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Keywords = polyploid genome

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13 pages, 1803 KiB  
Article
Characterization of Polyploid Embryoid Lines Induced via Unfertilized Ovule Culture of Loquat (Eriobotrya japonica Lindl.)
by Shuming Wang, Tingting Ruan, Yin Zhang, Peng Wang, Jiangbo Dang, Yan Xia, Danlong Jing, Guolu Liang, Qigao Guo and Qiao He
Horticulturae 2025, 11(7), 727; https://doi.org/10.3390/horticulturae11070727 - 23 Jun 2025
Viewed by 355
Abstract
Polyploidy plays a significant role in loquat breeding, particularly in triploid breeding for seedless fruit production. Currently, loquat polyploid breeding primarily relies on natural seedling selection and sexual hybridization approaches. In this study, unfertilized ovules from four loquat varieties were in vitro cultured. [...] Read more.
Polyploidy plays a significant role in loquat breeding, particularly in triploid breeding for seedless fruit production. Currently, loquat polyploid breeding primarily relies on natural seedling selection and sexual hybridization approaches. In this study, unfertilized ovules from four loquat varieties were in vitro cultured. Gynogenesis and embryoid regeneration were achieved in ‘Xingning 1’ and ‘Huabai 1’, with ‘Xingning 1’ demonstrating the highest gynogenesis efficiency (21.63%). Flow cytometry and chromosome counting revealed that the obtained embryoid lines included haploid, diploid, tetraploid, hexaploid, and chimeric ploidy types. Further characterization of ‘Xingning 1’-derived embryoid lines through SSR markers and whole-genome resequencing confirmed that the haploid, diploid, tetraploid, and hexaploidy embryoid originated from haploid–somatic chimeras, diploid, doubled diploid and tripled diploid, respectively. Metabolic analysis showed a positive correlation between ploidy level and the content of both soluble sugars and organic acids. This study explored a novel platform for polyploid induction in loquat and may provide methodological insights for improvement of other perennial fruit trees. Full article
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13 pages, 1975 KiB  
Article
Leaf Organogenesis Improves Recovery of Solid Polyploid Shoots from Chimeric Southern Highbush Blueberry
by Emily Walter, Akshaya Biswal, Peggy Ozias-Akins and Ye Chu
BioTech 2025, 14(2), 48; https://doi.org/10.3390/biotech14020048 - 12 Jun 2025
Viewed by 628
Abstract
Interspecific and intersectional crosses have introduced valuable genetic traits for blueberry (Vaccinium sect. Cyanococcus) cultivar improvement. Introgression from Vaccinium species at the diploid, tetraploid, and hexaploid levels has been found in cultivated blueberries. Continued efforts to integrate wild blueberry genetic resources [...] Read more.
Interspecific and intersectional crosses have introduced valuable genetic traits for blueberry (Vaccinium sect. Cyanococcus) cultivar improvement. Introgression from Vaccinium species at the diploid, tetraploid, and hexaploid levels has been found in cultivated blueberries. Continued efforts to integrate wild blueberry genetic resources into blueberry breeding are essential to broaden the genetic diversity of cultivated blueberries. However, performing heteroploid crosses among Vaccinium species is challenging. Polyploid induction through tissue culture has been useful in bridging ploidy barriers. Mixoploid or chimeric shoots often are produced, along with solid polyploid mutants. These chimeras are mostly discarded because of their genome instability and the difficulty in identifying periclinal mutants carrying germline mutations. Since induced polyploidy in blueberries often results in a low frequency of solid mutant lines, it is important to recover solid polyploids through chimera dissociation. In this study, two vegetative propagation methods, i.e., axillary and adventitious shoot induction, were evaluated for their efficiency in chimera dissociation. Significantly higher rates of chimera dissociation were found in adventitious shoot induction compared to axillary shoot induction. Approximately 89% and 82% of the adventitious shoots induced from mixoploid lines 145.11 and 169.40 were solid polyploids, respectively, whereas only 25% and 53% of solid polyploids were recovered through axillary shoot induction in these lines. Effective chimera dissociation provides useful and stable genetic materials to enhance blueberry breeding. Full article
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18 pages, 6079 KiB  
Article
Integrative Genomic and Cytogenetic Analyses Reveal the Landscape of Typical Tandem Repeats in Water Hyacinth
by Liqing Feng, Ying Zhuang, Dagang Tian, Linwei Zhou, Jinbin Wang and Jingping Fang
Horticulturae 2025, 11(6), 657; https://doi.org/10.3390/horticulturae11060657 - 10 Jun 2025
Viewed by 331
Abstract
Tandem repeats in eukaryotic genomes exhibit intrinsic instability that drives rapid evolutionary diversification. However, their evolutionary dynamics in allopolyploid species such as the water hyacinth (Pontederia crassipes or Eichhornia crassipes) remain largely unexplored. Our study used integrated genomic and cytogenetic analyses [...] Read more.
