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Flow cytometry is a versatile tool used for cell sorting, DNA content imaging, and determining various cellular characteristics. With the possibility of high-throughput analyses, it combines convenient labelling techniques to serve rapid, quantitative, and qualitative workflows. The ease of sample preparation and the broad range of applications render flow cytometry a preferred approach for many scientific questions. Yet, we lack practical adaptations to fully harness the quantitative and high-throughput capabilities of most cytometers for many organisms. Here, we present simple and advanced protocols for the analysis of total DNA content, de novo DNA synthesis, and protein association to chromatin in budding yeast and human cells. Upon optimization of experimental conditions and choice of fluorescent dyes, up to four parameters can be measured simultaneously and quantitatively for each cell of a population in a multi-well plate format. Reducing sample numbers, plastic waste, costs per well, and hands-on time without compromising signal quality or single-cell accuracy are the main advantages of the presented protocols. In proof-of-principle experiments, we show that DNA content increase in S-phase correlates with de novo DNA synthesis and can be predicted by the presence of the replicative helicase MCM2-7 on genomic DNA. [ABSTRACT FROM AUTHOR]

Modern 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. [ABSTRACT FROM AUTHOR]

The Festuca marginata group includes several taxa of glaucous, thick-leaved fescues with three sclerenchyma strands, first described by Hackel as Festuca ovina subsp. laevis, which are common in the Euro-Mediterranean area. Ten different taxa (species and subspecies) are currently recognized in this group. Nevertheless, taxonomical works typically have only a local (mostly national) scope, and the recognized taxa show only few or negligible morphological differences, making their separate taxonomic status doubtful. Here we provide morphological, karyological, and genetic analysis (ISSR, flow cytometry) of the Festuca marginata group based on populations in a larger geographical scale including Central and Southern-Eastern Europe. We found that most of the local taxa are difficult to definitively distinguish and we suggest the whole group be treated as a single species, Festuca marginata, with two subspecies reflecting major differences in morphology and genome size between the Greece subsp. heldreichii and the remaining European populations, referred to as subsp. marginata. [ABSTRACT FROM AUTHOR]

The genus Vigna (Leguminosae) comprises about 150 species grouped into five subgenera. The present study aimed to improve the understanding of karyotype diversity and evolution in Vigna, using new and previously published data through different cytogenetic and DNA content approaches. In the Vigna subgenera, we observed a random distribution of rDNA patterns. The 35S rDNA varied in position, from terminal to proximal, and in number, ranging from one (V. aconitifolia, V. subg. Ceratotropis) to seven pairs (V. unguiculata subsp. unguiculata, V. subg. Vigna). On the other hand, the number of 5S rDNA was conserved (one or two pairs), except for V. radiata (V. subg. Ceratotropis), which had three pairs. Genome size was relatively conserved within the genus, ranging from 1C = 0.43 to 0.70 pg in V. oblongifolia and V. unguiculata subsp. unguiculata, respectively, both belonging to V. subg. Vigna. However, we observed a positive correlation between DNA content and the number of 35S rDNA sites. In addition, data from chromosome-specific BAC-FISH suggest that the ancestral 35S rDNA locus is conserved on chromosome 6 within Vigna. Considering the rapid diversification in the number and position of rDNA sites, such conservation is surprising and suggests that additional sites may have spread out from this ancestral locus. [ABSTRACT FROM AUTHOR]

Copyright of Sultan Qaboos University Journal for Science is the property of Sultan Qaboos University and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Modern Food Science & Technology is the property of Editorial Office of Modern Food Science & Technology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Flow cytometry (FCM) is a widely used technique to study genome size (C-value), but recalcitrant metabolites in grapevines often hinder its efficiency in grapevine research. The aim of the present study was (i) to develop a novel buffer tailormade for the nuclei isolation of grapevines and (ii) to characterize a Cypriot germplasm collection based on C-values. A local cultivar "Xinisteri" was used as a pilot test to evaluate a Sorbitol-based buffer, while sprouting, young, and fully matured leaves were examined to evaluate the developmental parameter. The novel Sorbitol buffer was shown to have a coefficient of variation (CV) of 4.06%, indicating improved properties compared to other commonly used FCM buffers [WPB (7.69%), LB01 (6.69%), and LB (7.13%), respectively]. In addition, a significant variation in genome size between genotypes was found in a comprehensive application with 24 grape varieties. Nucleic content (2C) ranged from 0.577/1C pg for the "Assyrtiko" cultivar up to 0.597/1C pg for the "Spourtiko" cultivar, revealing a 17.6/1C Mbp difference. The lowest coefficient of variation (CV) across all entries was found in the variety "Ofthalmo" (2.29%), while the highest was observed in "Pinot Noir" (3.44%). Anova analysis revealed several distinct clusters, showing that in several cases, C-values can be used as a simple method to distinguish grapevine cultivars. [ABSTRACT FROM AUTHOR]

Due to the scarcity of literature data on the DNA content of different human tissues, this study aimed to isolate DNA from different tissues and fluids of the human body together with the determination of its content in the samples studied. Material was collected and tests were performed between 1990 and 2010, during autopsies performed for prosecutor's offices in the Department of Forensic Medicine. Goiter and thyroid cancer tissues were obtained from the Department of General Surgery, Gastroenterology and Endocrinology of Wroclaw Medical University. Isolated samples were measured spectrophotometrically, yielding an R 260/280 nm between 1.5 and 1.6. In some cases (when a sufficiently pure preparation could not be obtained), isolation was continued using the silica-based commercial QIAquick PCR Purification Kit (Qiagen). If the sampling tissues showed signs of decomposition such as bad odour or colour, the results were calibrated by Real-Time PCR, using the Quantifiler DNA assay (Thermo Fisher Scientific, Applied Biosystems). The results have shown that the maximum amount of genetic material was obtained from hair roots, adrenal glands, gonads and lymph nodes. The lowest DNA content per gram or milliliter of tissue or body fluid was found in adipose tissue, blood, saliva, bile, sweat, tears and the vitreous body of the eye. The presented findings indicate the best sources of high-quality DNA from the human body: gonads, kidneys, muscle (including heart), blood and bones (after decalcification). [ABSTRACT FROM AUTHOR]

