Ethyl Methanesulfonate Research Articles

Elemental Composition, Antioxidant, Anti-inflammatory and Anti-genotoxic Properties of Nitophyllum punctatum

ObjectivesMarine algae are important sources of nutritionally and pharmacologically important products. Here in the study, nutritional composition and biological activities of a marine red algae Nitophyllum punctatum were assessed. The present study aimed to analyze the nutritional composition and biological effects of the N. punctatum. MethodsThe marine algae N. punctatum was collected, shade dried and extracted using Soxhelet apparatus. The elemental composition was estimated using Atomic Absorption Spectroscopy; the anti-inflammatory effect was determined using Raw 264.7 macrophages. Genotoxicity was assessed by standard methods using Allium cepa root model. ResultsThe elemental composition analysis revealed the presence of essential minerals such as calcium, magnesium, potassium, and iron, indicating the potential nutritional value of the algae. The extract demonstrated significant inhibition of Nitric oxide (NO) production and downregulated the expression of pro-inflammatory cytokines such as TNF-α and IL-6, suggesting its potential as an anti-inflammatory agent. Furthermore, the anti-genotoxic potential of the methanol extract was evaluated using the Allium cepa model, a well-established system for assessing genotoxicity. Onion root tips were treated with various concentrations of the extract, and the chromosomal changes were performed as indicators of genotoxicity. Results indicated a protective effect against Ethyl methyl sulfonate (EMS) induced genotoxicity in onion roots. ConclusionsThe findings suggest that the methanol extract of N. punctatum possesses potent anti-inflammatory and anti-genotoxic properties, which may be attributed to the presence of bioactive compounds. The study highlights the potential pharmacological importance of N. punctatum as a source of natural compounds with anti-inflammatory and anti-genotoxic activities.

Multi-Endpoint Toxicological Assessment of Chrysin Loaded Oil-in-Water Emulsion System in Different Biological Models.

Chrysin is hypothesized to possess the ability to prevent different illnesses, such as diabetes, cancer, and neurodegenerative disorders. Nonetheless, chrysin has a low solubility under physiological conditions, resulting in limited bioavailability. In a previous study, we utilized an oil-in-water emulsion system (chrysin-ES or chrysin-NE) to encapsulate chrysin, thereby increasing its bioaccessibility and preserving its antioxidant and anti-Alzheimer's properties. To promote the chrysin-ES as a supplementary and functional food, it was obligatory to carry out a safety assessment. Cytotoxicity testing showed that chrysin-ES was harmless, with no killing effect on 3T3-L1 (adipocytes), RAW 264.7 (macrophages), HEK293 (kidney cells), and LX-2 (hepatic stellate cells). The acute toxicity evaluation demonstrated that the 50% lethal dose (LD50) for chrysin-ES was greater than 2000 mg/kg BW. Genotoxicity assessments found that chrysin-ES did not induce DNA mutations in vitro or in vivo. Furthermore, chrysin and chrysin-ES exhibited anti-mutagenic properties against PhIP-induced and IQ-induced mutagenesis in the Ames test, while they inhibited urethane-, ethyl methanesulfonate-, mitomycin C-, and N-nitrosomethylurea-mediated mutations in Drosophila. The present study illustrates the safety and anti-genotoxicity properties of chrysin-ES, allowing for the further development of chrysin-based food supplements and nutraceuticals.

Oxyfluorfen: a novel metabolic inhibitor to select microalgal chlorophyll-deficient mutant strains for nutritional applications

Nowadays, there is an increasing demand for novel feedstocks and alternative protein sources to meet global needs. Because of their rich nutritional profiles and high protein contents, microalgae-based food products and supplements are being developed. Nonetheless, these products present organoleptic characteristics such as taste, smell and colour that are often considered unpleasant by human and animal consumers. To address this constraint, strain improvement approaches such as random mutagenesis have been used, which combined with the right selection strategy, lead to more appealing microalgal biomass. In this work, a novel selection strategy using oxyfluorfen, an inhibitor of the chlorophyll synthesis pathway, was applied for the first time to isolate chlorophyll-deficient strains of Scenedesmus rubescens and Chlorella vulgaris upon treatment with ethyl methanesulfonate (EMS). With this approach, one S. rubescens brownish (37Y01) mutant strain, as well as two C. vulgaris mutant strains, one yellow (31Y15) and one white (31W62), were obtained. S. rubescens 37Y01 displayed a reduced protein content of 19.1% dry weight (DW) compared to that of the wildtype, which presented a protein content of 25.0% DW. C. vulgaris wildtype and mutants exhibited higher protein contents, in the 42.8‐44.3% DW range, compared to Scenedesmus rubescens (p < 0.05). The selective pressure of this inhibitor allowed the selection of S. rubescens and C. vulgaris mutants displaying 55% and 95% decrease in chlorophyll content, respectively. The reduced chlorophyll content greatly improves the sensory properties and consumer acceptance of established mutants, increasing the potential of both strains as feedstocks to develop novel food products.

