Browsing by Author "Neha Pandey"

Now showing 1 - 20 of 36

Accumulation of Secondary Metabolites and Improved Size of Glandular Trichomes in Artemisia annua
(Springer Science and Business Media B.V., 2020) Neha Pandey; Anupam Tiwari; Sanjay Kumar Rai; Shashi Pandey-Rai
Glandular trichomes are multicellular epidermal outgrowths that have characteristic globular head made up of secretory cells and store large quantities of specialized secondary metabolites. Artemisia annua is known for its medicinally important secondary metabolite “artemisinin” which is synthesized and stored in glandular trichomes. However, our understanding of morphological and transcriptional control related to glandular trichome development and accumulation of secondary metabolites in A. annua is available in scattered form. This chapter deals with the trichome biology including developmental and functional aspects along with their correlation with secondary metabolite accumulation in response to various biotic and abiotic signals of the environment using A. annua as model. This chapter also emphasizes the molecular mechanisms behind trichome development in A. annua and provides a glimpse of molecular players involved in this process. There are many environmental as well as intrinsic factors which directly or indirectly affect secondary metabolite synthesis and as a result determine the size of glandular trichomes. The compiled information available for A. annua trichome biology can further be utilized for exploring trichome engineering in many medicinal or aromatic plants which are less explored. © Springer Nature Switzerland AG 2020.
Biological and Pathogenic Blueprint of Chandipura Virus
(John Wiley and Sons Ltd, 2025) Neha Pandey; Sunit Kumar Singh
Chandipura virus (CHPV) is a single-stranded negative-sense RNA virus of the family Rhabdoviridae. CHPV is transmitted mainly through infected sandflies. CHPV paediatric encephalitis reported in 2003–2004 in central and south-western parts of India had a case fatality rate of ∼70%. CHPV infection leads to high-grade fever, vomiting, altered sensorium, generalised convulsions, decerebrate posture and coma. Neuroinflammation is the hallmark of CHPV infection and has a pronounced effect on cerebral and brainstem regions. Currently, there are no vaccines or treatments available for CHPV infection. Although previous studies have provided insights into the virus's pathology and host-pathogen interactions, the precise molecular mechanisms underlying CHPV pathogenesis are poorly understood. Understanding molecular pathogenesis is crucial for developing efficacious therapies and preventive measures. The review summarises CHPV epidemiology, transmission, genome structure, replication, pathogenesis and the latest antiviral therapies and vaccine developments. © 2025 John Wiley & Sons Ltd.
Biotechnological strategies for enhancing heavy metal tolerance in neglected and underutilized legume crops: A comprehensive review
(Academic Press, 2021) Krishna Kumar Rai; Neha Pandey; Ram Prasad Meena; Shashi Pandey Rai
Contamination of agricultural land and water by heavy metals due to rapid industrialization and urbanization including various natural processes have become one of the major constraints to crop growth and productivity. Several studies have reported that to counteract heavy metal stress, plants should be able to maneuver various physiological, biochemical and molecular processes to improve their growth and development under heavy metal stress. With the advent of modern biotechnological tools and techniques it is now possible to tailor legume and other plants overexpressing stress-induced genes, transcription factors, proteins, and metabolites that are directly involved in heavy metal stress tolerance. This review provides an in-depth overview of various biotechnological approaches and/or strategies that can be used for enhancing detoxification of the heavy metals by stimulating phytoremediation processes. Synthetic biology tools involved in the engineering of legume and other crop plants against heavy metal stress tolerance are also discussed herewith some pioneering examples where synthetic biology tools that have been used to modify plants for specific traits. Also, CRISPR based genetic engineering of plants, including their role in modulating the expression of several genes/ transcription factors in the improvement of abiotic stress tolerance and phytoremediation ability using knockdown and knockout strategies has also been critically discussed. © 2020 The Authors
CAMTA 1 regulates drought responses in Arabidopsis thaliana
(2013) Neha Pandey; Alok Ranjan; Poonam Pant; Rajiv K. Tripathi; Farha Ateek; Haushilla P. Pandey; Uday V. Patre; Samir V. Sawant
