Family Icacinaceae Research Articles

Chloroplast genome sequence of Nothapodytes nimmoniana (Icacinaceae), a vulnerable reservoir of camptothecin from Western Ghats

Nothapodytes nimmoniana belongs to family Icacinaceae and is a major source of compound Camptothecin. The global demand for Camptothecin has caused large-scale exploitation of N. nimmoniana from its wild habitat in Western Ghats of India, thereby making it vulnerable. The species is known to exhibit genetic diversity among the populations in Western Ghats. In this study, we report plastome sequence of N. nimmoniana, first for the genus. For the study, the species was collected from Western Ghats of Maharashtra. The plastome of N. nimmoniana was 150,726 bp in length and exhibited typical quadripartite structure with 83,771 bp LSC, 18,513 bp SSC and 24,221 IR region. The plastome was characterized by presence of 124 unique genes, 87 protein coding genes, 29 tRNA genes and four rRNA genes. Further, the plastome was compared with the available basal lamiid plastomes for gene order and composition. N. nimmoniana plastome exhibited SSC region in an inverted configuration. Phylogenomic study placed N. nimmoniana sister to Mappia mexicana. The SSR markers identified in this study, might help to distinguish genetically diverse populations, prioritizing the populations which need immediate conservation effects as well as for checking adulteration.

Gas Chromatography-Mass Spectrometry Analysis, Phytochemical Screening, Antioxidant and Antibacterial Activity of Methanol Root Extract of Rhaphiostylis beninensis (Hook.f.) Planch. [family ICACINACEAE] against Uropathogens

In Nigeria, urinary tract infections (UTIs) are amid the most prevalent bacterial contagions, with a higher incidence rate in women. Depending on the severity of the infection, different antibiotics are used to treat UTIs. Herbal remedies have, on the other hand, been seen as the paranormal means of treating UTIs while antibiotic resistance has emerged as a problem for medics. This study examined the methanol extract of Rhaphiostylis beninensis (MRB) for its phytochemical and bioactive components, antioxidant, and antibacterial activities. From the study, the methanol extract's antioxidant activities grew stronger as concentration increased. Alkaloids, saponin, tannins, cardiac glycosides, flavonoids, and reducing sugar—giving the extract its pharmacological potential—were among the phytochemicals identified from the phytochemical screening. The GC-MS Chromatogram showed that there are 42 bioactive compounds in the MRB with the most prominent compounds being (Z,Z)-9,12-Octadecadienoic acid 19.96%, Lupeol 18.96%, cis-13-Octadecenoic acid 9.96%, n-Hexadecanoic acid 9.87%, 3',5'-Dimethoxyacetophenone 6.67%, 4-((1E)-3-Hydroxy-1-propenyl)-2-methoxyphenol 3.21%, Octadecanoic acid 2.98%, Capsaicin 2.58%, α-Amyrin 2.53%, 9-Octadecenoic acid (Z)-, 2,3-dihydroxypropyl ester 2.49%, and Vitamin E 2.45%. The MRB showed significant minimum inhibitory concentration values against Pseudomonas putida PA25T, Escherichia coli EC3AT, Pseudomonas aeruginosa PA129AT and Serratia marcescens and a significant MBC value against Escherichia coli EC3AT. This study emphasizes the medicinal efficacy of R. beninensis root and indicated that the plant could make a promising candidate for the isolation of bioactive compounds that could be synthesized as medications for the treatment of bacteria in uropathogenic disorders.

Comprehensive insight into the extract optimization, phytochemical profiling, and biological evaluation of the medicinal plant Nothapodytesfoetida