Tandem repeats in eukaryotic genomes exhibit intrinsic instability that drives rapid evolutionary diversification. However, their evolutionary dynamics in allopolyploid species such as the water hyacinth (Pontederia crassipes or Eichhornia crassipes) remain largely unexplored. Our study used integrated genomic and cytogenetic analyses of this allotetraploid species to characterize five representative tandem repeats, revealing distinct genomic distribution patterns and copy number polymorphisms. The highly abundant centromeric tandem repeat, putative CentEc, was co-localized with the centromeric retrotransposon CREc, indicating conserved centromeric architecture. Remarkably, putative CentEc sequences showed high sequence conservation (91–100%) despite subgenome divergence, indicative of active concerted evolution. Fluorescence in situ hybridization (FISH) analysis showed ubiquitous telomeric repeats across all chromosomes, while an interstitial chromosome region tandem repeat (ICREc) displayed chromosome-specific localization, both exhibiting copy number variation. Furthermore, differential rDNA organization was observed. 5S rDNA was detected on a single chromosome pair, whereas 35S rDNA exhibited multichromosomal distribution with varying intensities. A comparative analysis of subgenome-specific rDNA sequences revealed substantial heterogeneity in both 5S and 35S rDNA units, suggesting subgenome-biased evolutionary trajectories. Collectively, these findings elucidate the structural and evolutionary significance of tandem repeats in shaping the water hyacinth genome, highlighting mechanisms of concerted evolution and subgenome-biased adaptation in invasive polyploids. Full article
(This article belongs to the Special Issue Latest Advances and Prospects in Germplasm of Tropical Fruits)
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14 pages, 962 KiB  
Review
Haplotype-Resolved Assembly in Polyploid Plants: Methods, Challenges, and Implications for Evolutionary and Breeding Research
by Zhenning Zhao and Tao Shi
Genes 2025, 16(6), 636; https://doi.org/10.3390/genes16060636 - 27 May 2025
Viewed by 744
Abstract
Polyploidization has been one of the key drivers of plant evolution, profoundly influencing plant adaptation in nature and crop traits in agriculture. Deciphering polyploid genomes is a crucial step for understanding evolutionary history and advancing agricultural applications. However, the inherent complexity of polyploid [...] Read more.
