WorldWideScience

Sample records for acid-elicited catharanthus roseus

  1. Metabolic changes of salicylic acid-elicited Catharanthus roseus cell suspension cultures monitored by NMR-based metabolomics

    Mustafa, Natali Rianika; Kim, Hye Kyong; Choi, Young Hae; Verpoorte, Robert

    2009-01-01

    The effect of salicylic acid (SA) on the metabolic profile of Catharanthus roseus suspension cells throughout a time course (0, 6, 12, 24, 48 and 72 h after treatment) was investigated using NMR spectroscopy and multivariate data analysis. When compared to control cell lines, SA-treated cells showed a high level of sugars (glucose and sucrose) up to 48 h after treatment, followed by a dynamic change in amino acids, phenylpropanoids, and tryptamine. Additionally, one compound—2,5-dihydroxybenz...

  2. Jasmonate-responsive transcriptional regulation in Catharanthus roseus

    Zhang, Hongtao

    2008-01-01

    Plants produce a variety of secondary metabolites. In Catharanthus roseus, several have pharmaceutical applications, including the monomeric alkaloids serpentine and ajmalicine, which are used as a tranquillizer and to reduce hypertension, respectively, and the dimeric alkaloids vincristine and vinb

  3. Novel Plant Regeneration and Transient Gene Expression in Catharanthus roseus

    Abdullah Makhzoum; Anica Bjelica; Genevieve Petit-Paly; Bernards, Mark A

    2015-01-01

    Catharanthus roseus genetic transformation represents a real challenge due, in part, to the lack of regeneration capability and this species’ recalcitrance to genetic transformation. In the present work, we demonstrate the regeneration of C. roseus plants from hypocotyls and cotyledons, using specific growth regulator conditions. Plants derived from hypocotyls and cotyledons were successfully acclimated and grown in the greenhouse. Furthermore, C. roseus meristem tissues were shown to have hi...

  4. Mutation breeding in Catharanthus roseus (L. G. Don: An Overview

    Ashutosh Kumar Verma

    2013-05-01

    Full Text Available Induced mutagenesis is best approach for creation of genetic variability but most of interest is paid towards crop plants with only a few exceptions. But even widely used medicinal plants could receive a little attention in this direction. Catharanthus roseus is an important plant medicinal plant yielding antihypertensive alkaloid ajmalcine and anticancerous alkaloid vincristine and vinblastine. Despite of considerable work had been done on biotechnological aspect of C. roseus alkaloid production but relatively little efforts has been made for its improvement by mutation breeding approach. The aim of this review is to summarize the all efforts of mutation breeding which were made for the improvement of Catharanthus roseus.

  5. Terpenoid Indole Alkaloids Biosynthesis and Metabolic Engineering in Catharanthus roseus

    2007-01-01

    Catharanthus roseus L. (Madagascar periwinkle) biosynthesizes a diverse array of secondary metabolites including anticancer dimeric alkaloids (vinblastine and vincristine) and antihypertensive alkaloids (ajmalicine and serpentine). The multi-step terpenoid indole alkaloids (TIAs) biosynthetic pathway in C. roseus is complex and is under strict molecular regulation. Many enzymes and genes involved in the TIAs biosynthesis have been studied in recent decades. Moreover,some regulatory proteins were found recently to control the production of TIAs in C. roseus. Based on mastering the rough scheme of the pathway and cloning the related genes, metabolic engineering of TIAs biosynthesis has been studied in C.roseus aiming at increasing the desired secondary metabolites in the past few years. The present article summarizes recent advances in isolation and characterization of TIAs biosynthesis genes and transcriptional regulators involved in the second metabolic control in C. roseus. Metabolic engineering applications in TIAs pathway via overexpression of these genes and regulators in C. roseus are also discussed.

  6. Monoterpenoid Indole Alkaloids from Catharanthus roseus Cultivated in Yunnan.

    Wang, Bei; Liu, Lu; Chen, Ying-ying; Li, Qiong; Li, Dan; Liu, Va-ping; Luo, Xiao-dong

    2015-12-01

    A new monoterpenoid indole alkaloid, 15,20-dehydro-3α-(2-oxopropyl) coronaridine (1), along with sixteen analogues (2-17) were isolated from the leaves of Catharanthus roseus cultivated in Yunnan. The new alkaloid was elucidated on the basis of extensive spectroscopic analysis, and the known alkaloids were identified by comparison with the reported spectroscopic data. Among them, alkaloid 16 was isolated from Catharanthus for the first time. PMID:26882670

  7. Jasmonate-responsive transcriptional regulation in Catharanthus roseus

    Hongtao ZHANG

    2008-01-01

    Plants produce a variety of secondary metabolites. In Catharanthus roseus, several have pharmaceutical applications, including the monomeric alkaloids serpentine and ajmalicine, which are used as a tranquillizer and to reduce hypertension, respectively, and the dimeric alkaloids vincristine and vinblastine, which are potent antitumour drugs. Jasmonic acid (JA) is a key defense hormone, which controls the expression of several alkaloid biosynthesis genes in Catharanthus. The JA-responsive expr...

  8. Mutation breeding in Catharanthus roseus (L.) G. Don: An Overview

    Ashutosh Kumar Verma; Singh, R R; Seema Singh

    2013-01-01

    Induced mutagenesis is best approach for creation of genetic variability but most of interest is paid towards crop plants with only a few exceptions. But even widely used medicinal plants could receive a little attention in this direction. Catharanthus roseus is an important plant medicinal plant yielding antihypertensive alkaloid ajmalcine and anticancerous alkaloid vincristine and vinblastine. Despite of considerable work had been done on biotechnological aspect of C. roseus alkaloid produ...

  9. Metabolomic characteristics of Catharanthus roseus plants in time and space

    Qifang, Pan

    2014-01-01

    The thesis aims at combining metabolomics with other methods to investigate the regulation of the TIA biosynthesis and how this is connected with other pathways and the plant’s physiology and development. It reviews the biosynthesis studies of Catharanthus roseus. An HPLC method is described for both precursors and alkaloids which was validated to analyze alkaloid contents in different organs, its relationship with leaf age and growth of C. roseus plants. A metabolomics investigation of C. ro...

  10. Metabolomic characteristics of Catharanthus roseus plants in time and space

    Qifang, Pan; Qifang, Pan

    2014-01-01

    The thesis aims at combining metabolomics with other methods to investigate the regulation of the TIA biosynthesis and how this is connected with other pathways and the plant’s physiology and development. It reviews the biosynthesis studies of Catharanthus roseus. An HPLC method is described for bot

  11. Discovery and reconstitution of the secoiridoid pathway of Catharanthus roseus

    Dong, L.

    2014-01-01

      Terpene indole alkaloids (TIAs) are important plant-produced secondary metabolites for humans, because of their anti-cancer properties. The production of TIAs still fully relies on extraction from medicinal plants like Catharanthus roseus, which only contains extreme low amounts of these

  12. Effect of Aqueous Flower Extract of Catharanthus roseus on Alloxan Induced Diabetes in Male Albino Rats

    Natarajan, A.; K. Syed Zameer Ahmed; Sundaresan, S.; A. Sivaraj; Devi, K; B Senthil Kumar

    2012-01-01

    Catharanthus roseus is widely used in Indian Ayurvedic medicine for the treatment of diabetes mellitus. The present study was carried out to investigate the antidiabetic and hyperlipidemic potential of Catharanthus roseus on alloxan induced diabetes in male albino rats. Oral administration of aqueous extract of Catharanthus roseus at a dose of 250 mg, 350 mg, and 450 mg/kg body weight for 30 days to diabetic rats resulted in significant reduction in blood glucose, reduction in lipid profile a...

