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Endophytes, both bacterial and fungal, constitute an integral component of the leaf ecosystem. Here we argue that the respiratory metabolism of endophytes in the intercellular spaces of leaves could have a significant role in enhancing leaf photosynthesis by enriching the internal COsub2/subconcentration, especially in C3 plants.

Camptothecin the third most in demand alkaloid, is commercially extracted in India from the endangered plant, Nothapodytes nimmoniana. Endophytes, the microorganisms that reside within plants, are reported to have the ability to produce host–plant associated metabolites. Hence, our research aims to establish a sustainable and high camptothecin yielding endophyte, as an alternative source for commercial production of camptothecin. A total of 132 endophytic fungal strains were isolated from different plant parts (leaf, petiole, stem and bark) of N. nimmoniana, out of which 94 were found to produce camptothecin in suspension culture. Alternaria alstroemeriae (NCIM1408) and Alternaria burnsii (NCIM1409) demonstrated camptothecin yields up to 426.7 ± 33.6 µg/g DW and 403.3 ± 41.6 µg/g DW, respectively, the highest reported production to date. Unlike the reported product yield attenuation in endophytes with subculture in axenic state, Alternaria burnsii NCIM1409 could retain and sustain the production of camptothecin up to ~ 200 μg/g even after 12 continuous subculture cycles. The camptothecin biosynthesis in Alternaria burnsii NCIM1409 was confirmed using 13C carbon labelling (and cytotoxicity analysis on different cancer cell lines) and this strain can now be used to develop a sustainable bioprocess for in vitro production of camptothecin as an alternative to plant extraction.

Contemporary losses of biodiversity, sometimes referred to as the sixth mass extinction, continue to mount (1, 2). A recent assessment by the Intergovernmental Science-Policy Platform on Biodiversity and Ecosystem Services (IPBES) estimates that one million of approximately 10 million species that exist now are threatened with extinction along with the ecosystems they inhabit (3). Yet, in the post-coronavirus disease 2019 (COVID-19) world, investments in conservation are likely to decline further. To arrest biodiversity losses, much of the recent debate advocates two traditional approaches: Put more land under wilderness (4, 5), and mitigate drivers of change through improved governance and policies (2). India and other nations need new frameworks that integrate science with policies to enhance human well-being, restore and conserve nature, and build capacity. Image credit: Sandesh Kadur (photographer). However, in the populous tropics, where most biodiversity resides, conservation of our ecosystems will remain elusive until there is a clear demonstration of the relevance of biodiversity to people’s well-being, most of whom live below the poverty line. Although nature-based solutions for addressing our most pressing challenges such as climate change, water scarcity, agricultural intensification, and health are well known (3), the link between biodiversity and human well-being has remained elusive. A major constraint, at least in the tropics, has been the lack of investment in biodiversity science that can build human resources and demonstrate the potential of biodiversity in 1) tackling climate change, natural disasters, stagnating agricultural productivity, water shortages, polluted air, and newly emerging infectious diseases such as COVID-19, 2) increasing human well-being, and 3) fostering future socioeconomic development. Here, we outline a National Mission on Biodiversity and Human Well-being for India that calls for a considerable increase in investment for this new framework for integrating biodiversity science with human well-being. This will demonstrate the linkages … [↵][1]1To whom correspondence may be addressed. Email: kamal.bawa{at}gmail.com. [1]: #xref-corresp-1-1

Endophytes, both of bacterial and fungal origin, are ubiquitously present in all plants. While their origin and evolution are enigmatic, there is burgeoning literature on their role in promoting growth and stress responses in their hosts. We demonstrate that a salt-tolerant endophyte isolated from salt-adapted Pokkali rice, a Fusarium sp., colonizes the salt-sensitive rice variety IR-64, promotes its growth under salt stress and confers salinity stress tolerance to its host. Physiological parameters, such as assimilation rate and chlorophyll stability index were higher in the colonized plants. Comparative transcriptome analysis revealed 1348 up-regulated and 1078 down-regulated genes in plants colonized by the endophyte. Analysis of the regulated genes by MapMan and interaction network programs showed that they are involved in both abiotic and biotic stress tolerance, and code for proteins involved in signal perception (leucine-rich repeat proteins, receptor-like kinases) and transduction (Ca2+ and calmodulin-binding proteins), transcription factors, secondary metabolism and oxidative stress scavenging. For nine genes, the data were validated by qPCR analysis in both roots and shoots. Taken together, these results show that salt-adapted Pokkali rice varieties are powerful sources for the identification of novel endophytes, which can be used to confer salinity tolerance to agriculturally important, but salt-sensitive rice varieties.

Bacillus velezensis EB14, isolated from a leaf of Populus × jackii, possesses antagonistic activity against Sphaerulina musiva, a fungal pathogen of Populus sp. that causes leaf spots and stem cankers on poplars, limiting the utility of hybrid poplars as plantation trees. We sequenced the genome of B. velezensis EB14 to gain insights into the underlying basis of its antagonistic activity. Here, we report the complete genome sequence of B. velezensis EB14, a gram-positive bacterium of the family Bacillaceae. Through antiSMASH analysis, we predicted several gene clusters coding for the biosynthesis of antimicrobial compounds and several genes involved in plant bacterial interactions. These findings support the potential of developing B. velezensis EB14 as a biocontrol agent against S. musiva in poplar plantations. The genome of B. velezensis EB14 along with genome sequences of closely related B. velezensis species are invaluable for comparative genomic analyses to gain insights into bacterial, fungal, and host plant interactions.

