RESUMO
CIM-Saumya is an improved, methyl chavicol rich variety of Ocimum basilicum (Family-Lamiaceae), developed by Council of Scientific and Industrial Research-Central Institute of Medicinal and Aromatic Plants. This plant possesses analgesic, anti-ulcerogenic, anti-inflammatory, anti-oxidant, cardiac stimulant, Central Nervous System depressant, hepatoprotective and immunomodulator activities due to the presence of various phytoconstituents. Among them rosmarinic acid, caffeic acid and ferulic acid are the three major phenolic compounds responsible for its therapeutic utility. These compounds are produced in very low amounts in the in vivo plants. Therefore, the present study has been conducted for establishment of cell suspensions, optimization of inoculums size, growth kinetics and screening of elicitor and precursors for the accumulation of cell biomass and the production of the three important phenolic compounds in cell suspension of O. basilicum (CIM-Saumya). Leaf derived friable callus was used for establishing the cell suspension in liquid Murashige and Skoog's medium fortified with 1 g/L casein hydrolysate + 2.26 µM 2,4-dichlorophenoxyacetic acid + 0.465 µM kinetin + 2.68 µM naphthalene acetic acid. The growth kinetic analysis pattern of cell suspension revealed the maximum biomass increments (% BI = 486.7) and production of RA 8.086 mg/g dry weight was found in 30th day harvested cells. Whereas, the other two phenolic compounds i.e. ferulic acid (0.0125 mg/g dry weight) and caffeic acid (0.38 mg/g dry weight) was recorded highest on 25th day of growth cycle. In the present study, one biotic elicitor i.e. yeast extract and three precursors [peptone, tryptone and lactalbumin hydrolysate] were tested, among them, lactalbumin hydrolysate (100 mg/L; added at 16th day) treated cells recorded highest estimated phenolic compounds yield (251.5 mg/L; 6.81 fold compared to the control) and biomass increments i.e. % BI = 1207 with 1.85 fold compared to the control. The highest rosmarinic acid content i.e. 25.47 mg/g DW (4.4 fold compared to the control) and 24.42 mg/g dry weight (4.1 folds compared to the control) was noticed in 30th day harvested cells treated with yeast extract (1 g/L on 0 day) and lactalbumin hydrolysate (100 mg/L added on 16th day), respectively. While caffeic acid content (0.91 mg/g dry weight) showed 2.9 folds higher compared to the control in cells treated with peptone 200 mg/L added on 16th day of culture cycle. All the treated cells showed enhanced phenylalanine ammonia-lyase enzyme activity with highest specific activity in lactalbumin hydrolysate followed by tryptone, peptone, and yeast extract. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-023-01316-6.
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Ginseng, an oriental gift to the world of healthcare and preventive medicine, is among the top ten medicinal herbs globally. The constitutive triterpene saponins, ginsenosides, or panaxosides are attributed to ginseng's miraculous efficacy towards anti-aging, rejuvenating, and immune-potentiating benefits. The major ginsenosides such as Rb1, Rb2, Rc, Rd., Re, and Rg1, formed after extensive glycosylations of the aglycone "dammaranediol," dominate the chemical profile of this genus in vivo and in vitro. Elicitations have successfully led to appreciable enhancements in the production of these major ginsenosides. However, current research on ginseng biotechnology has been focusing on the enrichment or production of the minor ginsenosides (the less glycosylated precursors of the major ginsenosides) in ginseng preparations, which are either absent or are produced in very low amounts in nature or via cell cultures. The minor ginsenosides under current scientific scrutiny include diol ginsenosides such as Rg3, Rh2, compound K, and triol ginsenosides Rg2 and Rh1, which are being touted as the next "anti-neoplastic pharmacophores," with better bioavailability and potency as compared to the major ginsenosides. This review aims at describing the strategies for ginsenoside production with special attention towards production of the minor ginsenosides from the major ginsenosides via microbial biotransformation, elicitations, and from heterologous expression systems.
