Interphase nuclei, karyotypes and nuclear DNA amounts in five species of Oenocarpus (Arecaceae).

The genus Oenocarpus Martius, 1823 (Arecaceae) includes five species commonly used in Amazonia, especially for their fruits. Little is known about the cytogenetic characteristics and DNA amounts of these species, except for O.bataua (Martius, 1823). This study characterized and compared the types of interphase nuclei, the chromosome sets, and estimated the nuclear DNA amounts of Oenocarpusbacaba (Martius, 1823), O.bataua, O.distichus (Martius, 1823), O.mapora (H. Karsten, 1857) and O.minor (Martius, 1823). Standard cytogenetic analyses and estimates of the nuclear DNA amount by flow cytometry were carried out. These are the first reports of chromosome numbers and DNA amounts, except for O.bataua, as is the description of the chromatin distribution in interphase nuclei and karyotype for all species. All species presented 2n = 36, confirming the previous report for O.bataua. Differences between karyotype formulas and the positioning of secondary constrictions were observed. There were no significant differences for the nuclear DNA amounts among species. The constancy in chromosome number and variations in karyotype formulas suggest the occurrence of chromosome rearrangement as an important mechanism in Oenocarpus speciation.

Forage potential of Urochloa genotypes by using leaf anatomy / Avaliação do potencial forrageiro de genótipos de Urochloa por meio da anatomia foliar

ABSTRACT: The digestibility potential of leaves from forages depends on the amount of nutrition in their tissues, with low lignin deposition in the cell wall, mainly parenchyma and phloem. This research evaluated the leaf structure of different Urochloa genotypes and discussed its potential for evaluating digestibility. The cultivars U. brizantha, U. decumbens, U. ruziziensis and three clones of U. ruziziensis (1, 95 and 97), which are under development in breeding programs, were evaluated. Plants were grown under the recommended culture conditions for the Urochloa species. Plants were cut 60 days after sowing, and leaves were collected at 15 days of regrowth. Leaves were fixed in FAA 70 and further stored in 70% ethanol until being submitted to the usual microtechniques for the preparation of microscopy slides. The area of the tissues from the interveinal and midrib regions was measured using ImageJ software, and their proportions were calculated. In the interveinal region, the proportion of chlorophyll parenchyma was greater for U. decumbens and the Clone 1 genotypes. Urochloa brizantha and clones 95 and 97 showed a higher proportion of the vascular bundle compared to U. ruziziensis, U. decumbens and Clone 1. The proportion of the ground parenchyma in the midrib was greater in U. brizantha, Clone 95 and Clone 97. Thus, it can be concluded that the Clone 1 genotypes (from U. ruziziensis) showed leaf tissues (parenchyma and phloem) with higher digestibility potential; in addition, U. brizantha and U. decumbens showed a high percentage of xylem and sclerenchyma, which reduces their quality as forage.

RESUMO: O potencial de digestibilidade das folhas de forrageiras depende da quantidade de seus tecidos com baixa deposição de lignina nas paredes celulares, principalmente o parênquima e floema. O presente trabalho objetivou avaliar a estrutura foliar de diferentes genótipos de Urochloa e seu potencial de digestibilidade foliar. Foram avaliadas as cultivares U. brizantha, U. decumbens, U. ruziziensis e três clones de U. ruziziensis (1, 95 e 97). As plantas foram cultivadas em campo com condições de manejo recomendadas para a espécie, sendo cortadas aos 60 dias e as folhas coletadas aos 15 dias de rebrota. As folhas foram fixadas em FAA 70, armazenadas em etanol 70% e posteriormente submetidas à microtécnica usual para preparação de lâminas semipermanentes. Foram analisadas as regiões internervural e da nervura mediana com o auxílio do software Image J sendo mensurada a área de cada tecido da folha e depois calculada a sua proporção em relação à área total das secções. Na região internervural, a proporção de parênquima foi maior em U. decumbens e no Clone 1. Ainda nesta região, Urochloa brizantha e os Clones 95 e 97 apresentaram maiores médias para a proporção de feixes vasculares. Na nervura mediana, a proporção do parênquima foi maior em U. brizantha, Clone 95 e Clone 97. De maneira geral, o Clone 1 (proveniente de U. ruziziensis) apresentou parênquima e floema em maiores proporções, enquanto genótipos de U. brizantha e U. decumbens demonstram altas quantidades de xilema e esclerênquima que reduzem o seu potencial de digestibilidade.