Sclerotium rolfsii is a necrotrophic, soil-borne pathogenic fungus responsible for serious crop losses worldwide. It is one of the major phytopathogens infecting chilli and causes yield losses ranging from 16 to 80%. Among the various approaches to manage S. rolfsii, biological control is an environmentally acceptable alternative over other chemical methods. In this study, 120 endophytic fungal isolates obtained from a medicinal plant, Nothapodytes nimmoniana were evaluated for their in vitro and in vivo antagonism towards S. rolfsii. Among these, one isolate, Alternaria sp. exhibited 46.62% inhibition against S. rolfsii in dual culture assay on PDA. The inhibition was found to be mediated by the mycotoxin, tenuazonic acid, produced by the endophyte. Pre-germinated chilli seeds treated with mycelial suspension of Alternaria sp. reduced the seedling mortality due to S. rolfsii infection. Pretreatment with the endophyte also enhanced seedling growth and biomass compared to untreated seedlings under greenhouse conditions. These results suggest that the endophytic fungus, Alternaria sp. has potential implication for use as a biocontrol against S. rolfsii as well as for promoting growth of chilli.

Alternative splicing (AS) of transcripts is a well-recognized phenomenon in plants. Several studies have shown that the frequency of AS events are significantly enhanced in plants subjected to abiotic stresses, presumably as an adaptation to increase transcript diversity. In this study, we examine the effect of a salt tolerant endophytic fungus, Fusarium sp. on the AS events of IR-64 rice plants, under salinity stress. RNA seq data generated from rice plants, treated and not treated with Fusarium sp. respectively and subjected to 150 mM salinity stress was used to analyze the number and type of AS events and their chromosomal distribution using approriate bioinformatics pipeline. Besides, we also annotated the locus IDs of genes for gene enrichment and KEGG pathway analysis. Our results show an unequivocal decrease in the number of AS events under salinity stress as influenced by the endophyte. The average AS events per gene also decreased from 2.28 to 2.11 upon colonization by the endophyte. This is the first documented evidence of an endophyte-induced alteration in the frequency of alternative splicing event in plants subjected to salinity stress. The exact mechanisms through which the splicing activities are restrained, however need to be further investigated.

Colchicine, a well-known alkaloid, is a potent inhibitor of polymerization of tubulin leading to mitotic arrest. It is highly toxic to eukaryotic cells but also widely used in the field of medicine and plant breeding. Gloriosa superba (family: Colchicaceae) is an important natural source of colchicine. The seeds, tubers and leaves of this plant contain about 0.8, 1.2 and 0.014% colchicine by dry weight respectively. A noctuid moth, Polytela gloriosae (family: Noctuidae), feeds voraciously on leaves of G. superba without any adverse effect. However, the fate of colchicine and the mechanisms by which the insect is able to overcome the toxicity of the metabolite is not known. Here, we trace the fate of colchicine in both, the larva and moth of P. gloriosae. Colchicine was quantified in different body parts of the larvae and moth by high performance liquid chromatography, liquid chromatography mass spectrophotometry and nuclear magnetic resonance methods. Of the total colchicine taken in by the larva, a larger portion was excreted, while the rest was sequestered in its cuticle. In the moths however, the wings, legs and antennae were found to accumulate high amount of colchicine. The sequestered colchicine, in both the larva and adult, were chemically identical to that found in the plant. Negligible amounts of demethyl-(−)-colchicine, a less toxic derivative of colchicine was also detected. We discuss the probable adaptive significance of sequestration of colchicine by the insect.

Plant domestication, at least in cereals, is associated with the loss of the shattering allele. In such species, grains are manually harvested and threshed, leaving behind naked seeds. This could have led to the loss of maternally borne seed microbiomes and their associated benefits in modern-day domesticated species.

Camptothecin (CPT), a monoterpene indole alkaloid, is a potent inhibitor of eukaryotic topoisomerase I (Top 1). Because of this property, several derivatives of CPT are widely used as chemotherapeutic agents. The compound is produced by several plant species, including Nothapodytes nimmoniana (Family: Icacinaceae) presumably as a deterrent to insect pests. Here, we report, a lepidopteran larva, Lymantria sp. of Lymantriidae family which feeds voraciously on the leaves of N. nimmoniana, without any adverse consequences. Larval body weight and molting period were unaffected despite captive feeding of the larva with CPT enriched leaves. Mass spectrometric analysis indicated that nearly 46% of the ingested CPT was excreted while the rest was sequestered predominantly in the exuviae and setae (~35%). Although most of the CPT was in the parental form as found in the plant, traces of inactive, sulfated forms of CPT were recovered from the larva. Compared to that in plant, there were no critical mutations at the CPT binding domain of the insect's Top 1. The gut pH of the larva was alkaline (pH 10.0). The alkaline gut environment converts CPT from its active, lactone form to inactive, carboxylate form. It is likely that such conversion might help the larva to reduce the overall burden of CPT in its gut. We discuss the results in the context of the mechanisms of resistance adapted by insects to plant toxins.

