Variation In DNA Content Research Articles

FLOW CYTOMETRY ANALYSIS REVEALS NUCLEAR DNA CONTENT VARIATION IN PHALAENOPSIS YOUNG LEAF AND ROOT TIP CELLS

FLOW CYTOMETRY ANALYSIS REVEALS NUCLEAR DNA CONTENT VARIATION IN PHALAENOPSIS YOUNG LEAF AND ROOT TIP CELLS

Genome size and chromosome number variation in sugarcane (Saccharum spp.) families and phenotypically contrasting Saccharum genotypes

AbstractModern sugarcane (Saccharum spp.) cultivars originated from an intricate hybridization process, resulting in complex polyploid and aneuploid genomes. Flow cytometry (FCM), which has emerged as a precise and fast method for research on genome size in plants, has potential to unveil key aspects of sugarcane cytogenetics. In this study, FCM probes were conducted to estimate 2C DNA content in 175 Saccharum genotypes embracing phenotypically contrasting canes (commercial cultivars, energy‐cane hybrids, and a Saccharum spontaneum accession) and full‐sib sugarcane families. FCM‐mediated DNA content estimates exhibited high accuracy in predicting chromosome numbers, offering a time‐efficient alternative to classical cytogenetic procedure. Low‐magnitude correlations detected between DNA content and phenotypic traits suggest that indirect selection of agronomic traits using FCM‐mediated DNA content estimates may not be efficient. DNA content variation patterns in sugarcane families varied according to the crossing, and revealed transgressive variations associated with rampant aneuploidy. Altogether, the findings uncover that loss or gain of genetic material constitutes a source of high genetic variability that may substantially impact on crop breeding.

Expedient Bayesian prediction of subfossil bone protein content using portable ATR-FTIR data

Rapid and minimally destructive methods for estimating the endogenous organic content of subfossil bone save time, lab consumables, and valuable ancient materials. Fourier transform infrared (FTIR) spectroscopy is an established method to estimate bone protein content, and portable spectroscopes enable field applications. We review the ability of benchtop and portable FTIR indices to predict %N and %collagen from 137 bone specimens drawn from eight taxa. We also explore associations of these indices with the endogenous DNA content estimated for 105 specimens. Bulk bone elemental abundance and crystallinity index data reflect diagenetic alteration of these specimens, which come from a variety of depositional environments in four countries (Madagascar, Greece, Monaco, and Germany). Infrared (IR) indices from benchtop and portable units perform similarly well in predicting observed sample N content and collagen yields. Samples that include little collagen (0–5 wt%) tend to have similar IR index values, and we present a Bayesian approach for the prediction of collagen yields. Bone type best explains variation in target species DNA content (endogenous DNA being particularly abundant in petrosals), but low IR index values were consistently associated with minimal DNA content. We conclude that, although portable FTIR fails to distinguish collagen preservation among poorly preserved samples, a simple approach with minimal sample preparation can effectively screen bone from a variety of taxa, elements, and environments for the extraction of organics.

Determination of Ploidy Levels and Nuclear DNA Content in Cryptococcus neoformans by Flow Cytometry: Drawbacks with Variability.

Flow cytometry is commonly employed for ploidy determination and cell cycle analysis in cryptococci. The cells are subjected to fixation and staining with DNA-binding fluorescent dyes, most commonly with propidium iodide (PI), before undergoing flow cytometric analysis. In ploidy determination, cell populations are classified according to variations in DNA content, as evidenced by the fluorescence intensity of stained cells. As reported in Saccharomyces cerevisiae, we found drawbacks with PI staining that confounded the accurate analysis of ploidy by flow cytometry when the size of the cryptococci changed significantly. However, the shift in the fluorescence intensity, unrelated to ploidy changes in cells with increased size, could be accurately interpreted by applying the ImageStream system. SYTOX Green or SYBR Green I, reported to enable DNA analysis with a higher accuracy than PI in S. cerevisiae, were nonspecific for nuclear DNA staining in cryptococci. Until dyes or methods capable of reducing the variability inherent in the drastic changes in cell size or shape become available, PI appears to remain the most reliable method for cell cycle or ploidy analysis in Cryptococcus.