Background: Transcription factors (TF) play a crucial role in regulating gene expression and are fit to regulate diverse cellular processes by interacting with other proteins. A TF named calmodulin binding transcription activator (CAMTA) was identified in Arabidopsis thaliana (AtCAMTA1-6). To explore the role of CAMTA1 in drought response, the phenotypic differences and gene expression was studied between camta1 and Col-0 under drought condition.Results: In camta1, root development was abolished showing high-susceptibility to induced osmotic stress resulting in small wrinkled rosette leaves and stunted primary root. In camta1 under drought condition, we identified growth retardation, poor WUE, low photosystem II efficiency, decline in RWC and higher sensitivity to drought with reduced survivability. The microarray analysis of drought treated camta1 revealed that CAMTA1 regulates " drought recovery" as most indicative pathway along with other stress response, osmotic balance, apoptosis, DNA methylation and photosynthesis. Interestingly, majority of positively regulated genes were related to plasma membrane and chloroplast. Further, our analysis indicates that CAMTA1 regulates several stress responsive genes including RD26, ERD7, RAB18, LTPs, COR78, CBF1, HSPs etc. and promoter of these genes were enriched with CAMTA recognition cis-element. CAMTA1 probably regulate drought recovery by regulating expression of AP2-EREBP transcription factors and Abscisic acid response.Conclusion: CAMTA1 rapidly changes broad spectrum of responsive genes of membrane integrity and photosynthetic machinery by generating ABA response for challenging drought stress. Our results demonstrate the important role of CAMTA1 in regulating drought response in Arabidopsis, thus could be genetically engineered for improving drought tolerance in crop. © 2013 Pandey et al.; licensee BioMed Central Ltd.
Chandipura virus changes cellular miRNome in human microglial cells
(John Wiley and Sons Inc, 2022) Meghna Agrawal; Meghana Rastogi; Smriti Dogra; Neha Pandey; Anirban Basu; Sunit K. Singh
Chandipura virus (CHPV) is a neurotropic virus, known to cause encephalitis in humans. The microRNAs (miRNA/miR) play an important role in the pathogenesis of viral infection. The present study is focused on the role of miRNAs during CHPV (strain 1653514) infection in human microglial cells. The deep sequencing of CHPV-infected human microglial cells identified a total of 12 differentially expressed miRNA (DEMs). To elucidate the role of DEMs, the target gene prediction, Gene Ontology term (GO Term), pathway enrichment analysis, and miRNA-messenger RNA (mRNA) interaction network analysis was performed. The GO terms and pathway enrichment analysis provided 146 enriched genes; which were involved in interferon response, cytokine and chemokine signaling. Further, the WGCNA (weighted gene coexpression network analysis) of the enriched genes were discretely categorized into three modules (blue, brown, and turquoise). The hub genes in the blue module may correlate to CHPV induced neuroinflammation. Altogether, the miRNA-mRNA interaction network and WGCNA study revealed the following pairs, hsa-miR-542-3p and FAF1, hsa-miR-92a-1-5p and MYD88, and hsa-miR-3187-3p and TNFRSF21, which may contribute to neuroinflammation during CHPV infection in human microglial cells. © 2019 Wiley Periodicals, Inc.
Chandipura virus dysregulates the expression of hsa-miR-21-5p to activate NF-κB in human microglial cells
(BioMed Central Ltd, 2021) Neha Pandey; Meghana Rastogi; Sunit K. Singh
Background: Chandipura virus (CHPV) is a negative single-stranded RNA virus of the Rhabdoviridae family. CHPV infection has been reported in Central and Western India. CHPV causes acute encephalitis with a case fatality rate of 70 % and mostly affects children below 15 years of age. CHPV infection in brain leads to neuronal apoptosis and activation of the microglial cells. The microRNAs (miRNAs) are small endogenous non-coding RNA that regulate the gene expression. Viral infections perturb the expression pattern of cellular miRNAs, which may in turn affect the expression pattern of downstream genes. This study aims to investigate hsa-miR-21-5p mediated regulation of PTEN, AKT, NF-ĸBp65, IL-6, TNF-α, and IL-1β, in human microglial cells during CHPV infection. Methods: To understand the role of hsa-miR-21-5p in CHPV infection, the human microglial cells were infected with CHPV (MOI-0.1). Real-time PCR, western blotting, Luciferase assay, over-expression and knockdown techniques were used to understand the role of hsa-miR-21-5p in the regulation of PTEN, AKT and, NF-ĸBp65, IL-6, TNF-α, and IL-1β in this study. Results: The hsa-miR-21-5p was found to be upregulated during CHPV infection in human microglial cells. This led to the downregulation of PTEN which promoted the phosphorylation of AKT and NF-ĸBp65. Over-expression of hsa-miR-21-5p led to the decreased expression of PTEN and promoted further phosphorylation of AKT and NF-ĸBp65 in human microglial cells. However, the inhibition of hsa-miR-21-5p using hsa-miR-21-5p inhibitor restored the expression. Conclusions: This study supports the role of hsa-miR-21-5p in the regulation of pro-inflammatory genes in CHPV infected human microglial cells. © 2021, The Author(s).