Background and Aims Endoreduplication, the duplication of the nuclear genome without mitosis, is a common process in plants, especially in angiosperms and mosses. Accumulating evidence supports the relationship between endoreduplication and plastic responses to stress factors. Here, we investigated the level of endoreduplication in Ceratodon (Bryophyta), which includes the model organism Ceratodon purpureus. Methods We used flow cytometry to estimate the DNA content of 294 samples from 67 localities and found three well-defined cytotypes, two haploids and one diploid, the haploids corresponding to C. purpureus and Ceratodon amazonum , and the diploid to Ceratodon conicus , recombination occurring between the former two. Key Results The endoreduplication index (EI) was significantly different for each cytotype, being higher in the two haploids. In addition, the EI of the haploids was higher during the hot and dry periods typical of the Mediterranean summer than during spring, whereas the EI of the diploid cytotype did not differ between seasons. Conclusions Endopolyploidy may be essential in haploid mosses to buffer periods of drought and to respond rapidly to desiccation events. Our results also suggest that the EI is closely related to the basic ploidy level, but less so to the nuclear DNA content as previously suggested. [ABSTRACT FROM AUTHOR]

Flow cytometry has been widely used to estimate relative and absolute genome sizes (DNA contents) of plants for more than 50 years. However, the accuracy of these estimates can vary widely because of many factors, including errors in the genome size estimates of reference standards and various experimental methods. The objectives of this study were to reassess genome sizes of commonly used reference standards and quantify sources of variation and error in estimating plant genome sizes that arise from buffers, confounding plant tissues, tissue types, and plant reference standards using both 4′,6-diamidino-2-phenylindole (DAPI) and propidium iodide (PI) fluorochromes. Five separate studies were performed to elucidate these objectives. Revised estimates of genome sizes of commonly used plant reference standards were determined using human male leukocytes as a primary standard with an updated genome size (6.15 pg; 12.14% lower than that of earlier studies) using both DAPI and PI fluorochromes. Comparison of six different buffers (Galbraith’s, LB01, MB01, MgSO4, Otto’s, and Sysmex) resulted in variations in genome size estimates by as much as 18.1% for a given taxon, depending on the buffer–fluorochrome combination. The addition of different confounding plant tissues (representing 10 diverse taxa and associated secondary metabolites) resulted in variations in genome size estimates by as much as 10.3%, depending on the tissue–fluorochrome combination. Different plant tissue types (leaf color/exposure and roots) resulted in a variation in genome size estimates of 10.7%, independent of the fluorochrome. The selection of different internal reference standards introduced an additional variation in genome size estimates of 5.9%, depending on the standard–fluorochrome combination. The choice of fluorochrome (DAPI vs. PI) had one of the largest impacts on genome size estimates and differed by as much as 32.9% for Glycine max ‘Polanka’ when using human male leukocytes as an internal standard. A portion of this variation (∼10.0%) can be attributed to the base pair (bp) bias of DAPI and variations in Guanine-Cytosine (GC):Adenine-Thymine (AT) ratios between the sample and standard. However, as much as 22.9% of the variation in genome size estimates may result from how effectively these fluorochromes stain and report the genome. The combined variation/error from all these factors (excluding variation from bp bias for different fluorochromes, and assuming variations from confounding tissues and tissue types to both result from secondary metabolites) totaled 57.6%. Additional details of how selected factors impact accuracy, precision, and the interaction of these factors are presented. Overall, flow cytometry can be precise, repeatable, and extremely valuable for determining the relative genome size and ploidy of closely related plants when using consistent methods, regardless of fluorochrome. However, accurate determination of the absolute genome size by flow cytometry remains elusive, and estimates of genome size using flow cytometry should be considered gross approximations that may vary by ±29% or more as a function of experimental methods and plant environments. Additional recommendations of best practices are provided.

The Ferronikel smelter in Drenas is one of the main industrial areas in the Kosovo and pollution by heavy metals causes serious threat for all living organisms on this area. The objective of this study was to determine the concentration of some heavy metals (Fe, Cu, Mn, Cr, Cd, Ni and Pb) in agricultural soils and in maize plants, and their potential toxic effects on this plant through some sensitive biochemical and molecular markers. Maize seedlings growth in nine soil samples from different locations of this area. The highest concentrations of heavy metals in soils and maize leaves were conducted close to the Ferronikel smelter, and in some locations, the nickel and chromium concertation in soils exceeded 800 mg kg−1. A significant effects of heavy metals induced toxicity resulted in the, build-up aminolevulinic acid and reduced activity of δ-aminolevulinic acid dehydratase, and chlorophyll content in the maize leaves. In general, maize seedlings growth in polluted locations showed an increase in nuclear DNA content and in G2M phase. We concluded that locations close to the smelter are affected by soil heavy metals pollution and these biochemical and molecular analysis would be a powerful ecotoxicological tool in biomonitoring of heavy metal pollution. [ABSTRACT FROM AUTHOR]

Flow cytometry (FCM) is a widely used technique to study genome size (C-value), but recalcitrant metabolites in grapevines often hinder its efficiency in grapevine research. The aim of the present study was (i) to develop a novel buffer tailormade for the nuclei isolation of grapevines and (ii) to characterize a Cypriot germplasm collection based on C-values. A local cultivar “Xinisteri” was used as a pilot test to evaluate a Sorbitol-based buffer, while sprouting, young, and fully matured leaves were examined to evaluate the developmental parameter. The novel Sorbitol buffer was shown to have a coefficient of variation (CV) of 4.06%, indicating improved properties compared to other commonly used FCM buffers [WPB (7.69%), LB01 (6.69%), and LB (7.13%), respectively]. In addition, a significant variation in genome size between genotypes was found in a comprehensive application with 24 grape varieties. Nucleic content (2C) ranged from 0.577/1C pg for the “Assyrtiko” cultivar up to 0.597/1C pg for the “Spourtiko” cultivar, revealing a 17.6/1C Mbp difference. The lowest coefficient of variation (CV) across all entries was found in the variety “Ofthalmo” (2.29%), while the highest was observed in “Pinot Noir” (3.44%). Anova analysis revealed several distinct clusters, showing that in several cases, C-values can be used as a simple method to distinguish grapevine cultivars.

