Search

Essential oils were extracted by hydrodistillation from the traditional Australian medicinal plant Eremophila bignoniiflora, characterized chemically and then screened for bioactivity. Characterization and quantification were completed using gas chromatography-mass spectrometry (GC-MS) and GC-flame ionization detection, respectively. Antimicrobial capacity was assessed using disc diffusion and micro-titre plate broth dilution and further characterized using thin layer chromatography followed by bioautography to assign activity to separated individual active components. Antifungal capacity was investigated using micro-titre plate broth dilution against pathogenic Trichophyton species. Free radical scavenging ability was assessed using the diphenylpicrylhydradyl reaction in methanol. The predominant components of the essential oil were fenchyl-acetate and bornyl-acetate. However, bioautography indicated antimicrobial ability to be largely linked to the less abundant, more polar constituents. Oils displayed only modest antifungal ability against pathogenic Trichophyton species associated with dermatophytosis, but moderate to high antimicrobial activity, particularly against the yeast Candida albicans and the bacteria Staphylococcus epidermidis. Essential oils exhibited relatively low free radical scavenging ability. Speculation over the role of essential oils in the traditional medicinal applications of E. bignoniiflora follows, exploring correlations between traditional use and investigated bioactivities., (Copyright © 2012 John Wiley & Sons, Ltd.)

This paper reports on the isolation and identification of antibacterial constituents from the indigenous Australian medicinal plant Eremophila duttonii F. Muell. (Myoporaceae). Preparations derived from this plant are used by indigenous populations in the topical treatment of minor wounds, otitis and ocular complaints, and as a gargle for sore throat. Several authors have reported extracts of this plant to effect rapid bacteriolysis and inhibit growth of a wide range of Gram-positive micro-organisms. In other studies involving screening of native medicinal plants for antibacterial activity, extracts of Eremophila duttonii have been reported to consistently exhibit the highest potency amongst all species included. From a hexane extract, we identified two diterpenes of the serrulatane class, the principal constituents responsible for antibacterial activity and present as major constituents of the resinous leaf cuticle: serrulat-14-en-7,8,20-triol (1) and serrulat-14-en-3,7,8,20-tetraol (2). In addition, a hydroxylated furanosesquiterpene with mild antibacterial activity which appeared to be a novel compound was isolated from the extract and tentatively identified as 4-hydroxy-4-methyl-1-(2,3,4,5-tetrahydro-5-methyl[2,3'-bifuran]-5-yl) pentan-2-one. Minimum inhibitory concentrations for each of the compounds against three Gram-positive bacteria: Staphylococcus aureus (ATCC 29213), Staphylococcus epidermidis (ATCC 12228) and Streptococcus pneumoniae (ARL 10582), were determined using a micro-titre plate broth dilution assay.

The aerial parts of the endemic Australian plant Eremophila debilis (Myoporaceae) contain 3% dry weight of the biologically active 5,6,7,3',4',5'-hexamethoxyflavone, which had its structured confirmed using X-ray crystal crystallography. The presence of significant levels of the polypharmacologically active 5,6,7,3',4',5'-hexamethoxyflavone in the edible parts of the plant has potential implications for its use as a food and bush medicine., (Copyright © 2017 Elsevier B.V. All rights reserved.)

A detailed investigation of the wood, leaf, branch and root oil of Eremophila mitchellii (Benth.) was carried out by a combination of GC-FID, GC-MS and NMR. The wood oil was composed predominantly of eremophilanes, a rare class of biologically active, bicyclic sesquiterpenoids. The root oil was also found to contain the eremophilanes together with the zizaene sesquiterpene, sesquithuriferone. 9-Hydroxy-1,7(11),9-eremophilatrien-8-one and the known 1(10),11-eremophiladien-9-one (eremophilone), 9-hydroxy-7(11),9-eremophiladien-8-one (2-hydroxyeremophilone), 8-hydroxy-11-eremophilen-9-one (santalcamphor), 8-hydroxy-10,11-eremophiladien-9-one, sesquithuriferone and 8-hydroxy-1,11-eremophiladien-9-one were purified and elucidated by NMR. Three approaches to the purification of the major eremophilanes from the wood oil are described. (+) Spathulenol, α-pinene, globulol, viridiflorene were the major constituents of the leaf oil. All of the essential oils and the eremophilanes exhibited cytotoxicity against P388D(1) mouse lymphoblast cells in-vitro., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Volatile oils of Eremophila longifolia F. Muell. (Myoporaceae) leaves were obtained by hydrodistillation and analysed using GC-MS. A total of 33 compounds were identified in the oils examined and a high degree of intraspecific variability in chemical composition between specimens occurring in separate geographic localities was found. Multivariate statistical analysis of chemical composition of volatile oils enabled classification of three chemotypes in this species., ((c) 2010. Published by Elsevier Ltd. All rights reserved.)

