Phylogeny of Lantana, Lippia, and related genera (Lantaneae: Verbenaceae).

PREMISE: Lantana and Lippia (Verbenaceae) are two large Linnean genera whose classification has been based on associated fruit traits: fleshy vs. dry fruits and one vs. two seed-bearing units. We reconstruct evolutionary relationships and the evolution of the two fruit traits to test the validity of these traits for classification. METHODS: Previous studies of plastid DNA sequences provided limited resolution for this group. Consequently, seven nuclear loci, including ITS, ETS, and five PPR loci, were sequenced for 88 accessions of the Lantana/Lippia clade and three outgroups. RESULTS: Neither Lantana nor Lippia is monophyletic. Burroughsia, Nashia, Phyla, and several Aloysia species are included within the clade comprising Lantana and Lippia. We provide a hypothesis for fruit evolution and biogeographic history in the group and their relevance for classification. CONCLUSIONS: Fleshy fruits evolved multiple times in the Lantana/Lippia clade and thus are not suitable taxonomic characters. Several sections of Lantana and Lippia and the small genera are monophyletic, but Lippia section Zappania is broadly paraphyletic, making circumscription of genera difficult. Lippia sect. Rhodolippia is a polyphyletic group characterized by convergence in showy bracts. Species of Lantana sect. Sarcolippia, previously transferred to Lippia, are not monophyletic. The clade originated and diversified in South America, with at least four expansions into both Central America and the Caribbean and two to Africa. The types species of Lantana and Lippia occur in small sister clades, rendering any taxonomy that retains either genus similar to its current circumscription impossible.

Genetic relationships and polyploid origins in the Lippia alba complex.

PREMISE: Plant genomes vary in size and complexity due in part to polyploidization. Latitudinal analyses of polyploidy are biased toward floras of temperate regions, with much less research done in the tropics. Lippia alba has been described as a tropical polyploid complex with diploid, triploid, tetraploid, and hexaploid accessions. However, no data regarding relationships among the ploidal levels and their origins have been reported. Our goals are to clarify the relationships among accessions of Lippia alba and the origins of each ploidal level. METHODS: We investigated 98 samples representing all five geographical regions of Brazil and all ploidal levels using microsatellite (SSR) allelic variation and DNA sequences of ITS and trnL-F. Nine morphological structures were analyzed from 33 herbarium samples, and the chemical compounds of 78 accessions were analyzed by GC-MS. RESULTS: Genetic distance analysis, the alignment block pattern, as well as RAxML and Bayesian trees showed that accessions grouped by ploidal level. The triploids form a well-defined group that originated from a single group of diploids. The tetraploids and hexaploid grouped together in SSR and trnL-F analyses. The recovered groups agree with chemical data and morphology. CONCLUSIONS: The accessions grouped by ploidal level. Only one origin of triploids from a single group of diploids was observed. The tetraploid origin is uncertain; however, it appears to have contributed to the origin of the hexaploid. This framework reveals linkages among the ploidal levels, providing new insights into the evolution of a polyploid complex of tropical plants.

Isolated perfused udder model for transcriptome analysis in response to Streptococcus agalactiae.

