02494nas a2200277 4500008004100000022001400041245012300055210006900178260010600247300001200353490000800365520149500373653001301868653001801881653001001899653001701909653001201926653001601938100002001954700002601974700002602000700002402026700002002050700002102070856012502091 2015 eng d a0031-942200aMetabolic profiling of chickpea-Fusarium interaction identifies differential modulation of disease resistance pathways0 aMetabolic profiling of chickpeaFusarium interaction identifies d aTHE BOULEVARD, LANGFORD LANE, KIDLINGTON, OXFORD OX5 1GB, ENGLANDbPERGAMON-ELSEVIER SCIENCE LTDcAUG a120-1290 v1163 a

Chickpea is the third most widely grown legume in the world and mainly used as a vegetarian source of human dietary protein. Fusarium wilt, caused by Fusarium oxysporum f. sp. ciceri (Foc), is one of the major threats to global chickpea production. Host resistance is the best way to protect crops from diseases; however, in spite of using various approaches, the mechanism of Foc resistance in chickpea remains largely obscure. In the present study, non-targeted metabolic profiling at several time points of resistant and susceptible chickpea cultivars using high-resolution liquid chromatography-mass spectrometry was applied to better understand the mechanistic basis of wilt resistance or susceptibility. Multivariate analysis of the data (OPLS-DA) revealed discriminating metabolites in chickpea root tissue after Foc inoculation such as flavonoids, isoflavonoids, alkaloids, amino acids and sugars. Foc inoculated resistant plants had more flavonoids and isoflavonoids along with their malonyl conjugates. Many antifungal metabolites that were induced after Foc infection viz, aurantion-obstine beta-glucosides and querecitin were elevated in resistant cultivar. Overall, diverse genetic and biochemical mechanisms were operational in the resistant cultivar for Foc defense as compared to the susceptible plant. The resistant chickpea plants employed the above-mentioned metabolic pathways as potential defense strategy against Foc. (C) 2015 Elsevier Ltd. All rights reserved.

10aChickpea10aFusarium wilt10aLC-MS10aMetabolomics10aOPLS-DA10aPhytoalexin1 aKumar, Yashwant1 aDholakia, Bhushan, B.1 aPanigrahi, Priyabrata1 aKadoo, Narendra, Y.1 aGiri, Ashok, P.1 aGupta, Vidya, S. uhttp://library.ncl.res.in/content/metabolic-profiling-chickpea-fusarium-interaction-identifies-differential-modulation-002788nas a2200301 4500008004100000022001400041245015300055210006900208260000800277300000700285490000800292520177100300653002102071653001402092653001302106653001002119653001502129653001802144100002302162700002702185700002302212700002402235700002602259700002902285700002902314700002002343856012302363 2020 eng d a0032-093500aHigh-throughput metabolomic and transcriptomic analyses vet the potential route of cerpegin biosynthesis in two varieties of Ceropegia bulbosa Roxb.0 aHighthroughput metabolomic and transcriptomic analyses vet the p cJAN a280 v2513 a

Ceropegia bulbosa is an important medicinal plant, used in the treatment of various ailments including diarrhea, dysentery, and syphilis. This is primarily attributed to the presence of pharmaceutically active secondary metabolites, especially cerpegin. As this plant belongs to an endemic threatened category, genomic resources are not available hampering exploration on the molecular basis of cerpegin accumulation till now. Therefore, we undertook high-throughput metabolomic and transcriptomic analyses using different tissues from two varieties namely, C. bulbosa var. bulbosa and C. bulbosa var. lushii. Metabolomic analysis revealed spatial and differential accumulation of various metabolites. We chemically synthesized and characterized the cerpegin and its derivatives by liquid chromatography tandem-mass spectrometry (LC-MS/MS). Importantly, these comparisons suggested the presence of cerpegin and 5-allyl cerpegin in all C. bulbosa tissues. Further, de novo transcriptome analysis indicated the presence of significant transcripts for secondary metabolic pathways through the Kyoto encyclopedia of genes and genomes database. Tissue-specific profiling of transcripts and metabolites showed a significant correlation, suggesting the intricate mechanism of cerpegin biosynthesis. The expression of potential candidate genes from the proposed cerpegin biosynthetic pathway was further validated by qRT-PCR and NanoString nCounter. Overall, our findings propose a potential route of cerpegin biosynthesis. Identified transcripts and metabolites have built a foundation as new molecular resources that could facilitate future research on biosynthesis, regulation, and engineering of cerpegin or other important metabolites in such non-model plants.

