02116nas a2200313 4500008004100000022001400041245012800055210006900183260000800252300001600260490000800276520108500284653000801369653002001377653001301397653001901410653002101429100002301450700001801473700002001491700001801511700001901529700002201548700002501570700002401595700003301619700001801652856013201670 2019 eng d a0021-954100aEstrogen receptor activation in response to Azadirachtin A stimulates osteoblast differentiation and bone formation in mice0 aEstrogen receptor activation in response to Azadirachtin A stimu cDEC a23719-237350 v2343 a

The positive effectsof the sex hormone in sustaining bone homeostasis areexercised by maintaining the equilibrium betweencell activity and apoptosis. In this regard, the importance of estrogen receptors in maintaining the bone is that it is anattractive drug target,if devoid of known side effects. In this study, we show that a natural pure compound Azadirachtin A (Aza A) isolated from Azadirachta indica binds selectively to a site in the estrogen receptor, identifying itself to bea selective tissue modifier. Using computational and medicinal chemistry, we show that Aza A binds potentially and selectively to estrogen receptor-alpha (ER alpha) as compared with ER beta. This preferential binding of Aza A to ER alpha with good pharmacokinetic distribution in the body forms metabolites, showing that it is well absorbed. In in vivo estrogen deficiency models for osteoporosis, Aza A at a much lower dose enhances new bone formation at both sites of the trabecular and cortical bone with increased bone strength and presentswith no hyperplastic effect in the uterus.

10aALP10aGene expression10amicro-CT10amineralization10aosteoblast cells1 aKushwaha, Priyanka1 aAhmad, Naseer1 aDhar, Yogeshwar1 aVerma, Ashwni1 aHaldar, Saikat1 aMulani, Fayaj, A.1 aTrivedi, Prabodh, K.1 aMishra, Prabhat, R.1 aThulasiram, Hirekodathakallu1 aTrivedi, Ritu uhttps://library.ncl.res.in/content/estrogen-receptor-activation-response-azadirachtin-stimulates-osteoblast-differentiation-and02516nas a2200217 4500008004100000022001400041245008600055210006900141260000800210300000900218490000700227520175600234100002801990700001602018700003402034700001802068700002202086700003302108700003602141856012102177 2020 eng d a2045-232200aBasic limonoid modulates chaperone-mediated proteostasis and dissolve Tau fibrils0 aBasic limonoid modulates chaperonemediated proteostasis and diss cMAR a40230 v103 a

The Alzheimer's disease pathology is associated with accumulation of intracellular neurofibrillary tangles and extracellular senile plaques. The formation of initial nucleus triggers conformational changes in Tau and leads to its deposition. Hence, there is a need to eliminate these toxic proteins for proper functioning of neuronal cells. In this aspect, we screened the effect of basic limonoids such as gedunin, epoxyazadiradione, azadirone and azadiradione on inhibiting Tau aggregation as well as disintegration of induced Tau aggregates. It was observed that these basic limonoids effectively prevented aggregates formation by Tau and also exhibited the property of destabilizing matured Tau aggregates. The molecular docking analysis suggests that the basic limonoids interact with hexapeptide regions of aggregated Tau. Although these limonoids caused the conformational changes in Tau to beta-sheet structure, the cytological studies indicate that basic limonoids rescued cell death. The dual role of limonoids in Tau aggregation inhibition and disintegration of matured aggregates suggests them to be potent molecules in overcoming Tau pathology. Further, their origin from a medicinally important plant neem, which known to possess remarkable biological activities was also found to play protective role in HEK293T cells. Basic limonoids were non-toxic to HEK293T cells and also aided in activation of HSF1 by inducing its accumulation in nucleus. Western blotting and immunofluorescence studies showed that HSF1 in downstream increased the transcription of Hsp70 thus, aggravating cytosolic Hsp70 levels that can channel clearance of aberrant Tau. All these results mark basic limonoids as potential therapeutic natural products.

1 aGorantla, Nalini, Vijay1 aDas, Rashmi1 aChidambaram, Hariharakrishnan1 aDubey, Tushar1 aMulani, Fayaj, A.1 aThulasiram, Hirekodathakallu1 aChinnathambi, Subashchandrabose uhttps://library.ncl.res.in/content/basic-limonoid-modulates-chaperone-mediated-proteostasis-and-dissolve-tau-fibrils02560nas a2200313 4500008004100000022001400041245010100055210006900156260000800225300001100233490000800244520153800252653002601790653001201816653001201828653003301840653001401873653002801887653001701915653002901932653002001961100002001981700002502001700001902026700001802045700003302063700002002096856013002116 2020 eng d a0031-942200aBiosynthesis and tissue-specific partitioning of camphor and eugenol in Ocimum kilimandscharicum0 aBiosynthesis and tissuespecific partitioning of camphor and euge cSEP a1124510 v1773 a

