Ramanan Uma Shaanker

Dysoxylum binectariferum Hook.f (Meliaceae), a rich source of rohitukine

Rohitukine, a chromane alkaloid, is a precursor of flavopiridol, a promising anti-cancer compound. Currently in Phase III clinical trials, flavopiridol is a potent inhibitor of several cyclin-dependent kinases (CDKs). Rohitukine was first reported from Amoora rohituka (0.083% dry weight) followed by that in Dysoxylum binectariferum (0.9% dry weight), both belonging to the family Meliaceae. Here, we report incredibly high yields of rohitukine (7% dry weight) in trees of D. binectariferum from the Western Ghats, India. Crude extracts of the tree were found to be highly effective against ovarian and breast cancer lines tested.

Genetic structure and demographic history of the endangered tree species Dysoxylum malabaricum (Meliaceae) in Western Ghats, India: implications for conservation in a biodiversity hotspot.

The impact of fragmentation by human activities on genetic diversity of forest trees is an important concern in forest conservation, especially in tropical forests. Dysoxylum malabaricum (white cedar) is an economically important tree species, endemic to the Western Ghats, India, one of the worlds eight most important biodiversity hotspots. As D. malabaricum is under pressure of disturbance and fragmentation together with overharvesting, conservation efforts are required in this species. In this study, range-wide genetic structure of twelve D. malabaricum populations was evaluated to assess the impact of human activities on genetic diversity and infer the species’ evolutionary history, using both nuclear and chloroplast (cp) DNA simple sequence repeats (SSR). As genetic diversity and population structure did not differ among seedling, juvenile and adult age classes, reproductive success among the old-growth trees and long distance seed dispersal by hornbills were suggested to contribute to maintain genetic diversity. The fixation index (FIS) was significantly correlated with latitude, with a higher level of inbreeding in the northern populations, possibly reflecting a more severe ecosystem disturbance in those populations. Both nuclear and cpSSRs revealed northern and southern genetic groups with some discordance of their distributions; however, they did not correlate with any of the two geographic gaps known as genetic barriers to animals. Approximate Bayesian computation-based inference from nuclear SSRs suggested that population divergence occurred before the last glacial maximum. Finally we discussed the implications of these results, in particular the presence of a clear pattern of historical genetic subdivision, on conservation policies.

Forest trees in human modified landscapes: ecological and genetic drivers of recruitment failure in Dysoxylum malabaricum (Meliaceae).

Tropical agro-forest landscapes are global priority areas for biodiversity conservation. Little is known about the ability of these landscapes to sustain large late successional forest trees upon which much forest biodiversity depends. These landscapes are subject to fragmentation and additional habitat degradation which may limit tree recruitment and thus compromise numerous ecosystem services including carbon storage and timber production. Dysoxylum malabaricum is a large canopy tree species in the Meliaceae, a family including many important tropical timber trees. This species is found in highly fragmented forest patches within a complex agro-forest landscape of the Western Ghats biodiversity hot spot, South India. In this paper we combined a molecular assessment of inbreeding with ecological and demographic data to explore the multiple threats to recruitment of this tree species. An evaluation of inbreeding, using eleven microsatellite loci in 297 nursery-reared seedlings collected form low and high density forest patches embedded in an agro-forest matrix, shows that mating between related individuals in low density patches leads to reduced seedling performance. By quantifying habitat degradation and tree recruitment within these forest patches we show that increasing canopy openness and the increased abundance of pioneer tree species lead to a general decline in the suitability of forest patches for the recruitment of D. malabaricum. We conclude that elevated inbreeding due to reduced adult tree density coupled with increased degradation of forest patches, limit the recruitment of this rare late successional tree species. Management strategies which maintain canopy cover and enhance local densities of adult trees in agro-forest mosaics will be required to ensure D. malabaricum persists in these landscapes. Our study highlights the need for a holistic understanding of the incipient processes that threaten populations of many important and rare tropical tree species in human dominated agro-forest landscapes.

Pyrenacantha volubilis Wight, (Icacinaceae) a rich source of camptothecine and its derivatives, from the Coromandel Coast forests of India

Camptothecine, a potent eukaryotic topoisomerase inhibitor, is an important anticancer compound. The global demand for this compound was estimated to be

Translating Endophyte Research to Applications: Prospects and Challenges

1 billion in 2003 and is only further expected to increase. Partly to meet the expected increase in demand, in the recent past, several efforts have been made to discover newer and alternative plant and fungal sources of camptothecine. In this study we report a rich source of camptothecine and its natural derivatives, Pyrenacantha volubilis (Icacinaceae) from the eastern coast of peninsular India. Camptothecine and its derivatives were analyzed using high performance liquid chromatography (HPLC) coupled with electrospray mass spectrometry (ESI-MS) in all plant parts such as twigs, leaves, roots, seedling, ripened whole fruit, fruit coat, seed coat and cotyledons. Cotyledons and ripened whole fruits contained the highest amount of camptothecine (1.35% and 0.60% dry weight respectively). LC-MS and ESI-MS/MS analyses revealed besides camptothecine, other derivatives and precursors such as 10-hydroxycamptothecine, 9-methoxycamptothecine, 20-deoxycamptothecine, deoxypumiloside, strictosidine and strictosamide. Pure camptothecine was isolated from fruits and structurally confirmed using NMR. Seed extracts were found to be effective against breast cancer, ovarian, colon and carcinoma cell lines (with IC50 values of 4.0 μg/mL, 6.5 μg/mL, 25.0 μg/mL and 25.0 μg/mL respectively). We discuss the results in the context of exploring alternative sources of camptothecine.