Thirty-eight microsatellite loci were developed for the bird pollinated, autohexaploid, Eremophila glabra ssp. glabra. A genomic library was screened with dinucleotide and trinucleotide sequence repeats. Polymorphism ranged from one to 21 alleles per locus. Twenty-four loci exhibited null alleles, based on patterns of inheritance between maternal and progeny phenotypes. Cross-species amplification of nine Eremophila species was successful for most primers, indicating wide applicability across the genus. These microsatellites will be used to study the gene flow patterns of fragmented populations of E. glabra ssp. glabra., (© 2009 Blackwell Publishing Ltd.)

Epingaione (4-Methyl-1-(5-methyl-2, 3,4,5-tetrahydro-[2,3']bifuranyl-5-yl)-pentan-2-one) was isolated as one of the major lipophilic secondary metabolites from the leaves and stems of Bontia daphnoides L. The compound gave 79.24% and 50.83% anti-proliferation/cytotoxic activity on the human SH-SY5Y neuroblastoma and TE-671 sarcoma cells in vitro at 50 pg/mL, respectively. Epingaione was transformed into eleven derivatives under laboratory conditions using ethanol, some gave greater anti-proliferation/cytotoxic activity on the cancer cell lines tested. One of the derivatives (compound 2) with enhanced cytotoxic activity was elucidated as 5'-Ethoxy-5-methyl-5-(4-methyl-2-oxo-pentyl)-2,3,4,5-tetrahydro-5'H-[2,3']bifuranyl-2'-one. Both epingaione and compound 2 caused an accumulation of arrested or dead SH-SY5Y neuroblastoma in the m-phase of the cell cycle as revealed by the m-phase specific marker KE 67.

Eremophila is the largest genus in the plant tribe Myoporeae (Scrophulariaceae) and exhibits incredible morphological diversity across the Australian continent. The Australian Aboriginal Peoples recognize many Eremophila species as important sources of traditional medicine, the most frequently used plant parts being the leaves. Recent phylogenetic studies have revealed complex evolutionary relationships between Eremophila and related genera in the tribe. Unique and structurally diverse metabolites, particularly diterpenoids, are also a feature of plants in this group. To assess the full dimension of the chemical space of the tribe Myoporeae, we investigated the metabolite diversity in a chemo-evolutionary framework applying a combination of molecular phylogenetic and state-of-the-art computational metabolomics tools to build a dataset involving leaf samples from a total of 291 specimens of Eremophila and allied genera. The chemo-evolutionary relationships are expounded into a systematic context by integration of information about leaf morphology (resin and hairiness), environmental factors (pollination and geographical distribution), and medicinal properties (traditional medicinal uses and antibacterial studies), augmenting our understanding of complex interactions in biological systems., (© 2021 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.)

The secondary metabolites isolated from members of the Myoporaceae are reviewed. More than 200 compounds, whose structures have been elucidated, are listed together with their source in the Appendix.

The figwort family, Scrophulariaceae, comprises c. 2000 species whose evolutionary relationships at the tribal level have proven difficult to resolve, hindering our ability to understand their origin and diversification. We designed a specific probe kit for Scrophulariaceae, targeting 849 nuclear loci and obtaining plastid regions as by-products. We sampled c. 87% of the genera described in the family and use the nuclear dataset to estimate evolutionary relationships, timing of diversification, and biogeographic patterns. Ten tribes, including two new tribes, Androyeae and Camptolomeae, are supported, and the phylogenetic positions of Androya, Camptoloma, and Phygelius are unveiled. Our study reveals a major diversification at c. 60 million yr ago in some Gondwanan landmasses, where two different lineages diversified, one of which gave rise to nearly 81% of extant species. A Southern African origin is estimated for most modern-day tribes, with two exceptions, the American Leucophylleae, and the mainly Australian Myoporeae. The rapid mid-Eocene diversification is aligned with geographic expansion within southern Africa in most tribes, followed by range expansion to tropical Africa and multiple dispersals out of Africa. Our robust phylogeny provides a framework for future studies aimed at understanding the role of macroevolutionary patterns and processes that generated Scrophulariaceae diversity., (© 2023 The Authors. New Phytologist © 2023 New Phytologist Foundation.)

Traditional classification of speciation modes has focused on physical barriers to gene flow. Allopatric speciation with complete reproductive isolation is viewed as the most common mechanism of speciation. Parapatry and sympatry, by contrast, entail speciation in the face of ongoing gene flow, making them more difficult to detect. The genus Iberodes (Boraginaceae, NW Europe) comprises five species with contrasting morphological traits, habitats and species distributions. Based on the predominance of narrow and geographically distant endemic species, we hypothesized that geographical barriers were responsible for most speciation events in Iberodes. We undertook an integrative study including: (i) phylogenomics through restriction-site-associated DNA sequencing (RAD-seq), (ii) genetic structure analyses, (iii) demographic modelling, (iv) morphometrics, and (v) climatic niche modelling and niche overlap analysis. The results revealed a history of recurrent progenitor-derivative speciation manifested by a paraphyletic pattern of nested species differentiation. Budding speciation mediated by ecological differentiation is suggested for the coastal lineage, deriving from the inland widespread Iberodes linifolia during the Late Pliocene. Meanwhile, geographical isolation followed by niche shifts are suggested for the more recent differentiation of the coastland taxa. Our work provides a model for distinguishing speciation via ecological differentiation of peripheral, narrowly endemic I. kuzinskyanae and I. littoralis from a widespread extant ancestor, I. linifolia. Ultimately, our results illustrate a case of Pliocene speciation in the probable absence of geographical barriers and get away from the traditional cladistic perspective of speciation as producing two species from an extinct ancestor, thus reminding us that phylogenetic trees tell only part of the story., (© 2022 John Wiley & Sons Ltd.)