In Ocimum kilimandscharicum, the relative volatile composition of camphor in leaves was as high as 55%, while that of eugenol in roots was 57%. These metabolites were differentially partitioned between the aerial and root tissues. Global metabolomics revealed tissue-specific biochemical specialization, evident by the differential distribution of 2588 putative metabolites across nine tissues. Next-generation sequencing analysis indicated differential expression of 51 phenylpropanoid and 55 terpenoid pathway genes in aerial and root tissues. By integrating metabolomics with transcriptomics, the camphor biosynthesis pathway in O. kilimandscharicum was elucidated. In planta bioassays revealed the role of geranyl diphosphate synthase (gpps) and borneol dehydrogenase (bdh) in camphor biosynthesis. Further, the partitioning of camphor was attributed to tissue-specific gene expression of both the pathway entry point (gpps) and terminal (bdh) enzyme. Unlike camphor, eugenol accumulated more in roots; however, absence of the eugenol synthase gene in roots indicated long distance transport from aerial tissues. In silico co-expression analysis indicated the potential involvement of ATP-binding cassette, multidrug and toxic compound extrusion, and sugar transporters in eugenol transport. Similar partitioning was evident across five other Ocimum species. Overall, our work indicates that metabolite partitioning maybe a finely regulated process, which may have implications on plant growth, development, and defense.

10aBorneol dehydrogenase10aCamphor10aEugenol10aGeranyl diphosphate synthase10aLamiaceae10aMetabolite partitioning10aMetabolomics10aOcimum kilimandscharicum10aTranscriptomics1 aSingh, Priyanka1 aKalunke, Raviraj, M.1 aShukla, Anurag1 aTzfadia, Oren1 aThulasiram, Hirekodathakallu1 aGiri, Ashok, P. uhttps://library.ncl.res.in/content/biosynthesis-and-tissue-specific-partitioning-camphor-and-eugenol-ocimum-kilimandscharicum02952nas a2200349 4500008004100000022001400041245011300055210006900168260000800237300001100245490000800256520180300264653002202067653003202089653002702121653001402148653001402162653001602176653001802192100002202210700002102232700001802253700002902271700002202300700002602322700002402348700002602372700002102398700002202419700003302441856012802474 2021 eng d a0031-942200aLimonoid biosynthesis 3: functional characterization of crucial genes involved in neem limonoid biosynthesis0 aLimonoid biosynthesis 3 functional characterization of crucial g cAPR a1126690 v1843 a

Neem (Azadirachta indica L.) is well known for its medicinal, agricultural, and pesticidal applications since ages. The secondary metabolites, limonoids, confer these biological properties, wherein over 150 different limonoids have been reported from neem. To understand limonoid biosynthesis, we analyzed tissue-specific (kernel, pericarp, leaves, and flower) transcriptome that resulted in the identification of one farnesyl diphosphate synthase (AiFDS), one squalene synthase (AiSQS), three squalene epoxidases (AiSQE1, AiSQE2, and AiSQE3), two triterpene synthases (AiTTS1 and AiTTS2), cycloartenol synthase (AiCAS), two cytochrome P450 reductases, and ten cytochrome P450 systems. Comparative tissue-expression analysis indicated that AiFDS, AiSQS, AiSQE3, and AiTTS1 are expressed higher in the kernel than in the other tissues. Heterologously expressed recombinant AiTTS1 produced tirucalla-7,24-dien-3 beta-ol as the sole product. Expression profile data, phylogeny with triterpene synthases from Meliaceae and Rutaceae families, real-time PCR of different tissues, and transient transformation revealed the involvement of tirucalla-7,24-dien-3 beta-ol synthase (AiTTS1) in limonoid biosynthesis. Further, mutagenesis studies of AiTTS1 indicated that Y125 and F260 are probably involved in stabilization of dammarenyl cation. A 2.6-fold increase in production of tirucalla-7,24-dien-3 beta-ol was observed when AiSQE1 was coexpressed with mutant AiTTS1 in a yeast system. Furthermore, we functionally characterized the highly expressed cytochrome P450 reductases and cycloartenol synthase. This study helps in further analysis and identification of genes involved in limonoid biosynthesis in Meliaceae/Rutaceae and their production in a metabolically tractable heterologous system.

10a24-dien-3 beta-ol10aAzadirachta indica A. Juss.10aCytochrome P450 system10aLimonoids10aMeliaceae10aTirucalla-710aTranscriptome1 aPandreka, Avinash1 aChaya, Patil, S.1 aKumar, Ashish1 aAarthy, Thiagarayaselvam1 aMulani, Fayaj, A.1 aBhagyashree, Date, D.1 aShilpashree, H., B.1 aJennifer, Cheruvathur1 aPonnusamy, Sudha1 aNagegowda, Dinesh1 aThulasiram, Hirekodathakallu uhttps://library.ncl.res.in/content/limonoid-biosynthesis-3-functional-characterization-crucial-genes-involved-neem-limonoid02949nas a2200385 4500008004100000022001400041245017500055210006900230260000800299300001200307490000800319520168600327653001902013653001802032653001502050653001402065653001402079653002202093653001402115653000902129653001202138653001602150100003302166700002002199700002302219700002902242700002002271700003302291700002502324700002502349700002102374700001902395700002202414856012702436 2022 eng d a0981-942800aStacking herbicide detoxification and resistant genes improves glyphosate tolerance and reduces phytotoxicity in tobacco (Nicotiana tabacum L.) and rice (Oryza sativa L.)0 aStacking herbicide detoxification and resistant genes improves g cOCT a126-1380 v1893 a