A total of 507 endophytic fungal isolates belonging to 82 operational taxonomic units (OTUs) were isolated from fifteen plant species naturally occurring in the Thar Desert, Rajasthan, India. Aspergillus, Alternaria, Chaetomium, Penicillium and Nigrospora were among the dominant fungal genera. Thermotolerance of the isolates was evaluated by culturing the fungi at 40 °C and 45 °C in shake flasks. Growth of nine OTUs (ACJ-2, ACJ-5, ACL-2, ACT-2, ACT-3, LAS-4, LAS-6, SAP-3 and SAP-6) was unaffected at 40 °C. Of these, six endophytes, namely, ACJ-2, ACJ-5 (Aspergillus flavus), SAP-3 (Aspergillus sp.), SAP-6, LAS-4 (Aspergillus sp.) and LAS-6 (Chaetomium sp.), were tolerant to as high as 45 °C. Rest of the OTUs did not survive culture temperatures beyond 35 °C. We evaluated the ability of one thermotolerant endophyte, LAS-6 (Chaetomium sp.) to confer high temperature tolerance and three OTUs, namely, LAS-4 (Aspergillus sp.), SAP-3 (Aspergillus sp.) and SAP-6 to confer drought tolerance to a rice cultivar, IR-64, at early seedling stage. Seedlings treated with LAS-6 (Chaetomium sp.) showed a higher survival percentage as well as maintained a significantly higher shoot and root growth under high temperature stress compared to seedlings not treated with the fungus. Under drought stress, endophyte treated seedlings maintained a significantly higher root growth compared to untreated seedlings. The identified thermotolerant fungal sources could be potentially useful in alleviating abiotic stress in agriculturally important crops.

The global economy of the international trade of herbal products has been increasing by 15% annually, with the raw material for most herbal products being sourced from South and Southeast Asian countries. In India, of the 8000 species of medicinal plants harvested from the wild, approximately 960 are in the active trade. With increasing international trade in herbal medicinal products, there is also increasing concern about the widespread adulteration and species admixtures in the raw herbal trade. The adverse consequences of such species adulteration on the health and safety of consumers have only recently begun to be recognised and documented. We provide a comprehensive review of the nature and magnitude of species adulteration in the raw herbal trade, and identify the underlying drivers that might lead to such adulteration. We also discuss the possible biological and chemical equivalence of species that are used as adulterants and substitutes, and the consequences thereof to consumer health and safety, and propose a framework for the development of a herbal trade authentication service that can help regulate the herbal trade market.

Lawsone (2-hydroxy-1, 4-napthoquinone), also known as hennotannic acid, is an orange red dye used as a popular skin and hair colorant. The dye is produced in the leaves of Lawsonia inermis L, often referred to as the "henna" tree. In this study, we report the production of lawsone by an endophytic fungus, Gibberella moniliformis isolated from the leaf tissues of Lawsonia inermis. The fungus produced the orange-red dye in potato dextrose agar and broth, independent of the host tissue. Presence of lawsone was confirmed spectrometrically using HPLC and ESI-MS/MS analysis. The fragmentation pattern of lawsone was identical to both standard lawsone and that extracted from plant tissue. This is a first report of lawsone being produced by an endophytic fungus, independent of the host tissue. The study opens up interesting questions on the possible biosynthetic pathway through which lawsone is produced by the fungus.

Antagonism of plant pathogenic fungi by endophytic fungi is a well-known phenomenon. In plate assays, the antagonism could be due to mycoparasitism, competition for space or antibiosis, involving a chemical diffusate, or a volatile organic compound (VOC). In this study, we demonstrate that besides mycoparasitism, VOCs play a major role in antagonism of pathogenic fungi by four endophytic fungi belonging to the genus Trichoderma. Using a double-plate assay, we show that all the four endophytic Trichoderma species significantly inhibited mycelial growth of three of the four pathogens, (Sclerotinia sclerotiorum-TSS, Sclerotium rolfsii-CSR and Fusarium oxysporum-CFO), while that of Macrophomina phaseolina-CMP was not affected. GC-MS analysis of the pure cultures of one of the endophytic fungi studied, namely, Trichoderma longibrachiatum strain 2 (Acc. No. MK751758) and the pathogens, F. oxysporum-CFO and M. phaseolina-CMP revealed the presence of several VOCs including hydrocarbons, alcohols, ketones, aldehydes, esters, acids, ethers and different classes of terpenes. In mixed double plates, where the endophyte was grown along with either of the two plant pathogens, F. oxysporum-CFO or M. phaseolina-CMP, there was an induction of a number of new VOCs that were not detected in the pure cultures of either the endophyte or the pathogens. Several of these new VOCs are reported to possess antifungal and cytotoxic activity. We discuss these results and highlight the importance of such interactions in endophyte-pathogen interactions.

Rapid climate change threatens plant communities. While many studies address the impact of climate change on plants and mechanisms of their resilience to climate stressors, the role of the plant microbiome in aiding plants' adaptation to climate change has been less investigated. We argue here that fungal endophytes, an important constituent of the plant microbiome, may be key to the ability of plants to adapt to climatic stressors. The rapid adaptive response of endophytes coupled with their ability to ‘transfer’ resistance to their hosts may fast-track plants' adaptation to climate change. We briefly review the importance of Class 3 fungal endophytes of terrestrial plants and discuss how they may accelerate adaptations to climate change in crops and natural plant communities and call for efforts directed at improving the understanding of fungal endophyte-facilitated plant health. Such information could aid in devising improved strategies for mitigating climate change effects on plant communities.

