Shilpi K. Saikia

Beta-caryophyllene modulates expression of stress response genes and mediates longevity in Caenorhabditis elegans

Beta-caryophyllene (BCP) is a natural bicyclic sesquiterpene and is a FDA approved food additive, found as an active ingredient in essential oils of numerous edible plants. It possesses a wide range of biological activities including anti-oxidant, anti-inflammatory, anti-cancerous and local anesthetic actions. We used the well established Caenorhabditis elegans model system to elucidate the stress modulatory and lifespan prolonging action of BCP. The present study for the first time reports the lifespan extension and stress modulation potential of BCP in C. elegans. Upon evaluation, it was found that 50μM dose of BCP increased the lifespan of C. elegans by over 22% (P≤0.0001) and significantly reduced intracellular free radical levels, maintaining cellular redox homeostasis. Moreover, the results suggest that BCP modulates feeding behavior, pharyngeal pumping and body size effectively. Further, this compound also exhibited significant reduction in intestinal lipofuscin levels. In the present investigation, we have predicted possible biological molecular targets for BCP using molecular docking approaches and BCP was found to have interaction with SIR-2.1, SKN-1 and DAF-16. The prediction was further validated in vivo using mutants and transgenic strains unraveling underlying genetic mechanism. It was observed that BCP increased lifespan of mev-1 and daf-16 but failed to augment lifespan in eat-2, sir-2.1 and skn-1 mutants. Relative quantification of mRNA demonstrated that several genes regulating oxidative stress, xenobiotic detoxification and longevity were modulated by BCP treatment. The study unravels the involvement of multiple signaling pathways in BCP mediated lifespan extension.

Longevity-promoting effects of 4-hydroxy-E-globularinin in Caenorhabditis elegans.

In modern times, there has been a major increase in the use of plants or herbal constituents for the prevention of age-related disorders. 4-Hydroxy-E-globularinin (4-HEG) is an iridoid and a major component of Premna integrifolia. This investigation represents a breakthrough in geriatrics by showing the longevity-promoting activity of 4-HEG in the animal model Caenorhabditis elegans. 4-HEG (20μM) enhanced the mean life span of worms by over 18.8% under normal culture conditions and also enhanced their survival under oxidative stress. The longevity-promoting activity was associated with reduced reactive oxygen species (ROS) levels and fat accumulation in the worms. Gene-specific mutant studies verified the role of ROS detoxification pathways and simultaneous nuclear translocation of DAF-16 in the 4-HEG-mediated effects. Quantitative real-time PCR estimations and observations of transcriptional reporters indicated that 4-HEG was able to upregulate stress-inducible genes, viz., hsp-16.2 and sod-3. Thus, 4-HEG may serve as a lead compound of plant origin for the development of important nutraceuticals superseding the aging process.

Exploitation of microbes for enhancing bacoside content and reduction of Meloidogyne incognita infestation in Bacopa monnieri L

Despite the vast exploration of rhizospheric microbial wealth for crop yield enhancement, knowledge about the efficacy of microbial agents as biocontrol weapons against root-knot disease is scarce, especially in medicinal plants, viz., Bacopa monnieri. In the present investigation, rhizospheric microbes, viz., Bacillus megaterium, Glomus intraradices, Trichoderma harzianum ThU, and their combinations were evaluated for the management of Meloidogyne incognita (Kofoid and White) Chitwood and bacoside content enhancement in B. monnieri var CIM-Jagriti. A novel validated method Fourier transform near infrared was used for rapid estimation of total bacoside content. A significant reduction (2.75-fold) in root-knot indices was observed in the combined treatment of B. megaterium and T. harzianum ThU in comparison to untreated control plants. The same treatment also showed significant enhancement (1.40-fold) in total bacoside contents (plant active molecule) content using Fourier transform near-infrared (FT-NIR) method that analyses samples rapidly in an hour without solvent usage and provides ample scope for natural product studies.

