Migration of repetitive DNAs during evolution of the permanent translocation heterozygosity in the oyster plant (Tradescantia section Rhoeo).

Due to translocation heterozygosity for all chromosomes in the cell complement, the oyster plant (Tradescantia spathacea) forms a complete meiotic ring. It also shows Rabl-arrangement at interphase, featured by polar centromere clustering. We demonstrate that the pericentromeric regions of the oyster plant are homogenized in concert by three subtelomeric sequences: 45S rDNA, (TTTAGGG)n motif, and TSrepI repeat. The Rabl-based clustering of pericentromeric regions may have been an excellent device to combine the subtelomere-pericentromere sequence migration (via inversions) with the pericentromere-pericentromere DNA movement (via whole arm translocations) that altogether led to the concerted homogenization of all the pericentromeric domains by the subtelomeric sequences. We also show that the repetitive sequence landscape of interstitial chromosome regions contains many loci consisting of Arabidopsis-type telomeric sequence or of TSrepI repeat, and it is extensively heterozygous. However, the sequence arrangement on some chromosomal arms suggest segmental inversions that are fully or partially homozygous, a fact that could be explained if the inversions started to create linkages already in a bivalent-forming ancestor. Remarkably, the subterminal TSrepI loci reside exclusively on the longer arms that could be due to sharing sequences between similarly-sized chromosomal arms in the interphase nucleus. Altogether, our study spotlights the supergene system of the oyster plant as an excellent model to link complex chromosome rearrangements, evolution of repetitive sequences, and nuclear architecture.

Pericentromere clustering in Tradescantia section Rhoeo involves self-associations of AT- and GC-rich heterochromatin fractions, is developmentally regulated, and increases during differentiation.

A spectacular but poorly recognized nuclear repatterning is the association of heterochromatic domains during interphase. Using base-specific fluorescence and extended-depth-of-focus imaging, we show that the association of heterochromatic pericentromeres composed of AT- and GC-rich chromatin occurs on a large scale in cycling meiotic and somatic cells and during development in ring- and bivalent-forming Tradescantia spathacea (section Rhoeo) varieties. The mean number of pericentromere AT-rich domains per root meristem nucleus was ca. half the expected diploid number in both varieties, suggesting chromosome pairing via (peri)centromeric regions. Indeed, regular pairing of AT-rich domains was observed. The AT- and GC-rich associations in differentiated cells contributed to a significant reduction of the mean number of the corresponding foci per nucleus in relation to root meristem. Within the first 10 mm of the root, the pericentromere attraction was in progress, as if it was an active process and involved both AT- and GC-rich associations. Complying with Rabl arrangement, the pericentromeres preferentially located on one nuclear pole, clustered into diverse configurations. Among them, a strikingly regular one with 5-7 ring-arranged pericentromeric AT-rich domains may be potentially engaged in chromosome positioning during mitosis. The fluorescent pattern of pachytene meiocytes and somatic nuclei suggests the existence of a highly prescribed ring/chain type of chromocenter architecture with side-by-side arranged pericentromeric regions. The dynamics of pericentromere associations together with their non-random location within nuclei was compared with nuclear architecture in other organisms, including the widely explored Arabidopsis model.

Three phases of energy-dependent induction of [Formula: see text] and Chl a fluorescence in Tradescantia fluminensis leaves.

In plants, the short-term regulation (STR, seconds to minute time scale) of photosynthetic apparatus is associated with the energy-dependent control in the chloroplast electron transport, the distribution of light energy between photosystems (PS) II and I, activation/deactivation of the Calvin-Benson cycle (CBC) enzymes, and relocation of chloroplasts within the plant cell. In this work, using a dual-PAM technique for measuring the time-courses of P700 photooxidation and Chl a fluorescence, we have investigated the STR events in Tradescantia fluminensis leaves. The comparison of Chl a fluorescence and [Formula: see text] induction allowed us to investigate the contribution of the trans-thylakoid pH difference (ΔpH) to the STR events. Two parameters were used as the indicators of ΔpH generation: pH-dependent component of non-photochemical quenching of Chl a fluorescence, and pHin-dependent rate of electron transfer from plastoquinol (PQH2) to [Formula: see text] (via the Cyt b6f complex and plastocyanin). In dark-adapted leaves, kinetics of [Formula: see text] induction revealed three phases. Initial phase is characterized by rapid electron flow to [Formula: see text] (τ1/2 ~ 5-10 ms), which is likely related to cyclic electron flow around PSI, while the outflow of electrons from PSI is restricted by slow consumption of NADPH in the CBC. The light-induced generation of ΔpH and activation of the CBC promote photooxidation of P700 and concomitant retardation of [Formula: see text] reduction (τ1/2 ~ 20 ms). Prolonged illumination induces additional slowing down of electron transfer to [Formula: see text] (τ1/2 ≥ 30-35 ms). The latter effect is not accompanied by changes in the Chl a fluorescence parameters which are sensitive to ΔpH generation. We suggest the tentative explanation of the latter results by the reversal of Q-cycle, which causes the deceleration of PQH2 oxidation due to the back pressure of stromal reductants.