Regrowth age modifies the leaf anatomy of Brachiaria genotypes / A idade de regeneração modifica a anatomia foliar dos genótipos de Brachiaria

Changes in leaf anatomy were evaluated in genotypes of Brachiaria brizantha, Brachiaria decumbens, Brachiaria ruziziensis and three Brachiaria ruziziensis clones through tissue proportion in internerval and midrib regions at three regrowth ages. Plants were grown and cutting was performed after 60 days. Further, leaves were sampled at 8, 15 and 29 regrowth days and processed with usual plant microtechnique. The internerval region showed higher parenchyma percentage at 15 days for Clone 95 and similar values at 15 and 29 days for Clone 1. The proportion of vascular bundles was lower after 15 days in Clones 1 and 95 and 29 days in B. brizantha. In the midrib, the parenchyma proportion was higher at 29 days in B. brizantha and lower at 15 days in B. ruziziensis. The proportion of vascular bundles was higher at 8 days in B. decumbens, B. brizantha and Clone 1, and lower at 29 days for Clones 97 and 95 and at 8 days in B. ruziziensis. Therefore, the regrowth age modifies the percentage of leaf tissues in Brachiaria genotypes, in which the fibers and vascular bundles increase at 29 days and 8-day-old leaves are not fully developed.

A anatomia foliar foi avaliada em genótipos de Brachiaria brizantha, Brachiaria decumbens, Brachiaria ruziziensis e três clones de Brachiaria ruziziensis; quanto à proporção de tecidos foliares, em três idades de rebrota. As plantas foram cultivadas e cortadas após 60 dias. As folhas foram amostradas aos 8, 15, 29 dias de rebrota e analisadas pela metodologia de microtécnica vegetal. Na região internervural, a proporção de parênquima foi maior aos 15 dias para o Clone 95, e aos 15 e 29 dias para o Clone 1. Proporção de feixes vasculares foi menor aos 15 dias para os Clones 1 e 95 e aos 29 dias para genótipo B. brizantha. Na nervura central, a proporção de parênquima foi maior aos 29 dias para B. brizantha e menor, aos 15 dias, para B. ruziziensis. Proporção dos feixes vasculares foi maior aos 8 dias para genótipo B. decumbens, B. brizantha e para o Clone 1, e menor aos 29 dias para os Clones 97 e 95; aos 8 dias, para genótipo B. ruziziensis. Portanto, a idade de rebrota modifica a porcentagem de tecidos foliares em genótipos de Brachiaria; contudo, fibras e feixes vasculares aumentam aos 29 dias; aos 8 dias, as folhas não estão totalmente desenvolvidas.

Effects of long exposure to spent potliner on seeds, root tips, and meristematic cells of Allium cepa L.

Spent potliner (SPL) is a solid waste generated in the aluminum mining and processing industry. It is sometimes dumped into the environment and leach in contact with water, thereupon affecting living beings, which are likely to be exposed to the waste for long periods. Considering this, we aimed to evaluate the effects of extended exposure to SPL through bioassays using Allium cepa as plant model system. Seeds of A. cepa were either directly exposed to SPL (continuous exposure) or first germinated in water and then exposed to SPL (discontinuous exposure). The germination rate was determined from 24 to 192 h of exposure. The maximum effects of SPL on germination were observed after 96 h in both exposure approaches. For the parameter root elongation, the discontinuous treatment was more efficient in demonstrating differences among the applied SPL concentrations (60% of reduction). Microscopic analysis was carried out in root tip cells discontinuously exposed to SPL for 96 h. A mitodepressive effect was observed (above 50%), as well as increased rate of chromosome abnormalities (up to 100-fold) and induction of cell death. The consequences of exposure to SPL for longer periods are discussed.

Reliability of plant root comet assay in comparison with human leukocyte comet assay for assessment environmental genotoxic agents.

Comet assay is an efficient test to detect genotoxic compounds based on observation of DNA damage. The aim of this work was to compare the results obtained from the comet assay in two different type of cells extracted from the root tips from Lactuca sativa L. and human blood. For this, Spent Pot Liner (SPL), and its components (aluminum and fluoride) were applied as toxic agents. SPL is a solid waste generated in industry from the aluminum mining and processing with known toxicity. Three concentrations of all tested solutions were applied and the damages observed were compared to negative and positive controls. It was observed an increase in the frequency of DNA damage for human leukocytes and plant cells, in all treatments. On human leukocytes, SPL induced the highest percentage of damage, with an average of 87.68%. For root tips cells of L. sativa the highest percentage of damage was detected for aluminum (93.89%). Considering the arbitrary units (AU), the average of nuclei with high levels of DNA fragmentation was significant for both cells type evaluated. The tested cells demonstrated equal effectiveness for detection of the genotoxicity induced by the SPL and its chemical components, aluminum and fluoride. Further, using a unique method, the comet assay, we proved that cells from root tips of Lactuca sativa represent a reliable model to detect DNA damage induced by genotoxic pollutants is in agreement of those observed in human leukocytes as model. So far, plant cells may be suggested as important system to assess the toxicological risk of environmental agents.