Many invasive alien species (IAS) are used by local communities for a variety of subsistence and income generating purposes. This frequently poses a conflict of interest for their removal due to forest conservation and biodiversity concerns. However, if local use can simultaneously check or control specific IAS, the conflict can be avoided and both development and forest conservation perspectives accommodated in the short to medium term. We examine this for Lantana camara invasion in southern India through assessment of the demand for and impacts of harvesting on this globally problematic IAS. We interviewed local artisans regarding their knowledge and quantities of Lantana used, along with forest surveys to estimate Lantana densities and size classes in harvested and unharvested sites, and lastly we undertook controlled cutting in moist and dry deciduous forests in both the wet and dry seasons to examine rate of regrowth and mortality. Over the entire study area the abundance of Lantana far outweighed local demand, but at small scales around villages, density and size classes were significantly reduced through harvesting. The controlled cutting experiment showed marked seasonal differences, with the most severe cutting intensity resulting in significant mortality when Lantana plants were cut in the wet season, but with limited effect when cut in the dry season. We conclude that promoting local use of IAS may be a feasible approach in controlling them and thereby limiting their impacts in forests.

Climate change has the potential to have significant influence on distribution of many species and alter ecosystems. With the current estimates of rate of projected environmental change for the 21st century, urgent adaptation and mitigation measures are required to slow down the impact of climate change on biodiversity. A number of studies have shown that recent human-induced environmental changes have already triggered species range shifts, changes in phenology and species extinctions. However, accurate projections of species responses to future climate change are more difficult to ascertain. In recent years, a number of modeling tools are being used to predict the consequences of climate change on the distribution of species and also identify possible ecological niche of a species under different climate change scenarios. In this paper, using the distribution data of the species, we have attempted to (a) identify the current geographical distribution of five species of the family Myristicaceae and predict the possible sites of occurrence of these species in the Western Ghats, India using the niche modeling tools and (b) predict the impact of climate change on the potential distribution of these species for two scenarios (A1B and A2A). All the five species selected for the study are distributed in the Western Ghats with higher densities in the Central Western Ghats. The model predicted an overall increase in the suitable habitats for the non-swampy species. However, there is a shift in the habitat range of these species in response to climate change. Gymnocranthera canarica and Myristica fatua , which are obligate swampy species, tend to be affected in changing climate. For both these species, the overall habitat is reduced due to climate change. These species already have restricted habitat and therefore climate change could pose a serious threat for their future survival. Predicting how species distribution will change in the wake of future global climate change is important to develop effective conservation strategies for these species, which inhabit fragile ecosystems.

Highly populated tropical countries face tremendous pressures in reconciling the needs for improved economic security and the protection of declining biodiversity. India is no exception and its biodiversity is under severe pressure due to complex interactions among land use change, other human economic activities, and climate change. Preservation and restoration of biodiversity is perhaps the cheapest and least risky way to mitigate the impacts of threats such as climate change, diminishing food and nutritional security, declining economy, absence of affordable healthcare, rising zoonotic diseases and lack of capacity to address these issues. Here we describe a framework for biodiversity conservation – the National Mission on Biodiversity and Human Well-Being (NMBHWB) for India – which integrates biodiversity, ecosystem services, climate change, agriculture, health, bio-economy and capacity building in the realm of biodiversity science. We provide an overview of the seven Programs of the Mission which make it interdisciplinary, integrative, and comprehensive in its approach. The Mission explicitly links research with policy-making and implementation for effective management of biodiversity with sustainable development. With its emphasis on convergence and synergies among various goals, themes and project sites, the Mission will further develop new models for stakeholder consultations and co-production of knowledge. We posit that the NMBHWB will enable India to realize the United Nations Sustainable Development Goals and Targets while advancing India's commitments to the Paris Agreement on climate change and other international environmental conventions and treaties.

Medicinal plants and their products have been used virtually in all cultures as a source of medicine since time immemorial. In India, several plant species are used as a medicine and in recent years, the trade-in medicinal plants have increased several folds. However, with burgeoning demand for raw herbal products, there has been concern over the safety and efficacy of raw herbal material as well as herbal products that contain only the desired plant species or labeled plant's product. In the present investigation, an emphasis was made to evaluate the extent of species adulteration in the raw herbal trade of Momordica charantia which is commonly called “karela” and used as alternate herbal medicine in the treatment of type-2 diabetes in south India. DNA barcodes were developed for six authentic reference species of Momordica and for the eighteen samples purchased from major raw herbal material distributors of south India representing the raw herbal products of M. charantia sold as “karela” in India. The DNA barcodes were developed using the nuclear region of the internal transcribed spacer (ITS) and the chloroplast region matK of six Momordica species, the biological reference standard, and the raw herbal drug trade samples. The species-level authenticity of the herbal drug samples was assessed adopting two identification methods namely BLAST similarity-based identification and phylogenetic tree approach. Our results indicate that the market samples sold were mostly authentic M. charantia except three. The present study offers a novel and reliable DNA barcode sequence-based approach for screening the species authenticity of raw herbal drug market samples of M. charantia and we have discussed the implications of the study in the larger context of authentication in raw herbal trade.

In this article, we report results of a molecular docking analysis of commonly occurring natural product compounds against COVID-19 6LU7 and 6Y2E proteases. Our results show that several of these compounds have binding affinity against both the COVID-19 proteases, and compare well with a known anti-HIV drug, Saquinavir. Many of the compounds form an integral component of many cuisines, both Indian as well as others. We propose that some of these compounds could be easily and quickly positioned to hold fort against the COVID-19 virus, until of course newer therapies are discovered and detailed studies are taken to empirically validate some of the compounds for their ability to inhibit the virus.