  13. Biosynthesis and regulation of terpenoid indole alkaloids in Catharanthus roseus.

    Zhu, Jianhua; Wang, Mingxuan; Wen, Wei; Yu, Rongmin

    2015-01-01

    Catharanthus roseus produces a wide range of terpenoid indole alkaloids (TIA). Many of them, such as vinblastine and vincristine, have significant bioactivity. They are valuable chemotherapy drugs used in combination with other drugs to treat lymphoma and leukemia. The TIA biosynthetic pathway has been investigated for many years, for scientific interest and for their potential in manufacturing applications, to fulfill the market demand. In this review, the progress and perspective of C. roseus TIA biosynthesis and its regulating enzymes are described. In addition, the culture condition, hormones, signaling molecules, precursor feeding on the accumulation of TIA, and gene expression are also evaluated and discussed. PMID:26009689

  14. Biosynthesis and regulation of terpenoid indole alkaloids in Catharanthus roseus

    Jianhua Zhu

    2015-01-01

    Full Text Available Catharanthus roseus produces a wide range of terpenoid indole alkaloids (TIA. Many of them, such as vinblastine and vincristine, have significant bioactivity. They are valuable chemotherapy drugs used in combination with other drugs to treat lymphoma and leukemia. The TIA biosynthetic pathway has been investigated for many years, for scientific interest and for their potential in manufacturing applications, to fulfill the market demand. In this review, the progress and perspective of C. roseus TIA biosynthesis and its regulating enzymes are described. In addition, the culture condition, hormones, signaling molecules, precursor feeding on the accumulation of TIA, and gene expression are also evaluated and discussed.

  15. Antimicrobial Activity of Catharanthus roseus – A Detailed Study

    Prajakta J. Patil

    2010-06-01

    Full Text Available Catharanthus roseus (periwinkle is an important medicinal plant for novel pharmaceuticals since most of the bacterial pathogens are developing resistance against many of the currently available anti microbial drugs. Plants have proved to be significant natural resources for effective chemotherapeutic agents and offering a broad spectrum of activity with greater emphasis on preventive action. This study aims to investigate some of the anti microbial properties of this plant. The anticancer properties of Catharanthus roseus has been the major interest in all investigations. The antimicrobial activity has been checked against microorganisms like Pseudomonas aeruginosa NCIM 2036, Salmonella typhimurium NCIM 2501, Staphylococcus aureus NCIM 5021. The findings show that the extracts from the leaves of this plant can be used as prophylactic agent in many of the diseases, which sometime are of the magnitude of an epidemic.

  16. Antimicrobial Activity of Catharanthus roseus – A Detailed Study

    Prajakta J. Patil; Jai S. Ghosh

    2010-01-01

    Catharanthus roseus (periwinkle) is an important medicinal plant for novel pharmaceuticals since most of the bacterial pathogens are developing resistance against many of the currently available anti microbial drugs. Plants have proved to be significant natural resources for effective chemotherapeutic agents and offering a broad spectrum of activity with greater emphasis on preventive action. This study aims to investigate some of the anti microbial properties of this plant. The anticancer pr...

  17. Induced Dwarf Mutant in Catharanthus roseus with Enhanced Antibacterial Activity

    Verma, A.K.; Singh, R R

    2010-01-01

    Evaluation of an ethyl methane sulphonate-induced dwarf mutant of Catharanthus roseus (L.) G. Don revealed that the mutant exhibited marked variation in morphometric parameters. The in vitro antibacterial activity of the aqueous and alcoholic leaf extracts of the mutant and control plants was investigated against medically important bacteria. The mutant leaf extracts showed enhanced antibacterial activity against all the tested bacteria except Bacillus subtilis.

  18. The seco-iridoid pathway from Catharanthus roseus

    Miettinen, K.; Dong, L.; Navrot, N.; Burlat, V.; Schneider, T; Pollier, J.; Woittiez, L.S.; Krol, van der, S.; Lugan, R.; Llc, T.; Verpoorte, R.; Oksman-Caldentey, K.M.; Martinoia, E.; Bouwmeester, H.J.

    2014-01-01

    The (seco) iridoids and their derivatives, the monoterpenoid indole alkaloids (MIAs), form two large families of plant-derived bioactive compounds with a wide spectrum of high-value pharmacological and insect-repellent activities. Vinblastine and vincristine, MIAs used as anticancer drugs, are produced by Catharanthus roseus in extremely low levels, leading to high market prices and poor availability. Their biotechnological production is hampered by the fragmentary knowledge of their biosynth...

  19. Catharanthus roseus (L.) G. Don - plant regeneration and alkaloids content

    Mirosława Furmanowa; Hanna Olędzka; Joanna Józefowicz; Agnieszka Pietrosiuk

    2014-01-01

    We describe here a regeneration of plantlets of Catharanthus roseus (L.) G. Don from shoot tips and axillary buds. Shoot tips were excised from 7-day-old seedlings and were incubated in solid Nitsch and Nitsch (NN) medium supplemented with kinetin, benzyladenine (BA), indole-3-butyric acid (IBA) and β-indolylacetic acid (IAA) in various combinations. After two months in culture, regenerated rooted plantlets were cut and transferred to a new medium; the explants contained shoot tips or axillar...

  20. Discovery and reconstitution of the secoiridoid pathway of Catharanthus roseus

    Dong, L.

    2014-01-01

      Terpene indole alkaloids (TIAs) are important plant-produced secondary metabolites for humans, because of their anti-cancer properties. The production of TIAs still fully relies on extraction from medicinal plants like Catharanthus roseus, which only contains extreme low amounts of these compounds and new ways need to be found to efficiently produce these anticancer drugs at low cost. The common precursor for TIAs is strictosidine and in my PhD project I tried to produce strictosidine ...

  1. Hypoglycemic Activity of Aqueous Extracts from Catharanthus roseus

    Elisa Vega-Ávila; José Luis Cano-Velasco; Alarcón-Aguilar, Francisco J.; María del Carmen Fajardo Ortíz; Julio César Almanza-Pérez; Rubén Román-Ramos

    2012-01-01

    Introduction. Catharanthus roseus (L.) is used in some countries to treat diabetes. The aim of this study was to evaluate the hypoglycemic activity of extracts from the flower, leaf, stem, and root in normal and alloxan-induced diabetic mice. Methods. Roots, leaves, flowers, and stems were separated to obtain organic and aqueous extracts. The blood glucose lowering activity of these extracts was determinate in healthy and alloxan-induced (75 mg/Kg) diabetic mice, after intraperitoneal adminis...