Assuntos
Antineoplásicos/metabolismo , Bactérias/genética , Ginsenosídeos/biossíntese , Panax/metabolismo , Antineoplásicos/química , Bactérias/metabolismo , Biotransformação , Expressão Gênica , Ginsenosídeos/química , Ginsenosídeos/isolamento & purificação , Ginsenosídeos/uso terapêutico , Humanos , Panax/química , Panax/genética , Plantas Medicinais/química , Plantas Medicinais/genéticaRESUMO
Cobalt nitrate, nickel sulphate, hydrogen peroxide, sodium nitroprusside, and culture filtrates of Pseudomonas monteili, Bacillus circularans, Trichoderma atroviridae, and Trichoderma harzianum were tested to elicit ginsenoside production in a cell suspension line of Panax quinquefolius. Abiotic elicitors preferentially increased panaxadiols whereas biotic elicitors upregulated the panaxatriol synthesis. Cobalt nitrate (50 µM) increased total ginsenosides content by twofold (54.3 mg/L) within 5 days. It also induced the Rc synthesis that was absent in the control cultures. Elicitation with P. monteili (2.5 % v/v, 5 days) also supported 2.4-fold enhancement in saponin yield. Elicitation by T. atroviridae or hydrogen peroxide induced the synthesis of Rg3 and Rh2 that are absent in ginseng roots. The highest ginsenosides productivity (3.2-fold of control) was noticed in cells exposed to 1.25 % v/v dose of T. atroviridae for 5 days. Treating cells with T. harzianum for 15 days afforded maximum synthesis and leaching (8.1 mg/L) of ginsenoside Rh1.
Assuntos
Ginsenosídeos/biossíntese , Panax/química , Células Vegetais/efeitos dos fármacos , Bacillus/química , Cobalto/química , Meios de Cultura , Peróxido de Hidrogênio/química , Níquel/química , Nitroprussiato/química , Panax/citologia , Células Vegetais/metabolismo , Pseudomonas/química , Trichoderma/químicaRESUMO
Spontaneous mesenteric hematomas (SMH) are not a common entity. Here we describe a case of 64 year old woman who presented with a vague abdominal pain and diffuse tenderness. Her CT abdomen revealed an ill-defined hyperdense mass like lesion in the mesentery and she underwent exploratory laparotomy which revealed a large hematoma in the mesentery with inflammation of the adjoining small bowel loop. Histopathology revealed findings consistent with hematoma with no evidence of neoplastic lesion.
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We present a survey of the current state-of-the-art in breast cancer detection and prognosis. We analyze the evolution of Artificial Intelligence-based approaches from using just uni-modal information to multi-modality for detection and how such paradigm shift facilitates the efficacy of detection, consistent with clinical observations. We conclude that interpretable AI-based predictions and ability to handle class imbalance should be considered priority.
RESUMO
Breast cancer is a deadly disease with a high mortality rate among PAN cancers. The advancements in biomedical information retrieval techniques have been beneficial in developing early prognosis and diagnosis systems for cancer patients. These systems provide the oncologist with plenty of information from several modalities to make the correct and feasible treatment plan for breast cancer patients and protect them from unnecessary therapies and their toxic side effects. The cancer patient's related information can be collected using various modalities like clinical, copy number variation, DNA-methylation, microRNA sequencing, gene expression, and histopathological whole slide images. High dimensionality and heterogeneity in these modalities demand the development of some intelligent systems to understand related features to the prognosis and diagnosis of diseases and make correct predictions. In this work, we have studied some end-to-end systems having two main components : (a) dimensionality reduction techniques applied to original features from different modalities and (b) classification techniques applied to the fusion of reduced feature vectors from different modalities for automatic predictions of breast cancer patients into two categories: short-time and long-time survivors. Principal component analysis (PCA) and variational auto-encoders (VAEs) are used as the dimensionality reduction techniques, followed by support vector machines (SVM) or random forest as the machine learning classifiers. The study utilizes raw, PCA, and VAE extracted features of the TCGA-BRCA dataset from six different modalities as input to the machine learning classifiers. We conclude this study by suggesting that adding more modalities to the classifiers provides complementary information to the classifier and increases the stability and robustness of the classifiers. In this study, the multimodal classifiers have not been validated on primary data prospectively.