Rauvolfia tetraphylla L. (family Apocynaceae), often referred to as the wild snakeroot plant, is an important medicinal plant and produces a number of indole alkaloids in its seeds and roots. The plant is often used as a substitute for Ravuolfia serpentine (L.) Benth. ex Kurz known commonly as the Indian snakeroot plant or sarphagandha in the preparation of Ayurvedic formulations for a range of diseases including hypertension. In this study, we examine the spatial localization of the various indole alkaloids in developing fruits and plants of R. tetraphylla using desorption electrospray ionization mass spectrometry imaging (DESI-MSI). A semi-quantitative analysis of the various indole alkaloids was performed using UPLC-ESI/MS. DESI-MS images showed that the distribution of ajmalcine, yohimbine, demethyl serpentine and mitoridine are largely localized in the fruit coat while that for ajmaline is restricted to mesocarp of the fruit. At a whole plant level, the ESI-MS intensities of many of the ions were highest in the roots and lesser in the shoot region. Within the root tissue, except sarpagine and ajmalcine, all other indole alkaloids occurred in the epidermal and cortex tissues. In leaves, only serpentine, ajmalcine, reserpiline and yohimbine were present. Serpentine was restricted to the petiolar region of leaves. Principal component analysis based on the presence of the indole alkaloids, clearly separated the four tissues (stem, leaves, root and fruits) into distinct clusters. In summary, the DESI-MSI results indicated a clear tissue localization of the various indole alkaloids, in fruits, leaves and roots of R. tetraphylla. While it is not clear of how such localization is attained, we discuss the possible pathways of indole alkaloid biosynthesis and translocation during fruit and seedling development in R. tetraphylla. We also briefly discuss the functional significance of the spatial patterns in distribution of metabolites.

A number of studies have shown that there could be widespread substitution and/or adulteration (hereafter referred to as substitution) in raw herbal trade of medicinal plants. Substitution could potentially endanger the health and safety of the consumers. In this study, the extent of adulteration in raw herbal trade of 30 important medicinal plants in South India was analyzed. Biological reference material (BRM) consisting of taxonomically authenticated samples of each of the 30 species along with 14 other co-occurring and congeneric allied species that are likely to be used in adulteration was established. DNA barcode signatures of 124 BRM using two candidate regions, nr-ITS and psbA-trnH were identified. A total of 203 herbal trade samples representing the 30 medicinal plant species were collected from 34 locations in South India. Using the DNA barcode sequences of the BRM as reference, the analysis indicated that the substitution ranged from 20 to 100%. Overall, approximately 12% of the market samples were adulterated. Considering the potential health hazard that such adulteration can cause, the need for a national regulatory framework that can authenticate and regulate raw herbal trade in the country is discussed. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s13205-018-1169-3) contains supplementary material, which is available to authorized users.

In nature, plants and microorganisms interact constantly to modulate their growth and development. The interaction could be beneficial or hostile. Plants are constantly exposed to a range of pathogens, and have evolved mechanisms to fight the pathogen infections with the help of complex networking pathways. In addition to the pathogens, many beneficial organisms such as endophytes, of both, bacterial and fungal origin, are ubiquitously present in all plants. Unlike from their pathogenic counterparts, the endophytes do not cause any disease symptoms in their host plants. Some of the recent scientific discoveries and views concerning plant–microbe interactions are presented here.

The genus Sida L. (family: Malvaceae) is widely used in India and many other countries including China, South East Asia, Africa and South America for treating various neurological disorders and for improving general health and vigour. However, as with many other herbal medicines, it is believed that the Sida products sold in the market may be adulterated with other related or unrelated plant species. In this study, we investigate species adulteration in the raw herbal trade of Sida natural health products (NHPs) in southern India. DNA barcoding was used as a tool to identify the ingredients in the NHPs. A biological reference material (BRM) library for Sida and closely related species was developed using taxonomically authenticated species. DNA barcodes for the species were developed using one nuclear (ITS) and two chloroplast regions (matK and psbA-trnH). The psbA-trnH and ITS region were found to effectively discriminate all species with an interspecific distance of 0.133 and 0.149 and intra-species distance of 0.007 and 0.015 respectively. These DNA barcodes were used to identify the ingredients in raw Sida herbal products obtained from 10 markets in Southern India. Our study indicated that species adulteration in the market samples is rampant especially in case of Sida cordifolia, where all the market samples analyzed were Sida acuta. We discuss the results and the need for a robust herbal drug authentication system to regulate the quality in raw herbal trade market.

It is well established that non-timber forest products make significant contributions to rural incomes throughout most of the developing world. NTFP use frequently raises concerns about the sustainability of, or ecological impacts associated with, NTFP harvesting, as well as local contextual factors which may limit or reduce the impacts. Here we test the conceptual model first advanced by Uma Shaanker et al. (2004) relating to the factors that may limit or exacerbate the ecological impacts associated with NTFP harvesting. These were the extent of local dependence on NTFPs, the degree of marketing and the level of local ecological knowledge. Data were collected via household questionnaires and ecological surveys of woody plants from eight villages throughout South Africa. We found no significant relationships between measures of ecological impact with local ecological knowledge or market proximity and engagement. There was a strong positive relationship between ecological impacts and NTFP dependency as indexed through mean annual direct-use value for NTFPs. This indicates that the higher the dependency and demand for NTFPs, the greater is the possibility of high impacts to the local environment. However, other contextual drivers not included in the original Uma Shaanker et al. (2004) model may also play a role, particularly the strength of local resource governance institutions.