  2. Induced dwarf mutant in Catharanthus roseus with enhanced antibacterial activity

    Verma A

    2010-01-01

    Full Text Available Evaluation of an ethyl methane sulphonate-induced dwarf mutant of Catharanthus roseus (L. G. Don revealed that the mutant exhibited marked variation in morphometric parameters. The in vitro antibacterial activity of the aqueous and alcoholic leaf extracts of the mutant and control plants was investigated against medically important bacteria. The mutant leaf extracts showed enhanced antibacterial activity against all the tested bacteria except Bacillus subtilis.

  3. Biosynthesis and regulation of terpenoid indole alkaloids in Catharanthus roseus

    Jianhua Zhu; Mingxuan Wang; Wei Wen; Rongmin Yu

    2015-01-01

    Catharanthus roseus produces a wide range of terpenoid indole alkaloids (TIA). Many of them, such as vinblastine and vincristine, have significant bioactivity. They are valuable chemotherapy drugs used in combination with other drugs to treat lymphoma and leukemia. The TIA biosynthetic pathway has been investigated for many years, for scientific interest and for their potential in manufacturing applications, to fulfill the market demand. In this review, the progress and perspective of C. rose...

  4. Induced Dwarf Mutant in Catharanthus roseus with Enhanced Antibacterial Activity

    Verma, A. K.; Singh, R. R.

    2010-01-01

    Evaluation of an ethyl methane sulphonate-induced dwarf mutant of Catharanthus roseus (L.) G. Don revealed that the mutant exhibited marked variation in morphometric parameters. The in vitro antibacterial activity of the aqueous and alcoholic leaf extracts of the mutant and control plants was investigated against medically important bacteria. The mutant leaf extracts showed enhanced antibacterial activity against all the tested bacteria except Bacillus subtilis. PMID:21695004

  5. Elucidation of the secoiridoid pathway in Catharanthus roseus

    Miettinen, Karel

    2013-01-01

    The (seco)iridoids and their derivatives, the monoterpenoid indole alkaloids (MIAs), form two large families of plant-derived bioactive compounds with a wide spectrum of high-value pharmacological and insect-repellent activities. Vinblastine and vincristine, MIAs used as anti-cancer drugs, are produced by Catharanthus roseus in extremely low levels, leading to high market prices and poor availability. Their biotechnological production is hampered by the fragmentary knowledge of their biosynth...

  6. Antihyperglycemic activity of Catharanthus roseus leaf powder in streptozotocin-induced diabetic rats

    Karuna Rasineni; Ramesh Bellamkonda; Sreenivasa Reddy Singareddy; Saralakumari Desireddy

    2010-01-01

    Catharanthus roseus Linn (Apocynaceae), is a traditional medicinal plant used to control diabetes, in various regions of the world. In this study we evaluated the possible antidiabetic and hypolipidemic effect of C. roseus (Catharanthus roseus) leaf powder in diabetic rats. Diabetes was induced by intraperitoneal injection of streptozotocin (STZ, 55 mg/kg body wt) to male Wistar rats. The animals were divided into four groups: Control, control-treated, diabetic, and diabetic-treated group. Di...

  7. New Sesquiterpene Glycosides from Culture Hairy Roots of Catharanthus roseus

    CHUNG ILL-Min; All Mohd; CHUN Se-Chul; LEE Sun-Joo; SAN Ka-Yiu; CHRISTIE A. M. Peebles; AHMAD Ateeque

    2007-01-01

    Two new compounds cadin-2-en-1β-ol-1β-D-glucuronopyranoside (1), guaia-1,7-dien-3β,13-diol-13α-D-glucofuranoside (2) along with three known compounds have been isolated from the Culture hairy roots of Catharanthus roseus. Their structures have been elucidated with the help of 500 MHz NMR using 1D and 2D spectral methods: viz: 1H and 13C NMR, 1H-1H COSY, 1H-13C HETCOR and DEPT aided by ELMS, FAB-MS, HR-FABMS and IR spectroscopy.

  8. Effect of Aqueous Flower Extract of Catharanthus roseus on Alloxan Induced Diabetes in Male Albino Rats

    A. Natarajan

    2012-04-01

    Full Text Available Catharanthus roseus is widely used in Indian Ayurvedic medicine for the treatment of diabetes mellitus. The present study was carried out to investigate the antidiabetic and hyperlipidemic potential of Catharanthus roseus on alloxan induced diabetes in male albino rats. Oral administration of aqueous extract of Catharanthus roseus at a dose of 250 mg, 350 mg, and 450 mg/kg body weight for 30 days to diabetic rats resulted in significant reduction in blood glucose, reduction in lipid profile and also prevented a decrease in body weight. Histological observation demonstrated significant fatty changes and inflammatory cell infiltrates in pancreas of diabetic rats. But, supplementation with Catharanthus roseus to diabetic rats significantly reduced the fatty changes and inflammatory cell infiltrates.

  9. Evaluation of In-vitro Anthelminthic Activity of Catharanthus roseus Extract

    Swati Agarwal; Simi Jacob; Nikkita Chettri; Saloni Bisoyi; Ayesha Tazeen; A. B. Vedamurthy; V. Krishna; H. Joy Hoskeri

    2011-01-01

    Helminthes infections are chronic illnesses in human beings and in cattle. Pherithema posthuma a helminthes is commonly known as earth-worms. Although the use of alternate drugs has been as a remedial measure against the resistant strains of helminth parasites, and as a means of reducing the cost of controlling helminthic diseases. Catharanthus roseus is a medicinally valuable plant and possess various pharmacological properties. Catharanthus roseus has been traditionally used as an anthelmin...

  10. Antidiabetic and Antioxidant Properties of Alkaloids from Catharanthus roseus (L.) G. Don

    Won Fen Wong; Shiau-Chuen Cheah; Mohd Rais Mustafa; Mohammadjavad Paydar; Aditya Arya; Soon Huat Tiong; Chung Yeng Looi; Khalijah Awang; Hazrina Hazni

    2013-01-01

    Catharanthus roseus (L.) G. Don is a herbal plant traditionally used by local populations in India, South Africa, China and Malaysia to treat diabetes. The present study reports the in vitro antioxidant and antidiabetic activities of the major alkaloids isolated from Catharanthus roseus (L.) G. Don leaves extract. Four alkaloids—vindoline I, vindolidine II, vindolicine III and vindolinine IV—were isolated and identified from the dichloromethane extract (DE) of this plant’s leaves. DE and comp...

  11. Binary stress induces an increase in indole alkaloid biosynthesis in Catharanthus roseus

    Wei ZHU; Yang, Bingxian; Komatsu, Setsuko; Lu, Xiaoping; Li, Ximin; Tian, Jingkui

    2015-01-01

    Catharanthus roseus is an important medicinal plant, which produces a variety of indole alkaloids of significant pharmaceutical relevance. In the present study, we aimed to investigate the potential stress-induced increase of indole alkaloid biosynthesis in C. roseus using proteomic technique. The contents of the detectable alkaloids ajmalicine, vindoline, catharanthine, and strictosidine in C. roseus were significantly increased under binary stress. Proteomic analysis revealed that the abund...