Abstract The genus Passiflora is the largest of the Passifloraceae family with many species and great phenotypic variability. There are nineteen species of Passiflora native to Argentina, distributed in four taxonomic subgenera: Passiflora, Decaloba, Dysosmia and Tacsonioides. Unlike most species of the genus, Argentinean species could tolerate colder climates. For most Argentinean species, genome size information is unavailable. The objective of this work has been to estimate the genomic size of 36 genotypes of thirteen Passiflora taxa by flow cytometry and to make a contrast with phenotypic ornamental characteristics. The genomic sizes of P. tucumanensis, P. elegans and P. mooreana are being introduced for the first time. The DNA amount per basic genome varied between 0.54 and 2.52 picograms in P. capsularis and P. alata, respectively. In the species P. caerulea, P. elegans and P. edulis f. flavicarpa, intraspecific variation in genomic size has been registered. The high correlation of genome size with flower diameter, and of leaf area with flower diameter can contribute to the selection of parents for the ornamental plant improvement plan. Also, the data collected in this work will be very useful in the study of inter-specific hybrids.

Acellular matrices are mainly composed of mammalian tissues, and aquatic tissues with lower biological risks and less religious restrictions are considered alternatives to mammalian tissues. The acellular fish skin matrix (AFSM) has been commercially available. Silver carp has the advantages of farmability, high yield and low price, but there are few studies on the silver carp acellular fish skin matrix (SC‐AFSM). In this study, an acellular matrix with low DNA and endotoxin was prepared from the skin of silver carp. After treatment with trypsin/sodium dodecyl sulfate and Triton X‐100 solutions, the DNA content in SC‐AFSM reached 11.03 ± 0.85 ng/mg, and the endotoxin removal rate was 96.8%. The porosity of SC‐AFSM was 79.64% ± 0.17%, which is favorable for cell infiltration and proliferation. The relative cell proliferation rate of SC‐AFSM extract was 117.79% ± 15.26%. The wound healing experiment showed that SC‐AFSM had no adverse acute pro‐inflammatory response, which had a similar effect as commercial products in promoting tissue repair. Therefore, SC‐AFSM has great application potential in biomaterials. [ABSTRACT FROM AUTHOR]

Copyright of Acta Agriculturae Slovenica is the property of Biotechnical Faculty of the University of Ljubljana and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Relationships between genome size and environmental variables suggest that DNA content might be adaptive and of evolutionary importance in plants. The genus Larrea provides an interesting system to test this hypothesis, since it shows both intra- and interspecific variation in genome size. Larrea has an amphitropical distribution in North and South American deserts, where it is most speciose. Larrea tridentata in North America shows a gradient of increasing autopolyploidy; while three of the four studied South American species are diploids, Larrea divaricata, Larrea nitida, Larrea ameghinoi, and the fourth is an allopolyploid, Larrea cuneifolia. We downloaded available focal species’ georeferenced records from seven data reservoirs. We used these records to extract biologically relevant environmental variables from WorldClim at 30 arc seconds scale, to have a broad characterization of the variable climatic conditions of both regions, and a climatic envelope for each species. We estimated relative DNA content index and relative monoploid genome values, by flow cytometry, of four most abundant Larrea species throughout their respective ranges. Then we winnow the bioclimatic dataset down to uncorrelated variables and sampled locales, to analyse the degree of association between both intra- and interspecific relative DNA content and climatic variables that are functionally relevant in arid environments using Pearson correlations, general linear and mixed effects models. Within the genus Larrea, relative DNA content increases with rising temperature and decreases with rising precipitation. At the intraspecific level, all four species show relative DNA content variation across climatic conditions. Larrea is a genus that shows genome size variation correlated with climate. Our results are also consistent with the hypothesis that extreme environmental pressures may have facilitated repeated whole genome duplication events in North America, while in South America, reticulate evolution, as allopolyploidization, and speciation might have been climate-dependent since the Oligocene.

Dorstenia spp. are characterized by a herbaceous habit, the presence of rhizomes and inflorescences being coenanthia. They occur in different forest formations, and it is believed that there are two distribution centres for Dorstenia , one Palaeotropical (Africa) and one Neotropical (Central and South America). The present study aimed to undertake a comparative analysis of the karyotype diversity of Dorstenia , based on chromosome data and DNA C values of 13 Neotropical species, and an analysis of low-coverage genomic sequencing of three African and one Neotropical species. The most frequent chromosome number is 2 n  = 32, with variations for 2 n  = 28 and 30, suggesting an important role of polyploidy in the karyotype differentiation of Dorstenia. Karyotypes varied in the number of metacentric, submetacentric and acrocentric chromosomes, with evidence of rearrangements. Genome sizes (C values) varied almost two-fold, from 2C = 2.50 to 5.47 pg. There was no clear relationship between DNA C value variations and karyotype features, such as chromosome numbers and distribution of CMA/DAPI bands or rDNA sites. Proximal heterochromatin was most frequent, but CMA+ and DAPI+ bands predominated in relation to DAPI+ bands. 5S and 35S ribosomal DNA (rDNA) sites were also observed in the proximal regions in almost all species. Based on a chromosome modelling approach, our data suggest that dysploidy events may have been responsible for the chromosome number variations, whereas rearrangements involving repetitive fractions may have been associated with changes in genome size. There was great variability in the amount of rDNA between species, including a remarkable 5S rDNA amplification in D. bahiensis , and a diversity of transposable elements in the four genomes with genomic sequence data. The combination of data from karyotypes, genomes and phylogeny suggests that dysploidy has played a predominant role in the evolution of Neotropical Dorstenia spp. [ABSTRACT FROM AUTHOR]

The aim of the study was to verify whether the use of heparin as an anticoagulant can affect the results of genome size measurements using a flow cytometer. The values obtained by measuring blood samples with different concentrations of heparin were compared. The differences observed were highly significant, suggesting that the results of previous comparative studies, or those determining individual polyploidy, may need reinterpretation. We anticipate that our findings will increase the accuracy of flow cytometric analysis and will be useful for further surveys of genome size undertaken using blood samples. [ABSTRACT FROM AUTHOR]

DNA content is an important trait linked to the evolutionary routes of taxa and often connected to speciation. In the present study, we studied C-values variation across the Cypriot Fabeae gene pool. Several hundred plants (Vicia spp., Lens spp., Pisum spp.) were sampled across Cyprus. Accurate estimates were established by flow cytometry and propidium iodine staining for 155 discrete populations/accessions. A ten-fold variation was detected across lineages with 1C DNA content varying from 1.584 pg for V. cretica (ARI02420) to 13.983 pg for V. faba (ARI00187). In general, flow cytometry was precise for the characterization of species, even though there were instances of genome overlapping across taxa. Most analyses in the current work refer to species that have not been characterized before by flow cytometry (or any other DNA content estimation method). Still, a correlation to C-values previously reported in Kew Plant DNA C-values database was attempted. A high degree of correlation except for V. dalmatica was established. The evaluation of genome size trait in relation with the Fabeae phylogeny, revealed that Pisum and Lens genera were rather homogenous, but an astonishing fluctuation was shown for Vicia spp. Moreover, it was established that genome up- or down-scaling was not directly linked to speciation drivers. The genomic size measurements presented here could deliver extra quality control for the identification and characterization of taxa in germplasm collections, particularly in cases where species share morphological characters. [ABSTRACT FROM AUTHOR]