Divergence of 10 satellite repeats in Artemisia (Asteraceae: Anthemideae) based on sequential fluorescence in situ hybridization analysis: evidence for species identification and evolution.

Artemisia is a large genus encompassing about 400 diverse species, many of which have considerable medicinal and ecological value. However, complex morphological information and variation in ploidy level and nuclear DNA content have presented challenges for evolution studies of this genus. Consequently, taxonomic inconsistencies within the genus persist, hindering the utilization of such large plant resources. Researchers have utilized satellite DNAs to aid in chromosome identification, species classification, and evolutionary studies due to their significant sequence and copy number variation between species and close relatives. In the present study, the RepeatExplorer2 pipeline was utilized to identify 10 satellite DNAs from three species (Artemisia annua, Artemisia vulgaris, Artemisia viridisquama), and fluorescence in situ hybridization confirmed their distribution on chromosomes in 24 species, including 19 Artemisia species with 5 outgroup species from Ajania and Chrysanthemum. Signals of satellite DNAs exhibited substantial differences between species. We obtained one genus-specific satellite from the sequences. Additionally, molecular cytogenetic maps were constructed for Artemisia vulgaris, Artemisia leucophylla, and Artemisia viridisquama. One species (Artemisia verbenacea) showed a FISH distribution pattern suggestive of an allotriploid origin. Heteromorphic FISH signals between homologous chromosomes in Artemisia plants were observed at a high level. Additionally, the relative relationships between species were discussed by comparing ideograms. The results of the present study provide new insights into the accurate identification and taxonomy of the Artemisia genus using molecular cytological methods.

Foraminifera as a model of eukaryotic genome dynamism.

In contrast to the canonical view that genomes cycle only between haploid and diploid states, many eukaryotes have dynamic genomes that change content throughout an individual's life cycle. However, the few detailed studies of microeukaryotic life cycles render our understanding of eukaryotic genome dynamism incomplete. Foraminifera (Rhizaria) are an ecologically important, yet understudied, clade of microbial eukaryotes with complex life cycles that include changes in ploidy and genome organization. Here, we apply fluorescence microscopy and image analysis techniques to over 2,800 nuclei in 110 cells to characterize the life cycle of Allogromia laticollaris strain Cold Spring Harbor (CSH), one of few cultivable foraminifera species. We show that haploidy and diploidy are brief moments in the A. laticollaris life cycle and that A. laticollaris nuclei endoreplicate up to 12,000 times the haploid genome size. We find that A. laticollaris reorganizes a highly endoreplicated nucleus into thousands of haploid genomes through a non-canonical mechanism called Zerfall, in which the nuclear envelope degrades and extrudes chromatin into the cytoplasm. Based on these findings, along with changes in nuclear architecture across the life cycle, we believe that A. laticollaris uses spatio-temporal mechanisms to delineate germline and somatic DNA within a single nucleus. The analyses here extend our understanding of the genome dynamics across the eukaryotic tree of life.IMPORTANCEIn traditional depictions of eukaryotes (i.e., cells with nuclei), life cycles alternate only between haploid and diploid phases, overlooking studies of diverse microeukaryotic lineages (e.g., amoebae, ciliates, and flagellates) that show dramatic variation in DNA content throughout their life cycles. Endoreplication of genomes enables cells to grow to large sizes and perhaps to also respond to changes in their environments. Few microeukaryotic life cycles have been studied in detail, which limits our understanding of how eukaryotes regulate and transmit their DNA across generations. Here, we use microscopy to study the life cycle of Allogromia laticollaris strain CSH, an early-diverging lineage within the Foraminifera (an ancient clade of predominantly marine amoebae). We show that DNA content changes significantly throughout their life cycle and further describe an unusual process called Zerfall, by which this species reorganizes a large nucleus with up to 12,000 genome copies into hundreds of small gametic nuclei, each with a single haploid genome. Our results are consistent with the idea that all eukaryotes demarcate germline DNA to pass on to offspring amidst more flexible somatic DNA and extend the known diversity of eukaryotic life cycles.

Loss of mitochondrial DNA is associated with reduced DNA content variability in Saccharomyces cerevisiae.