  12. Binary Stress Induces an Increase in Indole Alkaloid Biosynthesis in Catharanthus roseus

    Wei eZhu; Bingxian eYang; Setsuko eKomatsu; Xiaoping eLu; Ximin eLi; Jingkui eTian

    2015-01-01

    Catharanthus roseus is an important medicinal plant, which produces a variety of indole alkaloids of significant pharmaceutical relevance. In the present study, we aimed to investigate the potential stress-induced increase of indole alkaloid biosynthesis in C. roseus using proteomic technique. The contents of the detectable alkaloids ajmalicine, vindoline, catharanthine, and strictosidine in C. roseus were significantly increased under binary stress. Proteomic analysis revealed that the abund...

  13. Characterization of a new potyvirus causing mosaic and flower variegation in Catharanthus roseus in Brazil

    Sheila Conceição Maciel; Ricardo Ferreira da Silva; Marcelo Silva Reis; Adriana Salomão Jadão; Daniel Dias Rosa; José Segundo Giampan; Elliot Watanabe Kitajima; Jorge Alberto Marques Rezende; Luis Eduardo Camargo

    2011-01-01

    Catharanthus roseus is a perennial, evergreen herb in the family Apocynaceae, which is used as ornamental and for popular medicine to treat a wide assortment of human diseases. This paper describes a new potyvirus found causing mosaic symptom, foliar malformation and flower variegation in C. roseus. Of 28 test-plants inoculated mechanically with this potyvirus, only C. roseus and Nicotiana benthamiana developed systemic mosaic, whereas Chenopodium amaranticolor and C. quinoa exhibited chlorot...

  14. Comparative studies of elemental composition in leaves and flowers of Catharanthus roseus growing in Bangladesh

    Shahin Aziz; Koushik Saha; Nasim Sultana; Husna Parvin Nur; Md. Aminul Ahsan; Shamim Ahmed; Md Kamal Hossain

    2016-01-01

    Objective: To investigate the elemental composition of the leaves and flowers of Catharanthus roseus (C. roseus) due to the plant's wide application in the indigenous medicinal system and its chemical constituents' importance. Methods: The atomic absorption spectrophotometer was used for quantitative analysis of various elements. Results: Total 13 important elements were analyzed in leaves and flowers of C. roseus. Results indicated the presence of Na, K, Ca, Mg, Cr, Fe, Zn, Al, Cu, Ni,...

  15. In vitro evaluation of crude extracts of Catharanthus roseus for potential antibacterial activity

    Goyal Pankaj; Khanna Arjun; Chauhan Abhishek; Chauhan Garima; Kaushik Purshotam

    2008-01-01

    Context: Catharanthus roseus (periwinkle) is an important medicinal plant, mentioned in Ayurveda, an ancient Indian Sanskrit literature. The plant is selected to evaluate the possibility for novel pharmaceuticals since most of the bacterial pathogens are developing resistance against currently available antibiotics. Aims: To determine the antibacterial activity of crude extracts from different parts of Catharanthus roseus against several bacterial species of clinical significance. Materials ...

  16. The seco-iridoid pathway from Catharanthus roseus.

    Miettinen, Karel; Dong, Lemeng; Navrot, Nicolas; Schneider, Thomas; Burlat, Vincent; Pollier, Jacob; Woittiez, Lotte; van der Krol, Sander; Lugan, Raphaël; Ilc, Tina; Verpoorte, Robert; Oksman-Caldentey, Kirsi-Marja; Martinoia, Enrico; Bouwmeester, Harro; Goossens, Alain; Memelink, Johan; Werck-Reichhart, Danièle

    2014-01-01

    The (seco)iridoids and their derivatives, the monoterpenoid indole alkaloids (MIAs), form two large families of plant-derived bioactive compounds with a wide spectrum of high-value pharmacological and insect-repellent activities. Vinblastine and vincristine, MIAs used as anticancer drugs, are produced by Catharanthus roseus in extremely low levels, leading to high market prices and poor availability. Their biotechnological production is hampered by the fragmentary knowledge of their biosynthesis. Here we report the discovery of the last four missing steps of the (seco)iridoid biosynthesis pathway. Expression of the eight genes encoding this pathway, together with two genes boosting precursor formation and two downstream alkaloid biosynthesis genes, in an alternative plant host, allows the heterologous production of the complex MIA strictosidine. This confirms the functionality of all enzymes of the pathway and highlights their utility for synthetic biology programmes towards a sustainable biotechnological production of valuable (seco)iridoids and alkaloids with pharmaceutical and agricultural applications. PMID:24710322

  17. Effect of thermal power plant emissions on Catharanthus roseus L

    Khan, A.M.; Pandey, V.; Shukla, J.; Singh, N.; Yunus, M.; Singh, S.N.; Ahmad, K.J. (National Botanical Research Institute, Lucknow (India))

    1990-06-01

    Most of the industrialized nations depend largely on the combustion of fossil fuels for their energy requirements. During the past few years in India quite a few thermal power plants have been commissioned to cater to the increasing energy requirements. As most of the power plants are coal-fired, a complex mixture of several pollutants is released in the atmosphere on the combustion of coal. Leaves by virtue of their unique position on plants and their functions, experience the maximum brunt of exposure and undergo certain changes in form, structure and function with the changes in surrounding environs, and such modifications are likely to serve as markers of environmental pollution. The present paper deals with the long term exposure effects of thermal power plant emissions on Catharanthus roseus L. - a common perennial shrub, with glossy leaves and white, mauve or pink colored flowers and of great medicinal value is grown as an ornamental plant all over the country.

  18. Hypoglycemic Activity of Aqueous Extracts from Catharanthus roseus

    Elisa Vega-Ávila

    2012-01-01

    Full Text Available Introduction. Catharanthus roseus (L. is used in some countries to treat diabetes. The aim of this study was to evaluate the hypoglycemic activity of extracts from the flower, leaf, stem, and root in normal and alloxan-induced diabetic mice. Methods. Roots, leaves, flowers, and stems were separated to obtain organic and aqueous extracts. The blood glucose lowering activity of these extracts was determinate in healthy and alloxan-induced (75 mg/Kg diabetic mice, after intraperitoneal administration (250 mg/Kg body weight. Blood samples were obtained and blood glucose levels were analyzed employing a glucometer. The data were statistically compared by ANOVA. The most active extract was fractioned. Phytochemical screen and chromatographic studies were also done. Results. The aqueous extracts from C. roseus reduced the blood glucose of both healthy and diabetic mice. The aqueous stem extract (250 mg/Kg and its alkaloid-free fraction (300 mg/Kg significantly ( reduced blood glucose in diabetic mice by 52.90 and 51.21%. Their hypoglycemic activity was comparable to tolbutamide (58.1%, . Conclusions. The best hypoglycemic activity was presented for the aqueous extracts and by alkaloid-free stem aqueous fraction. This fraction is formed by three polyphenols compounds.