Induction of genetic diversity in gepak kuning soybean cultivar and M.1.1.3 line using ethyl methane sulfonate in M1 generation

Abstract Genetic diversity is the main determining factor in plant breeding activities. Various breeding methods can be used to induce plant variability, including mutation breeding. The chemical mutagen commonly used for mutagenesis is Ethyl Methane Sulfonate (EMS). This study explores the effects of EMS mutagen on the morphological and agronomic diversity of gepak kuning cultivar and M.1.1.3 soybean line. The research was conducted in Tambon Tunong Village, North Aceh District, Indonesia, using a single-factor Randomized Block Completely Design (RCBD). Concentrations of 0%, 0.05%, 0.075%, and 0.1% EMS were tested on the gepak kuning cultivar and M.1.1.3 line. Treating gepak kuning soybeans with EMS significantly impacted various variables, including harvesting age, pod number, seed weight, leaf appearance, and production. Additionally, the treatment resulted in changes in the morphology of the plant, particularly in the shape of its branches. The EMS treatment on M.1.1.3 line also significantly affected various variables like plant height, flowering age, harvesting age, and yield components. However, it led to sterile plants in the line.

Phenotypic characterization of point mutations spanning FHA domain and C-terminal region of Dawdle gene in Arabidopsis

The screening analysis of loss-of-function alleles in Arabidopsis thaliana revealed a mutation in the At3G20550 gene, called DAWDLE (DDL). The mutation in the DDL gene causes pleiotropic phenotypes and reduced the levels of several microRNAs. The DAWDLE gene encodes a protein with a Fork Head-Associated (FHA) domain, found in large range of proteins with significant cellular processes in prokaryotes and eukaryotes. However, it is not completely known whether the FHA domain and C-terminal region of the DDL are necessary for its function. The aim of this study was to determine the function of both regions by conducting a phenotypic analysis of point mutations spanning the FHA domain and C-terminal region in DDL Targeted Induced Local Lesions IN Genome (Tilling) screen was performed in the Columbia erecta-105 background of Arabidopsis resulting in point mutations spanning both regions of DDL. The mutants were phenotypically characterized. Height of the plant, hypocotyl and root length, and fertility were measured. Phenotypic analyses of the mutants revealed ddl phenotypes of varying degrees in different organs. Reduction in fertility and shortening in root, hypocotyl and stem lengths of the Tiller mutant lines suggest that the FHA domain and C-terminal region may require for DDL function in Arabidopsis. Key words: Dawdle, Fork Head-Associated Domain, Targeted Induced Local Lesions in Genome, Ethyl Methane Sulfonate, Arabidopsis

BnaC03.BIN2 regulates plant height by affecting the main inflorescence length and first effective branch height in Brassica napus L.

Rapeseed (Brassica napus L.) is one of the main oil crops in the world, and increasing its yield is of great significance for ensuring the safety of edible oil. Presently, improving rapeseed plant architecture is an effective way to increase rapeseed yield with higher planting density. However, the regulatory mechanism of rapeseed plant architecture is poorly understood. In this study, a dwarf rapeseed mutant dwarf08 (df08) is obtained by ethyl methane sulfonate (EMS)-mutagenesis. The decrease in plant height of df08 is mainly caused by the reduction in main inflorescence length and first effective branch height and controlled by a single semi-dominant gene. The hybrid plants (F1) show a semi-dwarf phenotype. Through map-based cloning and transgenic assay, we confirm that the nonsynonymous single nucleotide variant (SNV) (C to T) in BnaC03.BIN2, which is homologous with Arabidopsis (Arabidopsis thaliana) BIN2, is responsible for the dwarfism of df08. BnaC03.BIN2 interacts with BnaBZR1/BES1 and involves in brassinosteroids (BRs) signal transduction. Proline to Leucine substitution in 284 (P284L) enhances the protein stability of BnaC03.bin2-D, disrupts BRs signal transduction and affects the expression of genes regulating cell division, leading to dwarfism of df08. This study provides a new insight for the mechanism of rapeseed plant height regulation and creates an elite germplasm that can be used for genetic improvement of rapeseed architecture.