Colchicine and oryzalin effects on tetraploid induction and leaf anatomy of Solanum commersonii ssp. / Colchicina e orizalina na indução de tetraploides e na anatomia foliar de Solanum commersonii ssp.

ABSTRACT: The aim of the present study was to evaluate the effect of the colchicine and oryzalin anti-mitotic substances on the induction of tetraploid plants and foliar anatomy of two diploids clones of Solanum commersonii ssp. Nodal segments of Solanum commersonii subsp. commersonii Dun. and Solanum commersonii subsp. malmeanum Bitt. were treated with colchicine (3.5, 5.0, and 6.5mM; 72h) or oryzalin (10, 30, and 50µM; 24h). After the treatment with anti-mitotic substances, nodal segments were inoculated in the MS culture medium and cultivated in vitro (60 days). After in vitro cultivation, the plants were transferred to vases with the substrate Plantmax(r) and kept in the greenhouse (45 days). Plant ploidy level was assessed by flow cytometry and leaf anatomy was assessed by anatomic cuts. An increase was observed in the polar and equatorial diameter of stomata ("gigas effect") of the Solanum commersonii subsp. commersonii (SCC) and Solanum commersonii subsp. malmeanum (SCM) clones, which was due to the use of the anti-mitotic substances. Treatments with colchicine (6.5mM; 72h) and oryzalin (50µM; 24h) caused death of the SCC plants cultured in vitro. In the others treatments of the SCC clone, the use of oryzalin and colchicine caused production of chimeric plants. The treatment of nodal segments of SCM with oryzalin (10-50µM; 24h) was effective on induction of tetraploid plants, which can be employed in genetic breeding programs in crossbreeding with cultured potato.

RESUMO: O objetivo deste estudo foi avaliar o efeito das substâncias antimitóticas colchicina e orizalina na indução de plantas tetraploides e na anatomia foliar de dois clones diploides de Solanum commersonii ssp. Segmentos nodais de Solanum commersonii subsp. commersonii Dun. e Solanum commersonii subsp. malmeanum Bitt. foram tratados com colchicina (3,5, 5,0 e 6,5mM; 72h) ou orizalina (10, 30 e 50µM; 24h). Após o tratamento com as substâncias antimitóticas, os segmentos nodais foram inoculados em meio de cultura MS e cultivados in vitro (60 dias). Após o cultivo in vitro, as plantas foram transferidas para vasos contendo o substrato Plantmax(r) e mantidas em casa de vegetação (45 dias). O nível de ploidia das plantas foi avaliado por citometria de fluxo e a anatomia foliar foi avaliada através de cortes anatômicos. Foi observado um aumento no diâmetro equatorial e polar dos estômatos ("efeito gigas") dos clones de Solanum commersonii subsp. commersonii (SCC) e Solanum commersonii subsp. malmeanum (SCM), que foi produzido pelo uso de substâncias antimitóticas. O tratamento com colchicina (6,5mM; 72h) e orizalina (50µM; 24h) causou morte nas plantas de SCC cultivadas in vitro. Nos demais tratamentos do clone de SCC, o uso de orizalina ou colchicina causou a produção de plantas quiméricas. O emprego de orizalina (10-50µM; 24h), nos segmentos nodais de SCM, resultou na obtenção de plantas tetraploides consistentes, as quais podem ser usadas nos programas para melhoramento genético em cruzamentos com a batata cultivada.

Cytotoxicity of Spent Pot Liner on Allium cepa root tip cells: A comparative analysis in meristematic cell type on toxicity bioassays.

