Chemical characterization, cytotoxicity, antimicrobial and antioxidant potential of Justicia pectoralis Jacq and Croton jacobinensis Baill extracts

Abstract In this research, aqueous and ethanolic extracts from Justicia pectoralis Jacq and Croton Jacobinensis Baill were characterized. The UPLC-QTOF-MSE analysis was performed on the extracts identified, predominantly, flavonoids, tannins and acids. The extracts did not indicate toxicity in human epithelial cells. C. jacobinensis presented a concentration of phenolics 60.5% higher than J. pectoralis in all scenarios evaluated and, for both samples, the hydroalcoholic extract at 70% exhibited the best efficiency in the extraction (14501.3 and 32521.5 mg GAE 100 g-1 for J. pectoralis and C. jacobinensis, respectively). The antioxidant activity presented a positive correlation with the concentration of phenolics, being 1.186,1 and 1.507,9 µM of Trolox for J. pectoralis and C. jacobinensis at 70% of ethanol; however, it was not verified statistical difference between the ethanolic solutions (p < 0.05). The antimicrobial activity of J. pectoralis extracts was highlighted once was the most effective against gram-positive bacteria. The results suggest that both J. pectoralis and C. jacobinensis extracts present the potential to be applied as natural additives due to their antioxidant and antimicrobial activity and safety. Thus, it is suggesting the development of studies that could investigate the interaction of these plant extracts with food matrices is required

Biologically Active Orbitides from the Euphorbiaceae Family.

A comprehensive overview of natural orbitides isolated from Euphorbiaceae species and their most relevant biological activities are presented. Euphorbiaceae is a large and diverse family, which comprises about 300 genera, and is known as an important source of medicines and toxins. Several classes of secondary metabolites have been described for this taxon, however, orbitides have been broadly reported in Jatropha and Croton genera. Additionally, the latex is documented as the main source of orbitides in this family. Based on their structural and functional diversity, orbitides present a large variety of biological activities described as cytotoxicity, antimalarial, antibacterial, antifungal, enzymatic inhibition, and immunosuppressive, although the mechanism of action still needs to be further investigated. In recent years, the discovery of bioactive cyclic peptides from different sources has grown exponentially, making them promising molecules in the search for new drug leads. This review also highlights the attempts made by many researchers to organize the orbitides nomenclature and amino acid numbering, as well the important progress recently achieved in the biosynthetic study area.

Natural infection of croton yellow vein mosaic virus and its cognate betasatellite in germplasm of different Crambe spp in India.

Crambe is an important crop grown worldwide for industrial oil and seed meal. Besides the fungal and bacterial diseases, the crop is reported to be infected by tobacco mosaic virus, beet western yellows virus and turnip mosaic virus under experimental condition. Till now, there was no report of natural infection of any begomovirus in this crop. In the present study, a leaf curl disease was observed in germplasm accessions of three species of Crambe (C. abyssinica, C. glabrata and C. hispanica). Based on the symptoms and presence of whitefly population in the field, begomovirus infection was suspected. Molecular characterization through RCA approach, indicated presence of croton yellow vein mosaic virus (CYVMV, KJ747958) and croton yellow vein mosaic betasatellite (CroYVMB, KM229762). Co-agroinoculation of partial dimeric construct of CYVMV with complete dimeric construct of CroYVMB, produced typical leaf curl symptoms in C. abyssinica, whereas, agroinoculation of partial dimeric construct of CYVMV alone could not produce symptoms in the same plant. In contrast, the CYVMV construct alone could produce symptom in Nicotiana benthamiana, a model host for plant virus studies. In N. benthamiana co-inoculation of CroYVMV with CYVMV construct develop more severe symptoms. However, neither the CYVMV construct alone nor the co-inoculation with CroYVMB produce any symptom in Arabidopsis thaliana even with different methods of inoculation. Inoculated Arabidopsis thaliana also did not yield any amplification of the virus as assessed through PCR and rolling circle amplification (RCA). Thus it confirmed that for successful infection in crambe, CYVMV requires betasatellite, while in N. benthamiana, it does not require betasatellite for symptom induction and in Arabidopsis thaliana CYVMV alone or in presence of betasatellite did not replicate and produce any symptom. This study constitutes the first confirmed record of natural infection of a begomovirus in crambe and further confirmed that cognate betasatellite of CYVMV has differential role in infectivity in different hosts.

Genetic diversity analysis of Croton antisyphiliticus Mart. using AFLP molecular markers.