Phenotypic characterization of ethyl methanesulfonate (EMS) induced bigger pod (bp) with multiple seed mutant in lentil (Lens culinaris Medik.)

Lentils (Lens culinaris Medik.) is a protein- and nutrient-rich crop with limited genetic diversity and climate adaptability, making it ideal for mutation breeding research. Hence, in this study, we aimed to select and characterize distinct mutations in the flower and pod traits of lentil crops for enhanced yield and yield stability. In this study, healthy and viable lentil seeds were mutagenized using different concentrations of ethyl methanesulfonate (EMS) (0.10%, 0.25%, 0.50%, 0.75%, and 1.0%). The mutagenized lentil populations were cultivated up to the third generation (M3) to identify stable mutations in the flower and pod structures with increasing plant height. The notably larger 'bigger pod’ (bp) mutant, characterized by its unusually large pods containing 6–7 seeds per pod, was meticulously studied and quantified for its morphological traits in subsequent generations. Morphological observations of the tall bp mutant revealed significant mutations induced by EMS (0.10%) treatment. The quantitative trait means and confidence intervals showed that the mutant and parent cultivar L-4076 exhibited considerable variation. Seed yield (g) and yield-related components, including pods per plant, pod length (cm), and seeds per pod, were significantly higher in the mutant. Seed yield exhibited a significant positive phenotypic correlation with pod length, followed by branches per plant, seeds per pod, plant height, and pods per plant. The novel bp mutant induced in the present study produced increased number of seeds per plant and ensured optimal resource utilization, thus possibly contributing to yield improvement and stability. Further, it also aided in understanding the genetic networks that control legume flower and pod architecture and in genomics-assisted breeding for the development of superior lentil cultivars.

Resilient Response to Combined Heat and Drought Stress Conditions of a Tomato Germplasm Collection, Including Natural and Ethyl Methanesulfonate-Induced Variants

Agricultural systems are currently facing significant issues, primarily due to population growth rates in the context of global climate change. Rising temperatures cause plant heat stress and impact crop yield, which in turn compromises global food production and safety. Climate change is also having a significant impact on water availability around the world, and droughts are becoming more frequent and severe in many regions. The combined effect of both heat and drought stresses increases plant damage, resulting in reduced plant development and productivity loss. Therefore, developing heat–drought-tolerant crop varieties is crucial for enhancing yield under these challenging conditions. Tomato (Solanum lycopersicum L.), a major vegetable crop highly appreciated for its nutritional qualities, is particularly sensitive to extreme temperatures, which have a significant negative impact on tomato fruit setting and cause male gametophyte abortion. In this work, a classical genetic approach was employed to identify tomato genotypes showing a resilient response to combined heat and drought stress conditions. A phenotype screening of a natural germplasm collection and an ethyl methanesulfonate (EMS) mutagenized population resulted in the identification of a significant number of tomato lines tolerant to combined heat and drought conditions, specifically 161 EMS lines and 24 natural accessions as tolerant. In addition, TILLING and Eco-TILLING analyses were used as proof-of-concept to isolate new genetic variants of genes previously reported as key regulators of abiotic stress responses in different species. The identification of these variants holds the potential to provide suitable plant material for breeding programs focused on enhancing tomato resilience to adverse climate conditions.

