» Interest

Molecular Biology, Cytogenetics, Reproductive Biology, Plant Tissue Culture and Transgenics

» Focus

Our research endeavour has been towards pathway engineering through systems biology approach in some of the prioritized medicinal plant species viz. withania somnifera, Picrorrhiza kurrooa, Rheum emodi andNothapodytes nimmoniana. Major focus is on cloning and characterization key pathway genes and elucidation of unknown pathways of some biologically active metabolites.Substrate pool diversion and validation of potential metabolic branch-point genes for pathway diversion/ intensification is also an area of priority. One of the recent research programmes has been to understand the stem cell biology in plants and role of plant steroids in cell proliferation and differentiation. Additionally, other areas of interest have been, genome analysis using molecular and cytogenetic analysis to understand  the synteny in Chlorophytum complex,.; Development of transgenic system for  over-expression studies of key regulatory genes in P. kurroa for pathway intensification; In-vitro mutagenesis to elicit retrotransposon activity in sexually sterile polyploidy species (Rose scented Geranium) and ethylene modulation via antisense transgenics using ACC synthase gene for the development of adaptive cultivars to environmental stresses; Development of cell culture strategies for metabolite production (Onosma echioides) ; Genetic amelioration of commercially important medicinal and aromatic plants via classical genetic approaches (breeding experiments) in Lavendula officinalis Cymbopogon jawarancusa, Andrographis paniculata, Sisymbrium irio and W. somnifera ; Detailed study of reproductive biology/diversity  for effective conservation of genetic and allelic variability in W. somnifera, A. paniculata and Grewia asiatica; Gene-environmental interaction by different biometrical methods and multilocational trials for enabling to screen genotypes/somaclonal variants for diverse agro-ecological niches of  Pleragonium graveloens (rose scented geranium) sp.

» Cloning and Functional Characterization of Three Branch Point Oxidosqualene Cyclases from Withania somnifera (L.) Dunal

Oxidosqualene cyclases (OSCs) positioned at a key metabolic sub-dividing junction execute indispensable enzymatic cyclization of 2, 3-oxidosqualene for varied triterpenoid biosynthesis. Such branch-points present favourable gene targets for redirecting metabolic flux towards specific secondary metabolites. However, detailed information regarding the candidate OSCs covering different branches and their regulation is necessary for desired genetic manipulation. The aim of the present study, therefore, was to characterize members of OSC superfamily from Withania somnifera (Ws), a medicinal plant of immense repute known to synthesize a large array of biologically active steroidal lactone triterpenoids called withanolides. Three full length OSC cDNAs, β-amyrin synthase (WsOSC/BS), lupeol synthase (WsOSC/LS) and cycloartenol synthase (WsOSC/CS) having open reading frames of 2289 bp, 2268 bp and 2277 bp were isolated. Heterologous expression in Schizosaccharomyces pombe, LC-MS analyses and kinetic studies confirmed their mono-functionality. The three WsOSCs were found to be spatially regulated at transcriptional level with WsOSC/CS being maximally expressed in leaf tissue. Promoter analysis of three WsOSCs genes resulted in identification of distinct cis-regulatory elements. Further, transcript-profiling under methyl jasmonate (MeJA), gibberellic acid (GA3) and yeast extract (YE) elicitations displayed differential transcriptional regulation of each of the OSCs. Changes were also observed in mRNA levels under elicitations and further substantiated with protein expression levels by western blotting. Negative regulation by YE resulted in significant increase in withanolide content. Empirical evidence suggests that repression of competitive branch OSCs like WsOSC/BS and WsOSC/LS possibly leads to diversion of substrate pool towards WsOSC/CS for increased withanolide production.

