Non-nodulated bacterial leaf symbiosis promotes the evolutionary success of its host plants in the coffee family (Rubiaceae).

Every plant species on Earth interacts in some way or another with microorganisms and it is well known that certain forms of symbiosis between different organisms can drive evolution. Within some clades of Rubiaceae (coffee family), a specific plant-bacteria interaction exists in which non-pathological endophytes are present in the leaves of their hosts. It is hypothesized that the bacterial endophytes, either alone or by interacting with the host, provide chemical protection against herbivory or pathogens by producing toxic or otherwise advantageous secondary metabolites. If the bacteria indeed have a direct beneficial influence on their hosts, it is reasonable to assume that the endophytes may increase the fitness of their hosts and therefore it is probable that their presence also has an influence on the long-term evolution of the particular plant lineages. In this study, the possible origin in time of non-nodulated bacterial leaf symbiosis in the Vanguerieae tribe of Rubiaceae is elucidated and dissimilarities in evolutionary dynamics between species with endophytes versus species without are investigated. Bacterial leaf symbiosis is shown to have most probably originated in the Late Miocene, a period when the savannah habitat is believed to have expanded on the African continent and herbivore pressure increased. The presence of bacterial leaf endophytes appears to be restricted to Old World lineages so far. Plant lineages with leaf endophytes show a significantly higher speciation rate than plant lineages without endophytes, while there is only a small difference in extinction rate. The transition rate shows that evolving towards having endophytes is twice as fast as evolving towards not having endophytes, suggesting that leaf symbiosis must be beneficial for the host plants. We conclude that the presence of bacterial leaf endophytes may also be an important driver for speciation of host plants.

Avaliação da atividade antioxidante e antimicrobiana do extrato etanólico bruto e frações orgânicas obtidas a partir da casca do caule da espécie Guettarda uruguensis Cham, & Scthdl, (Rubiaceae) / Assessment of the antioxidant and antimicrobial activity of the crude ethanolic extract and organic fractions obtained from the stem bark of the specie Guettarda uruguensis Cham. & Scthdl. (Rubiaceae)

Este trabalho teve como objetivo avaliar o potencial antioxidante e antimicrobiano do extrato bruto e frações obtidas das cascas do caule da espécie Guettarda uruguensis, Os ensaios antioxidantes indicaram alto potencial antioxidante, No ensaio de redução de fosfomolibdênio, a fração acetato de etila apresentou atividade antioxidante de 41,67% em relação ao padrão de ácido ascórbico e superou em 35,21% a atividade do padrão rutina, No ensaio de redução do DPPH (2,2-diphenyl-1-picrylhydrazyl), a fração acetato de etila apresentou um IC50 de 10,91 µg mL-1, valor próximo ao do ácido ascórbico (IC50 = 4,78 µg mL-1) e da rutina (IC50 = 6,62 µg mL-1), Pelo ensaio de TBA (acido tiobabitúrico) o extrato bruto (IA = 71,48%) e a fração hexano (IA = 47,85%) apresentaram índices superiores ao controle de BHT (butil hidroxi tolueno) (IA = 42,66), Através do ensaio de microdiluição em placas, foi observado que o extrato bruto e frações apresentaram atividade antimicrobiana, O estudo fitoquímico qualitativo revelou a presença de alcaloides, cumarinas, esteroides e/ou triterpenos, heterosídeos saponínicos, taninos e aminogrupos...

This main purpose of this study was to evaluate the antimicrobial and antioxidant effects of the crude extract and fractions obtained from the stem bark of the plant species. The antioxidant assays indicated high antioxidant capacity. In the reduction assay of the phosphomolybdenum, the ethyl acetate fraction showed antioxidant activity of 41.67% compared to standard ascorbic acid and exceeded in 35.21% the activity of the standard rutin. In the reduction assay of the DPPH (2,2-diphenyl-1-picrylhydrazyl), the ethyl acetate fraction showed an IC50 of 10.91 µg mL-1, equivalent to the ascorbic acid (IC50 = 4.78 µg mL-1) and rutin (IC50 = 6.62 µg mL-1). By the TBA (thiobarbituric acid) assay the crude extract (IA = 71.48%) and hexane fraction (IA = 47.85%) had an index higher than the control of BHT (butyl hydroxy toluene) (IA = 42.675). Through of assay of microdilution on plates was verified that the crude extract and fractions showed antimicrobial activity. The qualitative phytochemical study revealed the presence of alkaloids, coumarins, steroids and/or triterpenoids, saponin glycosides, tannins and amino groups...

Mycoleptones A-C and polyketides from the endophyte Mycoleptodiscus indicus.

Three new azaphilones with an unusual methylene bridge, named mycoleptones A, B, and C (2, 4, and 5), were isolated from cultures of Mycoleptodiscus indicus, a fungus associated with the South American medicinal plant Borreria verticillata. Additionally, four known polyketides, austdiol (1), eugenitin (3), 6-methoxyeugenin (6), and 9-hydroxyeugenin (7), were also isolated. The structural characterization of compounds was carried out by nuclear magnetic resonance spectroscopy, high-resolution mass spectrometry, electronic circular dichroism spectroscopy, time-dependent density functional theory calculations, and X-ray crystallography. Compounds 1-9 were weakly active when tested in antileishmanial and cytotoxicity assays.