Croton antisyphiliticus Mart. is a medicinal plant native to Cerrado vegetation in Brazil, and it is popularly used to treat urogenital tract infections. The objective of the present study was to assess the genetic variability of natural C. antisyphiliticus populations using AFLP molecular markers. Accessions were collected in the states of Minas Gerais, São Paulo, and Goiás. The genotyping of individuals was performed using a LI-COR® DNA Analyzer 4300. The variability within populations was found to be greater than the variability between them. The F(ST) value was 0.3830, which indicated that the populations were highly structured. A higher percentage of polymorphic loci (92.16%) and greater genetic diversity were found in the population accessions from Pratinha-MG. Gene flow was considered restricted (N(m) = 1.18), and there was no correlation between genetic and geographic distances. The populations of C. antisyphiliticus exhibited an island-model structure, which demonstrates the vulnerability of the species.

Refining DNA Barcoding Coupled High Resolution Melting for Discrimination of 12 Closely Related Croton Species.

DNA barcoding coupled high resolution melting (Bar-HRM) is an emerging method for species discrimination based on DNA dissociation kinetics. The aim of this work was to evaluate the suitability of different primer sets, derived from selected DNA regions, for Bar-HRM analysis of species in Croton (Euphorbiaceae), one of the largest genera of plants with over 1,200 species. Seven primer pairs were evaluated (matK, rbcL1, rbcL2, rbcL3, rpoC, trnL and ITS1) from four plastid regions, matK, rbcL, rpoC, and trnL, and the nuclear ribosomal marker ITS1. The primer pair derived from the ITS1 region was the single most effective region for the identification of the tested species, whereas the rbcL1 primer pair gave the lowest resolution. It was observed that the ITS1 barcode was the most useful DNA barcoding region overall for species discrimination out of all of the regions and primers assessed. Our Bar-HRM results here also provide further support for the hypothesis that both sequence and base composition affect DNA duplex stability.

Croton argenteus preparation inhibits initial growth, mitochondrial respiration and increase the oxidative stress from Senna occidentalis seedlings.

Senna ocidentalis is a weed, native to Brazil, considered to infest crops and plantations, and is responsible for yield losses of several crops, particularly soybean. The aim of this work was to evaluate if the Croton argenteus extract and fractions possess phytotoxic activity on S. ocidentalis. The crude ethanolic extract (CEE) and its hexanic (HF), chloroformic (CLF) and ethyl acetate (EAF) fractions were tested in germination, growth, oxidative stress increase, Adenosine triphosphate, L-malate and succinate synthesis. The crude extract and its fractions slowed down the germination of S. ocidentalis and decreased the final percentage of germination. Oxidative stress was also increased in the seedlings, by an increase of catalase, peroxidase, superoxide dismutase, glutathione reductase and lipid peroxidation; and it became clear that the ethyl acetate fraction was more phytotoxic. The results indicate that the crude extract and fractions of C. argenteus compromise the mitochondrial energy metabolism, by the inhibition of mitochondrial ATP production, with a decrease in the production of L-malate and succinate. The ethyl acetate fraction of C. argenteus showed high activity on germination and growth, and these effects take place by means of mitochondrial metabolism alterations and increase the oxidative stress, leading the seedling death.

Antibacterial activity of the alkaloid extract and isolated compounds from Croton bonplandianum Baill. (Euphorbiaceae) / Atividade anti-bacteriana do extrato alcaloide e de compostos isolados de Croton bonplandianum Baill. (Euphorbiaceae)

ABSTRACT The antibacterial activity of the alkaloid extract from the leaves of Croton bonplandianum Baill. and its main compounds, sparsiflorine and crotsparine, was tested against Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa by the resazurin microtitre-plate method. Pure compounds were identified by spectroscopic techniques, mainly 1D and 2D NMR. The alkaloid extract showed activity particularly against the S. aureus and P. aeruginosa. Regarding the pure compounds, the crotsparine was inactive against the microorganisms assayed, whereas the sparsiflorine indicated a moderate activity similar to the alkaloid extract. The Pseudomonas aeruginosa was the most sensitive of the tested microorganisms with MIC of 0.141 mg/mL. The results suggest that the activity of the extract may be credited mainly to the presence of the sparsiflorine. Although the activity of the sparsiflorine does not get close to the antimicrobial drugs in clinical use, it still could be a lead compound for the development of new antibacterial substances.