Polyploidization has been one of the key drivers of plant evolution, profoundly influencing plant adaptation in nature and crop traits in agriculture. Deciphering polyploid genomes is a crucial step for understanding evolutionary history and advancing agricultural applications. However, the inherent complexity of polyploid genomes has long hindered accurate assembly and annotation. Recent advances in sequencing technologies and improved assembly algorithms have significantly enhanced the resolution of complex polyploid genomes. These innovations have led to the successful assembly and public release of an increasing number of high-quality polyploid plant genomes. This review summarizes the mechanisms of polyploid formation and their evolutionary relevance, with a focus on recent technological progress in sequencing and genome assembly. On this basis, we further discuss the current key challenges of polyploid genome assembly and the ways to address them. Full article
(This article belongs to the Special Issue Gene and Genome Duplications in Plants)
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27 pages, 8811 KiB  
Article
Participation of Wild Species Genus Avena L. (Poaceae) of Different Ploidy in the Origin of Cultivated Species According to Data on Intragenomic Polymorphism of the ITS1-5.8S rRNA Region
by Alexander A. Gnutikov, Nikolai N. Nosov, Igor G. Loskutov, Alexander V. Rodionov and Victoria S. Shneyer
Plants 2025, 14(10), 1550; https://doi.org/10.3390/plants14101550 - 21 May 2025
Viewed by 425
Abstract
The possible origin of four cultivated species of the genus Avena of different ploidy and different subgenome composition (A. strigosa, A. abyssinica, A. byzantina, and A. sativa) from possible wild species was investigated. The region of the internal [...] Read more.
The possible origin of four cultivated species of the genus Avena of different ploidy and different subgenome composition (A. strigosa, A. abyssinica, A. byzantina, and A. sativa) from possible wild species was investigated. The region of the internal transcribed spacer ITS1 and the 5.8S rRNA gene in the cultivated species was studied with next-generation sequencing (NGS), and the patterns of occurrence and distribution of the ribotypes were compared among them and with those of the wild species. According to these data diploid, A. strigosa is more closely related to the diploid A. hirtula than to polyploid oats, and it could have evolved independently of polyploid cultivated species. The tetraploid Avena abyssinica could be a cultivated derivative of A. vaviloviana. Two hexaploid cultivated species, A. byzantina and A. sativa, could have a different origin; A. sativa could be the cultivated form of A. fatua, whereas A. byzantina could originate independently. It was found that the oat species with the A and C subgenomes, even with strong morphological and karyological differences, could intercross and pass the further stages of introgression producing a new stable combination of genomes. Our data show that almost all species of Avena could form an introgressive interspecies complex. Full article
(This article belongs to the Special Issue Plant Molecular Phylogenetics and Evolutionary Genomics III)
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19 pages, 4585 KiB  
Article
Origin of Polyploidy, Phylogenetic Relationships, and Biogeography of Botiid Fishes (Teleostei: Cypriniformes)
by Lei Yang, Richard L. Mayden and Gavin J. P. Naylor
Biology 2025, 14(5), 531; https://doi.org/10.3390/biology14050531 - 11 May 2025
Cited by 1 | Viewed by 695
Abstract
Botiidae is a small family of freshwater fishes distributed across Southeast Asia, South Asia, and East Asia. It comprises two subfamilies: the diploid Leptobotiinae and the tetraploid Botiinae. Whether species in the Botiinae are autotetraploids or allotetraploids and how many polyploidization events occurred [...] Read more.
Botiidae is a small family of freshwater fishes distributed across Southeast Asia, South Asia, and East Asia. It comprises two subfamilies: the diploid Leptobotiinae and the tetraploid Botiinae. Whether species in the Botiinae are autotetraploids or allotetraploids and how many polyploidization events occurred during the evolution of this subfamily remain open questions. The phylogenetic relationships and biogeography of the Botiidae also require further investigation. In the current study, we compared phylogenetic trees constructed using DNA sequences from the mitochondrial genome and five phased nuclear genes. We also performed whole genome sequencing for two tetraploid species: Chromobotia macracanthus and Yasuhikotakia modesta. Genome profiling of five botiine species suggests that they are likely of allotetraploid origin. Nuclear gene tree topologies indicate that the tetraploidization of the Botiinae occurred only once in the common ancestor of this subfamily. Although the possible maternal progenitor and paternal progenitor of the Botiinae cannot be determined, the subfamily Leptobotiinae can be excluded as a progenitor. The gene trees built in this study generally agree on the following sister group relationships: Leptobotiinae/Botiinae, Leptobotia/Parabotia, Chromobotia/Botia, Yasuhikotakia/Syncrossus, and Sinibotia/Ambastaia. Clades formed by the last two generic pairs are also sisters to each other. Timetree analyses and ancestral range reconstruction suggest that the family Botiidae might have originated in East Asia and Mainland Southeast Asia approximately 51 million years ago and later dispersed to South Asia and the islands of Southeast Asia. Full article
(This article belongs to the Special Issue Young Researchers in Conservation Biology and Biodiversity)
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24 pages, 10390 KiB  
Article
Detection of Different Patterns of Genome-Wide Gene Expression Disturbance in Three Nullisomy Lines in Allotetraploid Brassica napus
by Shaolin Lei, Bo Wei, Qi Hu, Lang Liu, Feng Yu, Tuo Zeng, Xuye Du, Lei Gu and Hongcheng Wang
Plants 2025, 14(10), 1434; https://doi.org/10.3390/plants14101434 - 10 May 2025
Viewed by 454
Abstract
Aneuploidy-related disruptions are generally tolerated in polyploid plants, which exhibit a greater capacity for genomic compensation. In this study, we utilize allotetraploid Brassica napus as a model and generated three aneuploid variants (NC1, NC2, and NC8) to investigate the phenotypic and transcriptional consequences [...] Read more.
