band 3 is aging prematurely in erythrocytes from the propositus, (ii) senescent cell antigen appears on the propositus' middle-aged red cells, and (iii) band 3 alterations observed in the propositus may have a genetic component.

Anion Exchange Protein 1, Erythrocyte