Trichothecene macrolides from the endophytic fungus Paramyrothecium roridum and their cytotoxic activity.

The chemical investigation of the secondary metabolites of Paramyrothecium roridum (homotypic synonym: Myrothecium roridum), an endophytic fungus isolated from the medicinal plant Morinda officinalis, led to the isolation of twelve cytotoxic trichothecene macrolides, including two new ones, named myrothecines H and I. The structures of the new macrolides were elucidated by extensive spectroscopic measurements analyses. In addition, the cytotoxic activities of these compounds were evaluated against SF-268, NCI-H460, and HepG-2 tumor cell lines, and all isolated compounds (1-12) exhibited significant cytotoxic activity with the IC50 ranging from 0.0002-16.2 µM. Moreover, the inhibitory activity of myrothecines H and I was evidenced by inducing phosphorylation of JNK (c-Jun N-terminal protein kinase) protein and the PARP (poly ADP-ribose polymerase) cleavage, and eventually induce apoptosis of HepG-2 cells. The results indicated that myrothecines H and I could be applied as chemotherapeutic agents.

Four new metabolites from the endophytic fungus Diaporthe lithocarpus A740.

One new benzophenone derivative, named tenllone I (1), two new eremophilane derivatives lithocarins B (2) and C (3), and a new monoterpentoid lithocarin D (4), together with two know compounds (5 and 6) were isolated from the endophytic fungus Diaporthe lithocarpus A740. All of the structures for these new compounds were fully characterized and established on the basis of extensive spectroscopic interpretation. In addition, all the isolated compounds were evaluated in vitro for their cytotoxic activity. Compounds 2, 3, and 5 showed weak inhibitory activities against tumor cell lines.

An endophytic strain of genus Paenibacillus isolated from the fruits of Noni (Morinda citrifolia L.) has antagonistic activity against a Noni's pathogenic strain of genus Aspergillus.

Endophytes are microbes capable of colonizing the tissues of healthy plants and subsequently establishing a harmonious relationship with their hosts. In this research, the endophytic strain Paenibacillus sp. NEB was isolated from fruits of healthy Noni (Morinda citrifolia L.). Strain NEB was identified as Paenibacillus polymyxa using MALDI-TOF Mass Spectrometry. Pathogenic fungal strain NP-1 was isolated from Noni fruits infected by smut, and was identified as Aspergillus aculeatus by polyphasic taxonomy basing on morphological identification, and ITS-5.8S rDNA and ß-tubulin gene phylogenetic analyses. Through the antagonistic test against the pathogenic strain Aspergillus aculeatus NP-1, the results showed that strain NEB had a good antagonistic activity against smut pathogen of Noni. By sequencing with Illumina HiSeq 2000, the draft genome of Paenibacillus sp. NEB was acquired, and 3 CDSs for glucanases were annotated and potentially correlated to the antagonistic activity of this strain. Using realtime-PCR method with specific primers to amplify the biocontrol gene, ß-1,3-1,4- glucanase gene (gluB), it was found in Paenibacillus polymyxa NEB. This study would provide a theoretical and microbial basis for the rationally developing and using Noni beneficial microbial inoculants against its pathogenic strain in the future.

Two New Metabolites from the Endophytic Fungus Alternaria sp. A744 Derived from Morinda officinalis.

Two new compounds isobenzofuranone A (1) and indandione B (2), together with eleven known compounds (3-13) were isolated from liquid cultures of an endophytic fungus Alternaria sp., which was obtained from the medicinal plant Morinda officinalis. Among them, the indandione (2) showed a rarely occurring indanone skeleton in natural products. Their structures were elucidated mainly on the basis of extensive spectroscopic data analysis. All of the compounds were evaluated with cytotoxic and α-glucosidase inhibitory activity assays. Compounds 11 and 12 showed significant inhibitory activities against four tumor cell lines; MCF-7, HepG-2, NCI-H460 and SF-268, with IC50 values in the range of 1.91-9.67 µM, and compounds 4, 5, 9, 10, 12 and 13 showed excellent inhibitory activities against α-glucosidase with IC50 values in the range of 12.05-166.13 µM.

Three new highly-oxygenated metabolites from the endophytic fungus Cytospora rhizophorae A761.