Aneuploidy-related disruptions are generally tolerated in polyploid plants, which exhibit a greater capacity for genomic compensation. In this study, we utilize allotetraploid Brassica napus as a model and generated three aneuploid variants (NC1, NC2, and NC8) to investigate the phenotypic and transcriptional consequences of chromosome loss. Significant phenotypic variations were observed, with the most notable being a marked dwarfing phenotype in the aneuploid materials compared to the euploid Oro. Transcriptomic analysis revealed widespread alterations in gene expression across the entire genome in the deficient variants. Notably, most of the differentially expressed genes (DEGs) were attributed to trans-acting effects resulting from the deletion of C chromosomes. Deletion of the C chromosomes induced gene expression changes not only on the corresponding chromosomes, but also on the affected genes across other chromosomes. Specifically, in the C1-deleted variant, the average gene expression of the A1 chromosome increased, while the number of expressed genes on other chromosomes decreased. In contrast, for C2 and C8 deletions, the average expression levels of homologous genes decreased, but the number of expressed genes on other chromosomes increased. These findings shed light on the complex compensatory mechanisms that underlie aneuploidy in polyploid plants and provide valuable insights into how plants maintain genomic stability despite chromosomal loss. Full article
(This article belongs to the Special Issue Advances in Molecular Genetics and Breeding of Brassica napus L.)
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22 pages, 5259 KiB  
Article
Genome-Wide Identification and Evolution-Profiling Analysis of Heat Shock Protein Gene Family in Poaceae Barnhart
by Xiaoyi Huang, Yue Liu, Xiao Yu, Yajun Cai, Lingyu Hou, Jingyuan Zhang and Hongchun Yang
Int. J. Mol. Sci. 2025, 26(9), 4269; https://doi.org/10.3390/ijms26094269 - 30 Apr 2025
Viewed by 667
Abstract
Heat shock proteins (HSPs) function as molecular chaperones to maintain protein homeostasis and repair denatured proteins, counteracting abiotic stresses. Despite their functional importance, a systematic bioinformatics analysis of the HSP gene family was lacking in Poaceae. In this study, we revealed that [...] Read more.