Camptothecin (CPT, 1) is a potent anticancer natural product which led to the discovery of two clinically used anticancer drugs topotecan and irinotecan. These two drugs are semisynthetic analogs of CPT, and thus the commercial production of CPT as a raw material from various plant sources and tissue culture methods is highly demanding. In the present study, the Dysoxylum binectariferum bark, was identified as an alternative source of CPT, through bioassay-guided isolation. The barks showed presence of CPT (1) and its 9-methoxy analog 2, whereas CPT alkaloids were not present in seeds and leaves. This is the first report on isolation of CPT alkaloids from Meliaceae family. An efficient chromatography-free protocol for enrichment and isolation of CPT from D. binectariferum has been established, which was able to enrich CPT up to 21% in the crude extract. The LCMS (MRM)-based quantification method revealed the presence of 0.105% of CPT in dry barks of D. binectariferum. The discovery of CPT from D. binectariferum bark will certainly create a global interest in cultivation of this plant as a new crop for commercial production of CPT. Isolation of anticancer drug CPT from this plant, indicates that along with rohitukine, CPT and 9-methoxy CPT also contributes significantly to the cytotoxicity of D. binectariferum.

Rohitukine, a chromone alkaloid, has gained considerable international attention in recent years because of its novel semi-synthetic derivative, flavopiridol and P-276-00. Both these molecules are in advanced stages of clinical development and trial for cancer treatment. Recently, flavopiridol was approved as an orphan drug for treatment of chronic lymphocytic leukemia cancer. The natural occurrence of rohitukine is restricted to only four plant species, Amoora rohituka and Dysoxylum binectariferum (both from the Meliaceae family) and from Schumanniophyton magnificum and Schumanniophyton problematicum (both from the Rubiaceae family). Recently, an endophytic fungi isolated from D. binectariferum was reported to produce rohitukine in culture. In this study, we report the production of rohitukine and its subsequent attenuation by endophytic fungi, Fusarium oxysporum (MTCC-11383), Fusarium oxysporum (MTCC-11384) and Fusarium solani (MTCC-11385), all isolated from D. binectariferum and Gibberella fujikuroi (MTCC-11382) isolated from Amoora rohituka. The fungal rohitukine which was analyzed by HPLC, LC-MS and LC-MS/MS was identical to reference rohitukine and that produced by the plant. The rohitukine content in the mycelial samples ranged from 192.78μg to 359.55μg100g(-1) of dry weight of and in broth it ranged from 14.10 to 71.90μg100ml(-1). In all the fungal cultures, the production declined from first to fourth sub-culture. Studies are underway to unravel the mechanism by which the fungi produce the host metabolite in culture.

Tropical agro-forest landscapes are potentially valuable reserves of forest genetic resources for forestry and restoration of degraded forests. The Dipterocarpaceae is a dominant Southeast Asian family of tree species of global significance for the tropical timber industry. Very little information exists about how effective human modified landscapes are for conserving genetic diversity in dipterocarp species. This study provides a baseline for understanding how fragmented agro-forest landscapes in India sustain forest genetic resources in an endemic dipterocarp tree. We compare genetic diversity and fine-scale spatial genetic structure (FSGS) in the threatened tree species Vateria indica within an isolated and a continuous forest site in the Western Ghats, South India. We place these results in the context of dipterocarps from both the Seychelles and Borneo. Parentage analysis of 694 progeny using twelve nuclear microsatellite markers is applied to estimate pollen and seed dispersal. Using a nursery trial we evaluate effects of inbreeding on growth performance. Our results show that levels of FSGS, and gene dispersal are comparable between a small isolated and a large continuous site of V. indica. Realized long-distance pollen flow into the isolated patch appears to help maintaining genetic diversity. The nursery experiment suggests that selection favours outbred progeny. Individuals of V. indica in close proximity appear less related to each other than in another highly fragmented and endangered dipterocarp species from the Seychelles, but more related than in three dipterocarp species studied in continuous forest in Borneo. We discuss the wider implications of our findings in the context of conservation and restoration of dipterocarp forest genetic resources in fragmented populations.

Lantana camara. L (hereafter Lantana) was first introduced by the British into India in 1807 as an ornamental plant. Since then the species has spread across the length and breadth of the country. Attempts to control Lantana in India have not been successful. In this study, we analysed the use of Lantana by local communities in southern India and identified the possible causes and consequences of its use through the use of a household survey of the socio-economic profile of the user and nonuser households and an analysis of the ecological history of the communities. Communities have been using Lantana for over 25–30 years and apparently such use was not prompted by external agencies. The characteristics of user and nonuser households were similar, except that Lantana users were more literate and had a greater number of occupations per household than nonusers. Per capita income was similar between user and nonuser groups. For nonuser groups, their main income sources were from trading (44 %), wage labour (32 %) and forest resources (23 %). In contrast, the Lantana user groups substituted their loss of income from forest resources (7 %) by income from Lantana (46 %). The ecological history revealed that Lantana was adopted as a resource at a time when it was increasing in the landscape and traditional bamboo resources were in decline because of overuse by commercial enterprises and mast flowering. This change in ecological resource availability prompted a major shift in livelihoods for some in the area.

The chromone alkaloid dysoline (1), a new regioisomer of rohitukine (2) along with rohitukine and rohitukine-N-oxide (3) were isolated from the stem barks of Dysoxylum binectariferum. The structure of dysoline (1) was determined by extensive 2D-NMR studies and the absolute configuration was established by NOESY and CD spectra. Dysoline (1) consisted of a 5,7-dihydroxy-2-methylchromone nucleus substituted with a 2′-hydroxylated N-Me piperidine ring at the C-6 position. Dysoline differs from rohitukine by the position of the piperidine ring on the chromone nucleus. Dysoline displayed promising cytotoxicity in HT1080 fibrosarcoma cells with an IC50 of 0.21 μM, and also displayed significant inhibition of proinflammatory cytokines TNF-α and IL-6.