10a5-Allyl cerpegin10aCeropegia10aCerpegin10aLC-MS10ametabolite10aTranscriptome1 aGharat, Sachin, A.1 aShinde, Balkrishna, A.1 aMule, Ravindra, D.1 aPunekar, Sachin, A.1 aDholakia, Bhushan, B.1 aJayaramaiah, Ramesha, H.1 aRamaswamy, Gopalakrishna1 aGiri, Ashok, P. uhttp://library.ncl.res.in/content/high-throughput-metabolomic-and-transcriptomic-analyses-vet-potential-route-cerpegin02728nas a2200253 4500008004100000022001400041245018000055210006900235260000800304300001400312490000700326520183900333653001802172653001002190653001502200653000902215653001602224653001402240100002002254700002602274700002402300700002402324856012602348 2021 eng d a0971-589400aZCTs knockdown using antisense LNA GapmeR in specialized photomixotrophic cell suspensions of Catharanthus roseus: Rerouting the flux towards mono and dimeric indole alkaloids0 aZCTs knockdown using antisense LNA GapmeR in specialized photomi cJUL a1437-14530 v273 aThe present study was carried out to silence the transcription factor genes ZCT1, ZCT2 and ZCT3 via lipofectamine based antisense LNA GapmeRs transfection into the protoplasts of established photomixotrophic cell suspensions. The photomixotrophic cell suspensions with a threshold of 0.5% sucrose were raised and established using two-tiered CO2 providing flasks kept under high light intensity. The photomixotrophic cell suspensions showed morphologically different thick-walled cells under scanning electron microscopic analysis in comparison to the simple thin-walled parenchymatous control cell suspensions. The LC-MS analysis registered the vindoline production (0.0004 +/- 0.0001 mg/g dry wt.) in photomixotrophic cell suspensions which was found to be absent in control cell suspensions. The protoplasts were isolated from the photomixotrophic cell suspensions and subjected to antisense LNA GapmeRs silencing. Three lines, viz. Z1A, Z2C and Z3G were obtained where complete silencing of ZCT1, ZCT2 and ZCT3 genes, respectively, was observed. The Z3G line was found to show maximum production of vindoline (0.038 +/- 0.001 mg/g dry wt.), catharanthine (0.165 +/- 0.008 mg/g dry wt.) and vinblastine (0.0036 +/- 0.0003 mg/g dry wt.). This was supported by the multifold increment in the gene expression of TDC, SLS, STR, SGD, d4h, dat, CrT16H and Crprx. The present work indicates the master regulation of ZCT3 knockdown among all three ZCTs transcription factors in C. roseus to enhance the terpenoid indole alkaloids production. The successful silencing of transcription repressor genes has been achieved in C. roseus plant system by using photomixotrophic cell cultures through GapmeR based silencing. The present study is a step towards metabolic engineering of the TIAs pathway using protoplast transformation in C. roseus.10aCatharanthine10aLC-MS10aProtoplast10aTIAs10aVinblastine10aVindoline1 aVerma, Priyanka1 aKhan, Shamshad, Ahmad1 aParasharami, Varsha1 aMathur, Ajay, Kumar uhttp://library.ncl.res.in/content/zcts-knockdown-using-antisense-lna-gapmer-specialized-photomixotrophic-cell-suspensions02905nas a2200313 4500008004100000022001400041245005000055210005000105260000900155300001100164490000800175520208100183653001202264653002102276653001402297653001002311653001502321653000702336653001502343100002202358700001802380700002102398700001802419700001702437700002002454700001802474700001802492856008102510 2022 eng d a1874-391900aProteomic profile of candida albicans biofilm0 aProteomic profile of candida albicans biofilm cAUG  a1046610 v2653 a