Assuntos
Neoplasias da Mama , Humanos , Feminino , Neoplasias da Mama/patologia , Variações do Número de Cópias de DNA , Aprendizado de Máquina , Mama/patologia , Prognóstico , Máquina de Vetores de SuporteRESUMO
The advancement of medical research in the field of cancer prognosis and diagnosis using various modalities has put oncologists under tremendous stress. The complexity and heterogeneity involved in multiple modalities and their significantly varied clinical outcomes make it difficult to analyze the disease and provide the correct treatment. Breast cancer is the major concern among all cancers worldwide, specifically for females. To help oncologists and cancer patients, research for breast cancer survival estimation has been proposed. It ranges from complex deep neural networks to simple and interpretable architectures. We propose a utility kernel for a support vector machine (SVM) in this article. It is a simple yet powerful function, which performs better than other popular machine learning algorithms and deep neural networks in the task of breast cancer survival prediction using the TCGA-BRCA dataset. This study validates the proposed utility kernel using four different modalities (gene expression, copy number variation, clinical, and histopathological tissue images) and their multi-modal combinations. The SVM based on our utility kernel empirically proves its efficacy by achieving the highest value on various performance measures, whereas advanced deep neural networks fail to train on small and highly imbalanced breast cancer data.
Assuntos
Neoplasias da Mama , Feminino , Humanos , Neoplasias da Mama/genética , Neoplasias da Mama/diagnóstico , Máquina de Vetores de Suporte , Variações do Número de Cópias de DNA , Algoritmos , Redes Neurais de ComputaçãoRESUMO
Twenty-three pharmaceutically important plants, namely, Elaeocarpus spharicus, Rheum emodi, Indigofera tinctoria, Picrorrhiza kurroa, Bergenia ciliata, Lavandula officinalis, Valeriana wallichii, Coleus forskohlii, Gentiana kurroo, Saussurea lappa, Stevia rebaudiana, Acorus calamus, Pyrethrum cinerariaefolium, Aloe vera, Bacopa monnieri, Salvia sclarea, Glycyrrhiza glabra, Swertia cordata, Psoralea corylifolia, Jurinea mollis, Ocimum sanctum, Paris polyphylla, and Papaver somniferum, which are at the verge of being endangered due to their overexploitation and collection from the wild, were successfully established in vitro. Collections were made from the different biodiversity zones of India including Western Himalaya, Northeast Himalaya, Gangetic plain, Western Ghats, Semiarid Zone, and Central Highlands. Aseptic cultures were raised at the morphogenic level of callus, suspension, axillary shoot, multiple shoot, and rooted plants. Synseeds were also produced from highly proliferating shoot cultures of Bacopa monnieri, Glycyrrhiza glabra, Stevia rebaudiana, Valeriana wallichii, Gentiana kurroo, Lavandula officinalis, and Papaver somniferum. In vitro flowering was observed in Papaver somniferum, Psoralea corylifolia, and Ocimum sanctum shoots cultures. Out of 23 plants, 18 plants were successfully hardened under glasshouse conditions.
Assuntos
Conservação dos Recursos Naturais/métodos , Espécies em Perigo de Extinção , Plantas Medicinais/crescimento & desenvolvimento , Técnicas de Cultura de Tecidos/métodos , Biodiversidade , Ambiente Controlado , Flores/crescimento & desenvolvimento , Geografia , Germinação , Índia , Morfogênese , Raízes de Plantas/crescimento & desenvolvimento , Brotos de Planta/crescimento & desenvolvimento , Plantas Medicinais/classificação , Sementes/crescimento & desenvolvimento , Especificidade da EspécieRESUMO
Artemisinin, an antimalarial compound, at 5 mg/40 ml, was transformed by cell suspension cultures of Catharanthus roseus (L.) G.Don and Lavandula officinalis L. into deoxyartemisinin with yields >78% (3.93 mg deoxyartemisinin from 5 mg artemisinin). Maximum conversion (78.6 and 78%) occurred after 6 and 7 days of adding artemisinin to 20 and 9 days old cultures of C. roseus and L. officinalis, respectively. The procedure was scaled up by and 500 mg artemisinin was transformed into 390 mg deoxyartemisinin. Addition of artemisinin at the beginning of the culture cycle resulted in >50% reduction in dry biomass production with no bioconversion. Conversion of artemisinin occurred intracellularly followed by leaching of the product into the medium.