DNA content measurement by fluorescence-assisted cell sorting (FACS) provides information on cell cycle progression and DNA content variability. Saccharomyces cerevisiae mutants with DNA content variability that was reduced relative to wild-type strains had defects in mitochondrial DNA (mtDNA) maintenance and mitochondrial gene expression and were correlated with strains found to lack mtDNA ([ rho 0 ] cells) by genome sequencing and fluorescence microscopy. In contrast, mutants with increased variability had defects in cell cycle progression, which may indicate a loss of coordination between mtDNA and nuclear DNA replication. Thus, FACS measurement of DNA content variability can provide insight into cell-to-cell heterogeneity in mtDNA copy number.

Effect of high-energy ionizing radiation on the DNA content and genetic variation in chrysanthemum plants regenerated from irradiated ovaries

This study aimed to evaluate the range of quantitative and qualitative genetic changes in chrysanthemum plants regenerated in vitro from ovaries irradiated with high-energy photons (5, 10, and 15 Gy) and high-energy electrons (10 Gy). The highest DNA loss (up to 11%) was observed in plants originating from ovaries treated with 10 Gy high energy. AMOVA revealed significant differences between populations of plants representing different mutagenic treatments (18% for RAPD-based and 22% for SCoT-based analysis). The incidence of genetic changes was strongly correlated with the applied type and dose of ionizing radiation. The highest genetic distances to reference plants were observed for plants regenerated from 15 Gy high-energy photon (5.8% with RAPD and 1.7% with SCoT) and 10 Gy high-energy electron-treated explants (6.0% with RAPD and 2.9% with SCoT). Considerable changes in the phenotype of mutants were not necessarily correlated with the extent of genetic alterations. Qualitative and quantitative methods of evaluating post-radiation genetic changes should be combined for reliable detection of variant plants at early developmental stages.

Ecotype variations in karyotypes and 2C DNA content in Drimia indica (Roxb.) Jossop: An important medicinal plant

Indian squill (Drimia indica) belonging to the family Asparagaceae is a highly medicinal herb with several therapeutic properties. Detailed karyotype analysis and 2C DNA contents were reported in twelve ecotypes of D. indica from different geographical locations in the state of Odisha, India. The diploid somatic chromosome 2n = 20 was recorded in four ecotypes ODi-16 (Kendrapada), ODi-23 (Tikarpada), ODi-24 (Daspalla), and ODi-26 (Odagaon) while tetraploid having 2n = 40 was newly reported in one ecotype from Nuagaon (Eco-ODi-06). Aneuploid chromosome number 2n = 26 was obtained in ecotypes from Similipal and Satkosia besides 2n = 32 was newly reported in ecotypes from Bitarkanika and Narasinghpur. The ecotypes ODi-22 (Nayagarh) and ODi-11 (Nilagiri) were found to be with 2n = 16 and 2n = 22 respectively. The 2C DNA content analysis showed a 2.19-fold increase which is confirmed by an increase in the number and chromosome length of ecotypes. The 2C DNA values were found highest at 36.26 p.g. (2n = 40) in Eco-ODi-06 (Nuagaon) and lowest 16.54 p.g. (2n = 16) in Eco-ODi-22 of Nayagarh that corresponds with ploidy and genome size. The asymmetry in the karyotypes showed non-significant clusters among the ecotypes except ecotypes ODi-26 (Odagaon) and ODi-2a (Bhitarkanika) which are most asymmetric having advanced karyotypic features as compared to the ecotypes ODi-01, ODi-08 and ODi-22 having symmetric karyotypes. The chromosome characteristics along with genome size analysis of D. indica can be very useful in establishing genome-specific cytotypes which can be used for breeding programs for crop improvement. Furthermore, enhancement of therapeutic phytoconstituents and cyto-taxonomic identification from adulterants and confirmation of allopolyploidy or autopolyploidy through genomic in situ hybridization could unravel the potential of this plant in relation to secondary metabolite production.