Candida albicans biofilms are characterized by structural and cellular heterogeneity that confers antifungal resistance and immune evasion. Despite this, biofilm formation remains poorly understood. In this study, we used proteomic analysis to understand biofilm formation in C. albicans related to morphophysiological and architectural features. LC-MS/MS analysis revealed that 64 proteins were significantly modulated, of which 31 were upregulated and 33 were downregulated. The results indicate that metabolism (25 proteins), gene expression (13 proteins), stress response (7 proteins), and cell wall (5 proteins) composition are modulated. The rate of oxidative phosphorylation (OxPhos) and biosynthesis of UDP-N-acetylglucosamine, vitamin B6, and thiamine increased, while the rate of methionine biosynthesis decreased. There was a significant modification of the cell wall architecture due to higher levels of Sun41, Pir1 and Csh1 and increased glycosylation of proteins. It was observed that C. albicans induces hyphal growth by upregulating the expression of genes involved in cAMP-PKA and MAPK pathways. This study is significant in that it suggests an increase in OxPhos and alteration of cell wall architecture that could be contributing to the recalcitrance of C. albicans cells growing in biofilms. Nevertheless, a deeper investigation is needed to explore it further.Significance: Candida sps is included in the list of pathogens with potential drug resistance threat due to the increased frequency especially colonization of medical devices, and tissues among the patients, in recent years. Significance of our study is that we are reporting traits like modulation in cell wall composition, amino acid and vitamin biosynthesis and importantly energy generation (OxPhos) etc. These traits could be conferring antifungal resistance, host immune evasion etc. and thus survival, in addition to facilitating biofilm formation. These findings are expected to prime the further studies on devising potent strategy against biofilm growth among the patients.

10aBiofilm10aCandida albicans10aCell wall10aLC-MS10aMetabolism10aMS10aproteomics1 aAbdulghani, Mazen1 aIram, Rasiqua1 aChidrawar, Priti1 aBhosle, Kajal1 aKazi, Rubina1 aPatil, Rajendra1 aKharat, Kiran1 aZore, Gajanan uhttp://library.ncl.res.in/content/proteomic-profile-candida-albicans-biofilm03040nas a2200337 4500008004100000022001400041245006500055210006400120260000800184300001200192490000700204520212500211653001302336653000602349653001002355653000702365653001102372653002102383653001102404653001202415100002202427700002202449700001902471700001902490700001702509700001902526700002002545700002302565700001802588856009602606 2023 eng d a1369-378600aOpaque cell-specific proteome of Candida albicans ATCC 102310 aOpaque cellspecific proteome of Candida albicans ATCC 10231 cJUL amyad0620 v613 a

Candida albicans, a polymorphic opportunistic pathogen of humans, can exist in different morphological forms like yeast, hyphae, pseudohyphae, chlamydospores, and white and opaque cells. Proteomic analysis of opaque form of C. albicans ATCC 10231 is carried out in the present study using microflow liquid chromatography-tandem mass spectrometry and validated using expression analysis of selected genes using reverse transcription quantitative real-time PCR and mitochondrial membrane potential assay. This is the first report identifying opaque cell-specific proteins of C. albicans. A total of 188 proteins were significantly modulated under opaque form compared to white cells, of which 110 were upregulated, and 78 were downregulated. It was observed that oxidative phosphorylation (OxPhos) and oxidative stress are enhanced in C. albicans cells growing under opaque form as proteins involved in OxPhos (Atp1, Atp3, Atp16, Atp7, Cox6, Nuc2, Qcr7, and Sdh12) and oxidative stress response (Gcs1, Gtt11, Gpx2, Sod1, Ccp1, and Lys7) were significantly upregulated. The maximum upregulation of 23.16- and 13.93-fold is observed in the cases of Ccp1 and Nuc2, respectively. The downregulation of proteins, namely Als1, Csh1, Sap9, and Rho1, determining cell surface chemistry indicates modulation in cell wall integrity and reduced adhesion of opaque cells compared to white cells. This study is significant as it is the first draft of the proteomic profile of opaque cells that suggests enhanced OxPhos, oxidative stress, and modulation in cell surface chemistry indicating reduced adhesion and cell wall integrity, which could be associated with reduced virulence in opaque form. However, a deeper investigation is needed to explore it further. Lay Summary Opaque form is one of the least studied morphological forms of Candida albicans. To the best of our knowledge, this is the first report providing opaque cell-specific proteome. It suggests enhanced oxidative phosphorylation, oxidative stress, and modulation in cell surface chemistry, which could be associated with reduced virulence in opaque form.

10aalbicans10aC10aLC-MS10aMS10aopaque10aOxidative stress10aOxPhos10aprotein1 aAbdulghani, Mazen1 aTelang, Saraswati1 aDesai, Manisha1 aKadam, Shivdas1 aKazi, Rubina1 aShelar, Amruta1 aPatil, Rajendra1 aMaurya, Radheshyam1 aZore, Gajanan uhttp://library.ncl.res.in/content/opaque-cell-specific-proteome-candida-albicans-atcc-10231