Nothapodytes foetida is an endemic medicinal plant belonging to the family Icacinaceae from the deciduous forests of the Western Ghats. The extract yield, nutritional and phytochemical composition, metabolite profiling of N. foetida leaf extract has been analysed, along with the assessment of its antioxidant, anti-inflammatory, anti-microbial and anti-cancer potentialities. The effect of different extracting solvents and techniques was optimized to obtain maximum extract yield, and ultrasonication-assisted extraction with aqueous methanol as the solvent was chosen. Appreciable amounts of total phenolics (67.59 mg GAE/g DW), total flavonoids (24.75 mg QCE/g DW), and total tannins (55.67 mg GAE/g DW) were detected. FT-IR spectroscopy also confirmed the presence of alcohols, phenols, alkanes, amino acids, carboxylic acids, nitro compounds and amines in the extract. Further, GC-MS analysis detected the presence of 33 volatile compounds that comprised of viminalol, α, β-amyrins, β, γ-sitosterol, 9-methoxy-camptothecin, lupeols, and various di and tri terpenes in significant quantities. 1H NMR spectra revealed well-resolved signals for flavonoids, amino acids (trp, his, tyr, phe, ala, ile, gly, gln, thr, val) and organic acids. LC-MS analysis of the methanolic N. foetida extract depicted a higher polyphenolic content followed by the ethanolic and aqueous extracts. Additionally, the aqueous methanolic extract of N. foetida exhibited significant in-vitro antioxidant, anti-inflammatory, anti-microbial activity and anti-cancer activity. The overall results of this work ascertain the potency of N. foetida in nutraceutical and biomedical applications.

Systematics and phylogeny of Oecopetalum (Metteniusaceae), a genus of trees endemic to North and Central America

The genus Oecopetalum Greenm. & C.H. Thomps. (Metteniusaceae) is distributed in the southeastern portion of Mexico to Central America (Guatemala, Nicaragua, and Costa Rica). Three species have been described and two have been reported as edible. We studied herbarium specimens of the genus Oecopetalum and performed a phylogenetic analysis based on the chloroplast genes matK and ndhF to answers to main questions: How many species are in the genus Oecopetalum? Is the genus Oecopetalum monophyletic? What are the generic relationships with other members of the family? Oecopetalum is a monophyletic genus with only two species. Pittosporosis is the sister group of Oecopetalum. The Trans-Atlantic relationship of Oecopetalum and Pittosporosis is a recurrent geographic pattern in the family Mettenuisaceae as well as in the family Icacinaceae. Our results, in concordance with fossil evidence and relationships of other groups, support the Boreotropical model of high-latitude terrestrial migrations of tropical taxa during the globally warm Paleocene–Eocene.

First record of an Icacinaceae Miers fossil flower from Le Quesnoy (Ypresian, France) amber

Flowers embedded in amber are rare. Only about 70 flowers or inflorescences have been described among which only one lamiid is known. Nevertheless, these fossils are important to our understanding of evolutionary process and past diversity due to the exceptional preservation of fragile structures not normally preserved. In this work, a new flower named Icacinanthium tainiaphorum sp. nov. from Le Quesnoy (Houdancourt, Oise, France) is described. Our phylogenetic analysis with extant species suggests that the affinity of this flower lies with the family Icacinaceae, close to Natsiatum or Hosiea. The fossil shows a combination of features unknown in extant Icacinaceae and we thus propose the description of a new fossil genus. It reveals a previously unknown diversity in the family and demonstrates the complementarity of different types of fossil preservation for a better understanding of past floral diversity.

Fluctuation in seed abundance has contrasting effects on the fate of seeds from two rapidly germinating tree species in an Asian tropical forest.

The seed predator satiation hypothesis states that high seed abundance can satiate seed predators or seed dispersers, thus promoting seed survival. However, for rapidly germinating seeds in tropical forests, high seed abundance may limit dispersal as the seeds usually remain under parent trees for long periods, which may lead to high mortality due to rodent predation or fungal infestations. By tracking 2 species of rapidly germinating seeds (Pittosporopsis kerrii, family Icacinaceae; Camellia kissi, family Theaceae), which depend on dispersal by scatter-hoarding rodents, we investigated the effects of seed abundance at the community level on predation and seed dispersal in the tropical forest of Xishuangbanna Prefecture, Southwest China. We found that high seed abundance at the community level was associated with delayed and reduced seed removal, decreased dispersal distance and increased pre-dispersal seed survival for both plant species. High seed abundance was also associated with reduced seed caching of C. kissi, but it showed little effect on seed caching of P. kerrii. However, post-dispersal seed survival for the 2 plant species followed the reverse pattern. High seed abundance in the community was associated with higher post-dispersal survival of P. kerrii seeds, but with lower post-dispersal survival of C. kissi seeds. Our results suggest that different plant species derive benefit from fluctuations in seed production in different ways.