Assuntos
Artemisininas/metabolismo , Biotransformação , Catharanthus/metabolismo , Lavandula/metabolismo , Artemisininas/química , Técnicas de Cultura de CélulasRESUMO
A thin, profusely branched, fast growing hairy root line of Panax quinquefolium (American ginseng) was established by co-culturing epicotyl explants with a wild type strain of Agrobacterium rhizogenes. The transformed roots grew by over 10-fold from the initial inoculum within 8 weeks. The crude ginsenosides content in the roots was about 0.2 g/g dry wt level up to the 10th week of culture. Ginsenosides Rb2, Rd, Re, Rf and Rg1 constituted 47-49% of the crude saponin fraction between 6 and 8 weeks of growth whereas, Rc ginsenoside was accumulated only after 9th weeks when the biomass started receding. PCR amplification analysis of the hairy roots confirmed their transgenic nature by showing the presence of Ri-TL DNA with rolA, rolB and rolC genes in their genome.
Assuntos
Ginsenosídeos/biossíntese , Panax/crescimento & desenvolvimento , Panax/genética , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/genética , Transformação Genética , Análise de Variância , Cinética , Plantas Geneticamente Modificadas , Reação em Cadeia da PolimeraseRESUMO
Quantification of habitability is a complex task. Previous attempts at measuring habitability are well documented. Classification of exoplanets, on the other hand, is a different approach and depends on quality of training data available in habitable exoplanet catalogs. Classification is the task of predicting labels of newly discovered planets based on available class labels in the catalog. We present analytical exploration of novel activation functions as consequence of integration of several ideas leading to implementation and subsequent use in habitability classification of exoplanets. Neural networks, although a powerful engine in supervised methods, often require expensive tuning efforts for optimized performance. Habitability classes are hard to discriminate, especially when attributes used as hard markers of separation are removed from the data set. The solution is approached from the point of investigating analytical properties of the proposed activation functions. The theory of ordinary differential equations and fixed point are exploited to justify the "lack of tuning efforts" to achieve optimal performance compared to traditional activation functions. Additionally, the relationship between the proposed activation functions and the more popular ones is established through extensive analytical and empirical evidence. Finally, the activation functions have been implemented in plain vanilla feed-forward neural network to classify exoplanets. The mathematical exercise supplements the grand idea of classifying exoplanets, computing habitability scores/indices and automatic grouping of the exoplanets converging at some level.
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Catharanthus roseus today occupies the central position in ongoing metabolic engineering efforts in medicinal plants. The entire multi-step biogenetic pathway of its very expensive anticancerous alkaloids vinblastine and vincristine is fairly very well dissected at biochemical and gene levels except the pathway steps leading to biosynthesis of monomeric alkaloid catharanthine and tabersonine. In order to enhance the plant-based productivity of these pharma molecules for the drug industry, cell and tissue cultures of C. roseus are being increasingly tested to provide their alternate production platforms. However, a rigid developmental regulation and involvement of different cell, tissues, and organelles in the synthesis of these alkaloids have restricted the utility of these cultures. Therefore, the present study was carried out with pushing the terpenoid indole alkaloid pathway metabolic flux towards dimeric alkaloids vinblastine and vincristine production by over-expressing the two upstream pathway genes tryptophan decarboxylase and strictosidine synthase at two different levels of cellular organization viz. callus and leaf tissues. The transformation experiments were carried out using Agrobacterium tumefaciens LBA1119 strain having tryptophan decarboxylase and strictosidine synthase gene cassette. The callus transformation reported a maximum of 0.027% dry wt vindoline and 0.053% dry wt catharanthine production, whereas, the transiently transformed leaves reported a maximum of 0.30% dry wt vindoline, 0.10% catharanthine, and 0.0027% dry wt vinblastine content.