(A&B) Transcript profiles of WsOSCs in response to elicitor treatments. (C) Western immunoblot of WsOSCs in response to elicitor treatments. (D) Scheme of proposed withanolide biosynthesis pathway. (E) Time-course effect of elicitor treatments on accumulation of withanolides. Transcript profiles of WsOSCs in response to elicitor treatments (F) Southern blot analysis of (A) WsOSC/BS, (B) WsOSC/LS and (C) WsOSC/CS

» NADPH-Cytochrome P450 Reductase: Molecular Cloning and Functional Characterization of Two Paralogs from Withania somnifera (L.) Dunal

Cytochrome P450 reductase is the most imperative redox partner of multiple P450s involved in primary and secondary metabolite biosynthesis. We describe here the cloning and characterization of two paralogs of cytochrome P450 reductase from W. somnifera. The full length paralogs of WsCPR1 and WsCPR2 have open reading frames of 2058 and 2142 bp encoding 685 and 713 amino acid residues, respectively. Phylogenetic analysis demonstrated that grouping of dual CPRs was in accordance with class I and class II of eudicotyledon CPRs. The corresponding coding sequences were expressed in Escherichia coli as glutathione-S-transferase fusion proteins, purified and characterized. Recombinant proteins of both the paralogs were purified with their intact membrane anchor regions and it is hitherto unreported for other CPRs which have been purified from microsomal fraction. Southern blot analysis suggested that two divergent isoforms of CPR exist independently in Withania genome. Quantitative real-time PCR analysis indicated that both genes were widely expressed in leaves, stalks, roots, flowers and berries with higher expression level of WsCPR2 in comparison to WsCPR1. Similar to CPRs of other plant species, WsCPR1 was un-inducible while WsCPR2 transcript level increased in a time-dependent manner after elicitor treatments. High performance liquid chromatography of withanolides extracted from elicitor-treated samples showed a significant increase in two of the key withanolides, withanolide A and withaferin A, possibly indicating the role of WsCPR2 in withanolide biosynthesis.

                                      PLoS ONE (2013), 8(2): e57068. doi:10.1371/journal.pone.0057068

(A) Sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS-PAGE) of heterologously expressed WsCPR1 and WsCPR2 (B) Southern blot analysis of genomic DNA (C) Time course effect of elicitor treatments on expression profiles of WsCPRs paralogs. (D) Time course effect of elicitor treatments on withanolides accumulation

» Molecular characterization of two A-type P450s, WsCYP98Aand WsCYP76A from Withania somnifera (L.) Dunal: expression analysis and withanolide accumulation in response to exogenous elicitations

Two A-types P450 WsCYP98A and WsCYP76A were isolated, sequenced and heterologously expressed in E. coli. Both P450s are spatially regulated at transcript level showing differential tissue specificity. Exogenous elicitors acted as both positive and negative regulators of mRNA transcripts. Methyl jasmonate and salicylic acid resulted in copious expression of WsCYP98A andWsCYP76A. Enhanced mRNA levels also corroborated well with the increased accumulation of withanolides in response to elicitations. The empirical findings suggest that elicitors possibly incite defence or stress responses of the plant by triggering higher accumulation of withanolides.

                                    BMC Biotechnology (2014) 14(1), 89-105

(A) HPLC analysis of withanolide A (WS-1), withanone (WS-2), withaferin A (WS-3) at 6 h, 48 h from methyl jasmonate(0.1mM), salicylic acid (0.1mM) and gibberellic acid (0.1mM) treated microshoots from Withania Somnifera. (B) Ramachandran plot of WsCYP98A and WsCYP76A 3D models of Withania somnifera. (C) Sodium dodecyl sulphate polyacrylamide gel electrophoresis pattern of proteins obtained from E. coli BL-21 (DE3) transformed with pGEX-WsCYP76A and pGEX-WsCYP98A. (D) Multiple sequence alignment of deduced amino acid sequences of WsCYP98A and WsCYP76A with their respective homologs using Multalign tool.

» Molecular characterization and promoter analysis of squalene epoxidase gene from Withania somnifera (L.) Dunal

Squalene epoxidase (SQE; EC. 1.14.99.7) is one of the rate limiting enzymes in the biosynthesis of triterpenoids, catalyzing the stereospecific epoxidation of squalene to 2,3-oxidosqualene. A full length SE gene (WsSQE) of 1,956 bp was cloned which contained an open reading frame of 1,596 bp, encoding a protein of 531 amino acids with a predicted molecular mass of 57.67 kDa and theoretical PI of 8.48. Full length WsSQE was cloned into pGEX4T-2 vector and expressed in E. coli. Phylogenetic analysis indicated a significant evolutionary relatedness of WsSQE with squalene epoxidases of other plant species and the degree of relatedness with deduced amino acid sequences showed a significant correlation with different plant species. Using genome walking approach, a promoter sequence of 513 bp of WsSQE was isolated which revealed several key cis-regulatory elements known to be involved in various biotic and abiotic plant stresses. Comparative expression analysis of tissue specific WsSQE done by quantitative-PCR demonstrated the highest transcript levels in leaves, as compared to stalk and root tissues. This is the first report of cloning and bacterial expression of SE from W. somnifera and may be of significant interest to understand the regulatory role of SE in the biosynthesis of withanolides.