Autopolyploidization has driven the successful invasion of Solidago canadensis in East Asia. However, it was believed that only diploid S. canadensis invaded Europe, whereas polyploids never did. Here, molecular identification, ploidy level, and morphological traits of ten S. canadensis populations collected in Europe were compared with previously identified S. canadensis populations from other continents and S. altissima populations. Furthermore, the ploidy-driven geographical differentiation pattern of S. canadensis in different continents was investigated. All ten European populations were identified as S. canadensis with five diploid and five hexaploid populations. Significant differences in morphological traits existed among diploids and polyploids (tetraploids and hexaploids), rather than between polyploids from different introduced ranges and between S. altissima and polyploidy S. canadensis. The invasive hexaploids and diploids had few differences in latitudinal distributions in Europe, which was similar to the native range but different from a distinct climate-niche differentiation in Asia. This may be attributed to the bigger difference in climate between Asia and Europe and North America. The morphological and molecular evidences proved the invasion of polyploid S. canadensis in Europe and suggest that S. altissima may be merged into a complex of S. canadensis species. Our study may be concluded that geographical and ecological niche differentiation of an invasive plant driven by ploidy depends on the degree of difference in the environmental factors between the introduced and native range, which provides new insight into the invasive mechanism. [ABSTRACT FROM AUTHOR]

Important changes in vegetation types occur along elevational gradients. The genus Gymnocalycium is endemic to southern South America, and its species are distributed along elevational gradients. In particular, Gymnocalycium quehlianum is a globular cactus endemic to the Sierras de Córdoba. Studying cytogenetic aspects and DNA content in populations throughout their distribution is key to understanding the species. DNA content and cytogenetic characteristics were analyzed in four populations of G. quehlianum (615, 744, 948 and 1257 masl). The genome size in the four populations varied between 3.55 and 4.30 pg. The populations were diploid (2n = 22). All populations showed the karyotype formula of 10 metacentrics (m) + 1 submetacentric (sm). The species presented symmetrical karyotypes and constitutive heterochromatin CMA+/DAPI- associated with nucleolar organizing regions, always found in the first pair of m chromosomes. The 18-5.8-26S rDNA locus is found in the terminal regions of the first pair of chromosomes m, and the 5S locus is adjacent to the 18-5.8-26S locus. A tendency for DNA content to decrease with increasing altitude was observed.

Ilex cornuta Lindl. & Paxton is a commercially important horticultural species worldwide, and extensive cultivation and hybridization have produced many varieties. Despite the considerable breeding, selection, widespread cultivation and domestication, which may have a significant role in the composition of genomes, there are no other previous reports of intraspecific genome size variation in the different cultivars or hybrids of this species. In the present work, genome size of 12 varieties of I. cornuta was assessed and analyzed through high-resolution flow cytometry (FCM). Nuclear DNA was analyzed using nuclei isolated from young leaves, which used propidium iodide (PI) staining, with rice (Oryza sativa cv. Nipponbare) as internal reference. As a result, statistically significant differences in genome size were detected among all diploid I. cornuta varieties considered. The estimated genome size (2C value) of I. cornuta varieties ranged from 1.47 to 1.80 pg, with 1.22-fold variation and an average size of 1.65 pg. The domestication and interspecific hybridization induced variation of genome size in I. cornuta, and the genome size of hybrids exhibited a wider range of variation compared with that of cultivars. In summary, flow cytometry is a useful tool to analyze the genome size of I. cornuta. The first report of the genome sizes of varieties of this species would provide useful data for further research on I. cornuta, and enrich the C value database of Ilex L. What’s more, our findings could be the foundation in the future of I. cornuta genome sequencing and breeding programs.

Premise: Whole‐genome duplication is considered a major mechanism of sympatric speciation due to the creation of strong and instantaneous reproductive barriers. Although postzygotic reproductive isolation between diploids and polyploids is often expected, the extent of reproductive incompatibility must be empirically determined and compared to patterns of genetic isolation to fully characterize the reproductive dynamics between cytotypes. Methods: We investigated reproductive compatibility between diploid and tetraploid Lycium australe in two mixed‐cytotype populations using (1) controlled crossing experiments to evaluate fruit and seed production and (2) germination trials to test seed viability following homoploid and heteroploid crosses. We contrast these experiments with a single‐nucleotide polymorphism (SNP) data set to measure genetic isolation between cytotypes and explore whether cytotype or population origin better explains patterns of genetic variation. Finally, we explore mating patterns using the observed germination rates of naturally produced seeds in each population. Results: Although homoploid and heteroploid crosses resulted in similar fruit and seed production, reproductive isolation between co‐occurring diploids and tetraploids was nearly complete, due to low seed viability following heteroploid crosses. Of 191,182 total SNPs, 21,679 were present in ≥90% of individuals and replicate runs using unlinked SNPs revealed strong clustering by cytotype and differentiation of tetraploids based on population origin. Conclusions: As often reported, diploid and tetraploid L. australe experience strong postzygotic isolation via hybrid seed inviability. Consistent with this result, cytotype explained a greater amount of variation in the SNP data set than population origin, despite some evidence of historical introgression. [ABSTRACT FROM AUTHOR]

The genus Passiflora is the largest of the Passifloraceae family with many species and great phenotypic variability. There are nineteen species of Passiflora native to Argentina, distributed in four taxonomic subgenera: Passiflora, Decaloba, Dysosmia and Tacsonioides. Unlike most species of the genus, Argentinean species could tolerate colder climates. For most Argentinean species, genome size information is unavailable. The objective of this work has been to estimate the genomic size of 36 genotypes of thirteen Passiflora taxa by flow cytometry and to make a contrast with phenotypic ornamental characteristics. The genomic sizes of P. tucumanensis, P. elegans and P. mooreana are being introduced for the first time. The DNA amount per basic genome varied between 0.54 and 2.52 picograms in P. capsularis and P. alata, respectively. In the species P. caerulea, P. elegans and P. edulis f. flavicarpa, intraspecific variation in genomic size has been registered. The high correlation of genome size with flower diameter, and of leaf area with flower diameter can contribute to the selection of parents for the ornamental plant improvement plan. Also, the data collected in this work will be very useful in the study of inter-specific hybrids. [ABSTRACT FROM AUTHOR]