Aims: Microbial endophytes produce specialized metabolites, including antibiotics and other compounds of pharmaceutical and agricultural value. This study aimed to investigate the diversity and bioactivity of endophytes from medicinal plants used by the Dharawal People of Gamay (Botany Bay), Australia., Methods and Results: Of the 48 endophytes isolated, 19 tested positive for polyketide synthase or non-ribosomal peptide synthetase genes via a PCR incorporating degenerate primers. The biosynthetically talented endophytes were identified by 16S rRNA gene sequencing and included 4 bacteria species belonging to the orders Bacillales, Rhizobiales and Burkholderiales and 15 Ascomycota fungi species belonging to the orders Botryosphaeriales, Cladosporiales, Glomerellales, Microascales and Eurotiales. Antimicrobial testing using the disc diffusion assay demonstrated that 15 of the 19 isolates had broad-spectrum activity against a range of Gram-positive and Gram-negative bacteria., Conclusions: Taken together, these results suggest that Australian bush medicines harbour diverse biosynthetically talented microbial endophytes capable of producing broad-spectrum antibacterial compounds., Significance and Impact of the Study: This study suggests that compounds produced by microbial endophytes likely contribute to the collective medicinal properties of Australian bush medicines. Significantly, it highlights that Indigenous botanical knowledge and modern molecular approaches can be used in tandem to prioritize microorganisms that produce pharmaceutically relevant compounds., (© 2021 The Society for Applied Microbiology.)

The effect of a natural sesquiterpene ketone, 9,10-dehydrofukinone (DHF), on pathogenic Staphylococcus aureus and Pseudomonas aeruginosa strains isolated from chronic infectious processes, was the focus of the present study. Lipophilic DHF produced important antibacterial synergistic effects in association with ciprofloxacin (CPX) against two biofilm-forming strains of S. aureus HT1 (FIC=0.21) and P. aeruginosa HT5 (FIC=0.05). Hence, this mixture constitutes an excellent strategy to combat these biofilm-producing bacteria that overexpress drug efflux pumps as a resistance mechanism. Additionally, a substantial rise in beneficial Lactobacillus biofilm biomass was determined as a very significant finding of this association. Particularly, a non-pathogenic biofilm increment of 119 % was quantified when the mixture was added to a probiotic L. acidophilus ATCC SD-5212 culture. A surface activity enhanced in 71 % with respect to untreated L. acidophilus culture was also generated by the DHF and CPX association, and therefore, a glycoprotein synthesis induction mediated by the mixture is discussed. The results obtained could help in the development of new selective antibiotics. From an ecological standpoint, the present study strongly suggests that DHF is a polyfunctional organic molecule produced with a high yield in Senecio punae that exerts a positive impact on a non-pathogenic plant bacterium L. plantarum CE105., (© 2020 Wiley-VHCA AG, Zurich, Switzerland.)

The performance of first-generation hybrids determines to a large extent the long-term outcome of hybridization in natural populations. F 1 hybrids can facilitate further gene flow between the two parental species, especially in animal-pollinated flowering plants. We studied the performance of reciprocal F 1 hybrids between Rhinanthus minor and R. major, two hemiparasitic, annual, self-compatible plant species, from seed germination to seed production under controlled conditions and in the field. We sowed seeds with known ancestry outdoors before winter and followed the complete life cycle until plant death in July the following season. Germination under laboratory conditions was much lower for the F 1 hybrid formed on R. major compared with the reciprocal hybrid formed on R. minor, and this confirmed previous results from similar experiments. However, this difference was not found under field conditions, which seems to indicate that the experimental conditions used for germination in the laboratory are not representative for the germination behaviour of the hybrids under more natural conditions. The earlier interpretation that F 1 hybrid seeds formed on R. major face intrinsic genetic incompatibilities therefore appears to be incorrect. Both F 1 hybrids performed at least as well as and sometimes better than R. minor, which had a higher fitness than R. major in one of the two years in the greenhouse and in the field transplant experiment. The high fitness of the F 1 hybrids confirms findings from naturally mixed populations, where F 1 hybrids appear in the first year after the two species meet, which leads to extensive advanced-hybrid formation and introgression in subsequent generations., (© 2019 European Society For Evolutionary Biology. Journal of Evolutionary Biology © 2019 European Society For Evolutionary Biology.)