This study aimed to evaluate the transcriptional changes occurring in isolated perfused mammary alveolar tissue in response to inoculation with S. agalactiae and to identify the most affected biological functions and pathways after 3 h. Four udders taken at slaughter from cows with healthy mammary gland were perfused ex situ with warmed and gassed Tyrode's solution. Mammary alveolar tissue samples were taken from the left fore and rear quarters (IQ-inoculated quarters) before inoculation (hour 0) and at 3 h post inoculation (hpi) and at the same times from control right fore and rear quarters (not inoculated: NIQ). A total of 1756 differentially expressed genes (DEGs) were identified between IQ and NIQ at 3 hpi using edgeR package. Within this set of DEGs, 952 were up regulated and mainly involved with innate immune response and inflammatory response, e.g., CD14, CCL5, TLR2, IL-8, SAA3, as well as in transcriptional regulation such as FOS, STAT3 and NFKBIA. Genes down-regulated (804) included those involved with lipid synthesis e.g., APOC2, SCD, FABP3 and FABP4. The most affected pathways were chemokine signaling, Wnt signaling and complement and coagulation cascades, which likely reflects the early stage response of mammary tissue to S. agalactiae infection. No significant gene expression changes were detected by RNA-Seq in the others contrasts. Real time-PCR confirmed the increase in mRNA abundance of immune-related genes: TLR2, TLR4, IL-1ß, and IL-10 at 3 hpi between IQ and NIQ. The expression profiles of Casp1 and Bax for any contrasts were unaffected whereas Bcl2 was increased in IQ, which suggests no induction of apoptosis during the first hours after infection. Results provided novel information regarding the early functional pathways and gene network that orchestrate innate immune responses to S. agalactiae infection. This knowledge could contribute to new strategies to enhance resistance to this disease, such as genomic selection.

rDNA mapping, heterochromatin characterization and AT/GC content of Agapanthus africanus (L.) Hoffmanns (Agapanthaceae).

Agapanthus (Agapanthaceae) has 10 species described. However, most taxonomists differ respect to this number because the great phenotypic plasticity of the species. The cytogenetic has been an important tool to aid the plant taxon identification, and to date, all taxa of Agapanthus L'Héritier studied cytologically, presented 2n = 30. Although the species possess large chromosomes, the group is karyologically little explored. This work aimed to increase the cytogenetic knowledge of Agapanthus africanus (L.) Hoffmanns by utilization of chromosome banding techniques with DAPI / CMA3 and Fluorescent in situ Hybridization (FISH). In addition, flow cytometry was used for determination of DNA content and the percentage of AT / GC nitrogenous bases. Plants studied showed 2n = 30 chromosomes, ranging from 4.34 - 8.55 µm, with the karyotype formulae (KF) = 10m + 5sm. Through FISH, one 45S rDNA signal was observed proximally to centromere of the chromosome 7, while for 5S rDNA sites we observed one signal proximally to centromere of chromosome 9. The 2C DNA content estimated for the species was 2C = 24.4 with 59% of AT and 41% of GC. Our data allowed important upgrade for biology and cytotaxonomy of Agapanthus africanus (L.) Hoffmanns.

Lippia alba (Verbenaceae): A new tropical autopolyploid complex?

• Premise of the study: Tropical regions have high species diversity, and polyploidization is a major mechanism of speciation in plants. However, few cases of natural polyploidy have been reported in tropical regions. Lippia alba, is a tropical, aromatic shrub with a wide distribution, extensive morphological plasticity, and several chemotypes. The species has long been recognized as a diploid with 2n = 30 chromosomes. Recently, two variations in chromosome number (2n = 60; 2n = 12-60) have been reported, suggesting the occurrence of polyploidy within the species.• Methods: Flow cytometry was used to investigate the genome size in 106 accessions from 14 Brazilian States. Conventional and molecular cytogenetic techniques and pollen viability analysis were employed to characterize each chromosome number observed.• Key results: The DNA 1C-value varied from 1.17 to 3.45 pg, showing a large variation in genome size. Five distinct chromosome numbers were observed (2n = 30, 38, 45, 60, 90); three are cytogenetically described here for the first time. The 5S rDNA signals varied proportionally according to each chromosome number, but 45S rDNA sites did not. High rates of meiotic irregularity were observed, mainly in cytotypes with higher chromosome numbers.• Conclusions: The data provide new support for the occurrence of a polyploid series in Lippia alba. We provide a hypothesis for how this complex may have arisen. Other cryptic polyploid complexes may remain undiscovered in tropical regions.