Thielavins A, J and K: α-Glucosidase inhibitors from MEXU 27095, an endophytic fungus from Hintonia latiflora.

Bioassay-guided fractionation of the bio-active organic extract obtained from solid-media culture of MEXU 27095, an endophytic fungus isolated from the Mexican medicinal plant Hintonia latiflora (Rubiaceae), led to separation of three tridepsides which were identified as thielavins A, J and K. All three compounds inhibited Saccharomyces cerevisieae α-glucosidase (αGHY) in a concentration-dependent manner with IC50 values of 23.8, 15.8, and 22.1µM, respectively. Their inhibitory action was higher than that of acarbose (IC50=545µM), used as a positive control. Kinetic analysis established that the three compounds acted as non-competitive inhibitors with ki values of 27.8, 66.2 and 55.4µM, respectively (α=1.0, 1.2, 0.7, respectively); acarbose behaved as competitive inhibitor with a ki value of 156.1µM. Thielavin J inhibited the activity of α-glucosidase from Bacillus stearothermophilus (αGHBs) with an IC50 of 30.5µM, being less active than acarbose (IC50=0. 015µM); in this case, compound (2) (ki=20.0µM and α=2.9) and acarbose (ki=0.008µM and α=1.9) behaved as non-competitive inhibitors. Docking analysis predicted that all three thielavins and acarbose bind to homologated αGHBs and to αGHY (PDB: 3A4A) in a pocket close to the catalytic site for maltose and isomaltose, respectively. The α-glucosidase inhibitory properties of thielavin K (3) were corroborated in vivo since it induced a noted antihyperglycemic action during an oral sucrose tolerance test (3.1, 10.0 and 31.6mg/kg) in normal and nicotinamide-streptozotocin diabetic mice. In addition, at a dose of 10mg/kg, it provoked a moderate hypoglycemic activity in diabetic mice.

Bioactive metabolites produced by Penicillium sp. 1 and sp. 2, two endophytes associated with Alibertia macrophylla (Rubiaceae).

In the course of our continuous search for bioactive metabolites from endophytic fungi living in plants from the Brazilian flora, leaves of Alibertia macrophylla (Rubiaceae) were submitted to isolation of endophytes, and two species of Penicillium were isolated. The acetonitrile fraction obtained in corn from a culture of Penicillium sp. 1 afforded orcinol (1). On the other hand, Penicillium sp. 1 cultivated in potato-dextrose-broth furnished two different compounds, cyclo-(L-Pro-L-Val) (2) and uracil (3). The chromatographic fractionation of the acetonitrile fraction obtained from Penicillium sp. 2 led to three dihydroisocoumarins, 4-hydroxymellein (4), 8-methoxymellein (5) and 5-hydroxymellein (6). Compounds 5 and 6 were obtained from the Penicillium genus for the first time. Additionally, metabolites 1-6 were evaluated for their antifungal and acetylcholinesterase (AChE) inhibitory activities. The most active compounds 1 and 4 exhibited detection limits of 5.00 and 10.0 microg against Cladosporium cladosporioides and C. sphaerospermum, respectively. Compound 2 showed a detection limit of 10.0 microg, displaying potent AChE inhibitory activity.

An endophytic Neurospora sp. from Nothapodytes foetida producing camptothecin.

The medicinal plant, Nothapodytes foetida contains a number of important alkaloids like camptothecin (an anticancer drug molecule) but its concentration is less to meet the existing demand of this important molecule, so in an effort for accessible availability of camptothecin. An endophyte (designated ZP5SE) was isolated from the seed of Nothapodytes foetida and was examined as potential source of anticancer drug lead compound i.e. camptothecin, when grown in Sabouraud liquid culture media under shake flask conditions. The presence of anticancer compound (camptothecin) in this fungus was confirmed by chromatographic and spectroscopic methods in comparison with authentic camptothecin. Isolated endophyte (Neurospora crassa) producing camptothecin may become an easily accessible source for the production of precursor anticancer drug molecule in future at large scale.

Characterization of an extracellular endopolygalacturonase from the saprobe Mucor ramosissimus Samutsevitsch and its action as trigger of defensive response in tropical plants.

In recent years, interest in the ability of non-pathogenic microorganisms to induce resistance in plants has grown, particularly with respect to their use as environmentally safe controllers of plant disease. In this study, we investigated the capacity of Mucor ramosissimus Samutsevitsch to release pectinases able to degrade cell walls of Palicourea marcgravii St. Hil., a tropical forest native Rubiaceae on which the spores of this saprobic fungus have been found. The fungus was grown in liquid culture medium containing pectin as the sole carbon source and filtrates were analyzed for pectinase activity. An endopolygalacturonase was partially purified by ion exchange chromatography, gel filtration, and preparative isoelectrofocusing, and characterized. This enzyme was more active upon pectic substrates with a low degree of methyl esterification. The products of hydrolysis of different pectic substrates (including pectin from P. marcgravii) by the action of this endopolygalacturonase elicited to different extents the phytoalexin production in soybean cotyledons. Also, the enzyme itself and the products of its action on the pectic fraction of P. marcgravii elicited the production of defensive compounds in the leaves of the plant. These results suggest that, besides the role in recycling organic matter, saprobes may also play an important role in the induction of defensive mechanisms in wild plants by enhancing their non-specific resistance against pathogens. Furthermore, they set the stage for future studies on the role of saprobic fungi in inducing resistance of host plants to pathogens.