Myoporum bontioides is a traditional medicinal plant in Asia with various biological activities, including anti-inflammatory and anti-bacterial characteristics. To identify the bioactive constituents from M. bontioides, a newly-identified flavone, 3,4'-dimethoxy-3',5,7-trihydroxyflavone (compound 1), along with eight known compounds, were investigated in human MCF-7 breast cancer, SCC4 oral cancer, and THP-1 monocytic leukemia cells. Among these compounds, compound 1 exhibited the strongest antiproliferative activity with half-maximal inhibitory concentration (IC 50 ) values ranging from 3.3 μM (MCF-7) to 8.6 μM (SCC4). Flow cytometric analysis indicated that compound 1 induced G2/M cell cycle arrest in MCF-7 cells. Mechanistic evidence suggests that the G2/M arrest could be attributable to compound 1's modulatory effects on the phosphorylation and expression of numerous key signaling effectors, including cell division cycle 2 (CDC2), CDC25C, and p53. Notably, compound 1 downregulated the expression of histone deacetylase 2 (HDAC2) and HDAC4, leading to increased histone H3 acetylation and p21 upregulation. Together, these findings suggest the translational potential of compound 1 as a breast cancer treatment.

The angiosperm family Boraginaceae includes ca. 1600 species distributed among ca. 110 genera. Some floral features are constant within the family, but many vegetative, floral, pollen, and nutlet traits vary. Utilizing 224 species of Boraginaceae and related taxa, five matrices were constructed with various combinations of morphological characters, three chloroplast DNA regions, and one nuclear ribosomal DNA region. Phylogenetic analyses were conducted for these matrices, and patterns of character evolution were examined. Boraginaceae is resolved as monophyletic, with Wellstedia as its sister. Codon is sister to Boraginaceae + Wellstedia. Although most of the investigated morphological characters have a low consistency index, particular character states are synapomorphies for large clades in each of the tribes of the family. In Boraginaceae, the breeding system heterostyly evolved at least 12 times, which is the largest number of origins resolved in any family; therefore Boraginaceae can serve as a model for the evolution and development of heterostyly. Nutlet ornamentation is most diverse in Cynoglosseae and Trichodesmeae, while pollen and floral features are most variable in Boragineae and Lithospermeae. Phylogenetic relationships and patterns of character evolution identified in the present study set the stage for future work creating an updated taxonomic system of Boraginaceae., (© The Willi Hennig Society 2013.)

Chemical investigations of the aerial parts of the Australian plant Eremophila microtheca resulted in the isolation of three serrulatane diterpenoids, 3-acetoxy-7,8-dihydroxyserrulat-14-en-19-oic acid (1), 3,7,8-trihydroxyserrulat-14-en-19-oic acid (2) and 3,19-diacetoxy-8-hydroxyserrulat-14-ene (3) as well as the previously reported compounds verbascoside (4) and jaceosidin (5). Acetylation and methylation of the major serrulatane diterpenoid 2 afforded 3,8-diacetoxy-7-hydroxyserrulat-14-en-19-oic acid (6) and 3,7,8-trihydroxyserrulat-14-en-19-oic acid methyl ester (7), respectively. The antibacterial activity of 1-7 was assessed against a panel of Gram-positive and Gram-negative bacterial isolates. All of the serrulatane compounds exhibited moderate activity against Streptococcus pyogenes (ATCC 12344) with minimum inhibitory concentrations (MICs) ranging from 64-128 μg/mL. Serrulatane 1 demonstrated activity against all Gram-positive bacterial strains (MICs 64-128 μg/mL) except for Enterococcus faecalis and Enterococcus faecium. This is the first report of natural products from E. microtheca., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Lithospermum (Boraginaceae) includes ca. 60 species and exhibits a wide range of floral, palynological, and vegetative diversity. Phylogenetic analyses based on 10 chloroplast DNA regions and 22 morphological characters were conducted in order to (i) examine evolutionary relationships within Lithospermum and among related genera of Boraginaceae, and (ii) investigate patterns of morphological evolution. Several morphological features, such as long-funnelform corollas, faucal appendages, reciprocal herkogamy, and evident secondary leaf venation, have evolved multiple times within the genus. In contrast, other morphological features, including the presence of glands and the position and number of pollen pores, are less plastic and tend to characterize larger clades. Some features, including the presence of glands, are interpreted as symplesiomorphic for Lithospermum, while others, such as evident secondary leaf venation, appear to have originated repeatedly. The range of structural diversity that occurs among the species of Lithospermum suggests the potential utility of this genus as a model for integrative studies of evolution, development, and molecular biology.© The Willi Hennig Society 2011., (© The Willi Hennig Society 2011.)

