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PeerJ 2023Chemical mutagenesis has been successfully used for increasing genetic diversity in crop plants. More than 800 novel mutant types of rice ( L.) have been developed...
BACKGROUND
Chemical mutagenesis has been successfully used for increasing genetic diversity in crop plants. More than 800 novel mutant types of rice ( L.) have been developed through the successful application of numerous mutagenic agents. Among a wide variety of chemical mutagens, ethyl-methane-sulfonate (EMS) is the alkylating agent that is most commonly employed in crop plants because it frequently induces nucleotide substitutions as detected in numerous genomes.
METHODS
In this study, seeds of the widely consumed Basmati rice variety (Super Basmati, L.) were treated with EMS at concentrations of 0.25%, 0.50%, 0.75%, 1.0%, and 1.25% to broaden its narrow genetic base.
RESULTS
Sensitivity to a chemical mutagen such as ethyl methanesulfonate (EMS) was determined in the M1 generation. Results in M1 generation revealed that as the levels of applied EMS increased, there was a significant reduction in the germination percent, root length, shoot length, plant height, productive tillers, panicle length, sterile spikelet, total spikelet, and fertility percent as compared to the control under field conditions. All the aforementioned parameters decreased but there was an increase in EMS mutagens in an approximately linear fashion. Furthermore, there was no germination at 1.25% of EMS treatment for seed germination. A 50% germination was recorded between 0.50% and 0.75% EMS treatments. After germination, the subsequent parameters, viz. root length and shoot length had between 05.0% and 0.75% EMS dose levels. Significant variation was noticed in the photosynthetic and water related attributes of fragrant rice. The linear increase in the enzymatic attributes was noticed by the EMS mediated treatments. After the establishment of the plants in the M1 generation in the field, it was observed that for fertility percentage was at EMS 1.0% level, for the rice variety.
CONCLUSION
Hence, it is concluded that for creating genetic variability in the rice variety (Super Basmati), EMS doses from 0.5% to 0.75% are the most efficient, and effective.
Topics: Ethyl Methanesulfonate; Oryza; Mutation; Mutagens; Mutagenesis
PubMed: 37780391
DOI: 10.7717/peerj.15821 -
Heliyon Dec 2023Four hundred tubers of four genotypes, two brown and two black tiger nuts were subjected to Ethyl Methanesulfonate (EMS) and Colchicine treatments at concentrations of...
Four hundred tubers of four genotypes, two brown and two black tiger nuts were subjected to Ethyl Methanesulfonate (EMS) and Colchicine treatments at concentrations of 0 %, 0.1 %, 0.25 %, 0.5 % and 1.0 % for 24 h. Each genotype had twenty tubers treated with each of the five different concentrations and were planted using Complete Randomized Design (CRD) in a greenhouse. Quantitative data was collected and LD and RD were analysed using Excell 2016 and Genstat 11.2. A general decreasing trend in percentage germination and plant height was observed with increasing concentrations of mutagens applied. An EMS treatment had LD and RD values of 0.97 % and 1.49 % for black and 0.63 % and 1.63 % for brown genotypes. Similarly, the percentage colchicine treatment had LD and RD values of 1.65 % and 19.51 % concentrations for black and 0.91 % and 1.71 % concentrations for brown genotypes.
PubMed: 38046175
DOI: 10.1016/j.heliyon.2023.e22619 -
Advanced Science (Weinheim,... Nov 2023Here, a sucrose non-fermenting-1-related protein kinase alpha subunit (TaSnRK1α-1A) is identified as associated with cold stress through integration of genome-wide...
Here, a sucrose non-fermenting-1-related protein kinase alpha subunit (TaSnRK1α-1A) is identified as associated with cold stress through integration of genome-wide association study, bulked segregant RNA sequencing, and virus-induced gene silencing. It is confirmed that TaSnRK1α positively regulates cold tolerance by transgenes and ethyl methanesulfonate (EMS) mutants. A plastid-lipid-associated protein 6, chloroplastic-like (TaPAP6L-2B) strongly interacting with TaSnRK1α-1A is screened. Molecular chaperone DJ-1 family protein (TaDJ-1-7B) possibly bridged the interaction of TaSnRK1α-1A and TaPAP6L-2B. It is further revealed that TaSnRK1α-1A phosphorylated TaPAP6L-2B. Subsequently, a superior haplotype TaPAP6L-2B is identified and confirmed that both R30S and G38S are important phosphorylation sites that influence TaPAP6L-2B in cold tolerance. Overexpression (OE) and EMS-mutant lines verified TaPAP6L positively modulating cold tolerance. Furthermore, transcriptome sequencing revealed that TaPAP6L-2B-OE lines significantly increased jasmonic acid (JA) content, possibly by improving precursor α-linolenic acid contributing to JA synthesis and by repressing JAR1 degrading JA. Exogenous JA significantly improved the cold tolerance of wheat plants. In summary, TaSnRK1α profoundly regulated cold stress, possibly through phosphorylating TaPAP6L to increase endogenous JA content of wheat plants.