Cytosporaphenones A-C, one new polyhydric benzophenone and two new naphtopyrone derivatives, along with eight known ones, were isolated from Cytospora rhizophorae, an endophytic fungus from Morinda officinalis. Their structures were fully characterized by means of detailed spectroscopic analysis and X-ray single crystal diffraction. To our knowledge, the three new compounds were the most highly oxygenated metabolites of their families discovered in nature. Moreover, all of the compounds were evaluated for in vitro cytotoxic activities against MCF-7, NCI-H460, HepG-2 and SF-268 tumor cell lines, and the new compound 1 exhibited weak growth inhibitory activity against the tumor cell lines MCF-7 and HepG-2 with IC50 values of 70 and 60µM, respectively.

[Isolation and Identification of Bacteria and Mycete from Preservation of Morinda officinalis].

OBJECTIVE: In order to select antistaling agent which had obvious antibacterial effect on Morinda officinalis, the colony counts of microorganism from the surface of Morinda officinalis and the species of bacteria and mycete were analyzed. METHODS: Bacteria and moulds classification identification method were carried out. RESULTS: After using 10 g burdock oligosaccharide, 20 g chitosan and 0. 5 g potassium sorbate treated for 60 d, the total bacteria on the surface of Morinda officinalis was 3. 0 x 10(4) cfu/g, mycete total of 1. 5 x 10(4) cfu/g, preservation effect was better than the other treatments. CONCLUSION: The common advantage microorganisms group of Morinda officinalis on the surface are from Leuconostoc ,Aerococcus and Penicillium genera.

Brachybacterium hainanense sp. nov., isolated from noni (Morinda citrifolia L.) branch.

A Gram-stain-positive bacterial strain, designated as NR2T, isolated from noni (Morinda citrifolia L.) branch was investigated using a polyphasic taxonomic approach. The cells were small coccoid to ovoid, non-spore-forming and motile. Phylogenetic analysis based on 16S rRNA gene sequences showed that the strain was a representative of a member of the genus Brachybacterium, to which the most closely related neighbours were Brachybacterium squillarum M-6-3T (97.90 % similarity), Brachybacterium faecium DSM 4810T (97.50 %), Brachybacterium sacelli LMG 20345T (97.41 %), Brachybacterium phenoliresistens phenol-AT (97.36 %), Brachybacterium nesterenkovii DSM 9573T (97.36 %) and Brachybacterium rhamnosum LMG 19848T (97.32 %). The polar lipid profile of strain NR2T consisted of diphosphatidylglycerol, phosphatidylglycerol, unknown phospholipids and unknown glycolipids. The predominant respiratory quinone was MK-8, with MK-9 and MK-7 as minor components. The major fatty acids were anteiso-C15 : 0 and iso-C15 : 0. Strain NR2T was clearly distinguishable from the type strains of related species on the basis of phylogenetic analysis, DNA-DNA hybridization, fatty acid composition data analysis and a range of physiological and comparison of biochemical characteristics. It is evident from the genotypic and phenotypic data that strain NR2T represents a novel species of the genus Brachybacterium, for which the name Brachybacterium hainanense sp. nov. is proposed. The type strain is NR2T ( = DSM 29535T = CICC 10874T).

Sphingomonas morindae sp. nov., isolated from Noni (Morinda citrifolia L.) branch.

Two yellow bacterial strains, designated NBD5(T) and NBD8, isolated from Noni (Morinda citrifolia L.) branch were investigated using a polyphasic taxonomic approach. Cells were Gram-stain-negative, aerobic, non-spore-forming, non-motile and short rod-shaped. Phylogenetic analysis based on 16S rRNA gene sequences suggested that the strains were members of a novel species of the genus Sphingomonas, the seven closest neighbours being Sphingomonas oligoaromativorans SY-6(T) (96.9% similarity), Sphingomonas polyaromaticivorans B2-7(T) (95.8%), Sphingomonas yantingensis 1007(T) (94.9%), Sphingomonas sanguinis IFO 13937(T) (94.7%), Sphingomonas ginsenosidimutans Gsoil 1429(T) (94.6%), Sphingomonas wittichii RW1(T) (94.6%) and Sphingomonas formosensis CC-Nfb-2(T) (94.5%). Strains NBD5T and NBD8 had sphingoglycolipid, phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylglycerol and phosphatidylcholine as the major polar lipids, ubiquinone 10 as the predominant respiratory quinone, and sym-homospermidine as the major polyamine. Strains NBD5(T) and NBD8 were clearly distinguished from reference type strains based on phylogenetic analysis, DNA-DNA hybridization, fatty acid composition data analysis, and comparison of a range of physiological and biochemical characteristics. It is evident from the genotypic and phenotypic data that strains NBD5(T) and NBD8 represent a novel species of the genus Sphingomonas, for which the name Sphingomonas morindae sp. nov. is proposed. The type strain is NBD5(T) ( = DSM 29151(T) = KCTC 42183(T) = CICC 10879(T)).