Chikungunya virus modulates the miRNA expression patterns in human synovial fibroblasts
(John Wiley and Sons Inc., 2020) Meghna Agrawal; Neha Pandey; Meghana Rastogi; Smriti Dogra; Sunit K. Singh
Chikungunya virus (CHIKV) is an alphavirus transmitted by mosquitoes. CHIKV infection leads to polyarthritis and polyarthralgia among patients. The synovial fibroblasts are the primary target for CHIKV. The microRNAs (miRNAs) are the small endogenous noncoding RNAs which posttranscriptionally regulate the expression of genes by binding to their target messenger RNAs (mRNAs) through their 3′-untranslated regions. The miRNAs are the key regulators for various pathological processes including viral infection, cancer, cardiovascular disease, and neurodegeneration. This study was designed to dissect out the roles of miRNAs during CHIKV (Ross Strain E1: A226V) infection in primary human synovial fibroblasts. The miRNA microarray profiling was performed on the primary human synovial fibroblasts infected by CHIKV. The gene target prediction analysis, enrichment, and network analysis were performed by various bioinformatics analyses. The subset of 26 differentially expressed microRNAs (DEMs) were identified through microarray profiling and were further screened for gene predictions, Gene Ontology, pathway enrichment, and miRNA-mRNA network using various bioinformatics tools. The bioinformatics analysis indicates the role of DEMs by suppressing the immune response which may contribute to CHIKV persistence in human primary synovial fibroblasts. Our study provides the plausible roles of DEMs, miRNAs, and mRNA interactions and pathways involved in the molecular pathogenesis of CHIKV. © 2019 Wiley Periodicals, Inc.
Deciphering UV-B-induced variation in DNA methylation pattern and its influence on regulation of DBR2 expression in Artemisia annua L.
(Springer Verlag, 2015) Neha Pandey; Shashi Pandey-Rai
Main conclusion: UV-B-caused DNA hypomethylation and UV-B-mediated epigenetic activation of additional WRKY-binding site(s) in theDBR2promoter may contribute to the overexpression of theDBR2gene inArtemisia annua. DNA methylation is one of the key mechanisms behind stress-induced transcriptional switch off/on. Here, we evaluate the DNA methylation level in response to UV-B radiation in Artemisia annua which produces artemisinin, a sesquiterpene that has been recommended by WHO for the frontline treatment of malaria. However, the drug is facing serious shortage due to its low concentration in plants. UV-B treatment (3 h) enhanced artemisinin concentration up to 1.91-fold as compared to control. A key regulatory gene of artemisinin biosynthesis, DBR2 was upregulated under UV-B. This study presents observations regarding contributions of DNA methylation to the gene regulation using DBR2 as an example. Restriction digestion of genomic DNA by isoschizomers (MspI and HpaII) suggested UV-B involvement in DNA hypomethylation in A. annua. The global level of DNA methylation (R) was 3.4 and 5.9 % for UV-B treated and control plants, respectively, attesting hypomethylation of DNA in response to UV-B. Further bisulfite sequencing PCR showed demethylation at two CHG sites in 18S rRNA gene. Similarly, bisulfite sequencing of promoter region of DBR2 has demonstrated demethylation at 4 CG-, 4 CHH- and 2 CHG-sites. In silico analysis revealed UV-B-mediated demethylation at seven putative transcription factor binding sites including WRKY, which are positive regulators of artemisinin biosynthesis. UV-B treatment has resulted in activation of additional WRKY-binding site in UV-B-treated plants compared with single active WRKY-binding site in control and this could be the probable reason for overexpression of DBR2. It is suggested that DNA demethylation is an important epigenetic response to UV-B radiation in A. annua that surely will provide new horizons to further elucidate the mechanistic evidence of plant’s responses to UV-B radiation. © 2015, Springer-Verlag Berlin Heidelberg.