RESUMO A atividade antibacteriana do extrato alcaloidal das folhas de Croton bonplandianum Baill., além dos principais compostos isolados, esparsiflorina e crotsparina, foi testada contra Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae e Pseudomonas aeruginosa. Foi utilizado o método de microdiluição em placa empregando resazurina como indicador da viabilidade celular. Os compostos isolados foram identificados por técnicas espectroscópicas, principalmente RMN 1D e 2D. O extrato alcaloidal foi ativo principalmente contra S. aureus e P. aeruginosa. Crotsparina mostrou-se inativa contra todos os micro-organismos testados, enquanto esparsiflorina apresentou atividade moderada, a qual foi similar à do extrato bruto. Pseudomonas aeruginosa foi a mais sensível das bactérias testadas, com CIM de 0,141 mg/mL. Os resultados sugerem que a atividade do extrato pode ser devida em grande medida pela presença de esparsiflorina. Apesar de a CIM da esparsiflorina não ter se aproximado daquela apresentada pelos agentes antimicrobianos em uso clínico, tal composto ainda pode compor um protótipo interessante para o desenvolvimento de novas substâncias antibacterianas.

Comparison of methods for estimates of molecular genetic diversity in genus Croton: influence of coefficients, clustering strategies and data projection.

We investigated 10 similarity (and disimilarity) coefficients in a set of 40 wild genotypes of Croton linearifolius subjected to analyses using hierarchical grouping methods, grouping methods by optimization and data projection in two-dimensional space. Genotypes were characterized by analyzing DNA polymorphism with the use of 15 ISSR and 12 RAPD markers. The distance measurements were compared by the Spearman correlation test, projection in two-dimensional space and grouping efficiency evaluation. The Spearman correlation coefficients between the 10 coefficients evaluated were significant (P < 0.001) and indicated significant changes in genotype ranking due to type of coefficient used (0.76 ≤ rs ≤ 1). Wide variation was also observed in the efficiency of clustering methods, where the unweighted pair group method with arithmetic mean was the most suitable (0.3 ≤ D ≤ 1.5 ; 0.41 ≤ rc ≤ 0.77; 5.99 ≤ S ≤ 12.61). Projection efficiencies in two-dimensional space showed high-stress values (65 < S < 89%). Similar to the results observed for hierarchical clustering methods and for projection in two-dimensional space, the formation of groups with grouping methods by optimization showed variations when using different coefficients. We believe that the results confirm the influence of coefficients in studies of genetic diversity, showing the need to use criteria and standards for selecting appropriate methods for genetic studies of the genus Croton.

Cytogenetics and characterization of microsatellite loci for a South American pioneer tree species, Croton floribundus.

Despite the recent advances in plant population genetic studies, the lack of information regarding pedigree, ploidy level, or mode of inheritance for many polyploids can compromise the analysis of the molecular data produced. The aim of this study was to examine both microsatellite and cytogenetic characteristics of the pioneer tree Croton floribundus Spreng. (Euphorbiaceae) to test for the occurrence of polyploidy in the species and to evaluate its implications for the appropriate use of SSR markers. Seven microsatellite markers were developed and screened for 62 individuals from a semi-deciduous tropical forest in Brazil. Chromosome number, meiotic behavior, and pollen viability were evaluated from male flower buds. All SSR loci were highly polymorphic. The number of bivalents observed in meiosis n = 56 (2n = 8× = 112) and the maximum number of alleles per individual (Ni = 8) demonstrated the occurrence of polyploidy in C. floribundus. The normal meiotic pairing and the high pollen viability suggested that C. floribundus is a regular and stable polyploid, most likely an allopolyploid. The combined SSR and cytogenetic data provided new evidence on the origin and evolution of the species as well as assured the accurate use of SSR loci for population genetic studies of the polyploid pioneer species.

Molecular phylogenetics and character evolution of the "sacaca" clade: novel relationships of Croton section Cleodora (Euphorbiaceae).

Phylogenetic relationships of Croton section Cleodora (Klotzsch) Baill. were evaluated using the nuclear ribosomal ITS and the chloroplast trnL-F and trnH-psbA regions. Our results show a strongly supported clade containing most previously recognized section Cleodora species, plus some other species morphologically similar to them. Two morphological synapomorphies that support section Cleodora as a clade include pistillate flowers in which the sepals overlap to some degree, and styles that are connate at the base to varying degrees. The evolution of vegetative and floral characters that have previously been relied on for taxonomic decisions within this group are evaluated in light of the phylogenetic hypotheses. Within section Cleodora there are two well-supported clades, which are proposed here as subsections (subsection Sphaerogyni and subsection Spruceani). The resulting phylogenetic hypothesis identifies the closest relatives of the medicinally important and essential oil-rich Croton cajucara Benth. as candidates for future screening in phytochemical and pharmacological studies.