Spent Pot Liner (SPL) is a waste generated during the production of aluminum. It is comprised of a mixture of substances most of which, like cyanide, aluminum and fluoride, are toxic. Previous studies indicate the highly toxic nature of SPL. However studies using cells of the differentiation/elongation zone of the root meristem (referred as M2 cells in this study) after a proper recovery period in water were never considered. Using these cells could be useful to further understanding the toxicity mechanisms of SPL. A comparative approach between the effects on M2 cells and meristematic cells of the proximal meristem zone (referred as M1 cells in this study) could lead to understanding how DNA damage caused by SPL behaves on successive generations of cells. Allium cepa cells were exposed to 4 different concentrations of SPL (2.5, 5, 7.5 and 10gL(-1)) mixed with soil and diluted in a CaCl2 0.01M to simulate the ionic forces naturally encountered on the environment. A solution containing only soil diluted on CaCl2 0.01M was used as control. M1 and M2 cells were evaluated separately, taking into account four different parameters: (1) mitotic alterations (MA); (2) presence of condensed nuclei (CN); (3) mitotic index (MI); (4) presence of micronucleus (MCN). Significant differences were observed between M1 and M2 roots tip cells for these four parameters accessed. M1 cells was more prompt to reveal citogenotoxicity through the higher frequency of MA observed. Meanwhile, for M2 cells higher frequencies of MCN and CN was noticed, followed by a reduction of MI. Also, it was possible to detect significant differences between the tested treatments and the control on every case. These results indicate SPL toxic effects carries on to future cells generations. This emphasizes the need to properly manage this waste. Joint evaluation of cells from both M1 and M2 regions was proven valuable for the evaluation of a series of parameters on all toxicity tests.

Karyotype and genome size in Euterpe Mart. (Arecaceae) species.

Euterpe (Martius, 1823), a genus from Central and South America, has species with high economic importance in Brazil, because of their palm heart and fruits, known as açaí berries. Breeding programs have been conducted to increase yield and establish cultivation systems to replace the extraction of wild material. These programs need basic information about the genome of these species to better explore the available genetic variability. The aim of this study was to compare Euterpe edulis (Martius, 1824), Euterpe oleracea (Martius, 1824) and Euterpe precatoria (Martius, 1842), with regard to karyotype, type of interphase nucleus and nuclear DNA amount. Metaphase chromosomes and interphase nuclei from root tip meristematic cells were obtained by the squashing technique and solid stained for microscope analysis. The DNA amount was estimated by flow cytometry. There were previous reports on the chromosome number of Euterpe edulis and Euterpe oleracea, but chromosome morphology of these two species and the whole karyotype of Euterpe precatoria are reported for the first time. The species have 2n=36, a number considered as a pleisomorphic feature in Arecoideae since the modern species, according to floral morphology, have the lowest chromosome number (2n=28 and 2n=30). The three Euterpe species also have the same type of interphase nuclei, classified as semi-reticulate. The species differed on karyotypic formulas, on localization of secondary constriction and genome size. The data suggest that the main forces driving Euterpe karyotype evolution were structural rearrangements, such as inversions and translocations that alter chromosome morphology, and either deletion or amplification that led to changes in chromosome size.

Tissue-specific genome instability in synthetic interspecific hybrids of Pennisetum purpureum (Napier grass) and Pennisetum glaucum (pearl millet) is caused by micronucleation.

Genome instability is observed in several species hybrids. We studied the mechanisms underlying the genome instability in hexaploid hybrids of Napier grass (Pennisetum purpureum R.) and pearl millet (Pennisetum glaucum L.) using a combination of different methods. Chromosomes of both parental genomes are lost by micronucleation. Our analysis suggests that genome instability occurs preferentially in meristematic root tissue of hexaploid hybrids, and chromosome elimination is not only caused by centromere inactivation. Likely, beside centromere dysfunction, unrepaired DNA double-strand breaks result in fragmented chromosomes in synthetic hybrids.

Variation of karyotype and nuclear DNA content among four species of Plectranthus L' Héritier, 1788 (Lamiaceae) from Brazil.

Plectranthus is a genus which includes species of ornamental and medicinal potential. It faces taxonomic problems due to aggregating species previously belonging to the genus Coleus, a fact that has contributed to the existence of various synonymies. The species Plectranthus amboinicus, Plectranthus barbatus, Plectranthus grandis and Plectranthus neochilus are included in this context. Some authors consider Plectranthus barbatus and Plectranthus grandis as synonyms. The present work was carried out with the aim of comparing plants of the above-mentioned species, originating from different localities in Brazil, with regards to chromosome number and karyotypic morphology, correlated to the nuclear DNA content. There was no variation in chromosome number among plants of the same species. Plectranthus amboinicus was the only species to exhibit 2n=34, whereas the others had 2n=30. No karyotypic differences were found among the plants of each species, except for Plectranthus barbatus. The plants of the Plectranthus species revealed little coincidence between chromosome pairs. The nuclear DNA content allowed grouping Plectranthus amboinicus and Plectranthus neochilus, with the highest mean values, and Plectranthus grandis and Plectranthus barbatus with the lowest ones. Differences in DNA amount among the plants were identified only for Plectranthus barbatus. These results allow the inference that the populations of Plectranthus amboinicus and Plectranthus neochilus present coincident karyotypes among their plants, and Plectranthus grandis is probably a synonym of Plectranthus barbatus.