Molecular Biology Reports (2013), 40:905–916

(A) Heterologous protein expression in E. coli using 1Mm IPTG at 2hr, 4 hr, 6hr and 8 hr. (B) Nucleotide and deduced amino acid sequence of WsSQE from W. somnifera. (C) Deduced 3-D structure of WsSQE. (D) Comparative expression levels of WsSQE in various tissues of  W. somnifera

» Molecular characterization of UGT94F2 and UGT86C4, two glycosyltransferases from Picrorhiza kurrooa: Comparative structural insight and evaluation of substrate recognition

Uridine diphosphate glycosyltransferases (UGTs) are pivotal in the process of glycosylation for decorating natural products with sugars. It is one of the versatile mechanisms in determining chemical complexity and diversity for the production of suite of pharmacologically active plant natural products. Picrorhiza kurrooa is a highly reputed medicinal herb known for its hepato-protective properties which are attributed to a novel group of iridoid glycosides known as picrosides. Although the plant is well studied in terms of its pharmacological properties, very little is known about the biosynthesis of these important secondary metabolites. In this study, we identified twofamily-1 glucosyltransferases from P. kurrooa. The full length cDNAs of UGT94F4 and UGT86C4 contained open reading frames of 1455 and 1422 nucleotides, encoding polypeptides of 484 and 473 amino acids respectively. UGT94F2 and UGT86C4 showed differential expression pattern in leaves, rhizomes and inflorescence. To elucidate whether the differential expression pattern of the two Picrorhiza UGTs correlate with transcriptional regulation via their promoters and to identify elements that could be recognized by known iridoid-specific transcription factors, upstream regions of each gene were isolated and scanned for putative cis-regulatory elements. Interestingly, the presence of cis-regulatory elements within the promoter regions of each gene correlated positively with their expression profiles in response to different phytohormones. HPLC analysis of picrosides extracted from different tissues and elicitor-treated samples showed a significant increase in picroside levels, corroborating well with the expression profile of UGT94F2 possibly indicating its implication in picroside biosynthesis. Using homology modeling and molecular docking studies, we provide an insight into the donor and acceptor specificities of both UGTs identified in this study. UGT94F2 was predicted to be an iridoid-specific glucosyltransferase having maximum binding affinity towards 7-deoxyloganetin while as UGT86C4 was predicted to be a kaempferol-specific glucosyltransferase.

PloS one (2013) 8(9), e73804

(A) A schematic diagram of the proposed picroside pathway in Picrorhiza kurrooa. (B) Tissue-specific real-time expression analysis and time-course effect of elicitor treatments on UGT86C4 and UGT94F2. (C) Three dimensional models and conserved residue prediction for UGT86C4 and UGT94F2. (D) Nucleotide sequence of Picrorhiza UGT gene promoters

» Efficient plant regeneration via direct organogenesis and Agrobacterium tumefaciens-mediated genetic transformation of Picrorhiza kurroa: an endangered medicinal herb of the alpine Himalayas

Picrorhiza kurroa Royle ex. Benth. is a medicinal herb of immense therapeutic value with restricted geographic distribution. Efficient plant regeneration via direct organogenesis and Agrobacterium tumefaciens-mediated genetic transformation was developed for this plant. Multiple shoot bud induction was achieved from leaf explants cultured in Gamborg’s B5 medium containing 3% (w/v) sucrose, 3 mg/l kinetin and 1 mg/l indole-3-butyric acid. More than 90% of leaf explants formed shoot buds leading to whole plant regeneration. An Agrobacterium-mediated genetic transformation protocol was developed using A. tumefaciens strain GV3101 harboring binary vector pCAMBIA1302 containing the green fluorescent protein and hygromycin phosphotransferase genes. Leaf explants precultured for 2 d were the most suitable for co-cultivation with Agrobacterium and transformation efficiency was enhanced with 200 μM acetosyringone. Putative transformants were selected using media containing 15 mg/l hygromycin. Transformation was verified by detection of the green fluorescent protein using fluorescence microscopy and by polymerase chain reaction. Approximately 56% of the explants were transformed with an average of 3.4±0.4 transgenic plantlets per explant. An efficient regeneration and transformation protocol thus developed enabling a fresh perspective of metabolic engineering in P. kurroa using an Agrobacterium-mediated transformation. This is the first report of direct organogenesis from leaf explants and genetic transformation of P. kurroa.