This review uses proxies of past temperature and atmospheric CO(2) composition based on fossil leaves to illustrate the uncertainties in biologically based proxies of past environments. Most leaf-based proxies are geographically local or genetically restricted and therefore can be confounded by evolution, extinction, changes in local environment or immigration of species. Stomatal frequency proxies illustrate how genetically restricted proxies can be particularly vulnerable to evolutionary change. High predictive power in the modern world resulting from the use of a very narrow calibration cannot be confidently extrapolated into the past (the Ginkgo paradox). Many foliar physiognomic proxies of climate are geographically local and use traits that are more or less fixed for individual species. Such proxies can therefore be confounded by floristic turnover and biome shifts in the region of calibration. Uncertainty in proxies tends to be greater for more ancient fossils. I present a set of questions that should be considered before using a proxy. Good proxies should be relatively protected from environmental and genetic change, particularly through having high information content and being founded on biomechanical or biochemical principles. Some current and potential developments are discussed, including those that involve more mechanistically sound proxies and better use of multivariate approaches., (© 2011 The Author. New Phytologist © 2011 New Phytologist Trust.)

•  Leaf morphology varies predictably with altitude, and leaf morphological features have been used to estimate average temperatures from fossil leaves. The altitude-leaf morphology relationship is confounded by the two processes of acclimation and adaptation, which reflect environmental and genetic influences, respectively. •  Here we describe the relationship between altitude and leaf morphology for Southern beech, Nothofagus cunninghamii (Hook.) Oerst.. Cuttings from several trees from each of four altitudes were grown in a common glasshouse experiment, and leaf morphology related to both genotype and altitude of origin. •  Genotype had a significant impact on leaf morphology, but in the field there was also a significant, overriding effect of altitude. This altitude effect disappeared in glasshouse-grown plants for all morphological variables other than leaf thickness and specific leaf area. •  These results show that, while leaf length, width and area are partially controlled by genetic factors, these variables are plastic and respond to environmental influences associated with a particular altitude. Thus altitudinal trends in leaf size in N. cunninghamii are unlikely to be the result of adaptation.

Australia's endemic desert shrubs are commonly aromatic, with chemically diverse terpenes and phenylpropanoids in their headspace profiles. Species from the genus Eremophila (Scrophulariaceae ex. Myoporaceae) are the most common, with 215 recognised taxa and many more that have not yet been described, widely spread across the arid parts of the Australian continent. Over the years, our research team has collected multiple specimens as part of a survey to investigate the chemical diversity of the genus and create leads for further scientific enquiry. In the current study, the diversity of volatile compounds is studied using hydrodistilled essential oils and leaf solvent extracts from 30 taxa. Several rare terpenes and iridoids were detected in chemical profiles widely across the genus, and three previously undescribed sesquiterpenes were isolated and are assigned by 2D NMR- E -11(12)-dehydroisodendrolasin, Z -11-hydroxyisodendrolasin and 10-hydroxydihydro-α-humulene acetate. Multiple sampling from Eremophila longifolia , Eremophila arbuscular , Eremophila latrobei , Eremophila deserti , Eremophila sturtii , Eremophila oppositifolia and Eremophila alternifolia coneys that species in Eremophila are highly chemovariable. However, taxa are generally grouped according to the expression of (1) furanosesquiterpenes, (2) iridoids or oxides, (3) mixtures of 1 and 2, (4) phenylpropanoids, (5) non-furanoid terpenes, (6) mixtures of 4 and 5, and less commonly (7) mixtures of 1 and 5. Furthermore, GC-MS analysis of solvent-extracted leaves taken from cultivated specimens conveys that many heavier 'volatiles' with lower vapour pressure are not detected in hydrodistilled essential oils and have therefore been neglected in past chemical studies. Hence, our data reiterate that chemical studies of the genus Eremophila will continue to describe new metabolites and that taxon determination has limited predictive value for the chemical composition.

Background: Five plants used traditionally by Australian Aboriginals and two edible native Australian fruits have been investigated for anticancer activity. The aim was to identify native Australian herbal medicines which displayed anticancer activity, with cytotoxicity to cancer cells but sparing or even proliferating normal immunological cells, and subsequently provide potentially new anticancer drug leads., Methods: Extracts and derived fractions were assayed for cell viability against a multiple myeloma cell line, RPMI-8226, in comparison to the peripheral blood mononuclear cells (PBMC) representing normal human immunological cells., Results: None of the crude extracts exhibited the desirable differential activity; however, following further fractionation of the Eremophila duttonii F. Muell. (Myoporaceae) extract, one fraction (termed F01) exhibited a greater cytotoxicity to the cancer cell line than to the normal cells., Conclusions: One fraction may potentially contain valuable compounds which may be useful for further investigation. This may focus on the identification of the bioavailable purified compounds present within these fractions or by detailed delineation of the related mechanisms of action.