  19. Hypoglycemic Activity of Aqueous Extracts from Catharanthus roseus

    Vega-Ávila, Elisa; Cano-Velasco, José Luis; Alarcón-Aguilar, Francisco J.; Fajardo Ortíz, María del Carmen; Almanza-Pérez, Julio César; Román-Ramos, Rubén

    2012-01-01

    Introduction. Catharanthus roseus (L.) is used in some countries to treat diabetes. The aim of this study was to evaluate the hypoglycemic activity of extracts from the flower, leaf, stem, and root in normal and alloxan-induced diabetic mice. Methods. Roots, leaves, flowers, and stems were separated to obtain organic and aqueous extracts. The blood glucose lowering activity of these extracts was determinate in healthy and alloxan-induced (75 mg/Kg) diabetic mice, after intraperitoneal administration (250 mg/Kg body weight). Blood samples were obtained and blood glucose levels were analyzed employing a glucometer. The data were statistically compared by ANOVA. The most active extract was fractioned. Phytochemical screen and chromatographic studies were also done. Results. The aqueous extracts from C. roseus reduced the blood glucose of both healthy and diabetic mice. The aqueous stem extract (250 mg/Kg) and its alkaloid-free fraction (300 mg/Kg) significantly (P < 0.05) reduced blood glucose in diabetic mice by 52.90 and 51.21%. Their hypoglycemic activity was comparable to tolbutamide (58.1%, P < 0.05). Conclusions. The best hypoglycemic activity was presented for the aqueous extracts and by alkaloid-free stem aqueous fraction. This fraction is formed by three polyphenols compounds. PMID:23056144

  20. Biosynthetic pathway of terpenoid indole alkaloids in Catharanthus roseus.

    Zhu, Xiaoxuan; Zeng, Xinyi; Sun, Chao; Chen, Shilin

    2014-09-01

    Catharanthus roseus is one of the most extensively investigated medicinal plants, which can produce more than 130 alkaloids, including the powerful antitumor drugs vinblastine and vincristine. Here we review the recent advances in the biosynthetic pathway of terpenoid indole alkaloids (TIAs) in C. roseus, and the identification and characterization of the corresponding enzymes involved in this pathway. Strictosidine is the central intermediate in the biosynthesis of different TIAs, which is formed by the condensation of secologanin and tryptamine. Secologanin is derived from terpenoid (isoprenoid) biosynthetic pathway, while tryptamine is derived from indole biosynthetic pathway. Then various specific end products are produced by different routes during downstream process. Although many genes and corresponding enzymes have been characterized in this pathway, our knowledge on the whole TIA biosynthetic pathway still remains largely unknown up to date. Full elucidation of TIA biosynthetic pathway is an important prerequisite to understand the regulation of the TIA biosynthesis in the medicinal plant and to produce valuable TIAs by synthetic biological technology. PMID:25159992

  1. Evaluation of In-vitro Anthelminthic Activity of Catharanthus roseus Extract

    Swati Agarwal

    2011-07-01

    Full Text Available Helminthes infections are chronic illnesses in human beings and in cattle. Pherithema posthuma a helminthes is commonly known as earth-worms. Although the use of alternate drugs has been as a remedial measure against the resistant strains of helminth parasites, and as a means of reducing the cost of controlling helminthic diseases. Catharanthus roseus is a medicinally valuable plant and possess various pharmacological properties. Catharanthus roseus has been traditionally used as an anthelminthic agent. To justify the ethnomedical claims, the anthelminthic property of Catharanthus roseus was evaluated using Pherithema posthuma as an experimental model. Piperazine citrate was used as the standard reference. Among the various concentrations tested, ethanol extract at 200 mg/ml showed efficient paralysis effect (6.67 min than other treated groups, whereas ethanol extract 250 mg/ml showed significant anthelminthic activity with death time of 46.33 min. Standard drug at 50 mg/ml showed paralysis at 31.33 min and death time was 40.67 min. This investigation revealed that ethanol extract of Catharanthus roseus showed significant anthelminthic activity against Pheretima posthuma. Ethanol extract also proved to be efficient than the standard drug. This investigation supported the ethnomedical claims of Catharanthus roseus as an anthelminthic plant.

  2. Catharanthus roseus flower extract has wound-healing activity in Sprague Dawley rats

    Pinto Pereira Lexley; Nayak BS

    2006-01-01

    Abstract Background Catharanthus roseus L (C. roseus) has been used to treat a wide assortment of diseases including diabetes. The objective of our study was to evaluate the antimicrobial and wound healing activity of the flower extract of Catharanthus in rats. Methods Wound healing activity was determined in rats, after administration (100 mg kg-1 day-1) of the ethanol extract of C. roseus flower, using excision, incision and dead space wounds models. The animals were divided into two groups...

  3. Enhanced catharanthine and vindoline production in suspension cultures of Catharanthus roseus by ultraviolet-B light

    Ramani, Shilpa; Jayabaskaran, Chelliah

    2008-01-01

    Suspension cultures of Catharanthus roseus were used to evaluate ultraviolet-B (UV-B) treatment as an abiotic elicitor of secondary metabolites. A dispersed cell suspension culture from C. roseus leaves in late exponential phase and stationary phase were irradiated with UV-B for 5 min. The stationary phase cultures were more responsive to UV-B irradiation than late exponential phase cultures. Catharanthine and vindoline increased 3-fold and 12-fold, respectively, on treatment with a 5-min UV-...

  4. Radiation Processed Carrageenan Improves Plant Growth, Physiological Activities, and Alkaloids Production in Catharanthus roseus L.

    Naeem, M.; Mohd Idrees; Tariq Aftab; M. Masidur Alam; Khan, M. Masroor A.; Moin Uddin; Lalit Varshney

    2015-01-01

    Catharanthus roseus (L.) G. Don (Apocynaceae) is a medicinal plant that produces indole alkaloids used in cancer chemotherapy. Commercially important antineoplastic alkaloids, namely, vinblastine and vincristine, are mainly present in the leaves of C. roseus. Gamma-rays irradiated carrageenan (ICR) has been proven as plant growth promoting substance for a number of medicinal and agricultural plants. Considering the importance of ICR as a promoter of plant growth and alkaloids production in C....

  5. The Enhancement of Catharanthine Content in Catharanthus roseus Callus Culture Treated with Naphtalene Acetic Acid

    DINGSE PANDIANGAN

    2006-09-01

    Full Text Available The research aim was to examine the enhancement of catharanthine content in Catharanthus roseus callus culture added with different concentration of Naphtalene Acetic Acid (NAA. NAA treatment produced callus that formed hairy roots. Fresh and dry weight of callus increased as the increasing of NAA concentration. The catharanthine content of C. roseus callus culture was increased by adding NAA as well. The highest catharanthine content was found in 2.5 ppm NAA added callus.

  6. Radiolytically degraded sodium alginate enhances plant growth, physiological activities and alkaloids production in Catharanthus roseus L.

    Naeem, M.; Tariq Aftab; Abid A. Ansari; Mohd Idrees; Akbar Ali; Khan, M. Masroor A.; Moin Uddin; Lalit Varshney

    2015-01-01

    Catharanthus roseus (L.) G. Don (Family Apocynaceae) is a medicinal plant that produces indole alkaloids used in cancer chemotherapy. The anticancerous alkaloids, viz. vinblastine and vincristine, are mainly present in the leaves of C. roseus. High demand and low yield of these alkaloids in the plant has led to explore the alternative means for their production. Gamma irradiated sodium alginate (ISA) has proved as a plant growth promoting substance for various medicinal and agricultural crops...

  7. Indole Alkaloids from Catharanthus roseus: Bioproduction and Their Effect on Human Health

    Lorena Almagro; Francisco Fernández-Pérez; Maria Angeles Pedreño

    2015-01-01

    Catharanthus roseus is a medicinal plant belonging to the family Apocynaceae which produces terpenoid indole alkaloids (TIAs) of high medicinal importance. Indeed, a number of activities like antidiabetic, bactericide and antihypertensive are linked to C. roseus. Nevertheless, the high added value of this plant is based on its enormous pharmaceutical interest, producing more than 130 TIAs, some of which exhibit strong pharmacological activities. The most striking biological activity investig...