The effect of the essential oils from five different Lippia species on the viability of tumor cell lines

Several Lippia species have been used in folk medicine mainly for gastrointestinal and respiratory diseases. Their biological properties have been partially associated to the terpenoids found in their essential oils. According to the World Health Organization, cancer is the leading cause of death worldwide and is described as a complex group of diseases with several hallmarks. One of its acceptable defining features is the cell proliferation beyond their boundaries forming the tumors. Importantly, some drugs currently available were discovered by the investigation of plant secondary metabolites. Thus, this study aimed to evaluate in vitro cytotoxic effect of the essential oils extracted from five Lippia species against tumor cell lines. The results indicated that mouse colon carcinoma CT26.WT cell line viability was significantly reduced showing an IC50 of 19.05, 30.20 and 36.30 µg/ml when treated with the essential oils of L. sidoides, L. salviifolia and L. rotundifolia, respectively. Human lung carcinoma A549 cell line also had a compromised viability to the action of L. alba carvone chemotype essential oil. The tested essential oils did not compromise viability of the normal cell line CHO. These finds suggest that the studied Lippia essential oils might be good candidates for further in-depth studies.

Relationship between pollen morphology and chromosome numbers in Brazilian species of Lippia L. (Verbenaceae).

The pollen morphology and exine structure of 17 species of Lippia L. were investigated in this work using light and scanning electron microscopy. Among the species studied, 14 showed tricolporate pollen grains, two had tri- and tetracolporate pollen grains and a single species exhibited, only tetracolporate pollen. The amb ranged from triangular to square, and the shape varied from oblate-spheroidal to prolate-spheroidal. Three different types of exine ornamentation were observed: psilate, scabrate and perforate. In addition to morphological data, we found positive association between the chromosome numbers and size of pollen grains, and also between the length and width of the colpi. The results indicate that the characteristics of pollen grains in Lippia may be used as an additional taxonomic character of the genus.

Karyological studies in Brazilian species of Lippia L. (Verbenaceae).

The genus Lippia (Verbenaceae) comprise around 160 species spread out mainly in South and Central Americas with few African species, some of them with potential medicinal use. Brazil is one the most important centers of diversity with approximately 75% of the species described so far. Innumerous species are endemic and poorly studied especially at a cytological level. Here, chromosomal length, karyomorphology and chromosome asymmetry of twelve Brazilian species of Lippia were evaluated [L. alba (Miller) N.E.Brown, L. diamantinensis Glaz., L. florida Cham., L. hermanioides Cham., L. lacunosa Mart. & Schauer, L. lupulina Cham., L. pohliana Schauer, L. pseudothea (St. Hil) Schauer, L. rosella Moldenke, L. rotundifolia Cham., L. rubella Moldenke and L. sidoides Cham.]. The analysis suggested that the genus has a variable chromosome number (from 2n = 20 to 2n = 56) originated by dysploidy and polyploidy. This is the first description of chromosome morphology for 11 of the 12 Lippia species studied.

Oil production at different stages of leaf development in Lippia alba

The aim of this work was to analyze terpene oil production and terpene synthases (TPS) gene expression from leaves at different developmental stages of different chemotypes of Lippia alba (Mill.) N.E. Br. ex Britton & P. Wilson, Verbenaceae. Hydro-distilled essential oil were used for chemical analysis and gene expression of three monoterpene synthase genes called LaTPS12, LaTPS23 and LaTPS25 were used for analyses of gene expression associated to oil production. The putative genes were associated to TPS-b gene class. Semi-quantitative PCR and quantitative PCR (qPCR) analysis were used to investigate the expression profile of those three putative genes in different leaf stages and different chemotypes. Additionally, total oil production and gene expression of putative TPS genes cloned from L. alba chemotype linalool were evaluated at different stages of leaf development. The expression level of those three genes was higher when the highest oil production was observed, mainly in young leaves at the fourth nodal segment for all evaluated chemotypes. Total oil production was higher at leaves that had unopened trichomes. We also observed that the 1mM of MeJA treatment increased the gene expression in all chemotypes after 24 h elicitation.

Micropropagation, antinociceptive and antioxidant activities of extracts of Verbena litoralis Kunth (Verbenaceae).