Field evaluation of M4 plants obtained from induced mutation of tomato ‘Cobra’ variety with ethyl methane sulfonate (EMS)

Field evaluation of M4 plants obtained from induced mutation of tomato ‘Cobra’ variety with ethyl methane sulfonate (EMS)

The effect of exopolysaccharides from EMS-induced Porphyridium cruentum mutant on human epidermal and dermal layers

IntroductionMicroalgae biotechnology utilizes species like Porphyridium cruentum for their valuable phycobiliproteins and exopolysaccharides, which have potential industrial applications and health benefits, particularly in skin condition improvement.MethodsA mutant of P. cruentum LIMS-PS-1061 was developed through ethyl methanesulfonate mutagenesis and subsequent colony screening to study changes in its biomass production and pigment composition under different lighting conditions.Results and discussionThe mutant exhibited a 33.9% increase in dry weight under white light compared to the wild type. Despite maintaining the total pigment content, specific components changed significantly: chlorophyll content decreased 2.20- and 3.61-fold under white and blue light respectively, while phycobiliproteins increased 1.59- and 1.23-fold under the same conditions. These alterations suggest a compensatory mechanism for maintaining photosynthetic capacity. Furthermore, the exopolysaccharides of P. cruentum upregulated genes related to skin moisturization, barrier enhancement, and elasticity, and promoted wound healing through fibroblast migration. This supports the proposed mechanism of action for P. cruentum’s exopolysaccharides in improving human skin conditions by integrating the effects of aquaporin 3, filaggrin, involucrin, loricrin, elastin, and fibrillin-1.

Differential Resistance to Acetyl-CoA Carboxylase Inhibitors in Rice: Insights from Two Distinct Target-Site Mutations.

Weeds present a significant challenge to agricultural productivity, and acetyl-CoA carboxylase (ACCase)-inhibiting herbicides have proven to be effective in managing weed populations in rice fields. To develop ACCase-inhibiting herbicide-resistant rice, we generated mutants of rice ACCase (OsACC) featuring Ile-1792-Leu or Gly-2107-Ser substitutions through ethyl methyl sulfonate (EMS) mutagenesis. The Ile-1792-Leu mutant displayed cross-resistance to aryloxyphenoxypropionate (APP) and phenylpyrazoline (DEN) herbicides, whereas the Gly-2107-Ser mutants primarily exhibited cross-resistance to APP herbicides with diminished resistance to the DEN herbicide. In vitro assays of the OsACC activity revealed an increase in resistance to haloxyfop and quizalofop, ranging from 4.84- to 29-fold in the mutants compared to that in wild-type. Structural modeling revealed that both mutations likely reduce the binding affinity between OsACC and ACCase inhibitors, thereby imparting resistance. This study offers insights into two target-site mutations, contributing to the breeding of herbicide-resistant rice and presenting alternative weed management strategies in rice cultivation.

Influence of gamma radiation and EMS on chlorophyll and macro-mutation in mungbean (Vigna radiata)

Present study was carried during rainy (kharif) season of 2021 (M1 generation), summer 2022 (M2 generation) and rainy (kharif) seasons of 2022 (M3 generation) at College of Agriculture, Swami Keshwanand Rajasthan Agricultural University, Bikaner, Rajasthan for creating genetic variability in mungbean [Vigna radiata (L.) R. Wilczek]. The experiment was conducted with 26 treatments and 2 popular archaic varieties of mungbean (Ganga-1 and GM-4), solitary application of gamma radiation (200, 400 and 600 Gy), and ethyl methane sulphonate (EMS) (0.1 and 0.2%) as well as their combinations. A wide spectrum of chlorophyll mutants (Chlorina, Xantha, Albina, Viridis and complex type) and morphological mutants (plant type, flower colour, pod type, leaf variation and maturity period) were identified at various crop growth stages. All mutagen treatments including solitary application of gamma radiation and EMS as well as their combinations were found effective for creating genetic variability. The lower concentration of EMS (0.1%) was found to be relatively more effective as compared to 0.2%. Similarly, the lower dose of gamma radiation (200 Gy) was also more effective than 400 and 600 Gy. Among all the treatment combinations, combined application of 200 Gy + 0.1% EMS was found most effective in generating chlorophyll and morphological mutants. Among two varieties of mungbean, Ganga-1 was found to be more sensitive to mutagens as compared to GM-4. Numerous mutants identified in the mutagenized population of both varieties exhibited stability and superiority in terms of agro- morphological traits over Ganga-1 and GM-4. A mutant (S-92 of GM-4) identified during M1 in mutagen treatment 0.1% EMS was very deep lobbed which altered the plant morphology and appearance. This novel unique mutant identified in M1 generation maintained its stability in M2 and M3 generations also. The leaf colour of this mutant was darker than the original leaf colour of GM-4 and found relatively more tolerant to soil moisture-deficit stress.

BrBCAT1 mutation resulted in deficiency of epicuticular wax crystal in Chinese cabbage.