Development of SCAR based molecular markers for identification of different species of Azolla
(2013) G. Abraham; Neha Pandey; Vagish Mishra; Anis Ahmad Chaudhary; Altaf Ahmad; Rajendra Singh; P.K. Singh
Azolla is commonly found in rice paddy fields where it serves as a biofertilizer and green manure. The conventional methods of identification of Azolla based on the presence or absence of glochidia on the spores is difficult and cumbersome. In the present study, therefore, molecular markers based on specific sequence characterized amplified region (SCAR) primers were developed from the nucleotide sequence of specific RAPD markers. A single distinct and highly resolved band of 390, 182, 490 and 709 bp, respectively was obtained for Azolla rubra, A. pinnata, A. filiculoides and A. microphylla. This confirms the specificity of SCAR primer for these species. Thus, the development of SCAR marker may aid in the accurate and precise identification of different species of Azolla without ambiguity.
DNA methylation dynamics in plants
(Nova Science Publishers, Inc., 2017) Neha Pandey; Krishna K. Rai; Anjana Kumari; Shashi Pandey-Rai
DNA methylation is an important epigenetic mechanism that governs many vital biological activities in all living beings. In plants, DNA methylation is often correlated with the repression of gene expression. In addition, DNA cytosine methylation has been shown to play essential roles in plant development as well as in plant defence against a variety of abiotic and biotic stresses. The process of DNA cytosine methylation involves enzymatic addition of a methyl group to C-5 position of regular cytosine bases. This involves DNA methyltransferases (DNA MTase) family of enzymes that transfer methyl group to DNA using S-adenosine-L-methionine (AdoMet) as a ubiquitous methyl donor. The mechanism of DNA methylation takes place through siRNA-mediated pathways called as RNA-directed DNA methylation (RdDM). The DNA methylation status of a plant genome or any specific region of the plant genome may be estimated through a variety of reliable tools. Bisulfite sequencing is among the most reliable method to detect DNA methylation in plants. The present chapter summarizes the concept of DNA methylation event in plants, its mechanism in various plant biological processes and common methods to study the DNA methylation status in plants. © 2017 Nova Science Publishers, Inc.
Epigenetic control of UV-B-induced flavonoid accumulation in Artemisia annua L.
(Springer Verlag, 2019) Neha Pandey; Niraj Goswami; Deepika Tripathi; Krishna Kumar Rai; Sanjay Kumar Rai; Shilpi Singh; Shashi Pandey-Rai
Main conclusion: UV-B-induced flavonoid biosynthesis is epigenetically regulated by site-specific demethylation of AaMYB1, AaMYC, and AaWRKY TF-binding sites inAaPAL1promoter-causing overexpression ofAaPALgene inArtemisia annua. The present study was undertaken to understand the epigenetic regulation of flavonoid biosynthesis under the influence of ultraviolet-B radiation using Artemisia annua L. as an experimental model. In-vitro propagated and acclimatized plantlets were treated with UV-B radiation (2.8 W m −2 ; 3 h), which resulted in enhanced accumulation of total flavonoid and phenolics content as well as eleven individual flavonoids measured through HPLC-DAC. Expression of eight genes (phenylanaline ammonia lyase, cinnamate-4-hydroxylase, 4-coumarate: CoA ligase; chalcone synthase, chalcone isomerase, cinnamoyl reductase, flavonoid-3′-hydroxylase, and flavones synthase) from upstream and downstream flavonoid biosynthetic pathways was measured through RT-PCR and RT-Q-PCR and all were variably induced under UV-B irradiation. Among them, AaPAL1 transcript and its protein were most significantly upregulated. Global DNA methylation analysis revealed hypomethylation of genomic DNA in A. annua. Further epigenetic characterization of promoter region of AaPAL1 revealed cytosine demethylation at five sites, which in turn caused epigenetic activation of six transcription factor-binding sites including QELEMENT, EBOXBNNAPA/MYCCONSENSUSAT, MYBCORE, MYBCOREATCYCB1, and GCCCORE. MYB transcription factors are positive regulators of flavonoid biosynthesis. Epigenetic activation of transcription-enhancing cis-regulatory elements in AaPAL1 promoter and subsequent overexpression of AaMYB1 and AaMYC and AaWRKY transcription factors under UV-B irradiation may probably be the reason for higher AaPAL1 expression and hence greater biosynthesis of flavonoids in A. annua L. The present study is the first report that provides mechanistic evidence of epigenetic regulation of flavonoid biosynthesis under UV-B radiation in A. annua L. © 2018, Springer-Verlag GmbH Germany, part of Springer Nature.