This work describes an efficient micropropagation protocol for Verbena litoralis and the study of the antinociceptive and antioxidant activities in extracts of this species. For the establishment in vitro, surface-sterilization procedures and PVPP showed high efficiency in fungal-bacterial contamination and phenol oxidation controls. Nodal segments cultivation in MS medium supplemented with 6-benzyladenine (7.5 µM)/α-naphthaleneacetic acid (NAA; 0.005 µM) induced multiple shoots. Elongated shoots were rooted with IAA (0.2 µM). Acclimatization rates were elevated and the plants showed the typical features of this species. The hexanic fraction (HF) of powdered leaves presented a radical scavenging activity with IC(50) = 169.3 µg mL(-1). HF showed a non-dose dependent analgesic activity in the writhing test; its antinociceptive activity in the hot plate test was restricted to 500 mg kg(-1), which is the highest dose. The results of this study showed the potential of tissue culture on conservation and large scale multiplication and confirmed the traditional folk medicine use of V. litoralis.

Karyotype analysis, DNA content and molecular screening in Lippia alba (Verbenaceae)

Cytogenetic analyses, of pollen viability, nuclear DNA content and RAPD markers were employed to study three chemotypes of Lippia alba (Mill.) (Verbenaceae) in order to understand the genetic variation among them. Different ploidy levels and mixoploid individuals were observed. This work comprises the first report of different chromosome numbers (cytotypes) in L. alba. The chromosome numbers of La2-carvone and La3-linalool chemotypes suggested that they are polyploids. Flow cytometric analysis showed an increase of nuclear DNA content that was not directly proportional to ploidy level variation. A cluster analysis based on RAPD markers revealed that La3-linalool shares genetic markers with La1-citral and La2-carvone. The analysis showed that the majority of genetic variation of La3-linalool could be a consequence of ixoploidy. ur data indicates that sexual reproduction aong those three chemotypes is unlikely and suggests the beginning of reproductive isolation. The results demonstrated that chromosome analysis, nuclear DNA content estimation and RAPD markers constitute excellent tools for detecting genetic variation among L. alba chemotypes.

Análises citogenéticas, de viabilidade do pólen, do conteúdo de DNA nuclear e marcadores RAPD foram empregadas no estudo de três quimiotipos de Lippia alba (Mill.) (Verbenaceae) visando contribuir para o entendimento da variação genética entre os mesmos. Diferentes níveis de ploidia e indivíduos mixoploides foram observados. Este trabalho compreende o primeiro relato de diferentes números cromossômicos (citótipos) em L. alba. Os números cromossômicos dos quimiotipos La2-carvona e La3-linalol sugere que eles seja poliploides. A análise da citometria de fluxo mostrou um aumento do conteúdo de DNA nuclear que não foi diretamente proporcional à variação no nível de ploidia. A análise de agrupamento baseada nos marcadores RAPD demonstrou que La3-linalol compartilha marcadores genéticos com La1-citral e La2-carvona. A análise mostrou que a maior parte da variação genética de La3-linalol pode ser consequência da mixoploidia. Nossos dados indicam que a reprodução sexual entre os três quimiotipos parece improvável, sugerindo o início de isolamento reprodutivo. Os resultados demonstraram que a análise cromossômica, a quantificação do DNA nuclear estimado e os marcadores RAPD constituem excelentes ferramentas para detecção de variação genética entre quimiotipos de L. alba.

Karyotype analysis, DNA content and molecular screening in Lippia alba (Verbenaceae).

Cytogenetic analyses, of pollen viability, nuclear DNA content and RAPD markers were employed to study three chemotypes of Lippia alba (Mill.) (Verbenaceae) in order to understand the genetic variation among them. Different ploidy levels and mixoploid individuals were observed. This work comprises the first report of different chromosome numbers (cytotypes) in L. alba. The chromosome numbers of La2-carvone and La3-linalool chemotypes suggested that they are polyploids. Flow cytometric analysis showed an increase of nuclear DNA content that was not directly proportional to ploidy level variation. A cluster analysis based on RAPD markers revealed that La3-linalool shares genetic markers with La1-citral and La2-carvone. The analysis showed that the majority of genetic variation of La3-linalool could be a consequence of ixoploidy. ur data indicates that sexual reproduction aong those three chemotypes is unlikely and suggests the beginning of reproductive isolation. The results demonstrated that chromosome analysis, nuclear DNA content estimation and RAPD markers constitute excellent tools for detecting genetic variation among L. alba chemotypes.