Bioconsortia Augments Antioxidant and Yield in Matricaria recutita L. Against Meloidogyne incognita (Kofoid and White) Chitwood Infestation

Plant-associated rhizospheric microflora has been vastly explored for its vital function in maintaining plant health. A better understanding however, for its impact on the root-knot disease management and enhancement of antioxidant potentials, especially in medicinal plants viz. Matricaria recutita L. (Chamomile) is yet seldom investigated. The present investigation evaluates selected potential of rhizospheric microbes viz. Bacillus megaterium,Trichoderma harzianum ThU and Glomus intraradices, singly as well as in consortia for the management of Meloidogyne incognita (Kofoid and White) Chitwood and augmentation of phytochemical contents in Chamomile. A significant reduction (2.50 fold) in root-knot indices was observed in the triple microbe treatment of B. megaterium, T. harzianum and G. intraradices as compared to control plants. Antioxidant potential at various levels viz. total phenolics and flavonoids, free radical scavenging activity, ferric reducing antioxidant power, total antioxidant capacity and reducing power was significantly higher in consortia treated M. recutita plants as compared to their single application. The results from the present experimentation indicate a possible new function of compatible interaction among microbe’s that can effectively cope with the root-knot nematode menace and is capable of enhancing antioxidant values of M. recutita.

Genetic revelation of hexavalent chromium toxicity using Caenorhabditis elegans as a biosensor

The interaction of heavy metals such as hexavalent chromium, Cr (VI) with the environment drastically influences living organisms leading to an ecological imbalance. Caenorhabditis elegans, a saprophytic nematode having 60–80% homology with human genes offers a distinct advantage to be used as a biosensor for the appraisal of heavy metal-induced environmental toxicity and risk monitoring. The present study examines the toxicity effects of K2Cr2O7 as Cr (VI) on stress-related gene expression and morphometric parameters of C. elegans under in vitro conditions to identify genetic markers for environmental pollution. Alterations in growth and modified gene expression were observed in Cr (VI)-exposed N2 worms. The 24-h median lethal concentration for Cr (VI) was observed as 158.5 mgl−1. Use of the responses of stress-related gene expression suggests that C. elegans can be used as an efficient biosensor for figuring out the precise route of Cr (VI)-induced environmental toxicity in a quick, simple, and inexpensive manner.

Hexavalent Chromium Induced Histological Alterations in Bacopa monnieri (L.) and Assessment of Genetic Variance

Chromium, a transition metal exists in nature in several oxidation states, the most important being Cr (III) and Cr (VI) species. The two forms display quite different chemical properties as Cr (VI) is highly toxic to most organisms whereas Cr (III) is relatively innocuous. Cr (VI) exposures are responsible for carcinogenic and mutagenic effects in humans also. The present study investigates the changes in cortical and vascular tissues of Bacopa monneiri after short-term exposure to Cr (VI). K 2 Cr 2 O 7 (0.05M) was added to Murashige and Skoog’s (MS) media to which Bacopa explants were introduced and the detrimental effects were studied after 21 days of incubation under different stress conditions. Histological results revealed a time-dependent noxious effect of Cr (VI) on cortical cells. The amount of damage in vascular region was observable but not much defined. Changes included enlargement of the cortical cells and accumulation of chromium within the live tissues. The molecular interpretation of damage due to Cr (VI) was done by PCR amplification of different antioxidant genes and analysing the products on 1% agarose gel. The genes were unable to show expression with cDNA template from explants exposed to Cr (VI) while with cDNA from explants exposed to bacterial isolates and chromium, expression of antioxidant genes was observed. The study will provide glimpses of Cr (VI) induced damage in plant tissue at cellular and genetic levels.

3β-Hydroxy-urs-12-en-28-oic Acid Modulates Dietary Restriction Mediated Longevity and Ameliorates Toxic Protein Aggregation in C. elegans