Desorption Electrospray Ionization (DESI) Mass Spectrometric Imaging of the Distribution of Rohitukine in the Seedling of Dysoxylum binectariferum Hook. F.

The horizontally transmitted fungal endophytes cause symptomless infections of plants. Endophytes synthesize a diverse array of metabolites and enzymes, an attribute that reflects their milieu: the host plant and co-occurring microbes. We illustrate the versatility and utility of endophytes by highlighting examples of how they enhance plant tolerance to abiotic/biotic stressors, produce biomolecules with unique molecular architectures, and elaborate novel enzymes of industrial importance. After considering the challenges that have retarded the translation of these findings to payoffs, we suggest future directions, including the need to understand the ecology and in planta community structure of endophytes, to help fully realize their technological potential.

Authentication of Garcinia fruits and food supplements using DNA barcoding and NMR spectroscopy

Ambient ionization mass spectrometric imaging of all parts of the seedling of Dysoxylum binectariferum Hook. f (Meliaceae) was performed to reconstruct the molecular distribution of rohitukine (Rh) and related compounds. The species accumulates Rh, a prominent chromone alkaloid, in its seeds, fruits, and stem bark. Rh possesses anti-inflammatory, anti-cancer, and immuno-modulatory properties. Desorption electrospray ionization mass spectrometry imaging (DESI MSI) and electrospray ionization (ESI) tandem mass spectrometry (MS/MS) analysis detected Rh as well as its glycosylated, acetylated, oxidized, and methoxylated analogues. Rh was predominantly distributed in the main roots, collar region of the stem, and young leaves. In the stem and roots, Rh was primarily restricted to the cortex region. The identities of the metabolites were assigned based on both the fragmentation patterns and exact mass analyses. We discuss these results, with specific reference to the possible pathways of Rh biosynthesis and translocation during seedling development in D. binectariferum.

Role of endophytes in early seedling growth of plants: a test using systemic fungicide seed treatment

Garcinia L. (Clusiaceae) fruits are a rich source of (−)-hydroxycitric acid, and this has gained considerable attention as an anti-obesity agent and a popular weight loss food supplement. In this study, we assessed adulteration of morphologically similar samples of Garcinia using DNA barcoding, and used NMR to quantify the content of (−)-hydroxycitric acid and (−)-hydroxycitric acid lactone in raw herbal drugs and Garcinia food supplements. DNA barcoding revealed that mostly G. gummi-gutta (previously known as G. cambogia) and G. indica were traded in Indian herbal markets, and there was no adulteration. The content of (−)-hydroxycitric acid and (−)-hydroxycitric acid lactone in the two species varied from 1.7% to 16.3%, and 3.5% to 20.7% respectively. Analysis of ten Garcinia food supplements revealed a large variation in the content of (−)-hydroxycitric acid, from 29 mg (4.6%) to 289 mg (50.6%) content per capsule or tablet. Only one product contained quantifiable amounts of (−)-hydroxycitric acid lactone. Furthermore the study demonstrates that DNA barcoding and NMR could be effectively used as a regulatory tool to authenticate Garcinia fruit rinds and food supplements.

Fusarium proliferatum, an endophytic fungus from Dysoxylum binectariferum Hook.f, produces rohitukine, a chromane alkaloid possessing anti-cancer activity

Systemic fungicide seed treatments are routinely used in conventional agriculture to control soil and seed-borne diseases, but little is known about their unintended adverse effects on non-target beneficial fungal endophytes that are known to be involved in plant growth and development. This study evaluated the seed treatment effect of a broad spectrum systemic fungicide, carbendazim (bavistin) on symbiotic association of fungal endophytes in rice and on early seedling growth of rice, green gram, soybean, and cowpea. Seeds were surface sterilized with sodium hypochlorite followed by 0.2% bavistin treatment. Growth of fungal endophytes was significantly affected by the seed treatment with fungicide in rice seedlings, while shoot and root growth was suppressed in all the crops. Quantitative real time PCR showed that the level of expression of two basal transcriptional regulator genes, OsBTF3 and OsNF-YC1 that are required for seed germination and seedling growth significantly decreased in bavistin treated rice seedlings. Re-inoculation of consortia of fungal endophytes onto bavistin treated rice seedlings significantly recovered seedling growth and development. These results suggest that fungicide treatment of seeds affects early seedling growth and has negative impact on beneficial fungal endophytes that are involved in plant growth and development. This study provides information on possible ill effects of fungicide on beneficial fungal endophytes that play key roles in early seedling growth of plants and also open up the prospect to additional research on different crops in vitro and field conditions to determine the consequences of fungicide effects and optimise fungicide application strategies to develop sustainable disease control methods.