Poa pratensis L. is a perennial turfgrass with high regeneration and fertility, resistance to cold and drought, and quick colonization. By facultative apomixis, this plant can create a wide range of ploidy levels (2n = 22 to 2n = 154), resulting in a wide range of chromosomal numbers and sexual and apomictic reproductive diversity. The plant materials included fifty accessions from Iran's Center, South, North, North-East, North-West, and West ecoregions. UPOV standards were used to measure the qualities that were researched. The squash technique of chromosome counting revealed that Iranian Kentucky bluegrass accessions had chromosomal counts ranging from 24 to 87. The relative sizes of the 2C genomes were measured using laser flow cytometry. The range of DNA content was fairly wide, ranging from 4.92 to 11.52 pg. DNA content has a strong positive correlation with elevation, a moderately positive correlation with flag leaf length and leaf sheath width, and a negative correlation with inflorescence anthocyanin color and leaf anthocyanin color. The genotypes and ecological zones of this plant in Iran were distinguished based on morphological diversity and DNA content. The results from this study could be useful in identifying and studying wild Kentucky bluegrass genotypes. It aids in predicting the location of rare genotypes used as breeding materials. It can also increase the plant's variability for future generations by introducing new ecotypes, with particular genomic and morphological traits, to previously cultivated populations. We expect that the findings of this study will aid in understanding the evolution of this plant in the context of Iran's climatic variety. [ABSTRACT FROM AUTHOR]

In this study, an efficient, reproducible, and genetically stable regeneration protocol has been developed in Artemisia maritima L. The experiments were conducted for callus induction, plant regeneration, and somatic embryogenesis using stem and leaf of A. maritima as explants. The optimal callus induction (81.3%) was observed on 2.5 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.5 mg L−1 6-benzylaminopurine (BAP). The shoot regeneration was observed on different concentrations of BAP, α-naphthaleneacetic acid (NAA), and thidiazuron (TDZ), using nodal segments and microshoot tips as explants. The microshoot tips were more responsive compared to nodal segments with the highest induction frequency (90.33%) obtained on 1.5 mg L−1 BAP. Maximum root induction frequency (74.36%) was obtained on 1.5 mg L−1 NAA. The somatic embryogenesis was induced on Murashige and Skoog (MS) medium amended with TDZ and indole-3-butyric acid (IBA) with maximum embryogenic induction frequency on 1.0 mg L−1 TDZ and 2.5 mg L−1 IBA. The somatic embryos developed into globular, heart-shaped, and bipolar plantlet stages on BAP and NAA as revealed through scanning electronic microscopy (SEM) and histological studies. The fully developed plants were acclimatized (75% survival rate) and transferred to natural photoperiod conditions. The DNA content and genetic stability of direct regenerated and somatic embryo–derived plants were analyzed by flow cytometry. The 2C DNA content of in vivo plants, direct regenerated, and somatic embryo–derived plants was 14.89, 14.61, and 14.37 pg, respectively. The genetic stability was maintained in in vitro cultures in comparison to field-grown plants of A. maritima. This study for the first time tried to formulate regeneration protocol via direct and indirect organogenesis and somatic embryogenesis for A. maritima. This paper was also the first report for comparing the 2C DNA content of A. maritima grown in vivo to in vitro cultured plants. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the organogenic potential of Brazilian sugarcane varieties, in addition to verifying the in vitro multiplication rate and genetic stability by flow cytometry over monthly and consecutive subcultures. For that, stem apexes of twenty-two varieties were collected in field conditions and taken to the laboratory where external layers of leaves were removed. After surface sterilization, the innermost portion of the stem segment was sectioned and placed in a MS medium supplemented with 5.0 mg L−1 of NAA and 0.5 mg L−1 of KIN. For the multiplication rate, ten varieties were selected and inoculated in MS medium of liquid or semi-solid consistency plus 0.10 mg L−1 of KIN and 0.20 mg L−1 of BAP. Subcultures were performed every 30 days for a period up to 8 months. Genetic stability was verified by flow cytometry every two subcultures. At the end of the experiment, the sprouts were rooted and acclimatized in a greenhouse. As a result, it was observed that the regeneration occurred both by direct and indirect organogenic pathway. The varieties of sugarcane differed significantly regarding the regeneration capacity and amount of adventitious shoots formed. In multiplication, a significant interaction was observed between variety, consistency of the culture medium and number of subcultures. In general, in the first subcultures, the liquid consistency medium presented similar or superior results when compared with the semi-solid medium, however, from the fourth subculture, the semi-solid medium was superior. Morphological variations were verified from the fourth subculture. In addition, in some varieties, small changes in the relative amount of DNA were detected by flow cytometry. Sprouts of normal-looking sugarcane were successfully rooted and plantlets acclimatized after the eighth subculture. [ABSTRACT FROM AUTHOR]

Ploidy analysis is the fundamental method of measuring DNA content. For decades, the principal way of conducting ploidy analysis was through flow cytometry. A flow cytometer is a specialized tool for analyzing cells in a solution. This is convenient in laboratory environments, but prohibits measurement reproducibility and the complete detachment of sample preparation from data acquisition and analysis, which seems to have become paramount with the constant decrease in the number of pathologists per capita all over the globe. As more open computer-aided systems emerge in medicine, the demand for overcoming these shortcomings, and opening access to even more (and more flexible) options, has also emerged. Image-based analysis systems can provide an alternative to these types of workloads, placing the abovementioned problems in a different light. Flow cytometry data can be used as a reference for calibrating an image-based system. This article aims to show an approach to constructing an image-based solution for ploidy analysis, take measurements for a basic comparison of the data produced by the two methods, and produce a workflow with the ultimate goal of calibrating the image-based system. [ABSTRACT FROM AUTHOR]

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·2C21. 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. [ABSTRACT FROM AUTHOR]