Genome size, chromosome number variation and its correlation with stomatal characters for assessment of ploidy levels in a core subset of mulberry (Morus spp.) germplasm

The large size of the germplasm collection along with scanty information on their cytological and genome constitution have hindered well-planned breeding schemes in mulberry. To address the issue, a study was undertaken to investigate the variability in DNA content and genome size, chromosome number, ploidy and its relation with important stomatal characteristics among 162 mulberry germplasm collection. These germplasm comprise a core subset of 150 collections along with a representative collection of different mulberry species including the wild. Among the germplasm belonging to 16 species, we identified 122 diploids (2n = 28), 4 aneuploids (2n = 30), 13 triploids (2n = 42), 15 tetraploids (2n = 56), 7 hexaploids (2n = 84) and 1 dodecosaploid (2n = 308) based on the chromosome count. Most of the cultivated mulberries are found to be diploids. The mean nuclear 2C DNA content estimated by Flow cytometry, varied from 0.723 ± 0.006 pg (M. australis, 2n = 2x) to 7.732 pg (M. nigra, 2n = 22x). The 2C DNA content positively correlated with the ploidy status and stomatal length (r = 0.814, p < 0.001). Based on the 1Cx value, the study also suggests that the majority of the polyploid species have experienced genome downsizing in relation to their diploid progenitors. This study provides the most essential information on chromosome number, ploidy and DNA content to facilitate the utilization of a core subset of germplasm in the mulberry breeding program.

Extreme genome diversity and cryptic speciation in a harmful algal-bloom-forming eukaryote

Harmful algal blooms of the toxic haptophyte Prymnesium parvum are a recurrent problem in many inland and estuarine waters around the world. Strains of P.parvum vary in the toxins they produce and in other physiological traits associated with harmful algal blooms, but the genetic basis for this variation is unknown. To investigate genome diversity in this morphospecies, we generated genome assemblies for 15 phylogenetically and geographically diverse strains of P.parvum, including Hi-C guided, near-chromosome-level assemblies for two strains. Comparative analysis revealed considerable DNA content variation between strains, ranging from 115 to 845 Mbp. Strains included haploids, diploids, and polyploids, but not all differences in DNA content were due to variation in genome copy number. Haploid genome size between strains of different chemotypes differed by as much as 243 Mbp. Syntenic and phylogenetic analyses indicate that UTEX 2797, a common laboratory strain from Texas, is a hybrid that retains two phylogenetically distinct haplotypes. Investigation of gene families variably present across the strains identified several functional categories associated with metabolic and genome size variation in P.parvum, including genes for the biosynthesis of toxic metabolites and proliferation of transposable elements. Together, our results indicate that P.parvum comprises multiple cryptic species. These genomes provide a robust phylogenetic and genomic framework for investigations into the eco-physiological consequences of the intra- and inter-specific genetic variation present in P.parvum and demonstrate the need for similar resources for other harmful algal-bloom-forming morphospecies.

Variability of morphological features and nuclear DNA content in haploids and doubled haploids of androgenic callus lines of rice (&lt;I&gt;Oryza sativa&lt;/I&gt; L.)

The work is relevant for understanding evolutionary processes in plant species. Twelve callus lines with multiple regeneration of haploids and doubled haploids were obtained in F1 hybrids of Oryza sativa L. through in vitro androgenesis. Intracallus variability of the morphological features of haploids was often accompanied by a decrease in the values of morphological features with an increase in the serial number (p &lt; 0.05). The number of panicles on a plant and the number of flowers on a panicle on two callus lines in the second or third group were increased. No variability was detected in five callus lines, i.e., such a phenomenon was not a rule. The nuclear DNA content of doubled haploids in four groups of the same callus line was 1.03– 1.09 pg, and for haploids it was 0.53–0.58 pg. Intracallus variability of nuclear DNA content was detected between groups of haploids of the same line and among doubled haploids of the same line. Significant differences were found between the haploids of one callus line and the three other callus lines of the Sadko × Kuboyar hybrid towards an increase of nuclear DNA content (p &lt; 0.0015). The theoretical possibility of the appearance of intraspecific variability among plants with a small number of chromosomes is considered. A scheme of genomic reorganization is proposed for such species: initial plant (2n) → aneuhaploid plants (n + 1) → megasporogenesis and microsporogenesis of the 0-n type, formation of fertile pollen (n + 1) → diploid plant (2n + 2). Aneuhaploid evolution explains the intraspecific variability of chromosome numbers among plant species with low ploidy. Aneuploid technologies can help in the artificial formation of new polyploid crops, and rice is given a primary role.