In Vitro Cellular & Developmental Biology (2012), 48:295-303

(A) Plant regeneration in P. kurroa. Shoot bud differentiation and regeneration on B5 minimal medium containing 3 mg/l kinetin and 1 mg/l indole-3-butyric acid(regeneration medium) after 4 wk in culture (B) Regenerated shoots showing profuse rooting induced on RM containing 1% activated charcoal. (C) Potted tissue culture-raised plantlets (D) Transient green fluorescent protein and β-glucuronidase expression shown by infected leaf explants after 3 days of cocultivation (E) Putative transgenic plants potted for hardening (F) Transgenic P. kurroa plants grown under containment conditions. (G) PCR detection of hpt and gfp in wild type (WT) and transformed cultures (T1-T4) of P. kurroa

» An inducible NADPH–cytochrome P450 reductase from Picrorhiza kurroa—an imperative redox partner of cytochrome P450 enzymes

Picrorhiza kurroa synthesizes a large array of pharmacologically important mono terpenoid iridoid glycosides called picrosides. Although chemical profile and pharmacological activities of P. kurrooa have been extensively studied, limited attempts have been made to decipher the biosynthetic route and to identify the key regulatory genes involved in picroside biosynthesis. In the present study, NADPH–cytochrome P450 reductase, a key enzyme involved in electron transfer to cytochrome P450s was identified from P. kurroa. The full length cDNA (2679 bp) contained an open reading frame of 2133 bp, corresponding to 710 amino acids. PkCPR was heterologously expressed in Escherichia coli and the kinetic parameters of the recombinant enzyme were determined. Specific activity, Vmax and Km of PkCPR were found to be 5.8±0.05 μmol min−1 mg−1, 8.1±0.12 μmol min−1 mg−1 and 7.8 μM, respectively. PkCPR was found to be spatially regulated at transcript level, being maximally expressed in leaf tissues. Altitude was found to have a positive effect on the picroside concentration and the picroside content positively correlated with the PkCPR transcript levels in samples collected

at varied altitudes. Further, transcript profiling under methyl jasmonate, salicylic acid, 2,4-dicholorophenoxy acetic acid and UV-B elicitations displayed differential transcriptional regulation of PkCPR that fully corroborated with the identified cis-elements within the PkCPR promoter. Expression of PkCPR was inducible by UV-B and phytohormone elicitation, indicating that the PkCPR is possibly related to defence reactions, including biosynthesis of secondary metabolites. Present study is so far the only report of identification and functional characterization of CPR ortholog from P. kurroa.

Functional & Integrative Genomics (2014) 14, 381-399

(A) Overview of the proposed picroside pathway in Picrorhiza kurrooa (B) Multiple sequence alignment of deduced amino acid sequences of P. kurrooa from NCBI database. (C) Expression characteristics of PkCPR in different tissues such as leaves, rhizome, and inflorescence. (D) Differential accumulation pattern of picrosides I, II, apocynin, androsin and feruloylcatalpol in response to various altitudinal gradients.

» Evaluation of anthraquinones from Himalayan rhubarb (Rheum emodi Wall. ex Meissn.) as antiproliferative agents