Flow cytometry (FCM) is currently the most widely‐used method to establish nuclear DNA content in plants. Since simple, 1‐3‐parameter, flow cytometers, which are sufficient for most plant applications, are commercially available at a reasonable price, the number of laboratories equipped with these instruments, and consequently new FCM users, has greatly increased over the last decade. This paper meets an urgent need for comprehensive recommendations for best practices in FCM for different plant science applications. We discuss advantages and limitations of establishing plant ploidy, genome size, DNA base composition, cell cycle activity, and level of endoreduplication. Applications of such measurements in plant systematics, ecology, molecular biology research, reproduction biology, tissue cultures, plant breeding, and seed sciences are described. Advice is included on how to obtain accurate and reliable results, as well as how to manage troubleshooting that may occur during sample preparation, cytometric measurements, and data handling. Each section is followed by best practice recommendations; tips as to what specific information should be provided in FCM papers are also provided. [ABSTRACT FROM AUTHOR]

Abstract Backgrounds Populus and Salix belong to Salicaceae and are used as models to investigate woody plant physiology. The variation of karyotype and nuclear DNA content can partly reflect the evolutionary history of the whole genome, and can provide critical information for understanding, predicting, and potentially ameliorating the woody plant traits. Therefore, it is essential to study the chromosome number (CN) and genome size in detail to provide information for revealing the evolutionary process of Salicaceae. Results In this study, we report the somatic CNs of seventeen species from eight genera in Salicaceae. Of these, CNs for twelve species and for five genera are reported for the first time. Among the three subfamilies of Salicaceae, the available data indicate CN in Samydoideae is n = 21, 22, 42. The only two genera, Dianyuea and Scyphostegia, in Scyphostegioideae respectively have n = 9 and 18. In Salicoideae, Populus, Salix and five genera closely related to them (Bennettiodendron, Idesia, Carrierea, Poliothyrsis, Itoa) are based on relatively high CNs from n = 19, 20, 21, 22 to n = 95 in Salix. However, the other genera of Salicoideae are mainly based on relatively low CNs of n = 9, 10, 11. The genome sizes of 35 taxa belonging to 14 genera of Salicaceae were estimated. Of these, the genome sizes of 12 genera and all taxa except Populus euphratica are first reported. Except for Dianyuea, Idesia and Bennettiodendron, all examined species have relatively small genome sizes of less than 1 pg, although polyploidization exists. Conclusions The variation of CN and genome size across Salicaceae indicates frequent ploidy changes and a widespread sharing of the salicoid whole genome duplication (WGD) by the relatives of Populus and Salix. The shrinkage of genome size after WGD indicates massive loss of genomic components. The phylogenetic asymmetry in clade of Populus, Salix, and their close relatives suggests that there is a lag-time for the subsequent radiations after the salicoid WGD event. Our results provide useful data for studying the evolutionary events of Salicaceae.

Abstract Polyploidy, a numerical alteration of the karyotype, is one of the most important mechanisms in plant speciation and diversification, but could also be detected among populations, the cytotypes. For example, Psidium cattleyanum, a polyploid complex, has chromosome numbers ranging from 2n=3x=33 to 2n=12x=132. Polyploidization causes an increase in DNA content, and both modifications may cause alteration in plant growth, physiology, and epigenetics. Based on this possibility, here we aim to verify the influence of the polyploidization on the production of P. cattleyanum essential oil chemotypes. Differences in the DNA contents, as a proxy to different ploidies, were observed and three distinct chemotypes were identified through the chromatographic profile analysis. The Psidium cattleyanum DNA content and qualitative and quantitative characteristics of the essential oils presented a positive relationship. Plants with higher DNA contents presented higher levels of oil production, which was mostly composed of hydrogenated sesquiterpenes, while plants with lower DNA contents produced lower amount of oil, which was mostly composed of hydrogenated monoterpenes. Based on the importance of essential oils, polyploid plants, which present higher DNA content, are recommended as possible matrices for the propagation of new plants with the potential to produce major compounds of agronomic and pharmacological interest.

Main conclusion: Coffea karyotype organization and evolution has been uncovered by classical cytogenetics and cytogenomics. We revisit these discoveries and present new karyotype data. Coffea possesses ~ 124 species, including C. arabica and C. canephora responsible for commercial coffee production. We reviewed the Coffea cytogenetics, from the first chromosome counting, encompassing the karyotype characterization, chromosome DNA content, and mapping of chromosome portions and DNA sequences, until the integration with genomics. We also showed new data about Coffea karyotype. The 2n chromosome number evidenced the diploidy of almost all Coffea, and the C. arabica tetraploidy, as well as the polyploidy of other hybrids. Since then, other genomic similarities and divergences among the Coffea have been shown by karyotype morphology, nuclear and chromosomal C-value, AT and GC rich chromosome portions, and repetitive sequence and gene mapping. These cytogenomic data allowed us to know and understand the phylogenetic relations in Coffea, as well as their ploidy level and genomic origin, highlighting the relatively recent allopolyploidy. In addition to the euploidy, the role of the mobile elements in Coffea diversification is increasingly more evident, and the comparative analysis of their structure and distribution on the genome of different species is in the spotlight for future research. An integrative look at all these data is fundamental for a deeper understanding of Coffea karyotype evolution, including the key role of polyploidy in C. arabica origin. The ‘Híbrido de Timor’, a recent natural allotriploid, is also in the spotlight for its potential as a source of resistance genes and model for plant polyploidy research. Considering this, we also present some unprecedented results about the exciting evolutionary history of these polyploid Coffea. [ABSTRACT FROM AUTHOR]

Gulf vervain (Verbena xutha) is a herbaceous perennial native to the southern United States. The species produces attractive spike inflorescences that provide a source of food for many pollinators. Besides botanical classifications, pollen morphology and cytological traits of this species have not been documented until now. Gulf vervain was found to have 2n = 42 chromosomes, with the first report of 2C nuclear DNA content of 2.95 pg. Convex-triangular pollen grains averaged 37.18 μm in diameter, with 84% stainability after cotton blue staining to estimate pollen viability. This information brings light to the genetic makeup of gulf vervain and may aid in future breeding programs.

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.