Glyphosate residues retained in the growing meristematic tissues or in grains of glyphosate-resistant crops affect the plants physiological functions and crop yield. Removing glyphosate residues in the plants is desirable with no penalty on crop yield and quality. We report a new combination of scientific strategy to detoxify glyphosate that reduces the residual levels and improve crop resistance. The glyphosate detoxifying enzymes Aldo-keto reductase (AKR1) and mutated glycine oxidase (mGO) with different modes of action were co-expressed with modified EPSPS, which is insensitive to glyphosate in tobacco (Nicotiana tabacum L.) and rice (Oryza sativa L.). The transgenic tobacco plants expressing individual PsAKR1, mGO, CP4-EPSPS, combinations of PsAKR1:CP4EPSPS, PsAKR1:mGO, and multigene with PsAKR1: mGO: CP4EPSPS genes were developed. The bio-efficacy studies of in-vitro leaf regeneration on different concentrations of glyphosate, seedling bioassay, and spray on transgenic tobacco plants demonstrate that glyphosate detoxification with enhanced resistance. Comparative analysis of the transgenic tobacco plants reveals that double and multigene expressing transgenics had reduced accumulation of shikimic acid, glyphosate, and its primary residue AMPA, and increased levels of sarcosine were observed in all PsAKR1 expressing transgenics. The multigene expressing rice transgenics showed improved glyphosate resis-tance with yield maintenance. In summary, results suggest that stacking genes with two different detoxification mechanisms and insensitive EPSPS is a potential approach for developing glyphosate-resistant plants with less residual content.

10aDetoxification10aGene stacking10aGlyphosate10aHerbicide10aMultigene10aResidual toxicity10aResistant10arice10aTobacco10aTransgenics1 aVennapusa, Amaranatha, Reddy1 aAgarwal, Subham1 aHm, Hanumanth, Rao1 aAarthy, Thiagarayaselvam1 aBabitha, K., C.1 aThulasiram, Hirekodathakallu1 aKulkarni, Mahesh, J.1 aMelmaiee, Kalpalatha1 aSudhakar, Chinta1 aUdayakumar, M.1 aVemanna, Ramu, S. uhttps://library.ncl.res.in/content/stacking-herbicide-detoxification-and-resistant-genes-improves-glyphosate-tolerance-and02549nas a2200277 4500008004100000022001400041245013400055210006900189260000800258300001400266490000800280520161000288653001201898653001301910653001401923653001401937653002001951653001901971100002201990700002702012700003002039700002402069700002502093700003302118856012002151 2022 eng d a1618-264200aUltra-high performance liquid chromatography Q-Orbitrap MS/MS-based profiling and quantification of limonoids in Meliaceae plants0 aUltrahigh performance liquid chromatography QOrbitrap MSMSbased  cAUG a6093-61060 v4143 a

Meliaceae plants have been extensively used in agriculture, folklore, and traditional medicine. They are the major storehouses for structurally diverse limonoids (meliacins) possessing various bioactivities like antifeedant, insecticidal, antimicrobial, etc. However accurate detection of these tetranortriterpenes from the vast pool of metabolites in plant tissue extracts or biological sample is a crucial challenge. Though the mass spectrum (MS) provides the molecular mass and the corresponding elemental composition, it cannot be relied precisely. The exact identification of a specific metabolite demands the MS/MS spectrum containing the signature product ions. In the present study, we have developed the UHPLC Q-Orbitrap-based method for identification, quantification, and characterization of limonoids in different plant tissue extracts requiring minimum plant material. Using this method, we carried out the limonoid profiling in different tissue extracts of sixteen Meliaceae plants and the identification of limonoids was performed by comparing the retention time (RT), ESI-(+)-MS spectrum, and HCD-MS/MS of the purified fifteen limonoids used as reference standards. Our results revealed that early intermediates of the limonoid biosynthetic pathway such as azadiradione, epoxyazadiradione, and gedunin occurred more commonly in Meliaceae plants. The MS/MS spectrum library of the fifteen limonoids generated in this study can be utilized for identification of these limonoids in other plant tissue extracts, botanical fertilizers, agrochemical formulations, and bio pesticides.

10aLC-HRMS10aLC-MS/MS10aLimonoids10aMeliaceae10aNatural product10aQuantification1 aMulani, Fayaj, A.1 aNandikol, Sharvani, S.1 aKajjihundi, Jagadeesh, S.1 aPathappa, Niranjana1 aPuttappa, Sharanappa1 aThulasiram, Hirekodathakallu uhttps://library.ncl.res.in/content/ultra-high-performance-liquid-chromatography-q-orbitrap-msms-based-profiling-and