Assessment of the genotoxic potential of water courses impacted by wastewater treatment effluents using micronucleus assay in plants from the species Tradescantia.

Water pollution and the increase in genotoxic consequences in aquatic environments are well documented indicating the necessity and importance of biomonitoring programs. The objective of the present study was to determine the environmental quality of water resources and genotoxic potential of materials present within water samples obtained from the Perdizes River and the Mumbuca Stream, located in a region of discharge of wastewater treatment effluents using Tradescantia micronucleus assay (Trad - MCN). Water samples were collected from different locations up and downstream of the wastewater treatment plant during rainy season and subsequently submitted to physico-chemical analysis and Trad-MCN bioassay. The spatial distribution of the physico-chemical parameters assessed suggested that discharges of wastewater treatment effluents reduced water quality at all sites examined. Further, exposure to wastewater treatment effluents produced genotoxic effects on tetrads of Tradescantia pallida. These results reinforce the sensitivity of the Trad-MCN bioassay and its potential application in water quality monitoring programs concomitant with physicochemical evaluation.

Genotoxicity of Water Contaminants from the Basin of Lake Sevan, Armenia Evaluated by the Comet Assay in Gibel Carp (Carassius auratus gibelio) and Tradescantia Bioassays.

Combination of bioassays and chemical analysis was applied to determine the genotoxic/mutagenic contamination in four different sites of the basin of Lake Sevan in Armenia. Water genotoxicity was evaluated using the single cell gel electrophoresis technique (comet assay) in erythrocytes of gibel carp (Carassius auratus gibelio), Tradescantia micronucleus (Trad-MCN) and Tradescantia stamen hair mutation (Trad-SHM) assays. Significant inter-site differences in the levels of water genotoxicity according to fish and Trad-MCN bioassays have been revealed. Two groups of locations with lower (south-southwest of the village Shorzha and Peninsula of Lake Sevan) and higher (estuaries of Gavaraget and Dzknaget rivers) levels of water genotoxicity were distinguished. Correlation analysis support the hypothesis that the observed genetic alterations in fish and plant may be a manifestation of the effects of water contamination by nitrate ions, Si, Al, Fe, Mn and Cu. Increase of DNA damage in fish also correlated with content of total phosphorus.

Genetic instability in plants associated with vehicular traffic and climatic variables.

To characterize the effect of vehicular traffic on air quality, the micronuclei of Tradescantia pallida tetrads were counted. Young inflorescences of T. pallida (Rose) D.R. Hunt var. purpurea were collected in 2010, 2011, and 2012, from three sites subjected to different intensities of vehicular traffic. The sites were located in the municipality of Dourados, in the State of Mato Grosso do Sul, Brazil. A standardized methodology was used to analyze the Tradescantia micronuclei, in order to evaluate the mutagenic potential of the local air pollutants. Statistical analyses using the Pearson's linear correlation were employed to determine the relationship between relative humidity and temperature, and the average number of micronuclei. In this study, an increase in the average number of passing vehicles was correlated with an increase in the frequency of micronucleus formation. Climatic factors also influenced micronucleus formation, although vehicular traffic remained the most important factor. Thus, the Tradescantia micronuclei assay may be a useful method of assessing air quality.

Assessing the genotoxicities of sparteine and compounds isolated from Lupinus mexicanus and L. montanus seeds by using comet assay.