Revision of Icacinaceae from the Early Eocene London Clay flora based on X-ray micro-CT

The Early Eocene (Ypresian) London Clay Formation contains one of the most important fruit and seed assemblages from the Paleogene, including a large diversity of taxa (>350 spp.) preserved as pyrite permineralizations retaining 3D structure as well as anatomical detail. Despite the importance of the flora for understanding angiosperm biogeographic and evolutionary history, the majority of the fossil material has not been revisited since the original taxonomic treatments by E.M. Reid and M.E.J. Chandler. Given subsequent advances in our understanding of angiosperm phylogeny and fruit morphology, coupled with technological advances in imaging/visualizing fossil material, many of the taxa represented in the flora deserve further study. Here we present a revision of the pantropical family Icacinaceae using X-ray micro-computed tomography (micro-CT) images of 21 species from the flora. Based on the results, four new combinations are made, a new fossil-genus is established for a distinctive species with affinities to the Phytocreneae, and emended diagnoses are provided for eight taxa. Of the seven genera recognized from the flora, only one, Iodes Blume, is extant. This study offers important insights on the biogeographic and evolutionary history of Icacinaceae, which is one of the most abundant and diverse components of the flora from the London Clay Formation.

Transcriptome analysis of stem wood of Nothapodytes nimmoniana (Graham) Mabb. identifies genes associated with biosynthesis of camptothecin, an anti-carcinogenic molecule.

Camptothecin (CPT), a monoterpene indole alkaloid, is a potent inhibitor of DNA topoisomerase I and has applications in treating ovarian, small lung and refractory ovarian cancers. Stem wood tissue of Nothapodytes nimmoniana (Graham) Mabb. (family Icacinaceae) is one of the richest sources of CPT. Since there is no genomic or transcriptome data available for the species, the present work sequenced and analysed transcriptome of stem wood tissue on an Illumina platform. From a total of 77,55,978 reads, 9,187 transcripts were assembled with an average length of 255 bp. Functional annotation and categorization of these assembled transcripts unraveled the transcriptome architecture and also a total of 13 genes associated with CPT biosynthetic pathway were identified in the stem wood tissue. Four genes of the pathway were cloned to full length by RACE to validate the transcriptome data. Expression analysis of 13 genes associated with CPT biosynthetic pathway in 11 different tissues vis-a-vis CPT content analysis suggested an important role of NnPG10H, NnPSLS and NnPSTR genes in the biosynthesis of CPT. These results indicated that CPT might be synthesized in the leaves and then perhaps exported to stem wood tissue for storage.

Evaluation of Cytotoxic and Antitumour Properties of Apodytes dimidiata and Characterisation of the Bioactive Component.

Apodytes dimidiata, belonging to the family Icacinaceae, is used for treating inflammation and various gastrointestinal ailments in Zulu traditional medicine. In the present study, significant cytotoxicity was exhibited by the methanolic extract of the A. dimidiata leaf against various cancer cell lines. The extract was purified partially through silica gel column by successive elution using various solvents of increasing polarity. Among these, the active methanolic fraction was found to be the most cytotoxic with IC50 values ranging from 0.92 to 3.95 µg/mL for Ehrlich's ascites carcinoma (a carcinoma cell line), Jurkat (human T lymphocyte cell line), and SK-BR-3 (mammary tumour cell line). The treated cells showed morphological alterations characteristic of apoptosis. Upon oral administration of active methanolic fraction at a dose of 250 mg/kg body weight, the solid tumour volume in mice was significantly reduced to 55.14% and the life span of the ascites tumour-bearing mice increased to 44.65% compared to untreated control. The active fraction with Rf value 0.56 was purified from the methanolic fraction by preparative thin-layer chromatography and was subjected to high-performance thin-layer chromatography, high-performance liquid chromatography, liquid chromatography-mass spectrometry, and nuclear magnetic resonance analysis. The iridoid glycoside genipin was identified as the active component.