Exogenous salicylic acid-mediated modulation of arsenic stress tolerance with enhanced accumulation of secondary metabolites and improved size of glandular trichomes in Artemisia annua L.
(Springer-Verlag Wien, 2018) Anjana Kumari; Neha Pandey; Shashi Pandey-Rai
The present study was undertaken to find out individual and interactive effects of arsenic (As) and salicylic acid (SA) on an important medicinal plant, Artemisia annua. As uptake and its accumulation was detected and found to be maximum in roots at higher As concentration (150 μM). Under As treatments, H2O2 and MDA content were induced. Biomass and chlorophyll content were negatively affected under As treatments. Furthermore, enzymatic (SOD, CAT, APX, and GR) and non-enzymatic antioxidants were also enhanced under As treatments. Exogenous application of SA reduced the extent of H2O2 and O2 − generation and lipid peroxidation, while reverted biomass and chlorophyll content to overcome oxidative stress. Simultaneous application of SA with As increased endogenous SA level, artemisinin, and dihydroartemisinic acid as compared with individual As treatment and pre-application of SA with As treatments. The expression of four key artemisinin biosynthetic pathway genes, i.e., ADS, CYP71AV1, DBR2, and ALDH1 were upregulated at a maximum in plants simultaneously treated with SA and As. Similar pattern of artemisinin accumulation and glandular trichome size was observed which attest that SA has a stimulatory impact on artemisinin biosynthesis under As stress. Our study suggests that exogenous application of SA and As together induced more tolerance in A. annua than a comparable dose of SA pre-treatment. The study may provide a platform with dual benefits by developing As-tolerant plants to be used for phytoremediation of arsenic from As-contaminated soil and obtaining high artemisinin-producing A. annua plants. © 2017, Springer-Verlag GmbH Austria.
Fake news - A manufactured deception, distortion and disinformation is the new challenge to digital literacy
(Amity University, 2018) Neha Pandey
In an era of information age, the defining variable of the age i.e. information is facing a tsunami in the form of fake news. Tsunami because it came all of a sudden, created havoc and is gargantuan in proportion. Facts have become a misnomer and reality an imagination. Confusion and chaos are the hurdle blocks for the one who wants to navigate through the digital landscape. The line between facts and fictions had been trounced. What is all the more alarming is that the fake-news tsunami is not a natural incidence but a whole business which blooms and grows on fabricated content and has become the new currency of globalized digitalization. Study on the spread of fake information, through social media and online networks, has become a significant object of scholarly research. Estimates suggest some fake news sites receive 50%-80% of their traffic via Facebook alone. This paper is a sincere attempt to map fake news into digital landscape by analysing its entry, varieties and survival. It also explores the various possibilities of duping users through a combination of intention and software algorithms. In the end, the researcher analyses how digital literacy and willingness on part of users can prove as a remedy in combating the fake news virus. If anything good has come from the recent furore over fake news and its menace, it is that fake news has highlighted the importance of making sure that the information one consumes and, especially, the information one shares is credible. It is like anti-virus potion which is created automatically. Or in simpler terms, it is like cultivating the habit to consume healthy, hygienic and balanced diet and toning the cognitive immune system. © 2018 Journal of Content, Community & Communication.