Discovery and Characterization of Novel Fertile Triploids and a New Chromosome Number in Caladium (Caladium × hortulanum)

Cultivated caladium (Caladium × hortulanum; 2n = 2x = 30) is an important ornamental aroid for the environmental horticulture industry. A better understanding of its genetic diversity is needed for continued improvement of caladium. The caladium germplasm maintained in the University of Florida’s caladium breeding program were surveyed for potential variation in chromosome number and nuclear DNA content to gain a better understanding of caladium genetic diversity at the cytogenetic level. For the first time caladium triploidy was discovered in two breeding lines, UF-15-414 and UF-4407 with 2n = 3x = 45 chromosomes and a nuclear DNA content of 13.86 pg·2C−1. In addition, a new chromosome number (2n = 2x = 34) was found in one cultivar, ‘White Wing’. Compared with their diploid parents or grandparents, the triploids showed a distinct, attractive leaf morphology with rounder and thicker leaves. The pollen stainability of UF-15-414 and UF-4407 was 63% and 73%, respectively, indicating potential male fertility, which was unexpected. Analysis of simple sequence repeat (SSR) marker banding patterns confirmed that UF-15-414 was a direct progeny of UF-4407 through hybridization with the diploid cultivar Aaron, whereas UF-4407 might result from fusion of an unreduced (female or male) gamete with a reduced gamete. Chromosome counting and SSR marker analysis of ‘White Wing’ and its progeny cultivars ‘White Wonder’ and ‘White Delight’ suggested that ‘White Wing’ possibly transmitted its 34 chromosomes to progeny during sexual reproduction. The discovery and characterization of these triploids revealed the occurrence of natural sexual polyploidization in caladium and indicated good potential for creating and selecting new triploids for future caladium breeding. The observed new chromosome number in ‘White Wing’ and its progeny cultivars implies that other chromosome variations may be present among cultivated caladiums. In summary, these results revealed two male-fertile triploid caladiums and a new chromosome number that can enrich the cytogenetic diversity in future caladium cultivar development.

Genome size and endoreplication in two pairs of cytogenetically contrasting species of Pulmonaria (Boraginaceae) in Central Europe.

Genome size is species-specific feature and commonly constant in an organism. In various plants, DNA content in cell nucleus is commonly increased in process of endoreplication, cellular-specific multiplication of DNA content without mitosis. This leads to the endopolyploidy, the presence of multiplied chromosome sets in a subset of cells. The relationship of endopolyploidy to species-specific genome size is rarely analysed and is not fully understood. While negative correlation between genome size and endopolyploidy level is supposed, this is species- and lineage-specific. In the present study, we shed light on this topic, exploring both genome size and endoreplication-induced DNA content variation in two pairs of morphologically similar species of Pulmonaria, P. obscura–P. officinalis and P. mollis–P. murinii. We aim (i) to characterize genome size and chromosome numbers in these species using cytogenetic, root-tip squashing and flow cytometry (FCM) techniques; (ii) to investigate the degree of endopolyploidy in various plant organs, including the root, stem, leaf, calyx and corolla using FCM; and (iii) to comprehensively characterize and compare the level of endopolyploidy and DNA content in various organs of all four species in relation to species systematic relationships and genome size variation. We have confirmed the diploid–dysploid nature of chromosome complements, and divergent genome sizes for Pulmonaria species: P. murinii with 2n = 2x = 14, 2.31 pg/2C, P. obscura 2n = 2x = 14, 2.69 pg/2C, P. officinalis 2n = 2x = 16, 2.96 pg/2C and P. mollis 2n = 2x = 18, 3.18 pg/2C. Endopolyploidy varies between species and organs, and we have documented 4C–8C in all four organs and up to 32C (64C) endopolyploid nuclei in stems at least in some species. Two species with lower genome sizes tend to have higher endopolyploidy levels than their closest relatives. Endoreplication-generated tissue-specific mean DNA content is increased and more balanced among species in all four organs compared to genome size. Our results argue for the narrow relationship between genome size and endopolyploidy in the present plant group within the genus Pulmonaria, and endopolyploidization seems to play a compensatory developmental role in organs of related morphologically similar species.