Background: Myoporum bontioides A. Gray, an evergreen shrub from the Myoporaceae family, is a commonly used medicinal plant. Many studies have been conducted on the biologically active constituents of whole parts of M. bontioides. However, the endophytes of M. bontioides have not been intensively investigated. A new chlorine-containing isocoumarin, named dichlorodiaportinol A (1) was isolated from the endophytic fungus Trichoderma sp. 09 isolated from the root of M. bontioides. Its cytotoxic activity against human breast cancer (MCF-7) and human liver cancer (HepG2) cell lines was evaluated., Materials and Methods: Different open silica gel column chromatographic techniques with different solvent systems were used for the separation of the constituents of the ethyl acetate extract of the culture broth of the endophytic fungus Trichoderma sp. 09. The structure of compound one was identified by analysis of spectroscopic data [one-dimensional (1D), two-dimensional (2D)-nuclear magnetic resonance (NMR), ultraviolet (UV), infrared (IR) and Mass spectrometry (MS)]. 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay method was used for the evaluation of cytotoxic activity of compound one against MCF-7 and HepG2 cell lines., Results: Compound one was identified as 3-(3,3-dichloro-2,3-dihydroxy-propyl)-8-hydroxy-6- methoxy-isochromen-1-one. It inhibited MCF-7 and HepG2 cell lines, with half maximal inhibitory concentration (IC50) values of 17.8 and 39.6 μg/mL, respectively., Conclusions: Compound one is a new chlorine-containing isocoumarin with moderate cytotoxic activity against MCF-7 and HepG2 cell lines. Thus, endophytes of M. bontioides are worthy of consideration for the development and research of antitumor agents.

Eremophila spp. (Myoporaceae family), endemic to Australia, are evergreen shrubs or small trees occurring in arid, semi-arid, tropical, or temperate regions. In Europe, Eremophila spp. are grown for their horticultural appeal. During 2009 and 2010, extensive wilting was observed on 2-month to 1-year-old potted plants of Eremophila laanii F. Muell., E. glabra subsp. carnosa Chinnock, and E. maculata (Ker Gawl.) F. Muell. grown in a commercial nursery near Catania (southern Italy). Internally, symptomatic plants had conspicuous vascular discoloration from the crown to the canopy. Diseased crown and stem tissues were surface disinfested for 30 s in 1% NaOCl, rinsed in sterile water, plated on potato dextrose agar (PDA) amended with 100 mg/liter of streptomycin sulfate, and incubated at 25°C. A Fusarium sp. was consistently isolated from affected plant tissues. Colonies with purple mycelia and violet reverse colors developed after 9 days. On carnation leaf agar, single-spore isolates produced microconidia on short monophialides, macroconidia that were three to five septate with a pedicellate base, and solitary and double-celled or aggregated chlamydospores. A PCR assay was conducted on two representative isolates (ITEM 12591 and ITEM 12592) by analyzing sequences of the partial CaM gene (coding calmodulin protein) and benA (coding beta-tubulin protein) using the primers as reported by O'Donnell et al. (1). Calmodulin sequences of ITEM 12951 and ITEM 12952 isolates (GenBank Nos. FR671157 and FR671158) exhibited 99.8 and 99.5% identity with Fusarium oxysporum strain ITEM 2367 (GenBank No. AJ560774), respectively, and had 99.5% homology between them. BenA gene sequences of ITEM 12951 (GenBank No. FR671426) exhibited an identity of 100% to F. oxysporum f. sp. vasinfectum strain CC-612-3 (GenBank No. AY714092.1), and benA gene sequences of ITEM 12952 (GenBank No. FR671427) exhibited an identity of 100% to F. oxysporum f. sp. vasinfectum strain LA 140 (GenBank No. FJ466740.1), whereas the homology between the two strains is 99.5%. Morphological characteristics, as well as CaM and benA sequences, identified the isolates as F. oxysporum Schlechtend:Fr. Pathogenicity tests were performed by placing 1-cm 2 plugs of PDA from 9-day-old mycelial cultures near the crown on potted, healthy, 3-month-old cuttings of E. laanii, E. glabra subsp. carnosa, and E. maculata. Twenty plants for each species were inoculated with each isolate. The same number of plants served as noninoculated controls. All plants were enclosed for 4 days in plastic bags and placed in a growth chamber at 24 ± 1°C. Plants were then moved to a greenhouse where temperatures ranged from 23 to 27°C. Symptoms identical to those observed in the nursery developed 20 days after inoculation with both strains. Crown and stem discoloration was detected in all inoculated plants after 45 days. Wilting was detected on 15% of plants. Control plants remained symptomless. F. oxysporum was consistently reisolated from symptomatic tissues and identified as previously above. To our knowledge, this is the first report of F. oxysporum causing disease of Eremophila spp. worldwide. Reference: (1) K. O'Donnell et al. Mycoscience 41:61, 2000.