Rheum emodi (Polygonaceae), a multipurpose medicinal herb is a rich repository of pharmacologically active secondary metabolites known as anthraquinones. The present study entails HPLC quantitation and in vitro activity of four major constituents and the extracts of R. emodi. The anthraquinone glycosides were more abundant than their aglycone constituents viz. emodin and chrysophanol. MTT assay was used to assess the in vitro antiproliferative activity of anthraquinones and extracts on four cancer cell lines namely MIAPaCa-2, HCT-116, MCF-7 and T47D. The cytotoxicity was more obvious on MIAPaCa-2. Further, the study of mechanism of action involving cell cycle analysis and determination of mitochondrial membrane potential (MMP) loss was also investigated. The extracts significantly reduced cell viability by inducing apoptosis/necrosis and cell cycle arrest with concurrent loss of MMP (Δψm) in a concentration dependent manner. The methanolic extract of rhizome (SPL5) with the least IC50 value (25 μg/ml) showed a significant increase in the percentage of apoptotic/necrotic cells (42.3% at 100 μg/ml) compared to that of vehicle treated cells (11.6%). These cellular manifestations corresponded remarkably with a decrease in the integrity of the mitochondrial membrane. In conclusion, SPL5 with emodin and chrysophanol as the preponderant constituents exhibited considerable antiproliferative activity possibly by reducing cell viability and stirring up Δψm loss.

South African Journal of Botany (2014), 95:1-8

(A) Anthraquinone concentrations in extracts. (a, b and c represent concentrations of major anthraquinones in methanolic & ethyl-acetate rhizome extracts and methanolic/ethyl acetate extracts of leaf. (B) MTT assay. Comparison of IC50 values of markers (SPL1-4) and extracts (SPL5-8) against various cancer cell lines. (C) Flow cytometric analysis of human cancer cell line (MIAPaCa-2). Cells were exposed to different concentrations of extracts (10, 50 and 100µg/ml) for 48 h and stained with PI (5μg) to determine DNA fluorescence and cell cycle phase distribution using FACS Aria II flowcytometer.