Cytotoxic and genotoxic activity of Achillea millefolium L., Asteraceae, aqueous extracts

The yarrow, Achillea millefoilum L. is an important species of Asteraceae family with common utilization in folk medicine of many countries. The aim of this study was to investigate the cytotoxic and genotoxic effects of aqueous extracts from yarrow leaves on Lactuca sativa (lettuce) root tip meristem cells by cytogenetic studies, since studies of this nature do not exist for the yarrow extracts. For this, lettuce seeds were treated for 72 h with different concentrations of yarrow aqueous extracts (5, 10, 20 and 30 mg/mL). The percentage of germination, root development and cellular behavior were analyzed and the results showed that the highest concentration of aqueous extracts reduced the mitotic index (MI), the seed germination and the root development of L. sativa. More yet, the extracts also induced chromosome aberrations (CA) and cellular death in the roots cells of L. sativa indicating precaution in the therapeutic use of A. millefolium and reinforcing the utilization of L. sativa in the screening of cytogenotoxic substances once that this species showed a good sensibility to the extract application.

Identification of botryticidal proteins with similarity to NBS-LRR proteins in rosemary pepper (Lippia sidoides Cham.) flowers.

Heavy agricultural losses are closely related to attacks by insect-pests and phytopathogens such as bacteria and fungi. Among them, the fungus Botrytis cinerea can cause gray mold in more than 200 different species of plants, and is considered a challenging problem for agribusiness. Fungicides are commonly used to control this pathogen because they are fast-working and easy to apply. However, the continuous use of fungicides may promote the selection of resistant fungi and can also cause profound contamination in ecosystems. Aiming to find alternative strategies to solve these problems, several studies have focused on searching for plant proteins and peptides with antifungal activities (AFPs). With this in mind, this report shows the isolation and characterization of two novels antifungal proteins from flowers of rosemary pepper (Lippia sidoides Cham.) with 10 and 15 kDa. Isolation was performed by using an Octyl-Sepharose hydrophobic column. In vitro bioassays indicated that isolated proteins were able to inhibit B. cinerea development, but were not effective against all bacteria tested. Moreover, N-termini sequences indicate that both proteins showed sequence homology with NBS-LRR R proteins with a lower molecular mass, suggesting possible protein fragmentation. Data reported here could help in the development of biotechnological products for crop protection against phytopathogenic fungi in the near future.

Botanical aspects of Heteropterys umbellata (Malpighiaceae): a cytological and palynological approach.

The genus Heteropterys is one of the major genera in Malpighiaceae. However, few cytological and palynological studies were reported. The present work described for the first time the chromosome number, heterochromatin pattern, meiotic behavior, pollen viability and palynological aspects of Heteropterys ubellata, a very spread species. One large Brazilian population was evaluated using conventional techniques for meiotic studies and acetolyse to access the pollen morphology. The species showed 2n = 20 chromosomes, normal meiotic development and viable pollens. Great blocks of heterochromatin were observed around the centromers. DAPI staining was positive for centroeric heterochroatin, hile CMA3 ark as observed just at terinal regions of one pair of hoologues chromosomes. This result and the presence of one chromosome pair attached to the nucleoli during the pachytene and diakinesis suggested the presence of only one pair of NORs. Palynological analysis revealed that pollen grains are apolar, 6 porate and with colpoids associated to all pores. The pollen content was positive for the starch test, and the exine was rugulate with little psilate regions.