  8. A virus-induced gene silencing approach to understanding alkaloid metabolism in Catharanthus roseus

    Liscombe, David K.; O’Connor, Sarah E.

    2011-01-01

    The anticancer agents vinblastine and vincristine are bisindole alkaloids derived from coupling vindoline and catharanthine, monoterpenoid indole alkaloids produced exclusively by Madagascar periwinkle (Catharanthus roseus) plants. Industrial production of vinblastine and vincristine currently relies on isolation from C. roseus leaves, a process that affords these compounds in 0.0003–0.01% yields. Metabolic engineering efforts to improve alkaloid content or provide alternative sources of the ...

  9. Cytogenetic characterization and genome size of the medicinal plant Catharanthus roseus (L.) G. Don

    Guimarães, Guilherme; Cardoso, Luísa; Oliveira, Helena; Santos, Conceição; Duarte, Patrícia; Sottomayor, Mariana

    2012-01-01

    Background and aims Catharanthus roseus is a highly valuable medicinal plant producing several terpenoid indole alkaloids (TIAs) with pharmaceutical applications, including the anticancer agents vinblastine and vincristine. Due to the interest in its TIAs, C. roseus is one of the most extensively studied medicinal plants and has become a model species for the study of plant secondary metabolism. However, very little is known about the cytogenetics and genome size of this species, in spite of ...

  10. Catharanthus roseus mitogen-activated protein kinase 3 confers UV and heat tolerance to Saccharomyces cerevisiae

    Raina, Susheel Kumar; Wankhede, Dhammaprakash Pandhari; Sinha, Alok Krishna

    2012-01-01

    Catharanthus roseus is an important source of pharmaceutically important Monoterpenoid Indole Alkaloids (MIAs). Accumulation of many of the MIAs is induced in response to abiotic stresses such as wound, ultra violet (UV) irradiations, etc. Recently, we have demonstrated a possible role of CrMPK3, a C. roseus mitogen-activated protein kinase in stress-induced accumulation of a few MIAs. Here, we extend our findings using Saccharomyces cerevisiae to investigate the role of CrMPK3 in giving tole...

  11. The Enhancement of Catharanthine Content in Catharanthus roseus Callus Culture Treated with Naphtalene Acetic Acid

    DINGSE PANDIANGAN; NELSON NAINGGOLAN

    2006-01-01

    The research aim was to examine the enhancement of catharanthine content in Catharanthus roseus callus culture added with different concentration of Naphtalene Acetic Acid (NAA). NAA treatment produced callus that formed hairy roots. Fresh and dry weight of callus increased as the increasing of NAA concentration. The catharanthine content of C. roseus callus culture was increased by adding NAA as well. The highest catharanthine content was found in 2.5 ppm NAA added callus.

  12. Somatic Embryos in Catharanthus roseus: A Scanning Electron Microscopic Study

    Junaid ASLAM

    2014-06-01

    Full Text Available Catharanthus roseus (L. G. Don is an important medicinal plant as it contains several anti-cancerous compounds, like vinblastine and vincristine. Plant tissue culture technology (organogenesis and embryogenesis has currently been used in fast mass propagating raw materials for secondary metabolite synthesis. In this present communication, scanning electron microscopic (SEM study of somatic embryos was conducted and discussed. The embryogenic callus was first induced from hypocotyls of in vitro germinated seeds on which somatic embryos, differentiated in numbers, particularly on 2,4-D (1.0 mg/L Murashige and Skoog (MS was medium. To understand more about the regeneration method and in vitro formed embryos SEM was performed. The SEM study revealed normal somatic embryo origin and development from globular to heart-, torpedo- and then into cotyledonary-stage of embryos. At early stage, the embryos were clustered together in a callus mass and could not easily be detached from the parental tissue. The embryos were often long cylindrical structure with or without typical notch at the tip. Secondary embryos were also formed on primary embryo structure. The advanced cotyledonary embryos showed prominent roots and shoot axis, which germinated into plantlets. The morphology, structure and other details of somatic embryos at various stages were presented.

  13. Catharanthus roseus (L. G. Don - plant regeneration and alkaloids content

    Mirosława Furmanowa

    2014-02-01

    Full Text Available We describe here a regeneration of plantlets of Catharanthus roseus (L. G. Don from shoot tips and axillary buds. Shoot tips were excised from 7-day-old seedlings and were incubated in solid Nitsch and Nitsch (NN medium supplemented with kinetin, benzyladenine (BA, indole-3-butyric acid (IBA and β-indolylacetic acid (IAA in various combinations. After two months in culture, regenerated rooted plantlets were cut and transferred to a new medium; the explants contained shoot tips or axillary buds. Four passages were done. We obtained about 200 rooting plantlets from one seedling. Then the plantlets were transferred to the soil and they grew under a foil tent. After five months of vegetation they were collected, dried and weighed. Chemical investigations of leaves of these plants were done. The vindoline and catharanthine were dominant alkaloids in the juvenile stage of plants (before blooming. Total amount of alkaloids, equal 2.95%, was gravimetrically determined in leaves of plants, after 4th passage, regenerated in vitro on NN medium supplemented with kinetin and IBA.

  14. Computational identification of microRNAs and their targets in Catharanthus roseus expressed sequence tags

    Pani, Alok; Mahapatra, Rajani Kanta

    2013-01-01

    No study has been performed on identifying microRNAs (miRNAs) and their targets in the medicinal plant, Catharanthus roseus. In the present study, using the comparative genomics approach, we have predicted two potential C. roseus miRNAs. Furthermore, twelve potential mRNA targets were identified in C. roseus genome based on the characteristics that miRNAs exhibit perfect or nearly perfect complementarity with their targeted mRNA sequences. Among them many of the targets were predicted to enco...

  15. Catharanthus roseus flower extract has wound-healing activity in Sprague Dawley rats

    Pinto Pereira Lexley

    2006-12-01

    Full Text Available Abstract Background Catharanthus roseus L (C. roseus has been used to treat a wide assortment of diseases including diabetes. The objective of our study was to evaluate the antimicrobial and wound healing activity of the flower extract of Catharanthus in rats. Methods Wound healing activity was determined in rats, after administration (100 mg kg-1 day-1 of the ethanol extract of C. roseus flower, using excision, incision and dead space wounds models. The animals were divided into two groups of 6 each in all the models. In the excision model, group 1 animals were topically treated with carboxymethyl cellulose as placebo control and group 2 received topical application of the ethanol extract of C. roseus at a dose of 100 mg/kg body weight/day. In an incision and dead space model group 1 animals were given normal saline and group 2 received the extract orally at a dose of 100 mg kg-1 day-1. Healing was assessed by the rate of wound contraction, period of epithelization, tensile strength (skin breaking strength, granulation tissue weight, and hydoxyproline content. Antimicrobial activity of the flower extract against four microorganisms was also assessed Results The extract of C. roseus significantly increased the wound breaking strength in the incision wound model compared with controls (P Pseudomonas aeruginosa and Staphylococcus aureus demonstrated sensitivity to C. roseus Conclusion Increased wound contraction and tensile strength, augmented hydroxyproline content along with antimicrobial activity support the use of C. roseus in the topical management of wound healing.