Structure and Absolute Configuration of the 11-Noriridoid “Chapingolide”

The natural product chapingolide {systematic name: (4aS, 7R, 7aR)-7hydroxy-7-(hydroxymethyl)-4,4a,7,7a-tetrahydrocyclopenta[c]pyran-3(1H)-one}, C9H12O4, (1), is a new 11-nor-iridoid isolated from Calatola mollis Standl. The colourless compound has a bicyclic structure with an endo double bond in the five-membered ring. The absolute configuration was established by mean of the anomalous dispersion of the oxygen scatters present in the structure. Calatola is a genus of trees in family Icacinaceae. To date, about seven Calatola species are recorded in America, and most of them are found in rain forest regions.1 Approximately, four species of Calatola plants were recorded in Mexico1 of which, Calatola mollis Standl is a tree up to 20 m tall. The seeds exhibited potently vomitive-purgative activity.2 At present Calatola mollis species had been poorly investigated with only botanical descriptions reported to date.1 Even thought, specimens of this species have been misidentified as Calatola costaricensis,1,3 and hence a phytochemical study of plants of Calatola genus will be useful to establish differences in secondary metabolites composition as a valuable systematic character. The chapingolide (1), a nor-iridoid, was obtained from the methanolic extract of the leaves of Calatola mollis Standl, as a crystalline colourless compound. Its bicyclic structure is closely related to the structures discussed in Valladares and Rios,4 and Franzyk and Stermitz.5 The main differences reside in the oxidation HETEROCYCLES, Vol. 91, No. 7, 2015 1417

Phylogenetic relationships of Icacinaceae focusing on the vining genera

Phylogenetic relationships of the pantropical and polyphyletic family Icacinaceae were investigated, focusing on the Old World vining genera. Plastid ndhF, rbcL and matK sequences from taxa representing 32 of the 36 currently recognized genera were analysed with maximum parsimony and Bayesian methods. As in previous studies, our results show that the family is divided into several poorly resolved groups. An evaluation of the traditional tribal classification revealed Iodeae as polyphyletic and the monogeneric Sarcostigmateae as sister to a monophyletic Phytocreneae (with the inclusion of Rhyticaryum). In Iodeae, the monospecific, eastern Malesian Polyporandra was embedded in the Old World Iodes. A strongly supported clade containing Phytocreneae plus Rhyticaryum was present in the Icacina group. The tropical African genera Chlamydocarya and Polycephalium were embedded in the Old World Pyrenacantha. Further relationships in the family and potential synapomorphic characters of the clades are discussed. New combinations are made for Polyporandra and Chlamydocarya/Polycephalium spp., which are formally synonymized with Iodes and Pyrenacantha, respectively. Conclusions about family-level relationships (and circumscription) cannot be reached with these data because of several weakly supported inter-relationships between some clades, such as Cassinopsis, Platea/Calatola and the Emmotum group.

Cytotoxic and growth inhibitory activity of aqueous extracts of root and leaf of <i>Rhaphiostylis beninensis</i> Planch ex Benth and <i>Pyrenacantha staudtii</i> Engl (Icacinaceae)

The aqueous extract of the root and leaf of Rhaphiostylis beninensis Planch ex Benth and Pyrenacantha staudtii Engl, both belonging to the family Icacinaceae were investigated for their claimed anti-tumour activity using simple bench top assays. The cytotoxic activity was evaluated within the concentration range of 1-20µg/ml using the tadpole mortality test, while the growth inhibitory activity was investigated within the concentration range of 1-30mg/ml using the Sorghum bicolor seed growth inhibitory test. At a concentration of 20µg/ml, aqueous extracts of the root and leaf of Rhaphiostylis beninensis was observed to effect a mortality of 100 and 80% respectively, while aqueous extract of the root and leaf of Pyrenacantha staudtii , at same concentration, effected mortality of 70 and 93% respectively. Nil mortality was observed in the control setup, while at other concentrations varying degrees of mortality was observed. All the extracts produced varying degrees of inhibition of growth in the emerging radicle of the seed of Sorghum bicolor. The results obtained suggest that these plants may probably have constituents with anti-tumour activities that merit further investigation using other test systems. Keywords : Rhaphiostylis beninensis; Pyrenacantha staudtii; Icacinaceae; Cytotoxic ; Growth inhibition