Botanical aspects of Heteropterys umbellata (Malpighiaceae): a cytological and palynological approach

The genus Heteropterys is one of the major genera in Malpighiaceae. However, few cytological and palynological studies were reported. The present work described for the first time the chromosome number, heterochromatin pattern, meiotic behavior, pollen viability and palynological aspects of Heteropterys ubellata, a very spread species. One large Brazilian population was evaluated using conventional techniques for meiotic studies and acetolyse to access the pollen morphology. The species showed 2n = 20 chromosomes, normal meiotic development and viable pollens. Great blocks of heterochromatin were observed around the centromers. DAPI staining was positive for centroeric heterochroatin, hile CMA3 ark as observed just at terinal regions of one pair of hoologues chromosomes. This result and the presence of one chromosome pair attached to the nucleoli during the pachytene and diakinesis suggested the presence of only one pair of NORs. Palynological analysis revealed that pollen grains are apolar, 6 porate and with colpoids associated to all pores. The pollen content was positive for the starch test, and the exine was rugulate with little psilate regions.

O gênero Heteropterys é um dos maiores gêneros entre as Malpighiaceae. Entretanto, poucos estudos citológicos e palinológicos foram relatados. O presente trabalho descreveu pela primeira vez o número cromossômico, o padrão heterocromático, o comportamento meiótico, a viabilidade polínica e aspectos palinológicos de Heteropterys umbellata, uma espécie muito disseminada. Uma grande população brasileira foi estudada utilizando técnicas convencionais para o estudo de cromossomos meióticos e acetólise para acessar a morfologia polínica. A espécie apresentou 2n = 20 cromossomos, desenvolvimento meiótico normal e pólens viáveis. Grandes blocos de heterocromatina foram observados ao redor dos cen-trômeros. A coloração com DAPI foi positiva para a hetero-cromatina centromérica, enquanto marcas com CMA3 foram observadas somente em um sítio terminal de um par de cromossomos homólogos. Este resultado e a presença de um par de cromossomos associados a um nucléolo durante o paquíteno e diacinese sugerem a presença de somente um par de RONs. A análise palinológica revelou que os grãos de pólen são apolar, com 6 poros e colpóides associados a todos eles. O conteúdo polínico foi positivo para o teste de amido e a exina mostrou-se rugulada com poucas regiões psiladas.

Chemical composition of the essential oil and hexanic fraction of Lippia and Lantana species / Composição química do óleo essencial e fração hexânica de espécies de Lippia e Lantana

A comparison between two extraction approaches of volatiles compounds from six species of Verbenaceae collected at Serra do Cipó, Minas Gerais, Brazil was done. The essential oil and hexanic fraction of leaves from two Lantana and four Lippia species collected in two different seasons were analyzed by GC/MS. Among various identified compounds from both extraction methods the majority of species showed major amounts of β-caryophyllene followed by germacrene D, bicyclogermacrene and α-pinene. Few differences were observed between the composition of essential oil and the hexanic fraction regarding the two studied genera. These results suggest that the analysis of hexanic fraction can be used, as an alternative way, to analyze the volatile compounds of the essential oil.

Foi realizada a comparação entre dois métodos de extração dos compostos voláteis obtidos de seis espécies de Verbenaceae, coletadas na Serra do Cipó, Minas Gerais, Brasil. Os óleos essenciais e as frações hexanica obtidos das folhas de duas espécies de Lantana e quatro espécies de Lippia, coletadas em duas diferentes estações do ano, foram analisados por CG/EM. Grande número de constituintes foi identificado nas amostras oriundas dos dois métodos de extração e o componente majoritário para a maior parte das espécies foi o β-cariofileno, seguido pelo germacreno D, biciclogermacreno e α-pineno. Para os dois gêneros estudados, foram observadas pequenas diferenças na composição do óleo essencial e fração hexânica. Esses resultados sugerem que a análise da fração hexânica pode ser usada para identificar os componentes voláteis majoritários dessas espécies, além de ser uma técnica alternativa para a análise dos compostos voláteis presentes no óleo essencial, uma vez que ambos mostraram composição similar.