BrBCAT1 encoding a branched-chain amino acid aminotransferase was responsible for the glossy trait, which was verified by allelic mutants in Chinese cabbage. The glossy characteristic, thanks to the epicuticular wax crystal deficiency, is an excellent commodity character for leafy vegetables. Herein, two allelic glossy green mutants, wdm11 and wdm12, were isolated from an ethyl methane sulfonate (EMS)-mutagenized population of Chinese cabbage, and the mutant phenotype was recessive inherited. Cryo-SEM detected that epicuticular wax crystal in the mutant leaves was virtually absent. MutMap and Kompetitive allele-specific PCR analyses demonstrated that BraA06g006950.3C (BrBCAT1), homologous to AtBCAT1, encoding a branched-chain amino acid aminotransferase was the candidate gene. A SNP (G to A) on the fourth exon of BrBCAT1 in wdm11 caused the 233rd amino acid to change from glycine (G) to aspartic acid (D). A SNP (G to A) on the second exon of BrBCAT1 in wdm12 led to the 112th amino acid change from glycine (G) to arginine (R). Both of the allelic mutants had genetic structural variation in the candidate gene, which indicated that the mutant phenotype was triggered by the BrBCAT1 mutation. The expression levels of BrBCAT1 and genes related to fatty acid chain extension were decreased significantly in the mutant compared to the wild-type, which might result in epicuticular wax crystal deficiency in the mutants. Our findings proved that the mutation of BrBCAT1 induced the glossy phenotype and provided a valuable gene resource for commodity character improvement in Chinese cabbage.

Identification and map-based cloning of an EMS-induced mutation in wheat gene TaSP1 related to spike architecture.

A candidate gene TaSP1 related to spike shape was cloned, and the gene-specific marker was developed to efficiently track the superior haplotype in common wheat. Spike shape, an important factor that affects wheat grain yield, is mainly defined by spike length (SPL), spikelet number (SPN), and compactness. Zhoumai32 mutant 1160 (ZM1160), a mutant obtained from ethyl methane sulfonate (EMS) treatment of hexaploid wheat variety Zhoumai32, was used to identify and clone the candidate gene that conditioned the spike shape. Genetic analysis of an F2 population derived from a cross of ZM1160 and Bainong207 suggested that the compact spike shape in ZM1160 was controlled by a single recessive gene, and therefore, the mutated gene was designated as Tasp1. With polymorphic markers identified through bulked segregant analysis (BSA), the gene was mapped to a 2.65-cM interval flanked by markers YZU0852 and MIS46239 on chromosome 7D, corresponding to a 0.42-Mb physical interval of Chinese spring (CS) reference sequences (RefSeq v1.0). To fine map TaSP1, 15 and seven recombinants were, respectively, screened from 1599 and 1903 F3 plants derived from the heterozygous F2 plants. Finally, TaSP1 was delimited to a 21.9Kb (4,870,562 to 4,892,493bp) Xmis48123-Xmis48104 interval. Only one high-confidence gene TraesCS7D02G010200 was annotated in this region, which encodes an unknown protein with a putative vWA domain. Quantitative reverse transcription PCR (qRT-PCR) analysis showed that TraesCS7D02G010200 was mainly expressed in the spike. Haplotype analysis of 655 wheat cultivars using the candidate gene-specific marker Xg010200p2 identified a superior haplotype TaSP1b with longer spike and more spikelet number. TaSP1 is beneficial to the improvement in wheat spike shape.

Effectiveness mutagenesis of Bacillus subtilis using EMS (Ethyl Methane Sulfonate) to increase xylanase enzyme activity

This research was conducted to determine the effect of mutagenizing Bacillus subtilis with Ethyl Methane Sulfonate (EMS) for xylanase production and evaluate the effect of different xylan concentrations from corn cobs. The xylanase enzyme is an enzyme that can reduce the xylan content which is an anti-nutrient in animal feed. The wild-type Bacillus subtilis was treated with 50 μg/ml and 100 μg/ml of EMS. The mutants generated were selected for xylanase production in a medium containing xylan from corn cobs as a carbon source. The parameters observed included: Total Plate Count (TPC) and reducing sugar (xylose). Protein concentration and xylanase enzymes were analyzed using the Bradford method and 3.5 Dinitro salicylic acid for reducing sugar according to Miller methods respectively. Five mutants developed from each of the EMS concentrations. Approximately 6 and 6.8 % of the mutants developed from 50 μg/ml and 100 μg/ml of EMS had higher xylanase activities than the wild type and protein and xylose concentrations were higher than the wild type. From the results of the research, it can be seen that the use of EMS compounds has the potential to increase enzyme activity so that it can be potential in reducing antinutrients in animal feedstuff.