Gist of Zika Virus pathogenesis
(Academic Press Inc., 2021) Utkarsh Bhardwaj; Neha Pandey; Meghana Rastogi; Sunit K. Singh
Zika virus (ZIKV) is a mosquito-borne neurotropic flavivirus. ZIKV infection may lead to microcephaly in developing fetus and Guillain-Barré Syndrome (GBS) like symptoms in adults. ZIKV was first reported in humans in 1952 from Uganda and the United Republic of Tanzania. Later, ZIKV outbreak was reported in 2007 from the Yap Island. ZIKV re-emerged as major outbreak in the year 2013 from French Polynesia followed by second outbreak in the year 2015 from Brazil. ZIKV crosses the blood-tissue barriers to enter immune-privileged organs. Clinical manifestations in ZIKV disease includes rash, fever, conjunctivitis, muscle and joint pain, headache, transverse myelitis, meningoencephalitis, Acute Disseminated Encephalomyelitis (ADEM). The understanding of the molecular mechanism of ZIKV pathogenesis is very important to develop potential diagnostic and therapeutic interventions for ZIKV infected patients. © 2021 Elsevier Inc.
Herbal cures for psoriasis, an autoimmune disorder
(Nova Science Publishers, Inc., 2018) Shashi Pandey Rai; Deepika Tripathi; Neha Pandey; Anjana Kumari; Krishna Kumar Rai; Apoorva; Sanjay Kumar Rai
Psoriasis is a persistent, long-lasting (chronic), immune-mediated inflammatory skin problem that changes the life cycle of skin cells There are several factors that contribute to the development and/or aggravation of the disease. The causes of psoriasis pathogenesis are generally a complex interaction among genetic, epigenetic, immunological, and environmental components that involve T cells (T lymphocytes) of the immune system which migrates to the dermis and trigger the release of several cytokines like interleukins (IL), tumor necrosis factor (TNF), and interferons (IF), thereby causing inflammation and rapid production of keratinocytes. Psoriasis also develops due to various stresses like oxidative stress, caused as a result of reactive oxygen species (ROS); psychological stress; and skin injuries. Lipoxygenases (LOXs), a class of enzymes producing inflammatory mediators, are also known to govern psoriasis pathogenesis. Excessive use of certain drugs such as lithium salt, beta blockers and the antimalarial drug chloroquininine can aggravate the disease as well. The bioactive compounds present in medicinal plants have remarkable antioxidant properties required for the protection of the cells against the damaging effects of ROS and LOXs. Thus, they prevent the body against the adverse effects caused by oxidative stress and/or inflammatory responses. These plants are therefore capable of preventing and/or curing pathological conditions of skin and inflammatory diseases. In this chapter, the plant extracts/phytocompounds that have efficient antioxidant, anti-inflammatory, and, immuno-modulatory properties (the significant attributes essential for combating psoriasis), and can be further exploited as potent drug candidates for curing psoriasis were described to prospect them for drug discovery. © 2018 Nova Science Publishers, Inc.
Heterologous expression of cyanobacterial PCS confers augmented arsenic and cadmium stress tolerance and higher artemisinin in Artemisia annua hairy roots
(Springer, 2021) Neha Pandey; Krishna Kumar Rai; Sanjay Kumar Rai; Shashi Pandey-Rai
The present study provides the first report of heterologous expression of phytochelatin synthase from Anabaena PCC 7120 (anaPCS) into the hairy roots of Artemisia annua. Transformed hairy roots of A. annua expressing anaPCS gene showed better tolerance to heavy metals, viz., arsenic (As) and cadmium (Cd) owing to 143 and 191% more As- and Cd-accumulation, respectively, as compared to normal roots with a bioconcentration factor (BCF) of 9.7 and 21.1 for As and Cd, respectively. Under As and Cd stresses, transformed hairy roots possessed significantly higher amounts of phytochelatins and thiols probably due to the presence of both AaPCS (Artemisia annua PCS) and anaPCS. In addition, artemisinin synthesis was also induced in transformed hairy roots under heavy metals stresses. In-silico analysis revealed the presence of conserved motifs in both AaPCS and anaPCS sequences as well as structural modelling of PCS functional domain was conducted. Interaction of AaPCS and anaPCS proteins with CdCl2 and sodium arsenate gene ontology analysis gave insights to anaPCS functioning in transformed hairy roots of A. annua. The study provides transformed hairy roots of A. annua as an efficient tool for effective phytoremediation with added advantages of artemisinin extraction from hairy roots used for phytoremediation. © 2021, Korean Society for Plant Biotechnology.