Topics: Triticum; Genome-Wide Association Study; Oxylipins; Cyclopentanes
PubMed: 37740426
DOI: 10.1002/advs.202303478 -
Plants (Basel, Switzerland) Dec 2023Somatic embryogenesis (SE) has many applications in grapevine biotechnology including micropropagation, eradicating viral infections from infected cultivars, mass...
Somatic embryogenesis (SE) has many applications in grapevine biotechnology including micropropagation, eradicating viral infections from infected cultivars, mass production of hypocotyl explants for micrografting, as a continuous source for haploid and doubled haploid plants, and for germplasm conservation. It is so far the only pathway for the genetic modification of grapevines through transformation. The single-cell origin of somatic embryos makes them an ideal explant for mutation breeding as the resulting mutants will be chimera-free. In the present research, two combinations of plant growth regulators and different explants from flower buds at two stages of maturity were tested in regard to the efficiency of callusing and embryo formation from the callus produced in three white grape cultivars. Also, the treatment of somatic embryos with the chemical mutagen ethyl methanesulfonate (EMS) was optimised. Medium 2339 supplemented with β-naphthoxyacetic acid (5 μM) and 6-benzylaminopurine (BAP-9.0 μM) produced significantly more calluses than medium 2337 supplemented with 2,4-dichlorophenoxyacetic acid (4.5 µM) and BAP (8.9 µM) in all explants. The calluses produced on medium 2337 were harder and more granular and produced more SEs. Although the stage of the maturity of floral bud did not have a significant effect on the callusing of the explants, calluses produced from immature floral bud explants in the premeiotic stage produced significantly more SEs than those from more mature floral buds. Overall, immature ovaries and cut floral buds exposing the cut ends of filaments, style, etc., tested for the first time in grapevine SE, produced the highest percentage of embryogenic calluses. It is much more efficient to cut the floral bud and culture than previously reported explants such as anthers, ovaries, stigmas and styles during the short flowering period when the immature flower buds are available. When the somatic embryos of the three cultivars were incubated for one hour with 0.1% EMS, their germination was reduced by 50%; an ideal treatment considered to obtain a high frequency of mutations for screening. Our research findings will facilitate more efficient SE induction in grapevines and inducing mutations for improving individual traits without altering the genetic background of the cultivar.
PubMed: 38140453
DOI: 10.3390/plants12244126 -
Frontiers in Plant Science 2023Polyphenol oxidase (PPO) activity is a major cause of the undesirable brown color of wheat-based products. , a major gene for PPO activity, was cloned based on sequence...
Polyphenol oxidase (PPO) activity is a major cause of the undesirable brown color of wheat-based products. , a major gene for PPO activity, was cloned based on sequence homology in previous studies; however, its function and regulation mechanism remain unclear. In this study, the function and genetic regulation of were analyzed using RNA interference (RNAi) and Targeting Induced Local Lesions IN Genomes (TILLING) technology, and superior mutants were identified. Compared with the control, the level of transcript in RNAi transgenic lines was drastically decreased by 15.5%-60.9% during grain development, and PPO activity was significantly reduced by 12.9%-20.4%, confirming the role of in PPO activity. Thirty-two mutants were identified in the ethyl methanesulfonate (EMS)-mutagenized population, including eight missense mutations, 16 synonymous mutations, and eight intron mutations. The expression of was reduced significantly by 6.7%-37.1% and 10.1%-54.4% in mutants M092141 (G311S) and M091098 (G299R), respectively, in which PPO activity was decreased by 29.7% and 28.8%, respectively, indicating that mutation sites of two mutants have important effects on PPO1 function. Sequence and structure analysis revealed that the two sites were highly conserved among 74 plant species, where the frequency of glycine was 94.6% and 100%, respectively, and adjacent to the entrance of the hydrophobic pocket of the active site. The M092141 and M091098 mutants can be used as important germplasms to develop wheat cultivars with low grain PPO activity. This study provided important insights into the molecular mechanism of and the genetic improvement of wheat PPO activity.
PubMed: 37583591
DOI: 10.3389/fpls.2023.1171839 -
International Journal of Molecular... Aug 2023Glucagon-like peptide-1 (GLP-1) receptor agonists are associated with reduced atrial fibrillation risk, but the mechanisms underlying this association remain unclear....