Assuntos
Catharanthus/química , Engenharia Genética/métodos , Triptaminas/metabolismo , Alcaloides de Vinca/síntese química , Alcaloides de Vinca/químicaRESUMO
Terpenoid indole alkaloid (TIA) biosynthetic pathway of Catharanthus roseus possesses the major attention in current metabolic engineering efforts being the sole source of highly expensive antineoplastic molecules vinblastine and vincristine. The entire TIA pathway is fairly known at biochemical and genetic levels except the pathway steps leading to biosynthesis of catharanthine and tabersonine. To increase the in-planta yield of these antineoplastic metabolites for the pharmaceutical and drug industry, extensive plant tissue culture-based studies were performed to provide alternative production systems. However, the strict spatiotemporal developmental regulation of TIA biosynthesis has restricted the utility of these cultures for large-scale production. Therefore, the present study was performed to enhance the metabolic flux of TIA pathway towards the biosynthesis of vinblastine by overexpressing two upstream TIA pathway genes, tryptophan decarboxylase (CrTDC) and strictosidine synthase (CrSTR), at whole plant levels in C. roseus. Whole plant transgenic of C. roseus was developed using Agrobacterium tumefaciens LBA1119 strain having CrTDC and CrSTR gene cassette. Developed transgenic lines demonstrated up to twofold enhanced total alkaloid production with maximum ninefold increase in vindoline and catharanthine, and fivefold increased vinblastine production. These lines recorded a maximum of 38-fold and 65-fold enhanced transcript levels of CrTDC and CrSTR genes, respectively.
Assuntos
Alcaloides/metabolismo , Descarboxilases de Aminoácido-L-Aromático/metabolismo , Carbono-Nitrogênio Liases/metabolismo , Catharanthus/metabolismo , Indóis/metabolismo , Terpenos/metabolismo , Catharanthus/enzimologia , Vimblastina/metabolismoRESUMO
Dual metabolite, i.e., ginsenoside and anthocyanin, co-accumulating cell suspensions of Panax sikkimensis were subjected to elicitation with culture filtrates of Serratia marcescens (SD 21), Bacillus subtilis (FL11), Trichoderma atroviridae (TA), and T. harzianum (TH) at 1.25% and 2.5% v/v for 1- and 3-week duration. The fungal-derived elicitors (TA and TH) did not significantly affect biomass accumulation; however, bacterial elicitors (SD 21 and FL11), especially SD 21, led to comparable loss in biomass growth. In terms of ginsenoside content, differential responses were observed. A maximum of 3.2-fold increase (222.2 mg/L) in total ginsenoside content was observed with the use of 2.5% v/v TH culture filtrate for 1 week. Similar ginsenoside accumulation was observed with the use of 1-week treatment with 2.5% v/v SD 21 culture filtrate (189.3 mg/L) with a 10-fold increase in intracellular Rg2 biosynthesis (31 mg/L). Real-time PCR analysis of key ginsenoside biosynthesis genes, i.e., FPS, SQS, DDS, PPDS, and PPTS, revealed prominent upregulation of particularly PPTS expression (20-23-fold), accounting for the observed enhancement in protopanaxatriol ginsenosides. However, none of the elicitors led to successful enhancement in in vitro anthocyanin accumulation as compared to control values.
Assuntos
Ginsenosídeos/genética , Ginsenosídeos/metabolismo , Panax/química , Raízes de Plantas/química , Meios de Cultura , SuspensõesRESUMO
Vasoactive intestinal peptide (VIP) receptors are expressed abundantly on many types of tumors and, hence, radiolabeled VIP analogues are being explored for tumor imaging and therapy. Here, we report synthesis of three VIP analogues and their radiolabeling with (99m)Tc via a novel tricarbonyl synthon. The radiolabeled product could be prepared in high yields (>95%) and stability. In vitro studies showed significant uptake of (99m)Tc(CO)((3))-VP05 in human colon carcinoma cells. Biodistribution studies in animal tumor model showed 0.4-1%ID/g tumor uptake.