Heat shock proteins (HSPs) function as molecular chaperones to maintain protein homeostasis and repair denatured proteins, counteracting abiotic stresses. Despite their functional importance, a systematic bioinformatics analysis of the HSP gene family was lacking in Poaceae. In this study, we revealed that HSPs are widely distributed from algae to eudicots, with varying numbers in Poaceae including Oryza, Triticum, and Panicum. Gene duplication events, particularly dispersed duplication (DSD), tandem duplication (TD), and genome polyploidization, have probably driven the increased number of HSP genes and the expansion of HSP family proteins. Gene Ontology (GO) annotation analyses suggested their conserved functions. Promoter cis-acting element analyses indicated that their expression levels were tightly regulated by abiotic stresses. We validated that many collinear HSP genes are indeed regulated by the cold stress by analyzing the published RNA-seq data in rice, maize, and wheat, and performing RT-qPCR in rice. Our findings shed light on the role of HSPs in the abiotic stress response, laying the groundwork for further exploration of HSP functions in Poaceae. Full article
(This article belongs to the Special Issue Plant Responses to Biotic and Abiotic Stresses)
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19 pages, 4516 KiB  
Article
The First Genome-Wide Survey Analysis of the Tibetan Plateau Tetraploid Schizothorax curvilabiatus Reveals Its Microsatellite Characteristics and Phylogenetic Relationships
by Bingjian Liu, Luxiu Gao, Yifan Liu, Kai He, Hongchi Li, Taobo Feng, Mingzhe Han and Chi Zhang
Genes 2025, 16(5), 491; https://doi.org/10.3390/genes16050491 - 25 Apr 2025
Viewed by 599
Abstract
Background/Objectives: Schizothorax curvilabiatus, a typical highland polyploid species within the subfamily Schizothoracinae, holds economic value and ecological research significance. Currently, there are no related genomic studies. To obtain its genetic information and lay the foundation for subsequent whole-genome map construction, this [...] Read more.
Background/Objectives: Schizothorax curvilabiatus, a typical highland polyploid species within the subfamily Schizothoracinae, holds economic value and ecological research significance. Currently, there are no related genomic studies. To obtain its genetic information and lay the foundation for subsequent whole-genome map construction, this study conducted a genome survey analysis, preliminary genome assembly, microsatellite identification, repeat sequence annotation, mitochondrial genome characterization, and phylogenetic relationship research. Methods: DNA was sequenced on a DNBSEQ-T7 platform to obtain paired-end genomic data. The genome was analyzed using GCE, and the draft genome was assembled with SOAPdenovo. Microsatellites were identified using MISA, and the mitochondrial genome was assembled with NOVOPlasty. Genome features were analyzed, and phylogenetic trees were constructed using PhyloSuite and MEGA. Results: The genome size was estimated at 2.53 Gb, with a heterozygosity of 6.55% and 47.66% repeat sequences. A 1.324 Gb preliminary genome draft was obtained, with repeat sequences comprising 47.17%, the majority being DNA transposons (24.64%). Dinucleotide repeats were most abundant (46.91%), followed by mononucleotide repeats (38.31%), with A/T and AC/GT being the most frequent. A complete mitochondrial genome of 16,589 bp was assembled, and a 939 bp D-loop was annotated. Phylogenetic relationships among genera in the Schizothoracinae subfamily were also clarified. Conclusions: This study provides the latest molecular data for analysis of the S. curvilabiatus genome and its related populations, and for the first time offers genomic resources for research on genomic adaptive evolution and polyploidization in high-altitude environments. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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24 pages, 314 KiB  
Review
Understanding the Regulation Activities of Transposons in Driving the Variation and Evolution of Polyploid Plant Genome
by Yafang Xiao and Jianbo Wang
Plants 2025, 14(8), 1160; https://doi.org/10.3390/plants14081160 - 8 Apr 2025
Viewed by 960
Abstract
Transposon is the main component of the eukaryotic genome, and more and more plant genome data show that transposons are diverse in regulating genome structure, variation, function and evolution, with different transposition mechanisms in the genome. Hybridization and polyploidy play an important role [...] Read more.