The genus Lupinus is widely distributed. Its seeds are used for animal and human food, and Lupinus possesses pharmacological potential because of its high content of quinolizidine alkaloids and flavonoids; however, there is little available information about its genotoxicity. We used the comet assay and staminal nuclei of Tradescantia (clone 4430) to evaluate the in vitro genotoxicity of 4 concentrations (0.01, 0.1, 0.5, and 1.0 mM) of alkaloid extracts of Lupinus mexicanus and Lupinus montanus, flavonoids of L. mexicanus, and commercial sparteine; nitrosodiethylamine was used as a positive control and untreated nuclei were used as a negative control. All concentrations of L. mexicanus and L. montanus showed significant genotoxic activity (P ≤ 0.05). A similar behavior was observed for flavonoid extracts of L. montanus except the 1.0 mM concentration. Sparteine showed genotoxic activity only at 0.5 mM. The order of genotoxicity of the compounds studied was as follows: L. mexicanus > L. montanus > flavonoids of L. montanus > sparteine. There is evident genotoxic activity in the compounds that were studied, particularly at lower concentrations (0.01 and 0.1 mM). Given the limited information about the genotoxicity of the compounds of L. mexicanus and L. montanus, further studies are necessary.

Assessment of genotoxicity of air pollution in urban areas using an integrated model of passive biomonitoring.

Atmospheric pollution is a major public health issue and has become increasingly critical for human health. Urban atmospheric pollution is typically assessed through physicochemical indicators aligned with environmental legislation parameters, providing data on air quality levels. While the effects of pollution on sensitive organisms serve as a warning for public health decision-makers, there remains a need to explore the interpretation of environmental data on pollutants. The use of species adapted to urban environments as sentinels enables continuous and integrated monitoring of environmental pollution implications on biological systems. In this study, we investigated the use of the plant species Tradescantia pallida as a biomonitor to evaluate the genotoxic effects of atmospheric pollution under diverse vehicular traffic conditions. T. pallida was strategically planted at the leading urban intersections in Uberlândia, Brazil. During COVID-19 pandemic lockdowns, we compared indicators such as physical, biological, and traffic data at different intersections in residential and commercial zones. The reduction in vehicular traffic highlighted the sensitivity of plant species to changes in air and soil pollutants. T. pallida showed bioaccumulation of heavy metals Cd and Cr in monitored areas with higher traffic levels. Additionally, we established a multiple linear regression model to estimate genotoxicity using the micronucleus test, with chromium concentration in the soil (X1) and particulate matter (PM) in the atmosphere (X2) identified as the primary independent variables. Our findings provide a comprehensive portrait of the impact of vehicular traffic changes on PM and offer valuable insights for refining parameters and models of Environmental Health Surveillance.

In situ biomonitoring of the genotoxic effects of vehicular pollution in Uberlândia, Brazil, using a Tradescantia micronucleus assay.

The growing number of cars in large cities is directly linked to changes in the chemical composition of urban air, which has increasingly high concentrations of potentially genotoxic chemicals. Therefore, discovering and monitoring the risks associated with exposure to atmospheric pollutants is indispensible for preventing environmental and health problems. Because of the lack of reliable data regarding the air quality in the city of Uberlândia, the present study sought to test whether the genotoxic risks in areas with different levels of vehicular traffic can be measured using the Tradescantia micronucleus assay (Trad-MN). Therefore, more than twenty inflorescences were exposed to locations with different amounts of vehicular traffic twice per year from the winter of 2006 to the summer of 2011. The inflorescences were then analysed to determine the micronucleus (MN) frequency. In addition, we sought to determine the influence of factors linked to city climate on the MN frequencies obtained at each monitored location. Our results show that, although low relative humidity positively influenced MN formation in Tradescantia pallida tetrads, the major determining factor for clastogenic events was the level of vehicular traffic at the locations monitored over the five-year study.

Assessment of the mutagenic potential of the water of an urban river by means of two Tradescantia-based test systems.