• Dhar, N.; Rana, S.; Razdan, S.; Bhat, W.W.; Hussain, A.; Dhar, R.S.; Vaishnavi, S.; Hamid, A.; Vishwakarma, R.A.; Lattoo, S.K. Cloning and functional characterization of three branch point oxidosqualene cyclases from Withania somnifera (L.) Dunal. The Journal of Biological Chemistry (2014), 289:17249-17267
• Rana, S.; Bhat, W.; Dhar, N.; Pandith, S.A.; Razdan, S.; Vishwakarma, R.; Lattoo, S.K. Molecular characterization of two A-type P450s, WsCYP98A and WsCYP76A from Withania somnifera (L.) Dunal: expression analysis and withanolide accumulation in response to exogenous elicitations. BMC Biotechnology (2014), 14(1): 89
• Bhat, W.W.; Rana, S.; Dhar, N.; Razdan, S.; Pandith, S.A.; Vishwakarma, R.A.; Lattoo, S.K. An inducible NADPH-cytochrome P450 reductase from Picrorhiza kurrooa- an imperative redox partner of cytochrome P450 enzymes. Functional and Integrative genomics (2014), 14(2), 381-399
• Bhat, W.W.; Razdan, S.; Rana ,S.; Dhar, N.; Wani, T.A.; Qazi, P.; Vishwakarma, R.; Lattoo, S.K. A phenylalanine ammonia-lyase ortholog (PkPAL1) from Picrorhiza kurrooa Royle ex. Benth: Molecular cloning, promoter analysis and response to biotic and abiotic elicitors. Gene (2014), 547(2): 245–256
• Bhat, W. W.; Dhar, N.; Razdan, S.; Rana, S.; Mehra, R.; Nargotra, A.; Dhar, R.S.; Ashraf, N.; Vishwakarma, R.; Lattoo, S.K. Molecular Characterization of UGT94F2 and UGT86C4, Two Glycosyltransferases from Picrorhiza kurrooa: Comparative Structural Insight and Evaluation of Substrate Recognition. PLoS ONE (2013), 8(9): e73804
• Rana, S.; Lattoo, S.K.; Dhar, N.; Razdan, S.; Bhat, W.W.; Dhar, R.S.; Vishwakarma, R. NADPH-Cytochrome P450 reductase: Molecular cloning and functional characterization of two paralogs from Withania somnifera (L.) Dunal. PLoS ONE (2013), 8(2): e57068
• Razdan, S.; Bhat, W.W.; Rana, S.; Dhar, N.; Lattoo, S.K.; Dhar, R.S.; Vishwakarma R.A. Molecular characterization and promoter analysis of squalene epoxidase gene from Withania somnifera (L.) Dunal. Mol Bio Rep. (2013), 40:905–916
• Dhar, N.; Rana, S.; Bhat, W. W.; Razdan, S.; Pandith, S.A.; Khan, S.; Dutt, P.; Dhar, R.S.; Vaishnavi, S.; Vishwakarma, R.; Lattoo, S.K. Dynamics of withanolide biosynthesis in relation to temporal expression pattern of metabolic genes in Withania somnifera (L.) Dunal: a comparative study in two morpho-chemovariants. Molecular Biology Reports (2013), 40(12):7007-16
• Farooq, U.; Pandith, S.A.; Saggoo, M. I. S.; Lattoo, S. K. Altitudinal variability in anthraquinone constituents from novel cytotypes of Rumex nepalensis Spreng-a high value medicinal herb of North western Himalayas. Industrial Crops  and  Products (2013), 50:112–117
• Rana, S.; Dhar, N.; Bhat,W.W.; Razdan, S.; Khan, S.; Dhar, R.S.; Dutt, P.; Lattoo, S.K. A 12-deoxywithastramonolide-rich somaclonal variant in Withania somnifera (L.) Dunal – molecular cytogenetic analysis and significance as a chemotypic resource. In Vitro Cell. Dev. Biol.—Plant (2012), 48:546–554
• Bhat,W.W.; Lattoo, S.K.; Razdan, S.; Dhar, N.; Rana, S.; Dhar, R.S.; Khan, S.; Vishwakarma, R.A. Molecular cloning, bacterial expression and promoter analysis of squalene synthase from Withania somnifera (L.) Dunal. Gene, (2012), 499: 25–36
• Bhat, W.W.; Lattoo, S.K.; Rana, S.; Razdan, S.; Dhar, N.; Dhar, R.S.; Vishwakarma R.A. An efficient plant regeneration via direct organogenesis and Agrobacterium tumefaciens-mediated genetic transformation of Picrorhiza kurrooa – an endangered medicinal herb of alpine Himalayas. In Vitro Cell. Dev. Biol.—Plant (2012), 48 (3): 295-303
• Rana, S.; Dhar, N.; Bhat, W.W.; Razdan, S.; Khan, S.; Dhar, R. S.; Dutt, P.; Lattoo, S. K. A 12-deoxywithastramonolide-rich somaclonal variant in Withania somnifera (L.) Dunal – molecular cytogenetic analysis and significance as a chemotypic resource. In Vitro Cell. Dev. Biol.—Plant (2012), 48:546–554
• Lattoo, S.K.; Bamotra, S.; Dhar, R.S.; Khan, S.; Dhar, A.K. Rapid plant regeneration and analysis of genetic fidelity of in vitro derived plants of Chlorophytum arundinaceum Baker – an endangered medicinal herb. Plant Cell Rep (2008), 25(6): 499-506
• Lattoo, S.K.; Dhar, R.S.; Khan, S.; Bomotra, S.; Bhan, M.K.; Dhar, A.K.; Gupta, K.K. Comparative analysis of genetic diversity using molecular and morphometric markers in Andrographis panicualata (Burm. f.Nees). Genetic Resour crop Evol (2008), 55 (1): 33-43
• Lattoo, S. K.; Dhar, R. S.; Khan, S.; Bomotra, S.; Bhan, M.K.; Dhar, A.K.; Gupta, K. K. Comparative analysis of genetic diversity using molecular and morphometric markers in Andrographis panicualata (Burm. f.Nees). Genetic Resour crop Evol (2008), 55 (1): 33-43
• Lattoo, S.K.; Khan, S.; Bamotra, S.; Dhar, A.K. Cytomixis impairs meiosis and influences reproductive success in Chlorophytum comosum (Thumb) Jacq. – an additional strategy and possible implications. J. Biosci (2006), 31(5): 1-9
• Lattoo, S. K.; Khan, S.; Dhar, A.K.; Choudhary, D. K.; Gupta, K. K.; Sharma, P.R. Genetics and mechanism of induced male sterility in Andrographis paniculata (Burm. f.) Nees and its significance. Curr. Sci., (2006), 91 (4): 515-519
• Wani, T.A.; Pandith, S.A.; Rana, S.; Bhat, W.W.; Dhar, N.; Razdan, S.; Chandra, S.; Kitchlu, S.; Sharma, N.; Lattoo, S.K. Promiscuous breeding behaviour in relation to reproductive success in Grewia asiatica L. (Malvaceae). Flora-Morphology, Distribution, Functional Ecology of Plants (2015), 211:62-71
• Pandith, S.A.; Hussain, A.; Bhat, W.W.; Dhar, N.; Qazi, A. K.; Rana, S.; Razdan, S.; Wani, T.A.; Shah, M. A.; Bedi, Y. S.; Hamid, A.; Lattoo, S. K. Evaluation of anthraquinones from Himalayan rhubarb (Rheum emodi Wall. ex Meissn.) as antiproliferative agents. South African Journal of Botany, (2014), 95:1–8
• Lattoo, S. K.; Dhar, R. S.; Khan, S.; Bomotra, S.; Dhar, A.K. Temporal sexual maturation and incremental staminal movement encourages mixed mating in Withania somnifera–an insurance for reproductive success. Curr. Sci, (2007), 92(10): 1390-1399
• Lattoo, S. K.; Dhar, R. S.; Dhar, A.K.; Sharma, P. R.; Agarwal, S. G. Dynamics of essential oil biosynthesis in relation to inflorescence and glandular ontogeny in Salvia sclarea. Flavour Fragg. J (2005), 21: 817-821.
• Lattoo, S. K.; Khan, S.; Dhar, A.K. A new chromosome number in Polygonatum cirrhifolium Royle – an endangered liliaceous medicinal herb. Curr. Sci, (2005), 89 (7): 1080-1081
• Agnihotri, V. K; Lattoo, S. K.; Thappa, R. K.; Kaul, P.; Qazi, G. N.; Dhar, A. K.; Saraf, A.; Kapahi, B. K.; Saxena, R. K.; Agarwal, S. G. Chemical variability in the essential oil components of Achillea millefolium Agg. from different Himalayan habitats (India). Planta Medica, (2005), 71: 278-280
• Lattoo, S. K.; Koul, S.; Dhar, M. K.; Khajuria, R. K.; Gupta, D. K.; Dhar, A. K.; Qazi, G. N. Production of b-b,dimethylacrylshikonin in callus cultures of Onosma echioides var hispidum. J. Plant Biochemistry and Biotechnology (2005), 14: 193-196
• Lattoo, S. K.; Dhar, A. K.; Jasrotia, A. Epicotyl seed dormancy and phenology of germination in Polygonatum cirrhifolium Royle. Curr. Sci., (2005), 81(11): 1414-1417