Transposon is the main component of the eukaryotic genome, and more and more plant genome data show that transposons are diverse in regulating genome structure, variation, function and evolution, with different transposition mechanisms in the genome. Hybridization and polyploidy play an important role in promoting plant speciation and evolution, and recent studies have shown that polyploidy is usually accompanied by the expansion of transposons, which affect the genome size and structure of polyploid plants. Transposons can insert into genes and intergenic regions, resulting in great differences in the overall genome structure of closely related plant species, and it can also capture gene segments in the genome to increase the copy number of genes. In addition, transposons influence the epigenetic modification state of the genome and regulate the expression of the gene, while plant phenotype, biological and abiotic stress response are also regulated by transposons. Overall, transposons play an important role in the plant genome, especially polyploid plant genome, adaptation and evolution. Full article
(This article belongs to the Special Issue Plant Molecular Phylogenetics and Evolutionary Genomics III)
12 pages, 2316 KiB  
Article
Polyploid Advantage? Comparing Salt Stress Responses of Di- and Tetraploid Acacia senegal (L.) Willd. Seedlings
by Adja Madjiguene Diallo, Erik Dahl Kjær, Lene Rostgaard Nielsen and Anders Ræbild
Forests 2025, 16(3), 412; https://doi.org/10.3390/f16030412 - 25 Feb 2025
Viewed by 559
Abstract
Polyploidy is often associated with the ability to grow under harsh conditions. The objective of this study was to test whether genome doubling in Acacia senegal confers superiority for growth under saline conditions. For this purpose, we assessed the cytotype distribution across ten [...] Read more.
Polyploidy is often associated with the ability to grow under harsh conditions. The objective of this study was to test whether genome doubling in Acacia senegal confers superiority for growth under saline conditions. For this purpose, we assessed the cytotype distribution across ten natural stands in Senegal, representing different soil salinities. We further compared the growth of diploid and tetraploid A. senegal seedlings in a shared-pot greenhouse experiment, in which seedling pairs (diploids and polyploids) were irrigated with and without added NaCl. We found polyploids in six stands, and the proportion of polyploids in these stands was significantly and positively correlated with their level of soil salinity. Under experimental conditions, we found that the growth rate of diploid and tetraploid seedlings was equally depressed by salt stress, but the mortality rate for tetraploids was lower than that for diploids (34% versus 54%). In addition, the root-top ratio was higher in tetraploids compared to diploids, which may influence fitness under harsh conditions. Overall, this study provides support for polyploid superiority in A. senegal growing in saline soils. The findings of this study may have practical implications for the reforestation of saline soils with polyploid A. senegal, and we recommend further studies to elucidate their potential. Full article
(This article belongs to the Special Issue Genomic Analysis of Growth and Stress Adaptation in Forest Trees)
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20 pages, 1868 KiB  
Article
Chromosome Morphology and Heterochromatin Patterns in Paspalum notatum: Insights into Polyploid Genome Structure
by Ana I. Honfi, A. Verena Reutemann, Juan S. Schneider, Lucas M. Escobar, Eric J. Martínez and Julio R. Daviña
Genes 2025, 16(3), 242; https://doi.org/10.3390/genes16030242 - 20 Feb 2025
Cited by 1 | Viewed by 782
Abstract
Background/Objectives: Paspalum notatum is a key multipurpose species native to American grasslands. This study provides, for the first time, a detailed karyotype analysis of diploid (2n = 2x = 20) and tetraploid (2n = 4x = 40) accessions of [...] Read more.