Plant species of the genus Eremophila (Myoporaceae) are native to Australia and are known to produce a diverse range of unusual secondary compounds. The purpose of this research was to examine the antimicrobial activity of 72 Eremophila species most of which had not been the subject of any previous pharmacological testing. Organic extracts of Eremophila species were screened for antimicrobial activity against Gram-positive and Gram-negative bacteria and yeasts of medical importance. Extracts of a number of Eremophila species showed selective activity against Gram-positive bacteria with MICs for the most active species in the range of 16 to 62 microg/ml for Streptococcus species, and 62 to 250 microg/ml for standard strains of Staphylococcus aureus. Extracts with the greatest activity against standard strains were tested against 68 clinical isolates of multi-resistant methicillin-resistant S. aureus (mMRSA). The majority of the clinical isolates were susceptible to concentrations below 62.5 microg/ml for the extracts of E. drummondii, E. linearis, E. serrulata, E. acrida, E. neglecta, E. virens and a new undescribed species affiliated with E. prolata. The extract of E. virens inhibited growth of all 68 clinical mMRSA isolates at the minimum tested concentration of 31 microg/ml. This study has shown for the first time that a number of different Eremophila species manifest selective antibacterial activity against Gram-positive organisms which are important causes of human disease. It shows that there are several Eremophila species possessing interesting antibacterial activity besides those that have published traditional use. These may yield novel antibacterial compounds with potential to be used in biomedical applications.

Activity-guided fractionation was used to determine the antibacterial component of an ethanolic extract of the leaves of an Australian native medicinal plant, Eremophila duttonii F. Muell. (Myoporaceae). The extract, previously shown to have activity against Gram positive bacteria, was shown to have activity against additional Gram positive bacteria, including Clostridium perfringens, C. sporogenes and Listeria monocytogenes. Thin layer chromatography (TLC) was used to separate the extract into seven coloured fractions in visible light, one of which was shown by bioautography to contain antibacterial activity. Recovery of the component from the TLC plate and testing for antibacterial activity using a plate-hole diffusion assay supported this result. The purity of the component was verified by high-performance liquid chromatography and a time-kill experiment indicated that the purified component showed identical bactericidal activity to the whole extract. TLC spray reagents indicated that the component was a sterol, terpene or sugar but not a flavonoid, while the pigmented nature suggested a carotenoid., (Copyright (c) 2004 John Wiley & Sons, Ltd.)

Ethanolic extracts of five traditional Australian medicinal plants, previously shown to display antibacterial activity against laboratory strains of the Gram positive bacteria Staphylococcus aureus and Enterococcus faecalis, were investigated for their abilities to inhibit clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE). Using plate-hole diffusion assays, the following results were obtained: (a) extract from the leaves of Eremophila alternifolia (Myoporaceae) showed activity against MRSA; (b) extract from the leaves of Acacia kempeana (Mimosaceae) showed incomplete inhibition of VRE; (c) extracts from the leaves of Amyema quandong (Loranthaceae) and Eremophila duttonii (Myoporaceae) were active against both types of bacteria; (d) extract from the stem base of Lepidosperma viscidum (Cyperaceae) was active against MRSA and exhibited incomplete inhibition of VRE. All active extracts were evaluated using time-kill assays. Most of the extracts showed bactericidal effects and reduced the number of viable cells by 4-6 logs within four hours, while the extracts from Acacia kempeana leaves and Lepidosperma viscidum stem base exhibited bacteriostatic activity against VRE. The extract from the leaves of Eremophila duttonii was the most active and reduced the number of viable cells of MRSA and VRE to undetectable levels within 1 hour.

Fifty-six ethanolic extracts of various parts of 39 plants used in traditional Australian Aboriginal medicine were investigated for their antibacterial activities against four Gram-positive (Bacillus cereus, Enterococcus faecalis, Staphylococcus aureus and Streptococcus pyogenes) and four Gram-negative (Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and Salmonella typhimurium) bacterial species. In a plate-hole diffusion assay, 12 extracts inhibited the growth of one or more of the bacteria, with five extracts showing broad spectrum antibacterial activity against Gram-positive bacteria. B. cereus was the most susceptible bacterium, with all 12 extracts displaying activity against this organism. Extracts from the leaves of Eremophila species (Myoporaceae) were the most active, with Eremophila duttonii exhibiting the greatest activity (against Gram-positive bacteria). The antibacterial effects of E. duttonii were further investigated by time-course growth assays which showed that significant growth inhibition was observed in cultures incubated in the presence of the extract within 1 h for B. cereus, E. faecalis and S. aureus and 2 h for S. pyogenes.

Twenty-eight extracts from 17 species of Australian native plants traditionally used as general anti-inflammatory medicines by Australian Aboriginal people were examined for inhibition of the enzyme xanthine oxidase (XO). The extracts from nine species were found to have more than 25% inhibition at a concentration of 100 microg/ml in the assay mixture. Extracts from three species Clerodendrum floribundum R. Br. (Verbenaceae), Eremophila maculata (Ker Gawler) (Myoporaceae) and Stemodia grossa Benth. (Scrophulariaceae) showed the greatest activity with inhibitions of 84, 61 and 57%, respectively, at 50 microg/ml, with four other species having more than 40% inhibitory activity at this concentration.