Impact of integrated omics technologies for identification of key genes and enhanced artemisinin production in Artemisia annua L
(CRC Press, 2017) Shashi Pandey-Rai; Neha Pandey; Anjana Kumari; Deepika Tripathi; Sanjay Kumar Rai
Medicinal plants are the source of an enormous variety of bioactive secondary metabolites and have potential synergistic effects against a broad range of human diseases. According to the World Health Organization (WHO), more than 80% of the world’s population depend on medicinal plants for everyday healthcare. One of the most popular secondary metabolites with immense 222therapeutic potential is artemisinin (AN), present in a well-known Asteraceae family member, Artemisia annua L. The AN isolated from the leaves of A. annua by Chinese scientist You You Tu is acknowledged as an effective antimalarial compound (Barbacka and Baer-Dubowska 2011). AN and its bioactive derivatives isolated from A. annua are powerful medicines widely used for their ability to swiftly control Plasmodium malaria. AN-based combination therapies (ACTs), with their established safety record, are the first line of treatment recommended by WHO (2014) for malaria caused by Plasmodium falciparum. In addition to its antimalarial effects, AN has recently been evaluated for its potential antibacterial, antiviral, antitumor, antileishmanial, antischistosomiatic, anti-sleeping sickness, anticancer, and herbicidal properties (Efferth et al. 2011; Utzinger et al. 2001; Sen et al. 2007; Mishina et al. 2007). However, the low content of AN in plant tissue has resulted in poor yield/production of AN, which seems insufficient to fulfill the demand for 392 million courses of ACT each year (WHO 2014). Semisynthetic derivatives of AN, such as artemether and artesunate, are also commonly produced commercially, but they are not routinely available in remote rural areas. Moreover, these derivatives are very expensive, and low yields of AN result in relatively high costs for its extraction and purification. Further, A. annua requires a relatively long period of time for its agricultural cultivation, which results in wide swings in affordable, best-quality, robust supply of raw materials, and prices. Intensified efforts have been carried out to increase AN production (Liu et al. 2006). However, the routine metabolic engineering strategy, via overexpressing or downregulating key genes in AN biosynthetic pathways, has not proved very effective. Glandular secretory trichomes, sites of AN biosynthesis on the surface of A. annua, are the new target for increasing AN yield (Duke et al. 1994). In general, the population and morphology of glandular secretory trichomes in A. annua (AaGSTs) are positively correlated with AN content. Higher production of AN requires breeding of A. annua to optimize the biomass yield and trichome density. Various efforts have been made to breed high-trichome density cultivars of the plant for increased AN production. However, various approaches have been already taken into consideration for the semisynthesis of AN (Paddon et al. 2013). The production of AN is also challenging because A. annua remains relatively undeveloped as a crop. Therefore, there is a need to improve the varieties and cultivation strategies of A. annua for farmers in developing countries, because this would bring immediate benefits to the existing supply chain of AN. Major advancements in omics technologies such as genomics, proteomics, and metabolomics have enabled high-throughput monitoring of a variety of molecular and biochemical processes. These techniques have been widely applied to identify biological variants and complex biochemical pathways/systems. Many omics platforms target the comprehensive analysis of genes (genomics), mRNA (transcriptomics), proteins (proteomics), and metabolites (metabolomics). However, the interpretation of obtained data is challenging due to very complex biochemical and metabolic pathways.223. © 2018 by Taylor & Francis Group, LLC.