Ribosomal DNA in diploid and polyploid Setaria (Poaceae) species: number and distribution.

Setaria Beauvois, 1812 is a genus of economically important forage species, including Setaria italica (Linnaeus, 1753) Beauvois, 1812 and Setaria viridis (Linnaeus, 1753) Beauvois, 1812, closely related species and considered as model systems for studies of C4 plants. However, complications and uncertainties related to taxonomy of other species of the genus are frequent due to the existence of numerous synonyms for the same species or multiple species with the same name, and overlapping of morphological characteristics. Cytogenetic studies in Setaria can be useful for taxonomic and evolutionary studies as well as for applications in breeding. Thus, this study is aimed at locating 45S and 5S rDNA sites through fluorescent in situ hybridization (FISH) in Setaria italica, Setaria viridis and Setaria sphacelata (Schumacher, 1827) Stapf, Hubbard, Moss, 1929 cultivars (cvs.) Narok and Nandi. Setaria italica and Setaria viridis have 18 chromosomes with karyotype formulas 6m + 3sm and 9m, respectively. The location of 45S and 5S rDNA for these species was in different chromosome pairs among the evaluated species. Setaria viridis presented a more symmetrical karyotype, strengthening the ancestral relationship with Setaria italica. Setaria sphacelata cvs. Narok and Nandi have 36 chromosomes, and karyotype formulas 11m+7sm and 16m+2sm, respectively. The 45S rDNA signals for both cultivars were also observed in distinct chromosome pairs; however chromosomes bearing 5S rDNA are conserved. Karyotypic variations found among the studied species are evidence of chromosomal rearrangements.

Genomic homeology between Pennisetum purpureum and Pennisetum glaucum (Poaceae).

The genus Pennisetum (Richard, 1805) includes two economically important tropical forage plants: Pennisetum purpureum (Schumacher, 1827) (elephant grass), with 2n = 4x = 28 chromosomes and genomes A'A'BB, and Pennisetum glaucum (Linnaeus, 1753) (pearl millet), with 2n = 2x = 14 chromosomes and genomes AA. The genetic proximity between them allows hybrids to be obtained (2n = 3x = 21) that yield forage of higher quality in relation to the parents. The study of genomic relationships provides subsidies for the knowledge about phylogenetic relations and evolution, and is useful in breeding programs seeking gene introgression. Concerning elephant grass and pearl millet, the homeology between the genomes A and A', and between these and the genome B, has been reported by conventional cytogenetic techniques. The objective of the present study was to demonstrate the degree of homeology between these genomes by means of genomic in situ hybridization (GISH). The results confirmed the homeology between the genomes A of pearl millet and A'B of elephant grass, and showed that there are differences in the distribution and proportion of homologous regions after hybridization. Discussion regarding the evolutionary origin of P. purpureum and P. glaucum was also included.

Cytotoxic and phytotoxic effects of the main chemical components of spent pot-liner: a comparative approach.

Spent pot-liner (SPL) is a hazardous solid waste produced by the aluminum industry. Although its composition may vary, fluoride and cyanide salts as well as aluminum are predominant components. A seed-germination and root-elongation test was performed with Lactuca sativa seeds as a test system. SPL induced decrease of seed germination rate and root elongation. The concentration of 26.5g/L SPL was established from a regression curve as the IC50 (inhibition concentration 50%). Through chemical analyses, the concentrations of fluoride, cyanide and aluminum in SPL solutions of 26.5g/L (IC50), 39.75g/L (1.5IC50) and 13.25g/L (0.5IC50) were determined. Further, a cell-cycle test was conducted with root tips of L. sativa exposed to these same SPL solutions. All test chemicals presented toxic effects on meristematic cells of L. sativa. Aluminum was identified as the SPL component mainly responsible for reduction of the mitotic index. Chromosomal alterations resulted from the interactions among the three main chemical components of SPL, without a clear predominantly responsible agent. Induction of condensed nuclei was mainly due to effects of aluminum and fluoride, and may serve as an indicator of induced cell death.