Glucagon-like peptide-1 (GLP-1) receptor agonists are associated with reduced atrial fibrillation risk, but the mechanisms underlying this association remain unclear. The GLP-1 receptor agonist directly impacts cardiac Ca homeostasis, which is crucial in pulmonary vein (PV, the initiator of atrial fibrillation) arrhythmogenesis. This study investigated the effects of the GLP-1 receptor agonist on PV electrophysiology and Ca homeostasis and elucidated the potential underlying mechanisms. Conventional microelectrodes and whole-cell patch clamp techniques were employed in rabbit PV tissues and single PV cardiomyocytes before and after GLP-1 (7-36) amide, a GLP-1 receptor agonist. Evaluations were conducted both with and without pretreatment with H89 (10 μM, an inhibitor of protein kinase A, PKA), KN93 (1 μM, an inhibitor of Ca/calmodulin-dependent protein kinase II, CaMKII), and KB-R7943 (10 μM, an inhibitor of Na/Ca exchanger, NCX). Results showed that GLP-1 (7-36) amide (at concentrations of 1, 10, and 100 nM) reduced PV spontaneous activity in a concentration-dependent manner without affecting sinoatrial node electrical activity. In single-cell experiments, GLP-1 (7-36) amide (at 10 nM) reduced L-type Ca current, NCX current, and late Na current in PV cardiomyocytes without altering Na current. Additionally, GLP-1 (7-36) amide (at 10 nM) increased sarcoplasmic reticulum Ca content in PV cardiomyocytes. Furthermore, the antiarrhythmic effects of GLP-1 (7-36) amide on PV automaticity were diminished when pretreated with H89, KN93, or KB-R7943. This suggests that the GLP-1 receptor agonist may exert its antiarrhythmic potential by regulating PKA, CaMKII, and NCX activity, as well as modulating intracellular Ca homeostasis, thereby reducing PV arrhythmogenesis.
Topics: Animals; Rabbits; Atrial Fibrillation; Glucagon-Like Peptide-1 Receptor; Calcium; Pulmonary Veins; Calcium-Calmodulin-Dependent Protein Kinase Type 2; Hormone Antagonists; Anti-Arrhythmia Agents; Amides; Bone Density Conservation Agents; Cyclic AMP-Dependent Protein Kinases; Glucagon-Like Peptide 1; Homeostasis
PubMed: 37685906
DOI: 10.3390/ijms241713100 -
Plant Biotechnology Journal Oct 2023Hexaploid wheat (Triticum aestivum), a major staple crop, has a remarkably large genome of ~14.4 Gb (containing 106 913 high-confidence [HC] and 159 840...
Hexaploid wheat (Triticum aestivum), a major staple crop, has a remarkably large genome of ~14.4 Gb (containing 106 913 high-confidence [HC] and 159 840 low-confidence [LC] genes in the Chinese Spring v2.1 reference genome), which poses a major challenge for functional genomics studies. To overcome this hurdle, we performed whole-exome sequencing to generate a nearly saturated wheat mutant database containing 18 025 209 mutations induced by ethyl methanesulfonate (EMS), carbon (C)-ion beams, or γ-ray mutagenesis. This database contains an average of 47.1 mutations per kb in each gene-coding sequence: the potential functional mutations were predicted to cover 96.7% of HC genes and 70.5% of LC genes. Comparative analysis of mutations induced by EMS, γ-rays, or C-ion beam irradiation revealed that γ-ray and C-ion beam mutagenesis induced a more diverse array of variations than EMS, including large-fragment deletions, small insertions/deletions, and various non-synonymous single nucleotide polymorphisms. As a test case, we combined mutation analysis with phenotypic screening and rapidly mapped the candidate gene responsible for the phenotype of a yellow-green leaf mutant to a 2.8-Mb chromosomal region. Furthermore, a proof-of-concept reverse genetics study revealed that mutations in gibberellic acid biosynthesis and signalling genes could be associated with negative impacts on plant height. Finally, we built a publically available database of these mutations with the corresponding germplasm (seed stock) repository to facilitate advanced functional genomics studies in wheat for the broad plant research community.
Topics: Triticum; Exome Sequencing; Mutation; Mutagenesis; Genomics; Ethyl Methanesulfonate; Genome, Plant
PubMed: 37401008
DOI: 10.1111/pbi.14111 -
Antibiotics (Basel, Switzerland) Jun 2023Antibiotic-resistant bacteria represent a serious public health threat. For that reason, the development of new and effective antibiotics to control pathogens has become...