Protection of soybean by orange peel extract and its nanocapsules against ethyl methanesulfonate damages

Citrus fruits are one of the most important sources of phenolic substances, known as antioxidants and protector agents against ethyl methanesulfonate (EMS). Orange peels were used to extract bioactive compounds such as phenols, in addition to evaluate antioxidant activity of the extracts. Soybean plant was used to study the effect of several concentrations (0.01, 0.1 and 1%) of orange peel water extract as protector material to which EMS (0.7%) was added, as well as nanoencapsulated extract at 0.1%. Several parameters were measured to evaluate the effect of these concentrations on soybean as germination rate, plant height, number of leaves, leaves characteristics, total number of flowers after 40 days from germination, number of pods per plant, number of seeds per plant, number of seeds per pod and weight of 100 seeds. The results showed that 1% of orange peel water extract had the highest protective effect, however no positive effect was detected when it was used without EMS. Using 0.01% of orange peels extract was indeed advantageous for plant growth. On the other hand, pectin-calcium nanoencapsulated extract at 0.1% showed better effectiveness when compared to non-encapsulated extract at the same concentration. These results revealed that using plant extracts could be a promising approach to protect plants from harmful substances existing in some mediums (tissue culture) and environments.

Effect of ethyl methane sulphonate on growth, yield and some physiological parameters of Bambara groundnut (Vigna subterranea (L.) verdc) in non - water stressed and water stressed conditions

Bambara groundnut (Vigna subterranea (L.) Verdc.) is a legume cultivatd majorly for it’s seeds and commonly grown by subsistence farmers in sub-Saharan Africa. In order to determine the effect of ethyl methane sulphonate (EMS) mutagen on the growth, yield and some physiological parameters of non-water stressed (NWS) and water stressed (WS) bambara groundnut. It’s seeds were treated with five concentrations (0.00 (control), 0.25, 0.50, 0.75 and 1.00% v/v) Ethyl methane sulphonate and were grown under two conditions: non-water stressed and water stressed. Analysis of variance (ANOVA) revealed significant differences (P ≤ 0.50) in the emergence percentage (6 DAS), germination percentage (7 DAS), plant height (PH), number of leaves per plant (NL), leaf length (LL), leaf width, (LW), relative water content (RWC), chlorophyll a, b and total chlorophyll contents in the non-water stressed and water stressed plants. After rewatering at 8 WAS, highest number of leaves in NWS and WS conditions were observed in 0.75% EMS concentration (229, 189), highest leaf length in NWS and WS plants were observed in 0.50% EMS concentration (9.47cm, 9.13cm). Highest leaf width in NWS and WS plants was observed in 0.25% EMS concentration (4cm, 3.63cm). Highest number of pods per plant (NP) and weight of pods per plant (WP) were observed in 0.75% EMS concentration only in NWS plants. Highest chlorophyll a, b and total chlorophyll content was observed in 1.00% EMS concentration. The result form this study showed that Ethyl methane sulphonate (EMS) mutagen concentration increased the growth, yield and physiological parameters of Bambara groundnut in non-water stressed condition at different doses, however there was no positive effect of EMS on yield in the water stressed plants.

Phenotypic characterization and candidate gene analysis of a short kernel and brassinosteroid insensitive mutant from hexaploid oat (Avena sativa).