Adaptive leaf anatomical characteristics of Brachiaria ruziziensis (Poaceae) genotypes in different environments / Características anatômicas foliares adaptativas de genótipos de Brachiaria ruziziensis (Poaceae) em diferentes ambientes

Studies on morphological plasticity are extremely important to plant breeding seeking to optimize genotype interactions with their environments. The present study evaluated 38 genotypes of Brachiaria ruziziensis cultivated during the rainy (October to March) and dry (April to September) seasons and examined variations in their anatomical characteristics. Fully expanded leaves were collected and fixed in FAA70 (formaldehyde: acetic acid: ethyl alcohol) for 72h and subsequently stored in 70% alcohol. Median sections of leaf blades were examined and evaluated according to traditional plant anatomical procedures. Genotypes 15, 27, 40, 53, and 90 demonstrated interactions with the environment in five of the anatomical characteristics evaluated; genotypes 1 and 33 presented no plasticity in any of the characteristics examined. Except for the thickness of abaxial (ABT) and adaxial (ADT) epidermal surfaces, the genotypes showed different behaviors for all the other characteristics evaluated: mesophyll thickness (MT), xylem diameter (XD), distance between bundles (DB), number of bundles (NB), number of bulliform cells (NBC), sclerenchyma tissue area (%L), and bulliform cell area (ABC).

O estudo da plasticidade morfológica é de grande importância para os programas de melhoramento que desejam obter materiais promissores quando apresentam a interação genótipo com ambiente. O objetivo deste trabalho foi avaliar 38 genótipos de Brachiaria ruziziensis cultivados na época das águas (outubro a março) e da seca (abril a setembro) e observar variações nas características anatômicas nas diferentes estações. Folhas completamente expandidas foram coletadas e fixadas em FAA70 por 72h e armazenadas em álcool 70%. Amostras foliares na parte mediana foram avaliadas de acordo com os procedimentos usuais de anatomia vegetal. Os genótipos 15, 27, 40, 53, e o 90 apresentaram interação com o ambiente em cinco características avaliadas. Os genótipos 1 e 33 não apresentaram plasticidade para nenhuma mensuração. Exceto para espessura das faces abaxial (EBA) e abaxial (EDA) da epiderme, os genótipos apresentaram comportamento diferente para as demais características avaliadas: espessura do mesofilo (EM), diâmetro do xilema (DX), distancia entre feixes (DF), número de feixes (NF) e de células buliformes (NCB), área do tecido esclerenquimático (%L) e das células buliforme (ACB).

Effects of Spent Pot Liner on mitotic activity and nuclear DNA content in meristematic cells of Allium cepa.

Industrial waste usually contains complex mixtures of mutagenic chemicals. Spent Pot Liner (SPL) is a complex solid waste from the aluminum industry, which is composed of organics, fluoride salts, inorganic cyanides, metals, and sodium. Due to the toxicity of these compounds, this study sought to use cytogenetics and flow cytometry to assess the effects of SPL on cell cycle parameters and DNA content in meristematic cells of Allium cepa. Three concentrations of leachates from SPL-soil mixtures were used for the study: 0, 10, and 25%. Roots were collected and analyzed after 4, 8, 12, 24, and 36 h of exposure to the above SPL leachates. The results showed an overall mitodepressive effect accompanied by an increased percentage of condensed nuclei and genomic instability as evidenced by the presence of cellular/chromosomal abnormalities. Terminal deoxynucleotidyl transferase dUTP nick end labeling revealed nuclei with fragmented DNA, a marker of programmed cell death. This study also addressed the question of reversibility of the effects of SPL and found that 36 h of exposure to 25% SPL seemed to be the point at which the effects on the induction of apoptosis became irreversible.

Application of Comet assay to assess the effects of white bean meal on DNA of human lymphocytes

This study was conducted to evaluate the potential induction of genotoxic effects of white bean flour using the Comet assay. The test was conducted with human lymphocytes present in whole blood immediately after collection, by incubation with white bean flour in three concentrations (3.92, 9.52 and 18.18 mg/mL) at 37 ºC for 4 h followed by preparation of slides. Samples were considered positive (above 20% damage) when the damage observed to cellular DNA was higher than the negative control. No genotoxic potential was found at the doses tested. However, it would be premature to suggest absence of risk to human health of DNA damage since the exposure of cells to the extract was restricted to four hours rather than a whole cell cycle. Additionally, further information on toxicology should be obtained in future studies.