A molecular systematic study of Scrophulariaceae sensu lato using DNA sequences of three plastid genes (rbcL, ndhF, and rps2) revealed at least five distinct monophyletic groups. Thirty-nine genera representing 24 tribes of the Scrophulariaceae s.l. (sensu lato) were analyzed along with representatives of 15 other families of Lamiales. The Scrophulariaceae s.s. (sensu stricto) include part or all of tribes Aptosimeae, Hemimerideae, Leucophylleae, Manuleae, Selagineae, and Verbasceae (= Scrophularieae) and the conventional families Buddlejaceae and Myoporaceae. Veronicaceae includes all or part of tribes Angelonieae, Antirrhineae, Cheloneae, Digitaleae, and Gratioleae and the conventional families Callitrichaceae, Globulariaceae, Hippuridaceae, and Plantaginaceae. The Orobanchaceae include tribes Buchnereae, Rhinantheae, and the conventional Orobanchaceae. All sampled members of Orobanchaceae are parasitic, except Lindenbergia, which is sister to the rest of the family. Family Calceolariaceae Olmstead is newly erected herein to recognize the phylogenetic distinctiveness of tribe Calceolarieae. The Calceolariaceae are close to the base of the Lamiales. The Stilbaceae are expanded by the inclusion of Halleria. Mimulus does not belong in any of these five groups.

Conspicuous air spaces in Leucophyllum (Scrophulariaceae; Leucophylleae) leaves have been suggested to be developmentally transformed secretory cavities. We reinvestigated air space development in Leucophyllum frutescens, using freehand sections of mature fresh leaves and paraffin sections of several leaf stages. Each of the numerous air spaces per leaf forms because greater separation occurs within a local group of spongy mesophyll cells than in the developing spongy mesophyll elsewhere. We found no anatomical evidence of transitory epithelial cells or lysis of cells in developing air spaces, thus the hypothesis that air spaces are transformed secretory cavities is not supported. However, an important finding was that all leaves had one pair of conspicuous true secretory cavities flanking the midvein at the apex, each lined by an epithelium and filled with oil. We also found conspicuous apical cavities in freehand sections of herbarium specimens of this and three other Leucophyllum species. Cavities were not seen in L. revolutum or in the related Eremogeton grandiflorus. This is the first report and description of a true internal secretory cavity in Scrophulariaceae. In the related family Myoporaceae, we found epithelium-lined cavities scattered abundantly in leaves of cleared samples of three genera.

Extracts of 40 different plant species used in the traditional medicine of the Australian Aboriginal people have been investigated for antiviral activity. The extracts have been tested for activity against one DNA virus, human cytomegalovirus (HCMV) and two RNA viruses, Ross River virus (RRV) and poliovirus type 1, at non-cytotoxic concentrations. The most active extracts were the aerial parts of Pterocaulon sphacelatum (Asteraceae) and roots of Dianella longifolia var. grandis (Liliaceae), which inhibited poliovirus at concentrations of 52 and 250 microg/ml, respectively. The extracts of Euphorbia australis (Euphorbiaceae) and Scaevola spinescens (Goodeniaceae) were the most active against HCMV. Extracts of Eremophila latrobei subsp. glabra (Myoporaceae) and Pittosporum phylliraeoides var. microcarpa (Pittosporaceae) exhibited antiviral activity against RRV.

Four iridoid glycosides isolated from different species of Eremophila (Myoporaceae) were tested for their effects on the Langendorff rat heart. Melampyroside (1), verminoside (2), ferruloylajugol (3) and catalpol (4) all significantly decreased contractile force in the isolated rat heart test. Within a minute after administration all, except catalpol, significantly increased heart rate and coronary perfusion rate. These results are compared to those of other cardioactive iridoid glycosides described in the literature., (Copyright © 1997 Gustav Fischer Verlag. Published by Elsevier GmbH.. All rights reserved.)

The isolation and identification of two cardioactive compounds from two Eremophila species (Myoporaceae) considered important in the pharmacopoeia of the Australian Aboriginal people is described. Verbascoside, isolated from the methanol and water extracts of E. alternifolia leaves, mediated significant increases in chronotropism, inotropism and coronary perfusion rate (CPR) in the Langendorff rat heart. Geniposidic acid, isolated from the methanol extract of E. longifolia leaves, mediated an inhibitory effect with significant negative chronotropism, negative inotropism and CPR.

An aqueous extract obtained from the leaves of the traditional Aboriginal medicinal plant Eremophila alternifolia R.Br. (Myoporaceae) was tested on isolated hearts of normotensive rats using the Langendorff heart preparation. A single injection of the extract into the retrograge perfusion solution induced cardioactivity, consisting of a short initial increase in force of contraction (positive inotropism), followed by a decrease in the force (negative inotropism) with simultaneous increase in heart rate (positive chronotropism) and in coronary perfusion rate. These effects were not mediated by alpha- or beta-adrenergic receptors.