Background/Objectives: Paspalum notatum is a key multipurpose species native to American grasslands. This study provides, for the first time, a detailed karyotype analysis of diploid (2n = 2x = 20) and tetraploid (2n = 4x = 40) accessions of P. notatum, the most common cytotypes within the species. Methods: The constitutive heterochromatin patterns revealed using CMA-DA-DAPI staining and genome size estimations are novel contributions to the understanding of the N genome in Paspalum. Results: Chromosomes were small (1.1–2.3 µm), with the diploid karyotype comprising nine metacentric pairs (one bearing microsatellites on the short arms, pair 6) and one submetacentric pair. In tetraploids, the diploid karyotype was duplicated. Heterochromatin analysis revealed two CMA++/DAPI bands located on the short arm and satellite of chromosome 6 in diploids, while tetraploids exhibited two to three CMA++/DAPI and one to two CMA++/DAPI0 bands. The proportion of GC-rich heterochromatin represented 2.8 and 3.47% of the total chromosome length in diploid and tetraploid cytotypes, respectively. Genome size analysis revealed a reduction in monoploid genome size in tetraploids (1Cx = 0.678 pg) compared to diploids (1Cx = 0.71 pg), consistent with the autopolyploid origin hypothesis. Conclusions: These findings provide essential cytogenetic insights and suggest only minor structural changes in the N genome following polyploidization, which could guide future studies integrating genomic and cytogenetic maps of P. notatum. Full article
(This article belongs to the Special Issue Genetics and Breeding of Polyploid Plants)
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67 pages, 32566 KiB  
Article
Advances in Understanding the Karyotype Evolution of Tetrapulmonata and Two Other Arachnid Taxa, Ricinulei and Solifugae
by Jiří Král, Alexandr Sember, Klára Divišová, Tereza Kořínková, Azucena C. Reyes Lerma, Ivalú M. Ávila Herrera, Martin Forman, František Šťáhlavský, Jana Musilová, Sabrina Torres Kalme, José G. Palacios Vargas, Magda Zrzavá, Iva Vrbová, Jairo A. Moreno-González, Paula E. Cushing, Alexander V. Gromov, Štěpánka Šebestiánová, Vendula Bohlen Šlechtová, Lorenzo Prendini and Tharina L. Bird
Genes 2025, 16(2), 207; https://doi.org/10.3390/genes16020207 - 8 Feb 2025
Cited by 2 | Viewed by 1869
Abstract
Background/Objectives: Arachnids are a megadiverse arthropod group. The present study investigated the chromosomes of pedipalpid tetrapulmonates (orders Amblypygi, Thelyphonida, Schizomida) and two arachnid orders of uncertain phylogenetic placement, Ricinulei and Solifugae, to reconstruct their karyotype evolution. Except for amblypygids, the cytogenetics of these [...] Read more.
Background/Objectives: Arachnids are a megadiverse arthropod group. The present study investigated the chromosomes of pedipalpid tetrapulmonates (orders Amblypygi, Thelyphonida, Schizomida) and two arachnid orders of uncertain phylogenetic placement, Ricinulei and Solifugae, to reconstruct their karyotype evolution. Except for amblypygids, the cytogenetics of these arachnid orders was almost unknown prior to the present study. Methods: Chromosomes were investigated using methods of standard (Giemsa-stained preparations, banding techniques) and molecular cytogenetics (fluorescence in situ hybridization, comparative genomic hybridization). Results and Conclusions: New data for 38 species, combined with previously published data, suggest that ancestral arachnids possessed low to moderate 2n (22–40), monocentric chromosomes, one nucleolus organizer region (NOR), low levels of heterochromatin and recombinations, and no or homomorphic sex chromosomes. Karyotypes of Pedipalpi and Solifugae diversified via centric fusions, pericentric inversions, and changes in the pattern of NORs and, in solifuges, also through tandem fusions. Some solifuges display an enormous amount of constitutive heterochromatin and high NOR number. It is hypothesized that the common ancestor of amblypygids, thelyphonids, and spiders exhibited a homomorphic XY system, and that telomeric heterochromatin and NORs were involved in the evolution of amblypygid sex chromosomes. The new findings support the Cephalosomata clade (acariforms, palpigrades, and solifuges). Hypotheses concerning the origin of acariform holocentric chromosomes are presented. Unlike current phylogenetic hypotheses, the results suggest a sister relationship between Schizomida and a clade comprising other tetrapulmonates as well as a polyploidization in the common ancestor of the clade comprising Araneae, Amblypygi, and Thelyphonida. Full article
(This article belongs to the Special Issue Commemorating the Launch of the Section "Cytogenomics")
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14 pages, 1609 KiB  
Article
Design of an Effective sgRNA for CRISPR/Cas9 Knock-Ins and Full Mutant Segregation in Polyploid Synechocystis sp. PCC 6803
by Maria Isabel Nares-Rodriguez and Esther Karunakaran
SynBio 2025, 3(1), 3; https://doi.org/10.3390/synbio3010003 - 27 Jan 2025
Viewed by 1048
Abstract
Synechocystis sp. PCC 6803 is a highly promising organism for the production of diverse recombinant compounds, including biofuels. However, conventional genetic engineering in Synechocystis presents challenges due to its highly polyploid genome, which not only results in low product yields but also compromises [...] Read more.