Antibiotic-resistant bacteria represent a serious public health threat. For that reason, the development of new and effective antibiotics to control pathogens has become necessary. The current study aims to search for new microorganisms expressing antibiotic production capacity. Fifteen sites covering a wide range of harsh environmental conditions in Egypt were investigated. Two hundred and eighty bacterial isolates were obtained and then tested against pathogenic bacteria using the agar disk diffusion technique. Fifty-two (18.6% of the total) of the isolates exhibited antagonistic properties, which affected one or more of the tested pathogens. The isolate 113 was identified as and isolate 10 was identified as using the 16S rRNA technique. The strain was stronger in antibiotic production against , and whereas the strain was more efficient against and sp. The sensitivity of the strains to commercial antibiotics showed that was highly sensitive to seven commercial antibiotics, whereas was sensitive to nine antibiotics. The two strains were subjected to ethyl methanesulfonate (EMS) mutagenesis to obtain mutants with a higher antibiotic production. The total bacterial count was measured after treatment with EMS mutagen and showed a significant gradual increase in the antimicrobial activity, which was achieved via shaking in the presence of EMS for 60 min. High antimicrobial activities were noted with 17 and 14 mutants from the and strains, respectively. The mutant (M15/Amo) was more active than the parent strain against (212.5%), while the mutant (B7/Neo) was more effective against (83.3%). The present study demonstrates the possibility of obtaining potent antibiotic-producing bacteria in hot spring waters and further improving the indigenous bacterial capacity to produce antibiotics by using EMS mutagenesis.
PubMed: 37508191
DOI: 10.3390/antibiotics12071095 -
ABIOTECH Sep 2023Induced mutations are important for genetic research and breeding. Mutations induced by physical or chemical mutagenesis are usually heterozygous during the early...
Induced mutations are important for genetic research and breeding. Mutations induced by physical or chemical mutagenesis are usually heterozygous during the early generations. However, mutations must be fixed prior to phenotyping or field trials, which requires additional rounds of self-pollination. Microspore culture is an effective method to produce double-haploid (DH) plants that are fixed homozygotes. In this study, we conducted ethyl methanesulfonate (EMS)-induced mutagenesis of microspore cultures of barley () cultivar 'Hua30' and landrace 'HTX'. The EMS concentrations were negatively correlated with the efficiency of callus induction and the frequency of mutant plant regeneration. The two genotypes showed different regeneration efficiencies. The phenotypic variation of the regenerated M plants and the presence of genome-wide nucleotide mutations, revealed by whole-genome sequencing, highlight the utility of EMS-induced mutagenesis of isolated microspore cultures for developing DH mutants. Genome-wide analysis of the mutation frequency in the regenerated plants revealed that a considerable proportion of mutations resulted from microspore culture (somaclonal variation) rather than EMS-induced mutagenesis. In addition to producing a population of 1972 homozygous mutant lines that are available for future field trials, this study lays the foundation for optimizing the regeneration efficiency of DH plants and the richness of mutations (mainly by fine-tuning the mutagen dosage).
PubMed: 37970468
DOI: 10.1007/s42994-023-00108-6 -
Scientific Reports Sep 2023Lipase is a kind of industrial enzyme preparation with various catalytic abilities and is widely used in food, energy, medicine and other fields. To increase lipase and...
Lipase is a kind of industrial enzyme preparation with various catalytic abilities and is widely used in food, energy, medicine and other fields. To increase lipase and enzyme cocktail activity through solid-state fermentation, the novel strain Penicillium sp. Y-21 was obtained through ethyl methanesulfonate (EMS) mutation from the novel strain Y, which was isolated from soils. Solid-state fermentation by strain Y-21 using agricultural byproducts was carried out in tray bioreactors. The optimum culture composition for enzyme cocktail fermentation was soybean meal 20 g, 3% (w/w) glucose, 1% (w/w) peptone, 5% (w/w) lard, 0.04% (w/w) CaCl, 0.04% (w/w) FeCl, 28 °C for 72 h. The enzyme cocktail produced by strain Y-21 is a kind of multienzyme complex, containing xylanase, glucanase, acidic protease, pectinase, cellulase and lipase, and their enzymatic activities (unit: U g) were 8000, 6000, 8000, 2000, 3000 and 120, respectively. During the fermentation process, the lipase coding genes pel, pha, and p12 were also studied and amplified from the RNA of Penicillium sp. Y-21 by RT-PCR. The results showed that the pel gene played an important role in enzyme production. Afterwards, an enzyme cocktail can be added to chicken feed as an additive, which improves animal growth and feed efficiency.
Topics: Animals; Lipase; Fermentation; Endopeptidases; Peptide Hydrolases; Bioreactors; Antibodies, Fungal
PubMed: 37667063
DOI: 10.1038/s41598-023-41912-w