Assuntos
Neoplasias do Colo/diagnóstico por imagem , Compostos de Organotecnécio/síntese química , Peptídeo Intestinal Vasoativo/análogos & derivados , Sequência de Aminoácidos , Animais , Humanos , Marcação por Isótopo , Camundongos , Compostos de Organotecnécio/farmacocinética , Cintilografia , Peptídeo Intestinal Vasoativo/síntese química , Peptídeo Intestinal Vasoativo/farmacocinéticaRESUMO
An artificial neural network (ANN)-based modelling approach is used to determine the synergistic effect of five major components of growth medium (Mg, Cu, Zn, nitrate and sucrose) on improved in vitro biomass yield in multiple shoot cultures of Centella asiatica. The back propagation neural network (BPNN) was employed to predict optimal biomass accumulation in terms of growth index over a defined culture duration of 35 days. The four variable concentrations of five media components, i.e. MgSO4 (0, 0.75, 1.5, 3.0 mM), ZnSO4 (0, 15, 30, 60 µM), CuSO4 (0, 0.05, 0.1, 0.2 µM), NO3 (20, 30, 40, 60 mM) and sucrose (1, 3, 5, 7 %, w/v) were taken as inputs for the ANN model. The designed model was evaluated by performing three different sets of validation experiments that indicated a greater similarity between the target and predicted dataset. The results of the modelling experiment suggested that 1.5 mM Mg, 30 µM Zn, 0.1 µM Cu, 40 mM NO3 and 6 % (w/v) sucrose were the respective optimal concentrations of the tested medium components for achieving maximum growth index of 1654.46 with high centelloside yield (62.37 mg DW/culture) in the cultured multiple shoots. This study can facilitate the generation of higher biomass of uniform, clean, good quality C. asiatica herb that can efficiently be utilized by pharmaceutical industries.
Assuntos
Biomassa , Centella/crescimento & desenvolvimento , Modelos Biológicos , Redes Neurais de Computação , Brotos de Planta/crescimento & desenvolvimento , Técnicas de Cultura de Tecidos/métodos , Análise de Regressão , Reprodutibilidade dos TestesRESUMO
The present study aims at developing an extraction protocol for efficient ginsenoside recovery from cell suspensions of Panax quinquefolius and P. sikkimensis. Methanol (100%, 70% and 30%), water (40°C, 90°C), water-saturated butanol and butanol-saturated water were compared for their ultrasonication-assisted ginsenoside retrieval efficacy. HPLC and HP-TLC analysis revealed 100% methanol as the best solvent for maximum retrieval of Rb (diol) and Rg (triol) ginsenosides (P. quinquefolius: Rb: 0.189, Rg: 3.163 mg/g DW; P. sikkimensis: Rb: 0.245, Rg: 4.073 mg/g DW), followed by water (90°C). Methanolic solutions, especially 70%, proved to be significant retrievers of Rg1 (1.812 and 1.327 mg/g DW in P. quinquefolius and P. sikkimensis), with poor Re recovery (0.328 and 0.342 mg/g DW). Water-saturated butanol also led to significant ginsenoside extraction (72.4% of content extracted by methanol), selectively in P. quinquefolius, with a less than 50% of total content extracted by methanol, in P. sikkimensis.
Assuntos
Ginsenosídeos/isolamento & purificação , Panax/química , Linhagem Celular , Cromatografia Líquida de Alta Pressão , Metanol , Solventes , Ultrassom , ÁguaRESUMO
The age-dependent production kinetics of ginsenosides and an anthocyanin pigment in a cell suspension line of Panax sikkimensis was followed in vitro. Highest total saponin content [7.37 mg/g dry weight (DW)] and biomass accumulation (% biomass increase = 209.67) in this line occurred after 3 and 5 weeks of culture, respectively. Accumulation of individual protopanaxatriol (Re, Rg1, and Rg2) and protopanaxadiol (Rb1, Rb2, and Rc) ginsenosides showed a variable pattern of accumulation independent of cell biomass buildup during the 7-week culture cycle. However, total content of triol ginsenosides was always significantly more than the diol group of ginsenosides, being 183.2-, 63.5-, and 72.1-folds at third, fourth, and fifth week stage of cell growth. Interestingly, in addition to these ginsenosides, the cell line also co-accumulated an anthocyanin pigment in vitro. The pigment content increased gradually from 8.66 to 14.29 mg/g DW after first to fifth week followed by a marginal fall to 12.79 and 10.95 mg/g DW during next 2 weeks. Therefore, in terms of total recovery of saponins (77.4 mg/l) and anthocyanin (199.16 mg/l), harvesting of cells after 3 and 5 weeks of growth was most profitable, respectively. The possible utility of this dual purpose cell line in nutraceutical industry is discussed.