In biotechnological processes such as chromosomal manipulation studies, semen has become a reference in the ploidy verification of the evaluated material. However, the use of fresh samples is limited to the use at field conditions because the analysis is performed under laboratory conditions. Thus, this study aimed to develop a simpler procedure for storing dry semen at 28 °C to reduce cold storage costs. For this, semen samples were evaluated according to established quality semen parameters, a protocol for dry, and 3 sterilization treatments of dry semen were applied to the store. The integrity of the DNA was evaluated every two months, using fresh semen, dry semen (untreated), and particles 3C to compare the peaks by flow cytometry. The results indicated that all samples evaluated before and after drying showed no significant difference in the DNA content. UV-treated semen showed a 1C peak in the histogram up to 180 days of storage and a non-significant difference (P > 0.05) from fresh control in the number of DNA particles up to 120 days and untreated only showed a 1C peak up to 120 days. The developed method may become an interesting procedure to serve as a reference peak for practical flow cytometric analysis, not only in the field of fish biology but also in biomedical and agricultural sciences. Furthermore, dried semen can become a tool for the preservation of genetic material and is a promising low-cost storage technique for biobanking. [ABSTRACT FROM AUTHOR]

Some species of Gentianaceae are an abundant source of valuable, much demanded pharmacologically active secondary metabolites. Embryogenic cell suspensions may provide both raw material for secondary metabolite extraction and somatic embryos (SE), protoplasts, and cells for plant biotechnology. Their cultures can be maintained for a long time with cryopreservation. Encapsulation-dehydration has been reported as a reliable technique for preserving the viability and recovery of embryogenic cells. Nonetheless, the elevated sucrose concentrations in the culture medium during preculture and dehydration may alter the embryogenic competence of cells, their genetic stability and metabolite biosynthesis. Therefore, the objectives of this study were to evaluate the effect of sucrose concentrations in the medium and cryopreservation on the growth kinetics, embryogenic potential and secondary metabolite production of cotyledon-derived cell suspensions of Gentiana capitata and Gentiana decumbens. The growth kinetics, the number of SE regenerated, and the 2 C DNA content were assessed for cell suspensions cultured on liquid Murashige and Skoog medium with 3% and 6% sucrose, either encapsulated in calcium alginate or non-encapsulated, either cryopreserved via encapsulation-dehydration or non-cryopreserved. The secondary metabolites were analyzed using UHPLC-DAD-IT-MS/MS. The biomass growth rate was always higher on medium with 3% sucrose than on medium with 6% sucrose irrespective of the gentian species, with the maximum growth rate of 146 ±7% for G. capitata and 116 ±22% for G. decumbens at 4 weeks. The growth rate of suspension culture from cryopreserved plant material was always lower and after 4 weeks being 28±5% and 33±3% for G. capitata and G. decumbens , respectively. Prior to encapsulation the SE productivity of G. capitata cell suspension was on average 7.5 times greater than that of G. decumbens for cultures from medium with 3% of sucrose and on average 4.6 times greater for cultures from medium with 6% sucrose. In both species cell suspensions cultured on medium with 6% of sucrose produced more somatic embryos (3.7±0.6 and 0.8±0.2 per 50 mg of cell suspension of G. capitata and G. decumbens , respectively) than those from MS3 medium (3.0±0.7 and 0.4±0.2 per 50 mg cell suspension of G. capitata and G. decumbens , respectively). Variation in the 2C DNA content was detected in G. decumbens tissues treated with higher sucrose concentrations. The secondary metabolite composition in cell suspensions differed from that in plants grown in vitro. Cell aggregate extracts did not include secoiridoids typical of Gentianaceae, but some benzophenone and isovitexin derivatives (flavonoids) previously undetected in these species. [Display omitted] • Cryopreservation maintains plant genetic stability, minimizes somaclonal variations. • Sucrose concentration in medium and cryopreservation affect growth kinetics and embryogenic potential. • The secondary metabolite composition in cell suspensions differed from that in plants grown in vitro. • The cells from suspensions culture contain benzophenone and flavonoids previously undetected in these species. [ABSTRACT FROM AUTHOR]

Autopolyploidization has driven the successful invasion of Solidago canadensis in East Asia. However, it was believed that only diploid S. canadensis invaded Europe, whereas polyploids never did. Here, molecular identification, ploidy level, and morphological traits of ten S. canadensis populations collected in Europe were compared with previously identified S. canadensis populations from other continents and S. altissima populations. Furthermore, the ploidy-driven geographical differentiation pattern of S. canadensis in different continents was investigated. All ten European populations were identified as S. canadensis with five diploid and five hexaploid populations. Significant differences in morphological traits existed among diploids and polyploids (tetraploids and hexaploids), rather than between polyploids from different introduced ranges and between S. altissima and polyploidy S. canadensis. The invasive hexaploids and diploids had few differences in latitudinal distributions in Europe, which was similar to the native range but different from a distinct climate-niche differentiation in Asia. This may be attributed to the bigger difference in climate between Asia and Europe and North America. The morphological and molecular evidences proved the invasion of polyploid S. canadensis in Europe and suggest that S. altissima may be merged into a complex of S. canadensis species. Our study may be concluded that geographical and ecological niche differentiation of an invasive plant driven by ploidy depends on the degree of difference in the environmental factors between the introduced and native range, which provides new insight into the invasive mechanism.

DNA content is an important trait linked to the evolutionary routes of taxa and often connected to speciation. In the present study, we studied C-values variation across the Cypriot Fabeae gene pool. Several hundred plants (Vicia spp., Lens spp., Pisum spp.) were sampled across Cyprus. Accurate estimates were established by flow cytometry and propidium iodine staining for 155 discrete populations/accessions. A ten-fold variation was detected across lineages with 1C DNA content varying from 1.584 pg for V. cretica (ARI02420) to 13.983 pg for V. faba (ARI00187). In general, flow cytometry was precise for the characterization of species, even though there were instances of genome overlapping across taxa. Most analyses in the current work refer to species that have not been characterized before by flow cytometry (or any other DNA content estimation method). Still, a correlation to C-values previously reported in Kew Plant DNA C-values database was attempted. A high degree of correlation except for V. dalmatica was established. The evaluation of genome size trait in relation with the Fabeae phylogeny, revealed that Pisum and Lens genera were rather homogenous, but an astonishing fluctuation was shown for Vicia spp. Moreover, it was established that genome up- or down-scaling was not directly linked to speciation drivers. The genomic size measurements presented here could deliver extra quality control for the identification and characterization of taxa in germplasm collections, particularly in cases where species share morphological characters.