• Lattoo, S.K.; Qazi, G.N.; Dhar, A.K.; Purohit, P.; Raina, R.K. Methods and composition for in vitro germination and propagation of Polygonatum cirrhifolium Royle. United State Patent 6, 905, 876; June 14, 2005. pp. 1-24. (PCT patent, PCT/IN2001/000200, Publication No. WO/2003/041492: 13.06.2003, South Africa ZA2004/3703, July 7, 2005, India 1N 225714, November 21, 2008)

• Lattoo, S. K.; Dhar, R. S.; Khan, S.; Bomotra, S.; Bhan, M.K.; Dhar, A.K.; Gupta, K. K. Molecular and morphometric diversity in relation to breeding system in Andrographis panicualata. In “Plant Tissue Culture, molecular markers and their role in crop productivity” (eds. Ashwani Kumar and N.S.Shekhawat). Publishers IK International Publishing House New Delhi (2008), pp. 434-449
• Lattoo, S. K.; Dhar, A.K.; Upadhya, R.N.; Ahmed, S. M.; Verma, A. Breeding behaviour, genetic system and reproductive effort in Sisymbrium irio Linn. under cultivation experiment. Scope and opportunities in Research of Medicinal and Aromatic Plants (eds. A.K.Mathur, S. Dwivedi, D.D.Patn, G.D.Bagchi, N.S.Khanuja) CIMAP – Lucknow, India, (2003), pp 106-113

Gulzar A. Rather

Niha Dhar

Aarti Sharma

Shahzad A. Pandith

Tariq A. Wani

• Genome Analyzer (Applied Biosystems 3130Xl)
• PCR/RT-PCR
• Gel Doc
• Light/Stereoscopic Microscope
• Microscope Freezer
• Centrifuges
• Laminar flows/Inoculation and Culture Chambers