Assuntos
Antocianinas/biossíntese , Ginsenosídeos/biossíntese , Panax/citologia , Extratos Vegetais/biossíntese , Antocianinas/isolamento & purificação , Linhagem Celular , Proliferação de Células , Cromatografia Líquida de Alta Pressão , Ginsenosídeos/isolamento & purificação , Panax/metabolismo , Extratos Vegetais/isolamento & purificaçãoRESUMO
Centella asiatica germplasm collected from north, north-eastern and southern parts of India was compared for biomass and centellosides productivity under uniform agro-climatic conditions of the Indo-Gangetic plains at Lucknow. The highest biomass accumulation (411.9 g FW/m2 area) was recorded in accession A from north India, followed by 284.0, 135.7 and 29.2 g FW/m2 in accessions M, B and E from southern, eastern and north-eastern regions, respectively. Accession M possessed the highest asiaticoside content (52.1 mg/gDW) that was 1.58, 2.34 and 21.7 folds more than accessions A, B and E, respectively. The madecassoside level in leaves of accessions B and M was comparable (28.9 and 25.7 mg/gDW) and two folds more than accession A (13.9 mg/gDW). The madecassic and asiatic acid content in leaf tissue of all four accessions remained low in Lucknow. Amplified fragment length polymorphism (AFLP) analysis with 23 primers yielded 696 fragments, 563 of which were polymorphic. Accession M out-grouped with genetic dissimilarity indices of 83, 85 and 95% from accessions A, E and B, respectively. Commercial cultivation of accessions M and A through a four months growth cycle (June to September) in agro-climatic conditions of the Indo-Gangetic plains is suggested.
Assuntos
Agricultura , Centella/genética , Centella/fisiologia , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , DNA de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Índia , Folhas de Planta/química , Folhas de Planta/metabolismo , Plantas MedicinaisRESUMO
Catharanthus roseus (The Madagaskar Periwinkle) plant is commercially valued for harbouring more than 130 bioactive terpenoid indole alkaloids (TIAs). Amongst these, two of the leaf-derived bisindole alkaloids-vinblastine and vincristine-are widely used in several anticancer chemotherapies. The great pharmacological values, low in planta occurrence, unavailability of synthetic substitutes and exorbitant market cost of these alkaloids have prompted scientists to understand the basic architecture and regulation of biosynthesis of these TIAs in C. roseus plant and its cultured tissues. The knowledge gathered over a period of 30 years suggests that the TIA biosynthesis is highly regulated by developmental and environmental factors and operates through a complex multi-step enzymatic network. Extensive spatial and temporal cross talking also occurs at inter- and intracellular levels in different plant organs during TIA biogenesis. A close association of indole, methylerythritol phosphate and secoiridoid monoterpenoid pathways and involvement of at least four cell types (epidermis, internal phloem-associated parenchyma, laticifers and idioblasts) and five intracellular compartments (chloroplast, vacuole, nucleus, endoplasmic reticulum and cytosol) have been implicated with this biosynthetic mechanism. Accordingly, the research in this area is primarily advancing today to address and resolve six major issues namely: precise localization and expression of pathway enzymes using modern in situ RNA hybridization tools, mechanisms of intra- and intercellular trafficking of pathway intermediates, cloning and functional validation of genes coding for known or hitherto unknown pathway enzymes, mechanism of global regulation of the pathway by transcription factors, control of relative diversion of metabolite flux at crucial branch points and finally, strategising the metabolic engineering approaches to improve the productivity of the desired TIAs in plant or corresponding cultured tissues. The present literature update has been compiled to provide a brief overview of some of the emerging developments in our current understanding of TIA metabolism in C. roseus.