Genetic and nuclear DNA content variation of stevia (Stevia rebaudiana Bertoni) accessions grown in Turkey

The objectives of this study were to make morphological and molecular characterization of Turkish stevia accessions. The field data showed significant variations (P &lt; 0.05) for all morphological parameters (plant height, number of main stems per plant, number of secondary stems, fresh and herb yields) exception with number of secondary stems. The primers of MtP5CS, AtNHX1, MtProDH, Mt-Actin MtSOS1and MtSOS2 genes were used as loci specific DNA markers. All markers were amplified in Stevia genome and provided a mean of 74.16% polymorphism rate. Flow cytometer analysis showed no statistically significant differences (P &lt; 0.16) for 2C DNA content with a mean of 1.62 pg. The UPGMA analysis revealed that Samsun accession was distinctly separated from the others and had the lowest 2C DNA content (1.52 pg). This study first time showed that the stevia accessions grown in Turkey have limited genetic and no nuclear DNA content variations. The results also revealed that gene targeted functional markers associated with salt and drought tolerance have a great potential to be used as DNA markers to determine the genetic variation in plants

Nuclear DNA Content Variation in Different Life Cycle Stages of Sugar Kelp, Saccharina latissima

Ploidy variants can be utilized to increase yield, introduce sterility, and modify specific traits with an economic impact. Despite economic importance of Saccharina species, their nuclear DNA content in different cell types and life stages remain unclear. The present research was initiated to determine the nuclear DNA content and intraindividual variation at different life cycle stages of the Laminarialean kelp Saccharina latissima. Nuclear DNA content in embryonic and mature sporophytes, released and unreleased zoospores, female, and male gametophytes from Sør-Trøndelag county in Norway were estimated by image analysis using the DNA-localizing fluorochrome DAPI and chicken’s red blood cells as a standard. DNA content of a total of 6905 DAPI-stained nuclei was estimated. This is the first study of nuclear DNA content which covered the life cycle of kelp. The lowest level of DNA content (1C) was observed in zoospores with an average of 0.76 pg. Male and female single spore gametophyte cultures presented higher average DNA content, more than double that of zoospores, suggesting the presence of polyteny. Female gametophyte nuclei were slightly larger and more variable in size than those of male gametophytes. The DNA content observed in embryonic sporophytes and in meristoderm cells from older sporophytes (1.51 pg) was 2C as expected and in the range of previously published studies of sporophytes of S. latissima. Mature sporophytes showed intra-plant variation with DNA content values ranging from 2-16C. The main difference was between meristoderm cells (mostly 2C) and cortical and medullary cells (2-16C).

The variability of nuclear DNA content of different Pelargonium species estimated by flow cytometry.

Pelargonium is a versatile genus mainly from the Cape Region, South Africa. The genus is divided into four subgenera and 16 sections characterized by several groups of chromosomes sizes and numbers. The DNA content of species from all subgenera and sections of Pelargonium, except for the sections Subsucculentia and Campylia was estimated using flow cytometry. Nuclei of Pelargonium samples (leaf or petal tissue) and an internal plant standard (leaf tissue) were isolated together and stained with propidium iodide. The DNA content was estimated providing that the 2C peaks of sample and standard be in linearity in the flow cytometer histograms. In total, 96 Pelargonium accessions of 60 species (22 Pelargonium species for the first time) were analyzed. The 2C DNA content ranged from 0.84 pg (P. longifolium, section Hoarea) to 6.69 pg (P. schizopetalum, section Magnistipulacea) and the corresponding 1Cx DNA content from 0.42 pg (P. longifolium) to 1.72 pg (P. transvaalense. This demonstrates the high plasticity within the genus Pelargonium. Some species, such as P. peltatum accessions revealed a pronounced endopolyploidization in leaves but not in petals underlining the importance to choose the right tissue as sample for the flow cytometry analysis. The reported genome sizes are a step forward towards the characterization of the Pelargonium collection within the German Gene Bank for Ornamental Plants and a valuable base for future sequencing programs of the Pelargonium genomes.

Karyological and nuclear DNA content variation of the genus Asparagus.