Actividad antimicrobiana y toxicidad frente a Artemia salina del extracto diclorometánico de raíces de Morinda royoc L / Antimicrobial action and toxicity against Artemia salina of the dichlormethane extract from Morinda royoc L roots

Introducción: las plantas son una fuente de diversidad natural por la gran variedad de compuestos que sintetizan. Particularmente las antraquinonas resultan un importante grupo de metabolitos secundarios con actividad antimicrobiana y antioxidante. Objetivos: evaluar la actividad antimicrobiana del extracto diclorometánico de raíces de Morinda royoc L, así como su toxicidad contra Artemia salina. Métodos: la actividad antimicrobiana se determinó utilizando el método de microdilución en placa de 96 pozos. Se evaluó la actividad del extracto frente a 7 aislados clínicos de Candida spp. y frente a las bacterias Staphylococcus aureus resistente a meticilina, Staphylococcus aureus ATCC 12598, Enterococus faecales, Escherichia coli, Acinetobacter baumanii, Pseudomonas aeruginosa y Klebsiella pneumoniae. La toxicidad del extracto se evaluó mediante el ensayo de letalidad con A salina. Resultados: el extracto crudo fue activo frente a todas las especies de Candida evaluadas. La concentración mínima inhibitoria más baja fue 1,95 µg/mL. El extracto mostró fuerte actividad inhibitoria contra S. aureus, E faecales, y E coli. El valor más bajo de concentración mínima inhibitoria obtenido fue 31,25 µg/mL. El extracto presentó una toxicidad moderada hacia A salina. Conclusiones: los resultados obtenidos demuestran el potencial del extracto diclorometánico de raíces de M. royoc L en el tratamiento de infecciones causadas por bacterias y hongos(AU)

Introduction: plants are a source of natural diversity because of the great variety of compounds that they synthesize. Anthraquinones in particular are an important group of secondary metabolites characterized by their antimicrobial and antioxidant action. Objectives: to evaluate the antimicrobial action of dichloromethane extract from Morinda royoc L roots as well as its toxicity against Artemia salina. Methods: the antimicrobial action was determined by using the brooth microdilution in 96-well plate. The extract action against 7 Candida spp clinical isolates and against bacteria such as methicillin-resistant Staphylococcus aureus, Staphylococcus aureus ATCC 12598, Enterococus faecales, Escherichia coli, Acinetobacter baumanii, Pseudomonas aeruginosa and Klebsiella pneumoniae was evaluated. The extract toxicity was measured using brine shrimp (Artemia salina) lethality test. Results: the crude extract proved to be active against all the tested Candida species. The lowest minimal inhibitory concentration was 1,95 µg/mL. The extract showed strong inhibitory action against S aureus, E. faecales, and E coli. The lowest minimal inhibitory concentration was 31.25 µg/mL. The extract presented moderate toxicity against A salina. Conclusions: the results showed the potentialities of dichloromethane extract from M. royoc L. roots for the treatment of bacterial and fungal infections(AU)

Actividad antimicrobiana y toxicidad frente a Artemia salina del extracto diclorometánico de raíces de Morinda royoc L / Antimicrobial action and toxicity against Artemia salina of the dichlormethane extract from Morinda royoc L roots