River pollution can be caused by anthropogenic or natural factors. When testing water quality for the presence of toxic substances, higher plants as bioindicators for the genotoxic effects of complex mixtures are effective and appropriate. Hence, in this work the Tradescantia (clone 02) stamen hair mutations (Trad-SHM) and Tradescantia micronuclei (Trad-MCN) were used to determine mutagenic and clastogenic potential of an urban river. A significant increase in the level of all studied endpoints as well as morphological changes, including pink cells (PC) and colorless cells (CC) in stamen hairs, stunted hairs (SH), tetrads with micronuclei (MN) and MN in tetrads of pollen microspores in the Tradescantia was observed compared to the negative control (tap water). As an example riverine system, part of the River Hrazdan (Armenia) flowing through a highly urbanized and industrial area was studied. The positive control (10 mM CrO3) showed the highest genotoxicity for the SHM assay (PC: 5.1 / 1000, CC: 17.9 / 1000) and for the MCN assay (12 MN / 100 tetrads and 9.4 ± 0.53 tetrads with MN). Genetic responses were analyzed in conjunction with the concentrations of select elements in the riverine water. Reasons for observing such a high level of genetic markers in the riverine water and applicability of the Tradescantia (clone 02) test-systems in environmental risk assessment and biomonitoring are discussed.

Structural heterozygosity, duplication of telomeric (TTTAGGG)(n) clusters and B chromosome architecture in Tradescantia virginiana L.

Fluorescent in situ hybridization, C-banding/DAPI, and CMA(3)-fluorescence were performed to reveal the cytomolecular constitution of the standard (A) and supernumerary (B) chromosomes of an autotetraploid Tradescantia virginiana L. The analyses show that translocations and/or inversions have occurred during the evolution of the T. virginiana karyotype, generating a significant level of structural heterozygosity. Regarding the structural level, the present paper confirms the occurrence of small subterminal duplications and/or inversions in T. virginiana already suggested by previous authors. Interestingly, many of the distal chromosome segments in T. virginiana possess duplicated subterminal telomere clusters, heterochromatin, 5S and 45S rDNAs orderly intermixed and share this complex cytomolecular architecture with the common type of a B chromosome. Based on the obtained results, it is proposed that in T. virginiana the B chromosome may have arisen via excision from the distal region of an A chromosome. The nascent B could have retained much of the ancestral sequence arrangement, including duplicated telomeric cluster(s), heterochromatin and rDNA, but developed a new centromere/kinetochore to successfully propagate through the cell cycle.

Genotoxic potential generated by biomass burning in the Brazilian Legal Amazon by Tradescantia micronucleus bioassay: a toxicity assessment study.

BACKGROUND: The Brazilian Amazon has suffered impacts from non-sustainable economic development, especially owing to the expansion of agricultural commodities into forest areas. The Tangará da Serra region, located in the southern of the Legal Amazon, is characterized by non-mechanized sugar cane production. In addition, it lies on the dispersion path of the pollution plume generated by biomass burning. The aim of this study was to assess the genotoxic potential of the atmosphere in the Tangará da Serra region, using Tradescantia pallida as in situ bioindicator. METHODS: The study was conducted during the dry and rainy seasons, where the plants were exposed to two types of exposure, active and passive. RESULTS: The results showed that in all the sampling seasons, irrespective of exposure type, there was an increase in micronucleus frequency, compared to control and that it was statistically significant in the dry season. A strong and significant relationship was also observed between the increase in micronucleus incidence and the rise in fine particulate matter, and hospital morbidity from respiratory diseases in children. CONCLUSIONS: Based on the results, we demonstrated that pollutants generated by biomass burning in the Brazilian Amazon can induce genetic damage in test plants that was more prominent during dry season, and correlated with the level of particulates and elevated respiratory morbidity.

Genotoxicity of SPL (spent pot lining) as measured by Tradescantia bioassays.

Spent Pot Liner (SPL) is a solid waste product generated in the process of aluminum production. Tradescantia micronuclei (Trad-MN) and stamen hair mutation (Trad-SHM) bioassays are very useful tests to assess genotoxicity of environmental pollutants. In the present study, we intended to investigate the genotoxicity of this waste with Tradescantia bioassays using leachates of SPL simulating the natural leachability of SPL in soil. The formation of micronuclei (MN) was found to be concentration dependent. MN frequency enhanced significantly with SPL treatment. In addition, SPL also appeared to increase the percentage of dyads and triads. Trad-SHM assay showed that SPL increases pink mutation events as SPL concentration increases. These results demonstrated that SPL is a cytogenotoxic agent that affects different genetic end-points (induction of micronuclei and point mutations) even at low concentration (2% and 3%).

Micronucleus study of the quality and mutagenicity of surface water from a semi-arid region.