New plant sources of the anti-cancer alkaloid, camptothecine from the Icacinaceae taxa, India

In this study, the production of camptothecine and its derivatives, in thirteen species of the family Icacinaceae, namely, Apodytes dimidiata, Codiocarpus andamanicus, Gomphandra comosa, Gomphandra coriacea, Gomphandra polymorpha, Gomphandra tetrandra, Iodes cirrhosa, Iodes hookeriana, Miquelia dentata, Miquelia kleinii, Natsiatum herpeticum, Pyrenacantha volubilis and Sarcostigma kleinii is reported. Seeds of M. dentata were found to produce the highest content of camptothecine (1.0–1.4% by dry weight of seeds). Full scan LC–MS and ESI-MS/MS analysis of M. dentata revealed, besides camptothecine, a number of other derivatives, namely, 10-hydroxycamptothecine, 9-methoxycamptothecine, 20-deoxycamptothecine. Crude extract preparations of the seeds of M. dentata were effective against a breast cancer cell line (IC50=3.82μg/ml for MDA MB273 cell lines) and two ovarian cancer cell lines (IC50=2.8μg/ml for NCI/ADR-RES and 4.5μg/ml for SKOV). These results are the first reports of camptothecine and its derivatives in these species and offer rich alternative plant sources for the anticancer compound, camptothecine.

Fruits of Icacinaceae from the Eocene of Southeastern North America and Their Biogeographic Implications

We describe three members of the pantropical family Icacinaceae from the Eocene Claiborne Group of Tennessee and Mississippi, providing the first recognition of this family from the Mississippi Embayment of southeastern North America. Natsiatum wilcoxiana (Berry) comb. nov. and Phytocrene densipunctata sp. nov. represent extant genera presently confined to Southeast Asia and Malesia. Croomiocarpon mississippiensis gen. et sp. nov. represents an extinct genus with affinities to the tribe Iodeae. These fossils increase the generic diversity of Icacinaceae’s fossil record and expand our understanding of the historical distribution of this family. Given the similarity of N. wilcoxiana and P. densipunctata with several fossils known from the Eocene of Europe, these records suggest a broader pattern of floristic connection between southeastern North America and Europe during the Eocene.

Sequestration of Camptothecin, an Anticancer Alkaloid, by Chrysomelid Beetles

Camptothecin (CPT), a monoterpene indole alkaloid, is a potent inhibitor of eukaryotic toposiomerase-I. Several derivatives of CPT are in clinical use against ovarian and lung cancers. CPT has been reported from several plant species belonging to the order Asterids, with the highest concentration in Nothapodytes nimmoniana (family Icacinaceae). In this paper, we report an intriguing observation of chrysomelid beetles (Kanarella unicolor Jacobby) feeding on the leaves of N. nimmoniana without any apparent adverse effect. LC-MS/MS analysis of the beetles indicated that 54.9% of the ingested CPT's was recovered from the wings, followed by lesser amounts in the head and abdomen. LC-HRMS analysis revealed that most of the CPT in the insect body was in the parental form available in the plants without any major metabolizable products, including sulfated and glucuronilated forms. The mechanism by which the beetles are able to tolerate substantially high levels of CPT in their body tissue is under investigation.