Este estudo foi realizado para avaliar o potencial de indução de efeitos genotóxicos da farinha de feijão branco utilizando o teste do Cometa. O ensaio foi realizado com linfócitos humanos presentes no sangue imediatamente após a coleta, por incubação com farinha de feijão branco em três concentrações (3,92, 9,52 e 18,18 mg/mL) a 37 ºC por 4 h, seguida de preparação das lâminas. As amostras foram consideradas positivas (acima de 20% de dano), quando os danos observados no DNA celular foram maiores do que o controle negativo. Verificou-se que as doses testadas não mostraram potencial genotóxico. No entanto, seria prematuro fazer recomendações sobre o padrão de riscos para a saúde humana resultantes de danos ao DNA já que exposição das células ao extrato foi restrito ao período de quatro horas e não durante um ciclo celular completo. Além disso, outras informações sobre a toxicologia devem ser obtidas no futuro.

Characterization of interphase nuclei from triploid hybrids between Pennisetum purpureum Schumach and Pennisetum glaucum (L.) R. Br. / Caracterização de núcleos interfásicos de híbridos tríploides entre Pennisetum purpureum Schumach e Pennisetum glaucum (L.) R. Br.

This study evaluated the structure and the volume of interphase nuclei from root meristems of the genotypes of napier grass (Pennisetum purpureum), pearl millet (Pennisetum glaucum) and hybrids resultant of such breeding. In napier grass, nuclei were areticulate. Both pearl millet and the triploid hybrid had semi-reticulate nuclei; also, the hybrid presented a small proportion (6 percent) of areticulate nuclei. Pearl millet had the highest averages of nuclear dimensions, such as volume, diameter and radius, followed by the interspecific hybrid and napier grass. There was no intraspecific variation for the type of nuclear structure, which indicates this feature is important for cytotaxonomic studies involving the genus Pennisetum. Results demonstrated that chromatin organization in these nuclei was influenced by the number and size of chromosomes, affecting the nucleus volume in the analyzed taxa.

Avaliou-se, neste estudo, a estrutura e o volume de núcleos interfásicos de meristemas radiculares de genótipos do capim-elefante (Pennisetum purpureum), milheto (Pennisetum glaucum) e do híbrido resultante deste cruzamento. O capim-elefante apresentou núcleos do tipo arreticulado. Tanto no milheto como no híbrido triplóide os núcleos apresentaram-se semi-reticulados, sendo que, no híbrido, foi observada uma pequena proporção (6 por cento) de núcleos arreticulados. As maiores médias para as dimensões nucleares como volume nuclear, diâmetro e raio foram obtidas para o milheto, seguidas do híbrido interespecífico e capim-elefante. Não houve variação intraespecífica para tipo de estrutura nuclear indicando que essa característica tem relevância para estudos citotaxonômicos no gênero Pennisetum. Os resultados demonstraram que a organização da cromatina nesses núcleos foi influenciada pelo número e tamanho dos cromossomos e essa afetou o volume nuclear dos táxons analisados.

Protoplast production from napier grass and pearl millet triploid hybrids / Obtenção de protoplastos de híbridos triplóides entre o capim-elefante e milheto

The objective of this work was to obtain protoplasts from napier grass and pearl millet triploid hybrids as a basis for future studies on chromosomal duplication. Explants were taken from mesophyll of in vitro- and in vivo-cultured plants or from calli of two triploid hybrids (H1 and H2), which were treated with enzymatic solutions containing different concentrations of cellulase R-10 (0.5, 1.0, 1.5 and 2.0 percent) with an additional 0.2 percent macerozyme and 0.1 percent driselase or 1.0 percent pectolyase Y-23 and 0.5 percent hemicellulase. Enzymatic digestion was monitored once every hour for five hours. Protoplasts were obtained from in vitro and in vivo leaflets of both triploid hybrids, and in vitro leaflets were the best explant sources. The quantity of produced protoplasts varied according to the hybrid, the enzymatic solution and the treatment time.