In vitro generation of high artemisinin yielding salt tolerant somaclonal variant and development of SCAR marker in Artemisia annua L
(Springer Netherlands, 2016) Neha Pandey; Ram Prasad Meena; Sanjay Kumar Rai; Shashi Pandey-Rai
Low levels of artemisinin in Artemisia annua is posing serious limitation in sustainable supply of this drug and ultimately affecting global struggle to cure malaria. The present study explores new ways for higher artemisinin production through generating somaclonal variant tolerant against salt stress. Here, through gamma-rays irradiation a total of 13 somaclonal variants (ASV1 to ASV13) were identified. They were characterized on the basis of morphological traits and further molecular characterization was carried out through RAPD analysis. Dendrogram, constructed on the basis of RAPD scores, showed six major clusters in which ASV12 was most distant from control to fallen alone in last cluster. Based on 16 metric traits examined among 13 somaclonal variants and wild type, correlation analysis was carried out. In addition, principle component analysis was conducted to attest the results. All statistical analysis suggested ASV12 as stable salt tolerant somaclonal variant of A. annua. ASV12 had higher artemisinin content as compared to wild type and under salt stress; expression of key artemisinin biosynthetic genes (ADS, CYP71AV1, DBR2 and ALDH1) was more in ASV12 as compared to wild type. Further SCAR marker (570 bp) linked with high artemisinin was developed and further tested on the low and high artemisinin yielding lines, F1 and F2 progenies. A single band was amplified in ASV12 and all high artemisinin-yielding plants. The study provides ample possibility of cultivation of ASV12 variant of A. annua on saline soil for dual benefit: high artemisinin production and utilization of waste saline land. © Springer Science+Business Media Dordrecht 2016.
In-vivo Studies and Molecular Docking of Modeled Mus musculas 8S Lipoxygenase Protein Using Some Natural Bioactive Compounds
(Springer, 2022) Shashi Pandey-Rai; Krishna Kumar Rai; Neha Pandey; Deepika Tripathi; Apoorva; Vinay Kumar Singh; Sanjay Kumar Rai
Mus musculus 8S Lipoxygenase (8SLOX) is expressed in suprabasal keratinocytes, and the massive accumulation of 8-HETE, the product of 8SLOX, plays a significant role in epidermal tumor development in papillomas and induces chromosomal alterations in basal keratinocytes. It has also been observed to be unregulated in inflammatory dermatoses. In an attempt to discover potent 8S LOX inhibitors, 5 bioactive compounds, viz., anthraquinone, celastrine, curcumin, gilloin and tinosporide, derived from different medicinal plants, were selected for the molecular docking study against 8S LOX protein. NDGA, a known lipoxygenase inhibitor was used as a standard. A 3D structure of Mus musculus Arachidonate 8S LOX protein (PMDB ID: PM0078979) was predicted using comparative homology modeling with homologous template (PDB ID: 3V98), retrieved from PDB resources. Conserved active site residues Gln109 and Asn174 were found to be involved in interaction with all drugs including NDGA. Docking studies revealed that celastrine has the best interaction followed by gilloin and curcumin, all of which performed much better than NDGA. Gene expression pattern in the skin of the control mice, IMQ-treated and phytocompound-treated psoriasis-like dermatitis inducedmice revealed that the curcumin and gillion compound were more influential in downregulating 8S lipoxygenase gene than the other compound used in this study. The results suggested that these potent compounds can efficiently inhibit 8S LOX protein and therefore can prove to be promising candidates in drug development for the treatment of inflammatory disorders of skin like psoriasis. © 2021, The National Academy of Sciences, India.
Influence of salicylic acid elicitation on secondary metabolites and biomass production in in-vitro cultured withania coagulans (l.) dunal
(Plant Archives, 2019) Bipin Maurya; Krishna Kumar Rai; Neha Pandey; Lakee Sharma; Niraj Kumar Goswami; Shashi Pandey Rai
Withania coagulans is an important medicinal plant possessing several secondary metabolites collectively termed as withanolides. Salicylic acid is widely known to stimulate plant growth and production under different conditions. Therefore, the present study was conducted to determine how SA regulates secondary metabolites and biomass production in tissue culture raised seedlings of W. coagulans. Among all the four treatments, exogenously supplemented salicylic acid at 150 and 200 μM concentrations exhibited maximum potential to modulate growth and physiological processes of the seedlings compared to 50 and 100 μM treatments. Exogenous application of SA at 150 and 200 μM significantly improved photosynthetic pigment contents, increased biomass, enhanced secondary metabolite contents viz., phenol, proline, and anthocyanin and strengthen antioxidative defense system for scavenging reactive oxygen species. Findings of the present study illustrated that exogenous supplementation of salicylic acid-regulated the mRNA level of genes involved in secondary metabolite biosynthetic pathway thus enhancing the production of the secondary metabolite, photosynthetic efficiency, improved plant growth and increased plant biomass of tissue culture raised seedlings of W. coagulans. © 2019 Plant Archives. All rights reserved.