Floral resources, pollinators and fruiting in a threatened tropical deciduous tree

Aims Nothapodytes nimmoniana (Family Icacinaceae) is a deciduous tree species distributed in Asia facing severe population decline. Wood chips from the tree are a source of camptothecin, a globally soughtafter alkaloid with cancer-treating properties, and are harvested unsustainably in natural forests. We studied the pollination ecology of the species and asked if there are constraints in pollination and fruiting success in its natural populations. We also discuss the potential effects of wood extraction on pollinators and reproductive success in the population. Methods The study was carried out during the flowering season in two populations with varying exploitation levels, both located in the Protected Area Network in the Western Ghats of Karnataka State. We assessed floral resource availability and measured pollinator diversity and activity in the canopy from the perspective of the biology of the species. We quantified pollinator abundance and percent fruit set as a function of population density. Important Findings Flowers belong to the fly pollination syndrome and are open to visits by generalists such as Apis dorsata and Trigona iridipennis. Fruiting success did not vary between exploited and unexploited sites, but there was a significant difference in pollinator assemblage. In a lean male flowering year, fruit set was low, suggesting pollen limitation in the population. No fruiting was recorded in the second year at the site where trees were felled soon after the flowering period. We argue that when male floral resources are altered in exploited populations, pollinators of generalist nature may show a shift in foraging pattern. Also, generalist plants may be as susceptible to pollination loss as are specialist plants. The need for outcrossing in the species would probably exacerbate this vulnerability.

An Acetylated Eudesmane Glucoside from Apodytes dimidiata Growing in Madagascar

A new eudesmane glucoside has been isolated from the aerial part of Apodytes dimidiata, an Icacinaceous plant widely used in the Southern African countries for its molluscicidal activities. The structure of the new compound was determined by spectroscopic methods. The chemosystematics of the genus belonging to the family Icacinaceae is discussed.

Multiple origin of the tropical forest tree family Icacinaceae

Analyses of DNA sequences from four genes (ndhF, rbcL, atpB, and 18S rDNA) and morphological data show that the members of the tropical forest tree family Icacinaceae do not have a common origin. All of the genera earlier placed in Icacinaceae are euasterids but placed in the following three different orders: Garryales, Aquifoliales, and Apiales. Icacina and related genera are members of Garryales and, pending more data, are still best treated as Icacinaceae (sensu stricto). The genera related to Aquifoliales are placed in Cardiopteridaceae and a new family, Stemonuraceae. The genus Pennantia is a member of Apiales and the family Pennantiaceae is recognized. Morphological characters delimiting these groups are discussed. Twenty-six new ndhF sequences were obtained for the study (25 from former Icacinaceae and 1 from Cardiopteris).

Jodes israeliisp. nov.: a huge phosphate-mineralized icacinacean fructification from the Late Cretaceous of the Negev, southern Israel

A beautifully preserved fossil fructification nearly 3 cm in maximum diameter was found in organic-rich Maastrichtian strata (Ghareb Formation) of the central Negev, southern Israel. The endocarp is entirely phosphate-mineralized, and was encountered embedded within a layer of phosphate concretions associated with a sediment-discontinuity surface in the lower part of the Maastrichtian Ghareb succession. The fossil endocarp is typical of those of the Recent family Icacinaceae, a pantropical group including trees, shrubs and climbers, distributed in primary and secondary rainforests, particularly in Malaysia and SE Asia. It is also comparable with similar taxa from the English Eocene. The attributes of the endocarp fit very well those of the genus Jodes.The ultrastructure of the phosphatic material and the state of preservation of the fructification suggest rapid, bacterially-induced phosphatization of the plant remains by dissolved pore phosphate, released by early decomposition of associated organic matter at, or near, the bottom sediment/water interface. A low net accretion rate related to gentle current activity on the sea floor maintained the Jodesendocarp at close proximity to this interface where phosphate precipitation took place. The structural configuration of the area during Campanian-Maastrichtian times suggests the nearby Ramon High as a possible source of the fructification. In terms of the climate system of Troll & Pfaffen, the Jodesfind also suggests tropical wet climate VI with humid summers during the Maastrichtian in Israel.

Study of antiulcer activity of aqueous extract of leaves of Pyrenacantha staudtii (family icacinaceae) using various models of experimental gastric ulcer in rats

The antiulcer activity of an aqueous extract of Pyrebnacantha staudtii was studied in rats and was compared with that of cimetidine. Ulcers were induced in rats by means of various experimental models: aspirin, indomethacin, serotonin and reserpine. The extract was found to have significant antiulcer activity against all the models studied. Cimetidine was found to be ineffective agianst serotonin- and reserpine-induced ulcers.