As espécies de Croton L. sect. Cyclostigma Griseb. e Croton L. sect. Luntia (Raf.) G. L. Webster subsect. Matourenses G. L. Webster (Euphorbiaceae s.s.) ocorrentes na Amazônia brasileira / Species of Croton L. sect. Cyclostigma Griseb. and Croton L. sect. Luntia (Raf.) G. L. Webster subsect. Matourenses G. L. Webster (Euphorbiaceae s.s.) occuring within Brazilian Amazon

Como parte de uma revisão taxonômica das espécies de Croton L. na Amazônia brasileira, estudou-se as seguintes espécies de Croton sect. Cyclostigma Griseb. e Croton sect. Luntia (Raf.) G. L. Webster subsect. Matourenses G. L. Webster: Croton urucurana Baill., C. draconoides Müll. Arg., C. trombetensis R. Secco, P. E. Berry & N.A. Rosa, C. sampatik Müll. Arg., C. palanostigma Kl., C. pullei Lanj. e C. matourensis Aubl. O estudo foi baseado em trabalho de campo realizado nos Estados do Pará e Maranhão, e em material depositado nos herbários IAN, INPA, MG e RB, incluindo tipos. Algumas dessas espécies, como C. urucurana, C. draconoides, C. palanostigma e C. sampatik , são frequentemente encontradas nos herbários com identificações equivocadas. São discutidas a posição taxonômica das espécies nas seções e suas afinidades, e uma chave dicotômica e ilustrações foram elaboradas para um melhor entendimento dos táxons.

As part of a taxonomic revision of Brazilian Amazonia species of Croton L., the following species of Croton sect. Cyclostigma Griseb. and Croton sect. Luntia (Raf.) G. L. Webster subsect. Matourenses G. L. Webster were studied: Croton urucurana Baill., C. draconoides Müll. Arg., C. trombetensis R. Secco, P. E. Berry & N.A. Rosa, C. sampatik Müll. Arg., C. palanostigma Kl., C. pullei Lanj. and C. matourensis Aubl. This study was based on field work in the States of Pará and Maranhão, and material deposited in herbaria IAN, INPA, MG e RB, including the types. Some of the species, such as C. urucurana, C. draconoides, C. palanostigma, and C. sampatik , are frequently found with misidentification in herbaria. The taxonomic position of the species within sections and their affinities are discussed, and a dychotomic key and illustrations were elaborated for a better understanding of the taxa.

Antibacterial activity of extracts of three Croton species collected in Mpumalanga region in South Africa.

The antibacterial activities of three Croton species were compared using bioautography and the serial microdilution methods. The methanolic extracts of all the species had low activity against Escherichia coli. The highest activity was observed with Croton megalobotrys against Enterococcus faecalis with a minimal inhibitory concentration (MIC) value of 0.02 mg/ml. Croton steenkapianus extracts were the least active of the species investigated, only managing an MIC value of 0.625 mg/ml against Pseudomonas aeruginosa. Croton megalobotrys leaf powder was serially extracted using solvents of various polarities. The lowest MIC value (0.06 mg/ml) of the serially extracted fractions was observed with acetone against Pseudomonas aeruginosa. The liquid-liquid fractions of the methanol extract of Croton megalobotrys were also tested. The lowest MIC value of 0.02 mg/ml was observed with n-hexane fraction against Enterococcus faecalis. The carbon tetrachloride fraction was further fractionated using column chromatography with silica as the immobile phase. The resulting seven fractions were tested for activity following the bioassay-guided practice, and it emerged that the first three fractions had active compounds against Staphylococcus aureus when the bioautography method was used.

Revisão taxonômica de Croton sect. Lamprocroton (Müll. Arg.) Pax (Euphorbiaceae s.s.) / Taxonomic revision of Croton sect. Lamprocroton (Müll. Arg.) Pax (Euphorbiaceae s.s.)