Species from lower invertebrates to a spectrum of mammals show antiaging health benefits of phytochemical(s). Here, we explored the pro-longevity effects of a natural triterpenoid, ursolic acid (3β-hydroxy-urs-12-en-28-oic acid; UA) in Caenorhabditis elegans with maximal life span being evident at 25 µM UA. Similar to eat-2 mutants, UA uptake by worm results in reduced fat storage and attenuation of reactive oxygen species (ROS), independent of superoxide dismutase(s) activation. The genetic requirements for UA-mediated longevity are quite similar to dietary restriction (DR) achieved through SKN-1/NRF-2 exhibiting upregulation of downstream target genes gcs-1 and daf-9. Longevity mechanism was independent of PHA-4/FOXA and attributed to partial dependence on sir-2.1. Altogether, our study suggests differential use of UA-elicited signaling cascades in nutrient sensing for longevity. Both the redox state and the proteostasis of an organism play critical role in aging and disease resistance. Interestingly, we observed a reduction of toxic protein aggregation in transgenic polyglutamine (polyQ) C. elegans model and UA-mediated JNK-1 (c-Jun-NH2-terminal kinase) activation in wild-type animals. Thus, our study demonstrates a small extent of prevention against proteotoxic stress by UA coupled with positive aspects of DR-mediated longevity.

3β-Hydroxy-urs-12-en-28-oic acid confers protection against ZnONPs induced adversity in Caenorhabditis elegans

Despite their well reported potent risk towards human health and environment Zinc oxide nanoparticles (ZnONPs) find an extensive commercial usage due to their antimicrobial properties. Here, we evaluated the efficacy of a natural triterpene ursolic acid (3β-hydroxy-urs-12-en-28-oic acid; UA) for overcoming the cytotoxic challenges of ZnONPs employing Ceanorhabditis elegans. The 24h LC50 of Zn-ONPs (<50nm TEM) was deduced as 4.75mgL-1. UA (25μM) was observed to defend against this toxicity by attenuating Reactive Oxygen Species (ROS) resulting into better survival at 2mgL-1 in a time dependent behavior. However, reproductive health remains compromised. Our study identifies UA as a natural inducer of metallothionein proteins along with antioxidant potential. We demonstrate that UA induces upregulation of predominantly antioxidant genes, including the superoxide dismutases (sod-1, sod-5 and sod-3), glutathione S-transferase 7 (gst-7), heat shock protein (hsp-16.2) along with the metal exposure responsive metallothionein (mtl-1 and mtl-2). Moreover, UA also restores elevated transcript levels of gst-4 during ZnONPs stress conditions to normal by directly scavenging ROS owing to its own antioxidant potential. Altogether, the toxic aspects of NPs that can be avoided compensated or postponed by supplementation of phytochemical(s) in biological system underscore their potential implications in near future.

Diseases of medicinal and aromatic plants, their biological impact and management

With increasing interest in natural products of plant origin for medicinal and health care benefits there is added emphasis on the quality of the source raw material. In most cases the vegetative tissues and organs are the source of the required raw material. However, such tissues/organs may become infested/susceptible to many diseases causing deterioration of the quality of the desired economic product and loss of genetic resources. Considerable progress has been made with respect to the identification of disease causing organisms, and their pathogenic impact at organ, cellular and biochemical level. This review focuses on various diseases caused by bacteria, fungi, nematodes, viruses and phytoplasmas threatening the yield, biomass, bioactive potential of medicinal and aromatic plants of the sub-tropics. Several studies suggest that management of diseases through biological measures is more effective to check multiple diseases. Necessary management strategies for efficient disease management to realize quality raw material and enhanced metabolite productive potential have been outlined.

Rhizospheric Microflora Escalating Aroma Constituents and Yield Attributes in Ocimum tenuiflorum (L.) cv. CIM-Ayu

The exploration of rhizospheric microbial flora for crop yield enhancement is well established. Rhizospheric microbes influence the plant physiology by imparting several beneficial effects, namely, Nitrogen fixation, increased nutrient uptake, and secondary metabolites production on their host plants. The present study investigates the response of Bacillus megaterium ATCC No. 13525, Pseudomonas fluorescens ATCC No. 14581, and Trichoderma viride MTCC No. 167 in alone and combined treatments for their effect on growth and yield parameters in a commercially important Ocimum tenuiflorum L. cv. CIM-Ayu. The plant is therapeutically important for its essential oil constituents, namely, eugenol, β-caryophyllene, and various monoterpenes. The combination treatments, T7 (B. megaterium