Introducción: las plantas son una fuente de diversidad natural por la gran variedad de compuestos que sintetizan. Particularmente las antraquinonas resultan un importante grupo de metabolitos secundarios con actividad antimicrobiana y antioxidante. Objetivos: evaluar la actividad antimicrobiana del extracto diclorometánico de raíces de Morinda royoc L, así como su toxicidad contra Artemia salina. Métodos: la actividad antimicrobiana se determinó utilizando el método de microdilución en placa de 96 pozos. Se evaluó la actividad del extracto frente a 7 aislados clínicos de Candida spp. y frente a las bacterias Staphylococcus aureus resistente a meticilina, Staphylococcus aureus ATCC 12598, Enterococus faecales, Escherichia coli, Acinetobacter baumanii, Pseudomonas aeruginosa y Klebsiella pneumoniae. La toxicidad del extracto se evaluó mediante el ensayo de letalidad con A salina. Resultados: el extracto crudo fue activo frente a todas las especies de Candida evaluadas. La concentración mínima inhibitoria más baja fue 1,95 µg/mL. El extracto mostró fuerte actividad inhibitoria contra S. aureus, E faecales, y E coli. El valor más bajo de concentración mínima inhibitoria obtenido fue 31,25 µg/mL. El extracto presentó una toxicidad moderada hacia A salina. Conclusiones: los resultados obtenidos demuestran el potencial del extracto diclorometánico de raíces de M. royoc L en el tratamiento de infecciones causadas por bacterias y hongos

Introduction: plants are a source of natural diversity because of the great variety of compounds that they synthesize. Anthraquinones in particular are an important group of secondary metabolites characterized by their antimicrobial and antioxidant action. Objectives: to evaluate the antimicrobial action of dichloromethane extract from Morinda royoc L roots as well as its toxicity against Artemia salina. Methods: the antimicrobial action was determined by using the brooth microdilution in 96-well plate. The extract action against 7 Candida spp clinical isolates and against bacteria such as methicillin-resistant Staphylococcus aureus, Staphylococcus aureus ATCC 12598, Enterococus faecales, Escherichia coli, Acinetobacter baumanii, Pseudomonas aeruginosa and Klebsiella pneumoniae was evaluated. The extract toxicity was measured using brine shrimp (Artemia salina) lethality test. Results: the crude extract proved to be active against all the tested Candida species. The lowest minimal inhibitory concentration was 1,95 µg/mL. The extract showed strong inhibitory action against S aureus, E. faecales, and E coli. The lowest minimal inhibitory concentration was 31.25 µg/mL. The extract presented moderate toxicity against A salina. Conclusions: the results showed the potentialities of dichloromethane extract from M. royoc L. roots for the treatment of bacterial and fungal infections

Phytophthora morindae, a new species causing black flag disease on noni (Morinda citrifolia L) in Hawaii.

A homothallic, papillate Phytophthora species causing foliar and fruit blight of noni (Morinda citrifolia var. citrifolia) in Hawaii was identified. The asexual phase of this species is characterized by the production of umbellate sporangiophores and papillate sporangia that are ellipsoid and obpyriform with conspicuously tapered bases and possess caducous, medium to long pedicels. The sexual phase is characterized by the production of oogonia with tapered bases, small amphigynous antheridia and thick-walled, plerotic oospores. The morphology of the taxon does not match any of the valid 95 Phytophthora species described to date. Phylogenetic analysis based on sequences of the internal transcribed spacer rDNA region (ITS) and the translation elongation factor 1 alpha (EF-1 alpha) of this taxon and those from other Phytophthora species from GenBank and the Phytophthora database indicates that the new taxon is most closely related to species in ITS clade 10, including P. kernoviae, P. boehmeriae and the recently described P. gallica. The most closely related species is P. kernoviae, an invasive plant pathogen causing bleeding stem lesions on forest trees (beech, Fagus sylvatica) and foliar necrosis of ornamentals (rhododendron, pieris and magnolia) in the UK, and isolated in New Zealand from necrotic cherimoya shoots and fruits and soil. Although the morphological characters of the sexual phase of P. morindae and P. kernoviae are similar, the umbellate sporangiophores produced by the new taxon marks the main morphological distinction. In this paper we describe the morphological characteristics, the phylogenetic relationships and pathogenicity characteristics that support the description of this taxon as a new species with the proposed name Phytophthora morindae sp. nov.

Anticancer activity of fungal taxol derived from Botryodiplodia theobromae Pat., an endophytic fungus, against 7, 12 dimethyl benz(a)anthracene (DMBA)-induced mammary gland carcinogenesis in Sprague Dawley rats.