Croton is the second bigger and more diverse genus in the family Euphorbiaceae, with about 1,200 species distributed in 40 sections, occurring in all tropical areas, most of them in Americas. In South America, Brazil is the country in which a larger number of taxa are found, ca. 356. According to recent classification, the genus belongs to the tribe Crotoneae, and despite the wide and morphological diversity, it would be a monophyletic taxon. However, a phylogenetic analysis using markers of ITS region from nuclear ribosomal DNA, and of trnL-F from plastidial DNA, showed that Croton, like traditionally circumscribed, is not a monophyletic taxon. A taxonomic revision of Croton section Lamprocroton (Müll. Arg.) Pax is presented here. It is a Neotropical group with most of its species occurring from Southeast and South Brazil to southern South America (Uruguay and Argentina). Morphologically, the members of Lamprocroton are characterized as monoecious or dioecious shrubs or subshrubs, with a lepidote indumentum at least in part of foliage, entire leaves with no glands. The staminate flowers have 9 to 16 stamens and the pistillate flowers may have equal or unequal sepals, reduced to absent petals, and styles once or twice bifid. Overall, are recognized 26 species in the group, three of them new to the science. Identification key, morphological descriptions, illustrations, phenological period, as well as data on geographic distribution and general comments of each species are presented. Four taxa were excluded from C. sect. Lamprocroton because they do not show the morphological features that are diagnostics of the section. Four species that are poorly known were not included in the taxonomic treatment.

O gênero Croton L. é o segundo maior e mais diverso da família Euphorbiaceae, possuindo cerca de 1.200 espécies, agrupadas em 40 seções, com distribuição pantropical, das quais a maioria ocorre nas Américas. Na América o Sul, o Brasil é o país que congrega o maior número de espécies, aproximadamente 356. De acordo com a mais recente classificação, o gênero pertence à tribo Crotoneae e, apesar do grande número de espécies e da grande diversidade morfológica, seria um táxon monofilético. Entretanto, uma análise filogenética recente, que utilizou dados moleculares das regiões ITS, do DNA nuclear ribossômico, e do fragmento trnL-F, do DNA plastidial, demonstrou que Croton, como tradicionalmente circunscrito, não é um táxon monofilético. Neste trabalho, é apresentada uma revisão taxonômica de Croton sect. Lamprocroton (Müll. Arg.) Pax. Trata-se de um grupo neotropical com a maioria das espécies ocorrendo nas regiões Sudeste e Sul do Brasil e sul da América do Sul. Seus representantes caracterizam-se por ser plantas arbustivas ou subarbustivas, monóicas ou dióicas, com indumento lepidoto presente em pelo menos parte da folhagem e folhas inteiras e sem glândulas. As flores estaminadas possuem androceu composto por 9 a 16 estames e as flores pistiladas apresentam sépalas iguais ou desiguais no tamanho, pétalas reduzidas ou ausentes e estiletes uma ou duas vezes bífidos. Neste trabalho são reconhecidas 26 espécies na seção sendo três novas para a ciência. Chave de identificação, descrições morfológicas, ilustrações, período fenológico, distribuição geográfica e comentários gerais de cada uma das espécies são apresentados. Quatro táxons foram excluídos de C. sect. Lamprocroton por não possuírem os caracteres morfológicos diagnósticos da seção. Quatro espécies não foram incluídas no tratamento taxonômico por falta de informação sobre as mesmas.

Bases químicas y farmacológicas de la utilización de la sangre de drago / No disponible

La sangre de drago es el látex rojo exudado al realizar incisiones en la corteza de diversas especies vegetales tropicales, especialmente de Croton lechleri Müll. Arg. Su uso en Medicina Tradicional está muy extendido en Sudamérica, sobre todo como cicatrizante. El látex de C. lechleri es rico en catequinas y proantocianidinas oligoméricas, y contiene 3',4-0-dimetilcedrusina (lignano) y taspina (alcaloide). Se ha demostrado su actividad cicatrizante, antiviral, antimicrobiana, antiinflamatoria, antiulcerosa, antidiarreica e inmunomoduladora. Muchas de estas actividades son coherentes con el uso tradicional y algunas ya han sido estudiadas clínicamente

Sangre de drago (dragon's blood) is the blood red latex that flows after tearing the bark of certain tropical plant species, especially Croton lechleri Müll. Arg. It is widely used in Traditional Medicine in South America, as wound healing. The latex of C. lechleri has catechins and oligomeric proanthocyanidins as major constituents, and it also contains 3'-4-0-dimethylcedrusin (lignan) and taspine (alkaloid). It has shown cicatrizing, antiviral, antimicrobial, anti-inflammatory, antiulcer, antidiarrhoeic and immunomodulatory activities. Many of these activities are in line with its traditional use and some of them have been clinically studied