The present study evaluated the mutagenic potential of surface water from the Lucrecia dam. The Tradescantia-micronucleus (Trad-MCN) test and CBMN assay in human peripheral blood lymphocytes were applied, corresponding to an in vivo and in vitro system, respectively. Heavy metals and some physicochemical properties were also measured. Water samples were collected in November 2009 (dry season) and May 2010 (rainy season) at three different points. Results of both assays for raw water showed positive responses for the points analyzed when compared to the negative control. The CBMN assay showed that diluted water was still able to induce a significant increase in micronucleus frequency. For both assays, the highest mean MN was observed in the dry season. Chemical analyses detected an increase in heavy metal levels at the sampling points and in the different seasons. These findings indicate the presence of genotoxins, such as heavy metals, in the water, which may be affecting the entire ecosystem, as well as human health. More prolonged monitoring is recommended in order to better characterize this public water supply.

Biomonitoring of air quality in the Bodoquena microregion, Mato Grosso Do Sul: mutagenic and morphoanatomical alterations in Tradescantia pallida (rose) D.R. Hunt var. purpurea.

The objective of this study was to assess air quality in relation to vehicular traffic flow in cities located at different elevations in the Bodoquena microregion, state of Mato Grosso do Sul, Brazil. To do so, a micronucleus test was carried out using the TRAD-MCN bioassay on young Tradescantia buds collected from February to November 2018 in seven cities of the microregion with different traffic flow intensities. Meteorological parameters were evaluated, and vehicular traffic was counted to determine traffic flow in each city. With data from the Shuttle Radar Topography Mission (SRTM) and processing in Esri ArcGIS® software version 10.5.1, the regions was mapped based on an Elevation Model. Morphoanatomical analyses were performed according to standard methodology. Measurements were taken of thickness, length and width of tissues and structures, including the upper and lower cuticle, upper and lower epidermis, hypodermis and mesophyll. The greatest traffic flow was found in the cities of Bodoquena, Guia Lopes da Laguna, Jardim, and Porto Murtinho, with the period from 5:00 to 6:00 p.m. showing the highest traffic flow. The greatest frequency of mutagenic alterations was found in the city of Guia Lopes da Laguna, although the results did not differ significantly from Bonito, Caracol, and Jardim. Throughout the biomonitoring, the summer and autumn seasons showed the greatest micronuclei frequencies in all evaluated cities. Variations in the tissue/structure thickness was observed across cities and seasons, but with a decrease in thickness during autumn. In general, the tissues/structures were smaller for the cities of Nioaque and Porto Murtinho, while the anatomical and morphological characteristics of leaf length and thickness showed no differences among cities. We found limited correlation between micronuclei frequency and traffic flow, supporting the hypothesis that although mutagenic alterations are observed in T. pallida, in this microregion the changes are numerically lower when compared to other regions of the state. In light of the genotoxic and morphoanatomical factors assessed herein, the Bodoquena microregion appears to be well preserved in terms of air quality, presenting low micronuclei frequency and a limited reduction in tissues and leaf structures, regardless of the season.

Soil-surface genotoxicity of military and urban territories in Lithuania, as revealed by Tradescantia bioassays.

The soil surface is a major natural system that accumulates pollutants and allows researchers to disclose the history and the present state of contamination of an area with toxic pollutants. This conclusion is based on the comparison of genotoxicity of aqueous extracts and DMSO extracts of topsoil from military territories in various locations left behind after the retreat of the Soviet Army from Lithuania, and several sites in the city of Vilnius, characterized by different history and current traffic intensity. The specific character of the soil-surface contamination was shown in a series of Tradescantia micronucleus (Trad-MN) and stamen-hair mutation (Trad-SHM) bioassays and tests. The most effective ones were the Trad-MN and, unexpectedly, the branched-hair tests. A preliminary result of the study is the somaclonal variation of individual Tradescantia plants revealed by the RAPD method of DNA analysis. A comparison of aqueous extracts and DMSO extracts of the soil showed the permanent character of the mobile forms of genotoxic pollutants in the soil surface, despite the fact that several military territories were already closed 20 years ago.

Ribosomal DNA, tri- and bi-partite pericentromeres in the permanent translocation heterozygote Rhoeo spathacea.