  16. Enhancing terpenoid indole alkaloid production by inducible expression of mammalian Bax in Catharanthus roseus cells

    XU MaoJun; DONG JuFang

    2007-01-01

    Bax, a mammalian pro-apoptotic member of the Bcl-2 family, triggers hypersensitive reactions when expressed in plants. To investigate the effects of Bax on the biosynthesis of clinically important natural products in plant cells, we generate transgenic Catharanthus roseus cells overexpressing a mouse Bax protein under the β-estradiol-inducible promoter. The expression of Bax in transgenic Catharanthus roseus cells is highly dependent on β-estradiol concentrations applied. Contents of catharanthine and total terpenoid indole alkaloid of the transgenic cells treated with 30 μmol/L β-estradiol are 5.0- and 5.5-fold of the control cells. Northern and Western blotting results show that expression of mammalian Bax induces transcriptional activation of Tdc and Str, two key genes in terpenoid indole alkaloid biosynthetic pathway of Catharanthus roseus cells, and stimulates the accumulation of defense-related protein PR1 in the cells, showing that the mouse Bax triggers the defense responses of Catharanthus roseus cells and activates the terpenoid indole alkaloid biosynthetic pathway. Thus, our data suggest that the mammalian Bax might be a potential regulatory factor for secondary metabolite biosynthesis in plant cells and imply a new secondary metabolic engineering strategy for enhancing the metabolic flux to natural products by activating the whole biosynthetic pathway rather than by engineering the single structural genes within the pathways.

  17. Enhancing terpenoid indole alkaloid production by inducible expression of mammalian Bax in Catharanthus roseus cells

    2007-01-01

    Bax,a mammalian pro-apoptotic member of the Bcl-2 family,triggers hypersensitive reactions when expressed in plants.To investigate the effects of Bax on the biosynthesis of clinically important natural products in plant cells,we generate transgenic Catharanthus roseus cells overexpressing a mouse Bax protein under the β-estradiol-inducible promoter.The expression of Bax in transgenic Catharanthus roseus cells is highly dependent on β-estradiol concentrations applied.Contents of catharanthine and total terpenoid indole alkaloid of the transgenic cells treated with 30 μmol/L β-estradiol are 5.0-and 5.5-fold of the control cells.Northern and Western blotting results show that expression of mammalian Bax induces transcriptional activation of Tdc and Str,two key genes in terpenoid indole alkaloid bio-synthetic pathway of Catharanthus roseus cells,and stimulates the accumulation of defense-related protein PR1 in the cells,showing that the mouse Bax triggers the defense responses of Catharanthus roseus cells and activates the terpenoid indole alkaloid biosynthetic pathway.Thus,our data suggest that the mammalian Bax might be a potential regulatory factor for secondary metabolite biosynthesis in plant cells and imply a new secondary metabolic engineering strategy for enhancing the metabolic flux to natural products by activating the whole biosynthetic pathway rather than by engineering the single structural genes within the pathways.

  18. First report of Tomato chlorotic spot virus on Annual Vinca (Catharanthus roseus) in the United States

    Tomato chlorotic spot virus was identified in the ornamental crop Catharanthus roseus (commonly known as vinca) in south Florida, the first report of this virus naturally infecting this species. Genetic diversity of the virus was characterized. This report provides an overview of this emerging vir...

  19. 7-O-methylpelargonidin glycosides from the pale red flowers of Catharanthus roseus.

    Tatsuzawa, Fumi

    2013-08-01

    Two new anthocyanidin glycosides were isolated from the pale red flowers of Catharanthus roseus 'Equator Apricot with Red Eye', and identified as 7-O-methylpelargonidin 3-O-[6-O-(alpha-rhamnopyranosyl)-beta-galactopyranoside] and 7-O-methylpelargonidin 3-O-(beta-galactopyranoside) by chemical and spectroscopic methods. PMID:24079176

  20. Analysis of Several Popular Cultivars of Madagascar Periwinkle (Catharanthus roseus (L. G. Don. using Biochemical Markers

    Owk ANIEL KUMAR

    2013-12-01

    Full Text Available Band designs of esterase (EST, peroxidase (PO and polyphenol oxidase (PPO isozymes in several selected cultivars of Catharanthus roseus by using native polyacrylamide gel electrophoresis (PAGE were investigated in this study. It was confirmed that cultivar differences in isozyme polymorphism can be revealed by applied electrophoretic patterns. Three isozyme systems produced a total of 16 bands with polymorphism ranged from 66.6-100%. Considering the patterns of isozyme variations in the five cultivars of Catharanthus roseus, it is evident that the cultivar ‘First kiss coral’ displayed crimson red petal with large white eye’ displayed demarked profiles of EST, PO and PPO isozymes than other cultivars. This is the first report on isozyme polymorphism in members of the Cathanarathus roseus (L. G. Don.

  1. Characterization of a new potyvirus causing mosaic and flower variegation in Catharanthus roseus in Brazil

    Sheila Conceição Maciel

    2011-12-01

    Full Text Available Catharanthus roseus is a perennial, evergreen herb in the family Apocynaceae, which is used as ornamental and for popular medicine to treat a wide assortment of human diseases. This paper describes a new potyvirus found causing mosaic symptom, foliar malformation and flower variegation in C. roseus. Of 28 test-plants inoculated mechanically with this potyvirus, only C. roseus and Nicotiana benthamiana developed systemic mosaic, whereas Chenopodium amaranticolor and C. quinoa exhibited chlorotic local lesions. The virus was transmitted by Aphis gossypii and Myzus nicotianae. When the nucleotide sequence of the CP gene (768nt was compared with other members of the Potyviridae family, the highest identities varied from 67 to 76 %. For the 3' UTR (286nt, identities varied from 16.8 to 28.6 %. The name Catharanthus mosaic virus (CatMV is proposed for this new potyvirus.

  2. Interaction Effects of Arbuscular Mycorrhizal Fungi and Different Phosphate Levels on Growth Performance of Catharanthus roseus Linn.

    Mohd AYOOB; Irfan AZIZ; Paramjit Kaur JITE

    2011-01-01

    Catharanthus roseus L. (Apocynaceae), a valuable medicinal plant with potential therapeutic value was inoculated with AM fungi Glomus fasciculatum under three different phosphate conditions. Catharanthus roseus plants raised in presence of the AM fungi showed increased growth in terms of (shoot length, root length, leaf number, fresh weight and dry weight). Total chlorophyll content and phosphate content of the shoot was found to be significantly higher in AM inoculated plants as compared to ...

  3. Influence of native arbuscular mycorrhizal fungi on growth, nutrition and phytochemical constituents of Catharanthus roseus (L.) G. Don.

    Rajendran Srinivasan; Chinnavenkataraman Govindasamy

    2014-01-01

    Objective: To study the isolation, identification, mass production and the effect of native arbuscular mycorrhizal fungi (AM fungi) on growth parameters of the Catharanthus roseus (C. roseus). Methods: A total of nine different AM fungi species such as Acaulospora scrobiculata, Acaulospora marrowae, Glomus aggregatum (G. aggregatum), Glomus fasciculatum, Glomus geosporum, Gigaspora margarita, Gigaspora nigra, Scutellospora heterogama and Scutellospora pellucida were isolated...