Breast cancer is the second most prevalent cancer worldwide and their incidence increases gradually. Taxol (paclitaxel), a potent anticancer drug, is naturally isolated from the bark of the Pacific yew. Taxol is widely used in the treatment of ovarian, lung and breast cancer. The increased demand for taxol, coupled with its limited availability from the protected Pacific yew, has had researchers scrambling for alternate sources. The purpose of the present study is to investigate chemopreventive effect of fungal taxol derived from a novel endophytic fungus Botryodiplodia theobromae Pat., isolated from a medicinal plant Morinda citrifolia Linn. The fungal taxol is found to be active against the 7, 12 dimethyl benz(a)anthracene (DMBA)-induced mammary gland carcinogenesis in Sprague dawley rats. The enzymic and non-enzymic antioxidants i.e. superoxide dismutase (SOD), catalase (CAT), glutatione peroxidase (GPx), glutatione-S-transferase (GST), reduced glutathione (GSH), vitamin C and vitamin E were evaluated in control and experimental groups. Lipid peroxides levels (LPO) were also tested. Histological analysis of breast tissue was analyzed by haematoxylin and eosin staining to assess the cytoprotective role of fungal taxol active against breast cancer. Immunohistochemical analyses were also performed to evaluate the effect of fungal taxol on the inflammatory marker such as Cyclooxygenase-2 (COX-2) in control and experimental groups. The results showed that the fungal taxol significantly suppresses the DMBA-induced breast cancer in Sprague dawley rats.

Probiotic potential of noni juice fermented with lactic acid bacteria and bifidobacteria.

The present study assesses the feasibility of noni as a raw substrate for the production of probiotic noni juice by lactic acid bacteria (Lactobacilluscasei and Lactobacillus plantarum) and bifidobacteria (Bifidobacteriumlongum). Changes in pH, acidity, sugar content, cell survival and antioxidant properties during fermentation were monitored. All tested strains grew well on noni juice, reaching nearly 109 colony-forming units/ml after 48 h fermentation. L.casei produced less lactic acid than B.longum and L. plantarum. After 4 weeks of cold storage at 4°C, B.longum and L. plantarum survived under low-pH conditions in fermented noni juice. In contrast, L.casei exhibited no cell viability after 3 weeks. Moreover, noni juice fermented with B.longum had a high antioxidant capacity that did not differ significantly (P <0.05) from that of lactic acid bacteria. Finally, we found that B.longum and L. plantarum are optimal probiotics for fermentation with noni juice.

Selection of lactic acid bacteria from fermented plant beverages to use as inoculants for improving the quality of the finished product.

Lactic Acid Bacteria (LAB) from fermented plant beverages were selected based on their antibacterial actions against potential food borne pathogenic bacteria (Staphylococcus aureus PSSCMI 0004, Escherichia coli PSSCMI 0001, Salmonella typhimurium PSSCMI 0034 and Vibrio parahaemolyticus VP 4). Antibacterial activities were measured using an agar spot method. The Lactobacillus plantarum W90A strain isolated from a wild forest noni (Morinda coreia Ham) beverage was used as an inoculant. Three different inoculation procedures were conducted with the fruit of wild forest noni fermentations to establish which one was the best for controlling the numbers of yeast in the finished product. A 5% inoculum of L. plantarum W90A (LAB set), initial cell density 8.6 log cfu mL(-1), produced a better product and inhibitory properties against the test organisms, particularly E. coli PSSCMI 0001 than one with no inoculum or with a 5% inoculum from a previous natural fermented product. An LAB inoculum resulted in a reduced total bacterial count and no yeast throughout fermentation period (90 days). The lower yeast resulted in a reduction of the ethanol content to 2.9 g L(-1) compared to 12.2 g L(-1) inthe culture with no inoculum. The highest acidity (1.3-1.4%) with the same pH (3.3) was observed in both sets of inoculated fermentations, whereas the uninoculated set gave a pH value of 3.7 (1.2% acidity).

Ensayo microbiológico en cápsulas de Morinda citrifolia L. (Noni) comercializadas ambulatoriamente en la ciudad de León en el mes de febrero del 2008 / Microbiological assay in capsules of Morinda citrifolia L. (Noni) commercialized ambulatory in the city of León in February 2008

El presente estudio trata de determinar el límite microbiano en las cápsulas de Noni comercializadas ambulatoriamente en la ciudad de León, así como, cuantificar la presencia de bacterias aerobias mesófilas e identificar presencia de microorganismos patógenos como Staphilococcus áureus, Pseudomona aeuruginosa, Escherichia coli y Salmonella spp. evidenciando la presencia de Hongos y Levaduras.