Synechocystis sp. PCC 6803 is a highly promising organism for the production of diverse recombinant compounds, including biofuels. However, conventional genetic engineering in Synechocystis presents challenges due to its highly polyploid genome, which not only results in low product yields but also compromises the reliability of recombinant strains for biomanufacturing applications. The CRISPR/Cas9 system, renowned for its precision, efficiency, and versatility across a wide range of chassis, offers significant potential to address the limitations posed by polyploid genomes. In this study, we developed and optimized an effective sgRNA for the targeted knock-in of nucleotide sequences of varying lengths into the neutral locus slr0168 of polyploid Synechocystis using CRISPR/Cas9. The gene encoding di-geranylgeranylglycerophospholipid reductase from Sulfolobus acidocaldarius and the methyl ketone operon from Solanum habrochaites were chosen as the exemplar nucleotide sequences for incorporation into the chromosome of Synechocystis. Our results demonstrate that the designed sgRNA effectively facilitated both knock-in events and that CRISPR/Cas9 enabled complete mutant segregation in a single round of selection and induction. Full article
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22 pages, 2771 KiB  
Article
Genomic and Transcriptomic Analysis of the Polyploidy Cyst Nematode, Heterodera trifolii, and Heterodera schachtii
by Parthiban Subramanian, Daegwan Kim, Hyoung-Rai Ko, Joon-Soo Sim, Vimalraj Mani, Chang-Muk Lee, Seon-Kyeong Lee, Soyoung Park, Dong-Gwan Kim, Yeisoo Yu and Bum-Soo Hahn
Int. J. Mol. Sci. 2025, 26(3), 948; https://doi.org/10.3390/ijms26030948 - 23 Jan 2025
Viewed by 1036
Abstract
Cyst nematodes remain a major threat to global agricultural production, causing huge losses. To understand the parasitism of the cyst nematodes Heterodera trifolii (HT) and Heterodera schachtii (HS), we constructed whole-genome assemblies using short- and long-read sequencing technologies. The nematode genomes were 379 [...] Read more.
Cyst nematodes remain a major threat to global agricultural production, causing huge losses. To understand the parasitism of the cyst nematodes Heterodera trifolii (HT) and Heterodera schachtii (HS), we constructed whole-genome assemblies using short- and long-read sequencing technologies. The nematode genomes were 379 Mb and 183 Mb in size, with the integrated gene models predicting 40,186 and 18,227 genes in HT and HS, respectively. We found more than half of the genes predicted in HT (64.7%) and HS (53.2%) were collinear to their nearest neighbor H. glycines (HG). Large-scale duplication patterns in HT and segmental duplications of more than half of the orthologous genes indicate that the genome of HT is polyploid in nature. Functional analysis of the genes indicated that 65.6% of the HG genes existed within the HT genome. Most abundant genes in HT and HS were involved in gene regulation, DNA integration, and chemotaxis. Differentially expressed genes showed upregulation of cuticle structural constituent genes during egg and female stages and cytoskeletal motor activity-related genes in juvenile stage 2 (J2). Horizontal gene transfer analyses identified four new vitamin biosynthesis genes, pdxK, pdxH, pdxS, and fabG, of bacterial origin, to be first reported in HT and HS. Mitogenomes of HT, HS, and HG showed similar structure, composition, and codon usage. However, rates of substitution of bases in the gene nad4l were significantly different between HT and HS. The described genomes, transcriptomes, and mitogenomes of plant-parasitic nematodes HT and HS are potential bio-resources used to identify several strategies of control of the nematode. Full article
(This article belongs to the Section Molecular Genetics and Genomics)
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