Asparagus wild relatives could be a promising possibility to extent the genetic variability of garden asparagus and for new cultivars with favorable traits such as high yield stability, disease resistance and stress tolerance. In order to achieve an efficient use in breeding, a detailed cytogenetic characterization of the accessions is necessary. This study worked on 35 Asparagus accessions, including A. officinalis cultivars (‘Darlise’, ‘Ravel’ and ‘Steiners Violetta’) and Asparagus wild relatives, for which the number of chromosomes, their size, the nuclear DNA content, and the genomic distribution of 5S and 45S rDNA were analyzed. Different ploidy levels (diploid, triploid, tetraploid, pentaploid and hexaploid) were found. Furthermore, the size of the chromosomes of all diploid Asparagus accessions was determined which led to differences in the karyotypic formula. A. plocamoides harbors the smallest chromosome with 1.21 μm, whereas the largest chromosome with 5.43 μm was found in A. officinalis. In all accessions one 5S rDNA locus per genome was observed, while the number of 45S rDNA loci varied between one (A. albus, A. plumosus, A. stipularis) to four (A. setaceus). In most Asparagus accessions, the 5S and 45S rDNA signals were located on different chromosomes. In contrast, the genomes of A. africanus, A. plocamoides, A. sp. (a taxonomically unclassified Asparagus species from Asia) and A. verticillatus (diploid accessions) have one 5S and one 45S rDNA signal on the same chromosome. The measured 2C DNA content ranges from 1.43 pg (A. plocamoides, diploid) to 8.24 pg (A. amarus, hexaploid). Intraspecific variations for chromosome number, karyotypic formula, signal pattern with 5S and 45s rDNA probes and DNA content were observed. Interspecific variations were also recognized in the genus Asparagus.

Integration of Genotoxic Biomarkers in Environmental Biomonitoring Analysis Using a Multi-Biomarker Approach in Three-Spined Stickleback (Gasterosteus aculeatus Linnaeus, 1758).

Water is impacted by a variety of increasing pressures, such as contaminants, including genotoxic pollutants. The proposed multi-biomarker approach at a sub-individual level gives a complementary indicator to the chemical and ecological parameters of the Water Framework Directive (WFD, 2000/60/EC). By integrating biomarkers of genotoxicity and erythrocyte necrosis in the sentinel fish species the three-spined stickleback (Gasterosteus aculeatus) through active biomonitoring of six stations of the Artois-Picardie watershed, north France, our work aimed to improve the already existing biomarker approach. Even if fish in all stations had high levels of DNA strand breaks, the multivariate analysis (PCA), followed by hierarchical agglomerative clustering (HAC), improved discrimination among stations by detecting an increase of nuclear DNA content variation (Etaing, St Rémy du Nord, Artres and Biache-St-Vaast) and erythrocyte necrosis (Etaing, St Rémy du Nord). The present work highlighted that the integration of these biomarkers of genotoxicity in a multi-biomarker approach is appropriate to expand physiological parameters which allow the targeting of new potential effects of contaminants.

The telomere-mitochondrial axis of aging in newborns.

Aging starts at the beginning of life as evidenced by high variability in telomere length (TL) and mitochondrial DNA content (mtDNAc) at birth. Whether p53 and PGC-1α are connected to these age-related markers in early life is unclear. In this study, we hypothesized that these hallmarks of aging are associated at birth.In 613 newborns from the ENVIRONAGE birth cohort, p53 and PGC-1α protein levels were measured in cord plasma, while TL and mtDNAc were measured in both cord blood and placental tissue. Cord blood methylation data of genes corresponding to the measured protein levels were available from the Human MethylationEPIC 850K BeadChip array. Pearson correlations and linear regression models were applied while accounting for selected covariates. In cord, a 10% increase in TL was associated with 5.22% (95% CI: 3.26 to 7.22; p < 0.0001) higher mtDNAc and −2.66% (95% CI: –5.04 to −0.23%; p = 0.032) lower p53 plasma level. In placenta, a 10% increase in TL was associated with 5.46% (95% CI: 3.82 to 7.13%; p < 0.0001) higher mtDNAc and −2.42% (95% CI: −4.29 to −0.52; p = 0.0098) lower p53 plasma level. Methylation level of TP53 was correlated with TL and mtDNAc in cord blood and with cord plasma p53 level.Our study suggests that p53 may be an important factor both at the protein and methylation level for the telomere-mitochondrial axis of aging at birth.