High- and low-stringency FISH and base-specific fluorescence were performed on the permanent translocation heterozygote Rhoeo spathacea (2n = 12). Our results indicate that 45S rDNA arrays, rDNA-related sequences and other GC-rich DNA fraction(s) are located within the pericentromeric regions of all twelve chromosomes, usually colocalizing with the chromomycin A(3)-positive bands. Homogenization of the pericentromeric regions appears to result from the concerted spread of GC-rich sequences, with differential amplification likely. We found new 5S rDNA patterns, which suggest a variability in the breakpoints and in the consequent chromosome reorganizations. It was found that the large 5S rDNA locus residing on each of the 8E and 9E arms consisted of two smaller loci. On each of the two chromosome arms 3b and 4b, in addition to the major subtelomeric 5S rDNA locus, a new minor locus was found interstitially about 40% along the arm length. The arrangement of cytotogenetic landmarks and chromosome arm measurements are discussed with regard to genome repatterning in Rhoeo.

Ability of Allium cepa L. root tips and Tradescantia pallida var. purpurea in N-nitrosodiethylamine genotoxicity and mutagenicity evaluation.

N-nitroso compounds, such as N-nitrosodiethylamine (NDEA), can be formed by the reaction of secondary amines with nitrosating agents, and are suspected to be involved in tumors in humans. NDEA has been considered a weak carcinogen in genotoxic assays probably due to the inefficient nitrosamine activation system that is used and/or to the efficient repair system. In this work, we evaluated the sensibility of Allium cepa L. root tips and Tradescantia stamen hair mutation assay (Trad-SH) using Tradescantia pallida var. purpurea for NDEA (0.1; 0.5; 5 and 25 mM) genotoxicity and mutagenicity induction. Allium cepa L. was treated with different NDEA concentrations for 3h, for 3 consecutive days, including negative control (distilled water) and positive control maleic hydrazide (MH 30 mg/mL). After treatment, the roots were hydrolyzed, squashed, and the mitotic index (MI) and cytological abnormalities were scored. The results revealed a cytostatic effect of NDEA (0.5 and 5mM), showing a significant reduction in the MI. Chromosome stickiness suggests a NDEA toxic effect. T. pallida purpurea did not respond to mutagens with a dose-dependent pattern. In conclusion, our study indicates that the root tips of Allium cepa L. have sensibility to detect NDEA genotoxicity, but not for Trad-SH test.

Analysis of genotoxic effects on plants exposed to high traffic volume in urban crossing intersections.

A biological assessment of environmental quality was performed using the tropical plant species Tradescantia pallida (Rose) D.R. Hunt. var. purpurea exposed to different levels of air contamination in urban intersections with high volume of vehicle traffic. Air quality (average daily levels of particulate material in the PM1, 2.5, 10 fractions) and traffic volume in crossing intersections were monitored for 30 days before the collection of plants. Frequency of micronuclei and pollen abortivity in inflorescences collected at different intersections with gradual levels of traffic volume were evaluated as biomarkers of genotoxicity. In addition, the concentrations of bioaccumulated heavy metals in the leaves of the collected plants were also investigated. The proposed biological assessment model found a positive association between the environmental variables (traffic volume; concentration of particulate material) and biological effects (leaf concentration of Cr and Cd; micronucleus frequencies and pollen abortivity).

Evaluation of the genotoxicity of treated urban sludge in the Tradescantia micronucleus assay.

The sludge produced in sewage treatment plants can contain toxic substances. Among these, the genotoxic substances are of great concern. The present paper aimed at evaluating the genotoxicity of treated sludge samples collected at four different sewage treatment plants (STP) located in the State of São Paulo, Brazil using the Trad-MN assay. Another objective of the study was to compare the responses of the Clone #4430 with the Tradescantia pallida. Sludge samples mixed with reference soil in concentrations of 10, 25 and 50% (v/v) were tested in experiments with 3 months exposure in the field. Negative and positive controls (arsenic trioxide) were also tested with both plants. In Clone #4430 two sludge samples induced genotoxicity while in T. pallida three were positive, although no clear dose-response were observed for both plants. Results with the negative and positive controls suggest that T. pallida presented similar results when compared to the Clone #4430. The protocol using plants chronically exposed to sludge mixed with soil seems to be a promising tool to assess the genotoxicity of sludge although time consuming.