In an ethyl methanesulfonate oat (Avena sativa) mutant population we have found a mutant with striking differences to the wild-type (WT) cv. Belinda. We phenotyped the mutant and compared it to the WT. The mutant was crossed to the WT and mapping-by-sequencing was performed on a pool of F2 individuals sharing the mutant phenotype, and variants were called. The impacts of the variants on genes present in the reference genome annotation were estimated. The mutant allele frequency distribution was combined with expression data to identify which among the affected genes was likely to cause the observed phenotype. A brassinosteroid sensitivity assay was performed to validate one of the identified candidates. A literature search was performed to identify homologs of genes known to be involved in seed shape from other species. The mutant had short kernels, compact spikelets, altered plant architecture, and was found to be insensitive to brassinosteroids when compared to the WT. The segregation of WT and mutant phenotypes in the F2 population was indicative of a recessive mutation of a single locus. The causal mutation was found to be one of 123 single-nucleotide polymorphisms (SNPs) spanning the entire chromosome 3A, with further filtering narrowing this down to six candidate genes. In-depth analysis of these candidate genes and the brassinosteroid sensitivity assay suggest that a Pro303Leu substitution in AVESA.00010b.r2.3AG0419820.1 could be the causal mutation of the short kernel mutant phenotype. We identified 298 oat proteins belonging to orthogroups of previously published seed shape genes, with AVESA.00010b.r2.3AG0419820.1 being the only of these affected by a SNP in the mutant. The AVESA.00010b.r2.3AG0419820.1 candidate is functionally annotated as a GSK3/SHAGGY-like kinase with homologs in Arabidopsis, wheat, barley, rice, and maize, with several of these proteins having known mutants giving rise to brassinosteroid insensitivity and shorter seeds. The substitution in AVESA.00010b.r2.3AG0419820.1 affects a residue with a known gain-of function substitution in Arabidopsis BRASSINOSTEROID-INSENSITIVE2. We propose a gain-of-function mutation in AVESA.00010b.r2.3AG0419820.1 as the most likely cause of the observed phenotype, and name the gene AsGSK2.1. The findings presented here provide potential targets for oat breeders, and a step on the way towards understanding brassinosteroid signaling, seed shape and nutrition in oats.

Functional analysis of a wheat class III peroxidase gene, TaPer12-3A, in seed dormancy and germination

BackgroundClass III peroxidases (PODs) perform crucial functions in various developmental processes and responses to biotic and abiotic stresses. However, their roles in wheat seed dormancy (SD) and germination remain elusive.ResultsHere, we identified a wheat class III POD gene, named TaPer12-3A, based on transcriptome data and expression analysis. TaPer12-3A showed decreasing and increasing expression trends with SD acquisition and release, respectively. It was highly expressed in wheat seeds and localized in the endoplasmic reticulum and cytoplasm. Germination tests were performed using the transgenic Arabidopsis and rice lines as well as wheat mutant mutagenized with ethyl methane sulfonate (EMS) in Jing 411 (J411) background. These results indicated that TaPer12-3A negatively regulated SD and positively mediated germination. Further studies showed that TaPer12-3A maintained H2O2 homeostasis by scavenging excess H2O2 and participated in the biosynthesis and catabolism pathways of gibberellic acid and abscisic acid to regulate SD and germination.ConclusionThese findings not only provide new insights for future functional analysis of TaPer12-3A in regulating wheat SD and germination but also provide a target gene for breeding wheat varieties with high pre-harvest sprouting resistance by gene editing technology.

The transcription factor CpMYB62 controls the genetic network that leads to the determination of female flowers in Cucurbita pepo.

In monoecious species, female flowering constitutes the developmental process that determines the onset and production of fruit and is therefore closely related to crop yield. This article presents the identification and phenotypic and molecular characterization of myb62, an ethylmethane sulfonate loss-of-function mutation that completely blocks the female floral transition, converting all female flowers into male flowers. BSA-seq analysis coupled with WGS showed that myb62 corresponds to a C>T transition in the coding region of the gene CpMYB62, generating a premature stop codon and a truncated transcription factor without its N-terminal effector domain. The myb62 phenotype was partially rescued by exogenous ethylene application, indicating that the function of CpMYB62 is mediated by ethylene. Different evidence supports this conclusion: first, the reduced ethylene production of the mutant, and second, the male flower productive phenotype of the double mutant between myb62 and the ethylene-insensitive mutant etr2b, which demonstrated that myb62 is epistatic over etr2b. Furthermore, transcriptomic analysis of WT and myb62 apical shoots confirmed that CpMYB62 regulates master sex-determining genes, upregulating those encoding the ethylene biosynthesis enzymes CpACO2B and CpACS27A and those encoding for transcription factors that promote the development of carpels(CpCRC), but downregulating those involved in the arrest of carpels (CpWIP1), In the gene network controlling sex determination in cucurbits, CpMYB62 occupies the most upstream position, activating ethylene and other sex determining genes involved in female flower determination in Cucurbita pepo.