Objetivou-se, neste trabalho, a obtenção de protoplastos de híbridos triplóides entre o capim-elefante e o milheto como base para futuros trabalhos de duplicação cromossômica. Foram utilizados explantes de mesofilo de plantas cultivadas in vitro e in vivo, ou de calos de dois híbridos triplóides (H1 e H2), os quais foram tratados com soluções enzimáticas em diferentes concentrações da enzima celulase R-10 (0,5; 1,0; 1,5 e 2,0 por cento), acrescidas de 0,2 por cento macerozyme e 0,1 por cento driselase ou 1,0 por cento pectolyase Y-23 e 0,5 por cento hemicelulase. A digestão enzimática foi monitorada a cada hora durante 5 horas. Obtiveram-se protoplastos a partir de folhas in vitro e in vivo dos dois híbridos triplóides, sendo as folhas in vitro as melhores fontes de explante. A quantidade de protoplastos variou em função do híbrido, da solução enzimática e do tempo de tratamento.

Pólen 2n e mecanismos meióticos de formação em Solanum commersonii ssp / 2n pollen and meiotic mechanisms of Formation in Solanum commersonii ssp

Solanum commersonii subsp. commersonii Dun. e Solanum commersonii subsp. malmeanum Bitt. (2n=2x=24 - 1EBN) não podem ser diretamente cruzadas com a batata cultivada de S. tuberosum subsp. tuberosum L. (2n=4x=48 -4 EBN) em razão da barreira da ploidia e do número de equilíbrio do endosperma (EBN). No entanto, a produção de pólen não reduzido por S. commersonii ssp. (2x=2n=24 - 1EBN), permite o cruzamento desta com dihaplóides de batata e com as espécies diplóides S. phureja Juz. & Bukasov. e S. chacoense Bitt., que possuem 2 EBN. Conduziu-se este trabalho, com o objetivo de determinar o mecanismo meiótico e a freqüência dos grãos de pólen não reduzidos em cinco clones de Solanum commersonii ssp. Foram avaliados três clones de S. commersonii subsp. commersonii (SCC) e dois de S. commersonii subsp. malmeanum (SCM). Nas avaliações, duzentos grãos de pólen viáveis foram corados (carmim acético 2%) e mensurados, sendo considerados não reduzidos aqueles com o diâmetro igual ou superior a 25 µm. O clone Solanum commersonii subsp. commersonii Dun. (SCC 100) produz 26.1% de polen não reduzido, por fusos fundidos e fusos paralelos, o qual pode ser utilizado em programas de melhoramento.

Solanum commersonii subsp. commersonii Dun. and Solanum commersonii subsp. malmeanum Bitt. (2n=2x=24 -1EBN) cannot be directly crossed with the cultivated Solanum tuberosum subsp. Tuberosum L. (2n=4x=48 - 4EBN) species due to ploidy barriers and endosperm balance number (EBN). However, non-reduced pollen production by the wild species S. commersonii ssp. (2x=2n=24 -1EBN), allows the crossing of these species with dihaploids of potato and with the diploids species, S. phureja Juz. & Bukasov. and S. chacoense Bitt., all with 2EBN. The aim of this work was to determine the meiotic mechanism and frequency of non-reduced pollen production of five clones of S. commersonii ssp. Three different clones of S. commersonii subsp. commersonii Dun. (SCC) and two of S. commersonii subsp. malmeanum Bitt. (SCM) were evaluated. In the evaluation, two hundred viable pollen grains were stained (2% acetic carmin) and measured. The pollen grains measuring 25 µm in diameter or more were considered nonreduced. The clone Solanum commersonii subsp. commersonii Dun. (SCC 100) produces 26.1% of non-reduced pollen grains caused by parallel spindles and fusion spindles, which may be used in breeding program.

Mitodepressive and clastogenic effects of aqueous extracts of the lichens Myelochroa lindmanii and Canoparmelia texana (Lecanorales, Parmeliaceae) on meristematic cells in plant bioassays

The cytotoxicity effect of aqueous extracts of the lichens species Myelochroa lindmanii and Canoparmelia texana (Lecanorales, Parmeliaceae) were evaluated using meristematic cells of lettuce (Lactuca sativa) and maize (Zea mays). The seasonal effect was also evaluated. Extracts of M. lindmanii and C. texana inhibited root growth and/or percentage germination, possibly due to alterations in the cell cycle. The M. lindmanii extract showed anti-mitotic effects and blocked the cell cycle in metaphase so that c-mitosis and cells with chromosome duplication were produced. The C. texana extract appeared to hinder cell division, increasing the number of interphase cells. In addition, both extracts caused an increase in percentage of cell death. Clastogenic effects were also observed, such as sticky chromosomes, bridges, fragments and later segregation. Both lichen species are thus potential sources of biologically active substances with possible applications in biology, medicine and agronomy.