Porterweed (Stachytarpheta spp.), a member of the verbena family, is frequently used in pollinator gardens to attract butterflies. This study was conducted to assess the morphological features, pollen stainability and morphology, nuclear DNA content, and chromosome number of five porterweed selections. Coral porterweed (S. mutabilis), ‘Naples Lilac’ porterweed (S. cayennensis × S. mutabilis ‘Violacea’), and nettleleaf porterweed (S. cayennensis) had the largest plant heights. Flower number was significantly higher in nettleleaf porterweed, jamaican porterweed (S. jamaicensis), and U*J3-2 porterweed (S. cayennensis × S. jamaicensis), with an average of 65–72 flowers per inflorescence. Internode length and flower width of jamaican porterweed had much lower values than the other selections. Coral porterweed recorded the lowest pollen stainability with only 10.6% stainability, but it had the largest relative pollen production. ‘Naples Lilac’ porterweed had the highest DNA content with an average of 3.79 pg/2C, like jamaican porterweed with 3.73 pg/2C. Ploidy levels varied between selections, and the basic chromosome number was x = 28. Coral, jamaican, and ‘Naples Lilac’ porterweed had 2n = 6x = 168 chromosomes, first reported in this genus. These results provide a guide and a new tool to distinguish native and non-native porterweed and may aid future breeding toward the production of noninvasive cultivars.

Abstract Background Up to now, diploid and triploid cultivars were reported for the ornamental crop Hydrangea macrophylla. Especially, the origin of triploids and their crossing behaviors are unknown, but the underlying mechanisms are highly relevant for breeding polyploids. Results By screening a cultivar collection, we identified diploid, triploid, tetraploid and even aneuploid H. macrophylla varieties. The pollen viability of triploids and tetraploids was comparable to that of diploids. Systematic crosses with these cultivars resulted in viable diploid, triploid, tetraploid and aneuploid offspring. Interestingly, crosses between diploids produced diploid and 0 or 1–94% triploid offspring, depending on the cultivars used as pollen parent. This finding suggests that specific diploids form unreduced pollen, either at low or high frequencies. In contrast, crosses of triploids with diploids or tetraploids produced many viable aneuploids, whose 2C DNA contents ranged between the parental 2C values. As expected, crosses between diploid and tetraploid individuals generated triploid offspring. Putative tetraploid plants were obtained at low frequencies in crosses between diploids and in interploid crosses of triploids with either diploid or tetraploid plants. The analysis of offspring populations indicated the production of 1n = 2x gametes for tetraploid plants, whereas triploids produced obviously reduced, aneuploid gametes with chromosome numbers ranging between haploid and diploid level. While euploid offspring grew normally, aneuploid plants showed mostly an abnormal development and a huge phenotypic variation within offspring populations, most likely due to the variation in chromosome numbers. Subsequent crosses with putative diploid, triploid and aneuploid offspring plants from interploid crosses resulted in viable offspring and germination rates ranging from 21 to 100%. Conclusions The existence of diploids that form unreduced pollen and of tetraploids allows the targeted breeding of polyploid H. macrophylla. Different ploidy levels can be addressed by combining the appropriate crossing partners. In contrast to artificial polyploidization, cross-based polyploidization is easy, cheap and results in genetically variable offspring that allows the direct selection of more robust and stress tolerant polyploid varieties. Furthermore, the generation of polyploid H. macrophylla plants will favor interspecific breeding programs within the genus Hydrangea.

Maize flowering time is an important agronomic trait, which has been associated with variations in the genome size and heterochromatic knobs content. We integrated three steps to show this association. Firstly, we selected inbred lines varying for heterochromatic knob composition at specific sites in the homozygous state. Then, we produced homozygous and heterozygous hybrids for knobs. Second, we measured the genome size and flowering time for all materials. Knob composition did not affect the genome size and flowering time. Finally, we developed an association study and identified a knob marker on chromosome 9 showing the strongest association with flowering time. Indeed, modelling allele substitution and dominance effects could offer only one heterochromatic knob locus that could affect flowering time, making it earlier rather than the knob composition. [ABSTRACT FROM AUTHOR]

Research Question: Is Raman spectroscopy an efficient and accurate method to detect sperm chromosome balance state by DNA content differences? Design: Semen samples were provided by diploid healthy men, and the analysis parameters met the current World Health Organization standards. The DNA content was assessed by analysis of the corresponding spectra obtained from a laser confocal Raman spectroscope. The sperm sex chromosome information was obtained by fluorescence in situ hybridization (FISH). Comparative analysis was performed between FISH results and Raman spectral analysis results. Results: Different parts of the sperm head showed different spectral signal intensities, which indicated that there were different chemical components. Standard principal component analysis (PCA) can preliminarily classify sperm with different DNA contents into two groups. Further analysis showed that there were significant differences in the 785 DNA backbone peaks and 714–1,162 cm−1 DNA skeleton regions among sperm with different DNA contents. The peak and regional peak of the DNA skeleton of X sperm were significantly higher than those of Y sperm (X vs. Y, p < 0.05). The above sperm types were confirmed by FISH. ROC curve analysis shows that there is a correlation between the Raman spectrum data and FISH results. Conclusion: Raman spectroscopy can identify X and Y sperms by analyzing the DNA content difference. However, the accuracy of the detection still needs to be improved. Nevertheless, Raman spectroscopy has a potential application value in the field of sperm aneuploidy detection and may even be used as a non-invasive predictor of sperm aneuploid state in preimplantation genetic testing (PGT-A). [ABSTRACT FROM AUTHOR]

The highly invasive Campuloclinium macrocephalum (Asteraceae; 'pompom weed'), native to South America, was introduced to South Africa where it has flourished and now threatens the native diversity of the grasslands it has invaded. The Argentine C. macrocephalum population is known to comprise at least three ploidy levels, and exhibit aneuploidy. Ploidy levels had not yet been assessed across South African populations of pompom weed and could influence the effectiveness of a biological control (biocontrol) programme. Therefore, we used flow cytometry to estimate ploidy levels of individuals from invaded regions in South Africa and Argentina. Diploids, triploids, tetraploids and aneuploids were detected in the sampled Argentine individuals. In South Africa, only triploid and tetraploid cytotypes were identified, with possible aneuploid variation. Mapping the distribution of these ploidy levels in South Africa revealed that the tetraploids are more widespread across all five sampled provinces, whereas the triploids occur only in Gauteng (the likely origin of the invasion) and neighbouring North West provinces and one sampled area in KwaZulu‐Natal province. Distinct phenotypic variation is evident between Argentine individuals of different ploidy levels. Polyploids have now been confirmed to occur in both the native and introduced ranges. Because triploids and tetraploids are present in both native and introduced ranges, we predict that polyploidy is not likely to be a constraint in effective biocontrol in South Africa as the natural enemies have probably co‐evolved with polyploids in the native range. However, other factors may impact the effectiveness of the biocontrol agents, such as feeding, oviposition (potentially affecting their reproduction), establishment, and spread, therefore, warranting further research. [ABSTRACT FROM AUTHOR]