We compared morphological and genetic attributes of progeny obtained from three seed production areas (SPAs) and three corresponding unimproved stands (UIS) of teak (Tectona grandis L.f) at three broad teak growing regions in the state of Karnataka, India. In general, seed morphological parameters such as seed weight, seed size and seed emptiness were significantly superior in SPAs compared to UISs. Seed germination percentage was also higher in seeds from SPAs. Seedling performance measured at two monthly intervals for six months was observed to be superior in SPAs at two of the three regions compared. Although the genetic diversity of progeny of SPAs was lower than UIS at all regions, the values were moderately high. Germination percent was positively correlated with fruit weight and kernel weight per seed, while diversity indices were negatively correlated with fruit weight, fruit diameter and kernel weight per seed. SPAs proved important as a source of moderately improved planting material with no severe threat to the genetic diversity of future plantations.

1Ashoka Trust for Research in Ecology and the Environment, Royal Enclave, Srirampura, Jakkur Post, Bangalore 560 064, India 2School of Ecology and Conservation, University of Agricultural Sciences, GKVK Campus, Bangalore 560 065, India 3Department of Crop Physiology, University of Agricultural Sciences, GKVK Campus, Bangalore 560 065, India 4Department of Forest Biology and Tree Improvement, College of Forestry, University of Agricultural Sciences, SIRSI campus, Dharwad 581 401, India

Camptothecine (CPT), a quinoline alkaloid, is a potent inhibitor of eukaryotic topoisomerase I. Because of this activity, several semi-synthetic derivatives of CPT are in clinical use against ovarian and small lung cancers. Together with its derivatives, CPT is the third largest anti-cancer drug in the world market. CPT is produced by several plant species belonging to the Asterid clade. In the recent past, several studies have reported the production of CPT by endophytic fungal associates of some of these plant species. In this paper, we report the production of CPT by endophytic bacteria isolated from Miquelia dentata Bedd. (Icacinaceae). Besides CPT, the bacteria also produced 9-methoxy CPT (9-MeO-CPT), in culture, independent of the host tissue. The chemical nature of CPT and 9-MeO-CPT was determined by LC-MS and ESI-MS/MS analysis, and was shown to be similar to that produced by the host tissue. One of the bacterial isolates examined, showed indications of attenuation of CPT production through sub-culture. This is the first report of production of CPT by endophytic bacteria. The identity of the bacteria was ascertained by Gram staining and 16s rRNA sequencing. We discuss the possible mechanisms that might be involved in the synthesis of CPT by endophytic bacteria.

Teak (Tectona grandis L. f.; family Verbanaceae) is an important plantation tree species in the tropics and in India one of the first species to be prioritized for improvement. Improvement efforts for the last 50 years have essentially concentrated on augmenting quality seed production by establishing seed production areas (SPA) and clonal seed orchards (CSO). Presently, these two form the main sources of quality planting material for teak throughout the country. However, there is no information on the genetic quality of such sources nor information on the progeny used in plantation programmes. Reports of studies based on coniferous and tropical species provide conflicting results on the impact of domestication on the genetic diversity of populations (Chaisurisri and El Kassaby 1994; Rajora 1999; Moran et al. 2000; Godt et al. 2001; Icgen et al. 2006). Also the impact of domestication on the genetic diversity of progeny populations is poorly understood (Stoehr and El-Kassaby 1997; Schmitdtling and Hiplins 1998). Such studies become pertinent not only for gauging the impact of selection on reforestation stock, but also for effective genetic conservation of existing breeding populations. We therefore address two issues in the present study: (i) the change in genetic diversity with increasing levels of improvement, and (ii) the impact of the above change on genetic diversity of progeny populations.

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.

This study sought to reconstruct the history of Lantana invasion and spread in India by considering two questions; (a) from where, by who, and when were Lantana species introduced into India? and (b) given its long history in the country, is it still spreading or more or less stable? We critically evaluated the archival and historical information on plant imports by the European powers into India during the period before and after British colonization. We then reconstructed the path of spread by analyzing the spatio-temporal patterns of occurrence and distribution of Lantana in India at both the national and local scale using a GIS platform. The spread of Lantana across the globe started as early as the 1690s. The European colonial powers moved the plants from Latin America to Europe and to their colonial countries in the early 1800s. Lantana species were introduced in India from 1807 onwards and thereafter the colonial powers moved this plant across the country. Following its introduction into India, the spread of Lantana across the country, either through subsequent multiple introductions from Europe to different British cantonments, or through moving the plants between cantonments within India, were reasonably rapid spanning only a few decades. In the absence of a rigorous control program, the spread of Lantana has gone on unabated and thereby impacting both wildlife and biodiversity.

Influence of genetic dissimilarity among teak (Tectona grandis Linn. f.) clones on flowering synchrony was studied in a Clonal Seed Orchard (CSO) of teak in Karnataka, Southern India. Flowering phenology was monitored for all the 24 teak clones of the CSO and flowering synchrony between clones was assessed adopting a novel ‘overlap index’. Genetic dissimilarity among these clones was assessed adopting DNA based ISSR (Inter Simple Sequence Repeats) analysis. Large variation in the time of ‘flower initiation’ and of ‘peak flowering initiation’ was observed among the clones belonging to diverse sources, suggesting large asynchronous flowering. Cluster analysis based on ISSR marker indicated that the clones originating from a same source clustered together and there was a clear segregation based on their origin. Correlation analysis revealed a significant negative association between the average Jaccards’s dissimilarity index between pairs of clones and average peak flowering overlap index. Clones from geographically diverse regions had high genetic dissimilarity and also showed high flowering asynchrony within them.