Cytogenotoxicity of Cymbopogon citratus (DC) Stapf (lemon grass) aqueous extracts in vegetal test systems.

The lemon grass, Cymbopogon citratus (DC) Stapf, is an important species of Poaceae family commonly used in the folk medicine in many countries. The aim of this study was to investigate the cytotoxic and genotoxic effects of aqueous extracts from C. citratus leaves on Lactuca sativa (lettuce) root tip meristem cells by cytogenetic studies that have never been done before for lemon grass extracts. For this, lettuce seeds were treated for 72h with different concentrations of lemon grass aqueous extracts (5; 10; 20 and 30 mg/mL). The percentage of germination, root development and cellular behavior were analyzed, and the results showed that the highest concentration of aqueous extracts reduced the mitotic index, the seed germination and the root development of lettuce. The extracts have also induced chromosome aberrations and cellular death in the roots cells of L. sativa.

Cytogenotoxicity of Cymbopogon citratus (DC) Stapf (lemon grass) aqueous extracts in vegetal test systems

The lemon grass, Cymbopogon citratus (DC) Stapf, is an important species of Poaceae family commonly used in the folk medicine in many countries. The aim of this study was to investigate the cytotoxic and genotoxic effects of aqueous extracts from C. citratus leaves on Lactuca sativa (lettuce) root tip meristem cells by cytogenetic studies that have never been done before for lemon grass extracts. For this, lettuce seeds were treated for 72h with different concentrations of lemon grass aqueous extracts (5; 10; 20 and 30 mg/mL). The percentage of germination, root development and cellular behavior were analyzed, and the results showed that the highest concentration of aqueous extracts reduced the mitotic index, the seed germination and the root development of lettuce. The extracts have also induced chromosome aberrations and cellular death in the roots cells of L. sativa.

O capim-limão, Cymbopogon citratus (DC) Stapf, é uma importante espécie da família Poaceae com uma comum utilização na medicina popular em vários países. O objetivo deste estudo foi investigar os efeitos citotóxicos e genotóxicos do extrato aquoso das folhas de C. citratus em células meristemáticas de Lactuca sativa (alface) por meio de estudos citogenéticos, uma vez que estudos desta natureza não existem para extratos aquosos de capim-limão. Para isso, sementes de alface foram tratadas por 72h com diferentes concentrações de extratos aquosos feitos das folhas de capim-limão (5, 10, 20 e 30 mg/mL). O percentual de germinação, desenvolvimento radicular e o comportamento celular foram avaliados e os resultados mostraram que as concentrações mais elevadas dos extratos aquosos reduziram o índice mitótico, o percentual de germinação das sementes e desenvolvimento radicular da alface. Os extratos também induziram aberrações cromossômicas e morte celular nas células das raízes de L. sativa.

Tissue-specific silencing of homoeologs in natural populations of the recent allopolyploid Tragopogon mirus.

Recent years have seen rapid advances in our knowledge of the transcriptomic consequences of allopolyploidy, primarily through the study of polyploid crops and model systems. However, few studies have distinguished between homoeologs and between tissues, and still fewer have examined young natural allopolyploid populations of independent origin, whose parental species are still present in the same location. Here, we examined the expression of 13 homoeolog pairs in seven tissues of 10 plants of allotetraploid Tragopogon mirus from two natural populations formed by independent polyploidizations between Tragopogon dubius and Tragopogon porrifolius c. 40 generations ago. We compare these with patterns of expression in the diploid parental species from the same locality. Of the 910 assays in T. mirus, 576 (63%) showed expression of both homoeologs, 63 (7%) showed no expression of either homoeolog, 186 (20%) showed nonexpression of one homoeolog across all tissues of a plant, and 72 (8%) showed non-expression of a homoeolog in a particular tissue within a plant. We found two cases of reciprocal tissue-specific expression between homoeologs, potentially indicative of subfunctionalization. Our study shows that tissue-specific silencing, and even apparent subfunctionalization, can arise rapidly in the early generations of natural allopolyploidy.