Tropical trees often display long-distance pollen dispersal, even in highly fragmented landscapes. Understanding how patterns of spatial isolation influence pollen dispersal and interact with background patterns of fine-scale spatial genetic structure (FSGS) is critical for evaluating the genetic consequences of habitat fragmentation. In the endangered tropical timber tree Dysoxylum malabaricum (Meliaceae), we apply eleven microsatellite markers with paternity and parentage analysis to directly estimate historic gene flow and contemporary pollen dispersal across a large area (216 km(2)) in a highly fragmented agro-forest landscape. A comparison of genetic diversity and genetic structure in adult and juvenile life stages indicates an increase in differentiation and FSGS over time. Paternity analysis and parentage analysis demonstrate high genetic connectivity across the landscape by pollen dispersal. A comparison between mother trees in forest patches with low and high densities of adult trees shows that the frequency of short-distance mating increases, as does average kinship among mates in low-density stands. This indicates that there are potentially negative genetic consequences of low population density associated with forest fragmentation. Single isolated trees, in contrast, frequently receive heterogeneous pollen from distances exceeding 5 km. We discuss the processes leading to the observed patterns of pollen dispersal and the implications of this for conservation management of D. malabaricum and tropical trees more generally.

The region of Kodagu, South India, comprises a fragmented landscape with a high density of remnant forest patches dispersed within a mosaic dominated by shaded coffee agro-forests. We evaluated the role of self, wind and insect pollination to Coffea canephora production in this landscape. The giant Asian honeybee, Apis dorsata, which nests in remnant forests, was the main pollinator of coffee (accounting for 58% of the floral visitors). The proportion of flowers that developed into fruits was highest when hand cross-pollinated (44%), followed by open- (insect and wind combined; 33%) and wind- (22.1%) pollination treatments. Pollination by bees therefore increases fruit production by 50% over that achieved by wind. Self-pollination (1.7%) and no pollination (1%) treatments produced very low fruit set, emphasizing the importance of cross-pollination in C. canephora. Unlike measures of pollination success, initial fruit set (five weeks after flowering) proved an unreliable proxy for final fruit set. Size of adjoining forest fragments (mostly 0.3–20 ha, with a few exceeding 200 ha) positively influenced pollinator visitation to coffee flowers, but distance to such fragments had no influence on pollination. This study demonstrates the importance of cross-pollination for crop production in C. canephora, the important contribution that pollinating insects make to coffee production, and the benefits of relatively large forest fragments within the landscape mosaic to support insect pollinators of coffee. A comparison of pollinator composition to that of 100 years ago indicated that coffee pollination services remained intact despite changes in pollinator community composition.

Phyllanthus (Euphorbiaceae) species are traditionally well-known for their medicinal properties including hepatoprotective activity.The study assessed the hepatoprotective and antioxidant activities of 11 Phyllanthus species, P. amarus Schumach., P. urinaria L., P. debilis Klein ex Willd, P. tenellus Roxb., P. virgatus G. Forst., P. maderaspatensis L., P. reticulatus Poir., P. polyphyllus Willd., P. emblica L., P. indofischerii Bennet. and P. acidus (L.) Skeels.The dried leaves and stems of each plant species were extracted in methanol and successively in water. The extracts were screened for hepatoprotective activity at a concentration of 50 µg/mL against tert-butyl hydroperoxide (t-BH) induced toxicity in HepG2 cells. Seven extracts from five species that showed hepatoprotective activity were assessed for their 50% effective concentration (EC₅₀) values and their antioxidant activity using a DPPH assay. Phyllanthin and hypophyllanthin contents were also determined in these Phyllanthus species.The methanol extracts of P. polyphyllus, P. emblica and P. indofischeri showed high levels of hepatoprotective activity with EC₅₀ values of 12, 19 and 28 µg/mL and IC₅₀ of 3.77, 3.38 and 5.8 µg/mL for DPPH scavenging activity respectively against an IC₅₀ of 3.69 µg/mL for ascorbic acid. None of these activities could be attributed to phyllanthin and hypophyllanthin.The hepatoprotective and antioxidant activities of P. indofischeri are demonstrated for the first time in literature. The study also confirms the hepatoprotective and antioxidant activities of leaves of P. emblica and P. polyphyllus. The molecule(s) responsible for the activities is being investigated.

Taxillus tomentosus is a mistletoe of significant management concern in southern India. This mistletoe affects the productivity of several nontimber forest products, most significantly Amla (Phyllanthus emblica and Phyllanthus indofischeri), whose fruits provide an important source of income for indigenous forest communities. Management interventions are required to secure the continued viability of Amla and thereby its role in livelihoods. To enhance the scientific basis for management, we characterized the distribution of T. tomentosus at three scales, examined potential mechanisms underlying this distribution, and compared both with those documented in other habitats. Mistletoes were aggregated at all three scales: on individual trees, within forest plots of 500 m2, and at scales of 3-4 km. Seed deposition patterns provide an initial distribution 'template' that largely determines patterns at tree and plot scales. This initial pattern is subsequently altered as variation in host suitability and mistletoe mortality from forest fires influence establishment success and postestablishment survival. Local prevalence of infection contributed most to infection spread, and areas of high infection prevalence are highlighted as appropriate foci for management intervention. Although novel underlying mechanisms and variation in the scale of aggregation have been identified, mistletoe distribution in a tropical forest appears to be shaped by similar forces as those documented previously in temperate systems.

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.