  4. Assessment of genetic diversity in a highly valuable medicinal plant Catharanthus roseus using molecular markers

    Ranjan Kumar Shaw

    2009-01-01

    Full Text Available Genetic diversity was evaluated among 14 cultivars of Catharanthus roseus using RAPD and ISSR markers.The RAPD primers resulted in the amplification of 56 bands, among which 46 (82% bands were polymorphic Four ISSRprimers amplified 31 loci out of which 17 were polymorphic and 14 are monomorphic. The Jaccard's similarity derived fromthe combined marker system showed that the varieties First Kiss Coral and Cooler Orchid were the most closely relatedcultivars, with 98% similarity. In the dendrogram constructed on the basis of both RAPD and ISSR data two clear clusterswere obtained. The smaller cluster included C. roseus Cv Blue Pearl and C. roseus Cv. Patricia White and the larger clusterwas subdivided into two sub clusters with C. roseus Cv. First Kiss Polka Dot isolated from the rest of the cultivars. This maybe useful for breeding for improved quality.

  5. Cytogenetic characterization and genome size of the medicinal plant Catharanthus roseus (L.) G. Don

    Guimarães, Guilherme; Cardoso, Luísa; Oliveira, Helena; Santos, Conceição; Duarte, Patrícia; Sottomayor, Mariana

    2012-01-01

    Background and aims Catharanthus roseus is a highly valuable medicinal plant producing several terpenoid indole alkaloids (TIAs) with pharmaceutical applications, including the anticancer agents vinblastine and vincristine. Due to the interest in its TIAs, C. roseus is one of the most extensively studied medicinal plants and has become a model species for the study of plant secondary metabolism. However, very little is known about the cytogenetics and genome size of this species, in spite of their importance for breeding programmes, TIA genetics and emerging genomic research. Therefore, the present paper provides a karyotype description and fluorescence in situ hybridization (FISH) data for C. roseus, as well as a rigorous characterization of its genome size. Methodology The organization of C. roseus chromosomes was characterized using several DNA/chromatin staining techniques and FISH of rDNA. Genome size was investigated by flow cytometry using an optimized methodology. Principal results The C. roseus full chromosome complement of 2n = 16 includes two metacentric, four subtelocentric and two telocentric chromosome pairs, with the presence of a single nucleolus organizer region in chromosome 6. An easy and reliable flow cytometry protocol for nuclear genome analysis of C. roseus was optimized, and the C-value of this species was estimated to be 1C = 0.76 pg, corresponding to 738 Mbp. Conclusions The organization and size of the C. roseus genome were characterized, providing an important basis for future studies of this important medicinal species, including further cytogenetic mapping, genomics, TIA genetics and breeding programmes. PMID:22479673

  6. Advances of Ornamental Catharanthus roseus%观赏用长春花研究进展

    石林; 何丽贞

    2013-01-01

    文章对观赏用长春花的种质资源、育种、基础生理研究及生产栽培等作了阐述,并对长春花研究方向和应用前景作了分析.%Catharanthus roseus is simultaneously an ornamental and medicinal plant. The researches on germplasm resources, breeding, basic physiology research and cultivation of C. roseus are discussed, and its research direction and application prospect were analyzed.

  7. PHOTOCHEMICAL INVESTIGATION OF VINBLASTINE IN 43 CULTIVARS OF CATHARANTHUS ROSEUS L.

    S.E AJADI; Verpoorte, R.

    2000-01-01

    Background. Vinblastine is one of the alkaloids extracted from Catharanthus roseus L. which is used for the treatment of Hodgkin's disease, lymphoma, and leukemia in children. In spite of the progresses in the synthesis of many drugs, synthesis of vinblastine is not feasible and it has been extracted from plant so far. In this study we used a screening method to facilitate selection for the best cultivar with a higher yield. Methods. The leaves of the 43 cultivars of C. roseus L was free...

  8. Enhanced catharanthine and vindoline production in suspension cultures of Catharanthus roseus by ultraviolet-B light

    Ramani, Shilpa; Jayabaskaran, Chelliah

    2008-01-01

    Suspension cultures of Catharanthus roseus were used to evaluate ultraviolet-B (UV-B) treatment as an abiotic elicitor of secondary metabolites. A dispersed cell suspension culture from C. roseus leaves in late exponential phase and stationary phase were irradiated with UV-B for 5 min. The stationary phase cultures were more responsive to UV-B irradiation than late exponential phase cultures. Catharanthine and vindoline increased 3-fold and 12-fold, respectively, on treatment with a 5-min UV-B irradiation. PMID:18439256

  9. Catharanthus roseus flower extract has wound-healing activity in Sprague Dawley rats

    Nayak, BS; Pinto Pereira, Lexley M

    2006-01-01

    Background Catharanthus roseus L (C. roseus) has been used to treat a wide assortment of diseases including diabetes. The objective of our study was to evaluate the antimicrobial and wound healing activity of the flower extract of Catharanthus in rats. Methods Wound healing activity was determined in rats, after administration (100 mg kg-1 day-1) of the ethanol extract of C. roseus flower, using excision, incision and dead space wounds models. The animals were divided into two groups of 6 each in all the models. In the excision model, group 1 animals were topically treated with carboxymethyl cellulose as placebo control and group 2 received topical application of the ethanol extract of C. roseus at a dose of 100 mg/kg body weight/day. In an incision and dead space model group 1 animals were given normal saline and group 2 received the extract orally at a dose of 100 mg kg-1 day-1. Healing was assessed by the rate of wound contraction, period of epithelization, tensile strength (skin breaking strength), granulation tissue weight, and hydoxyproline content. Antimicrobial activity of the flower extract against four microorganisms was also assessed Results The extract of C. roseus significantly increased the wound breaking strength in the incision wound model compared with controls (P < 0.001). The extract-treated wounds were found to epithelialize faster, and the rate of wound contraction was significantly increased in comparison to control wounds (P < 0.001), Wet and dry granulation tissue weights, and hydroxyproline content in a dead space wound model increased significantly (p < 0.05). Pseudomonas aeruginosa and Staphylococcus aureus demonstrated sensitivity to C. roseus Conclusion Increased wound contraction and tensile strength, augmented hydroxyproline content along with antimicrobial activity support the use of C. roseus in the topical management of wound healing. PMID:17184528

  10. Biochemical and Ultrastructural Changes in Sida cordifolia L. and Catharanthus roseus L. to Auto Pollution

    Verma, Vijeta; Chandra, Neelam

    2014-01-01

    Auto pollution is the by-product of our mechanized mobility, which adversely affects both plant and human life. However, plants growing in the urban locations provide a great respite to us from the brunt of auto pollution by absorbing the pollutants at their foliar surface. Foliar surface configuration and biochemical changes in plant species, namely, Sida cordifolia L. and Catharanthus roseus L. grown at roadside (polluted site 1, Talkatora; polluted site 2, Charbagh) in Lucknow city and in ...

  11. UV-B induced transcript accumulation of DAHP synthase in suspension-cultured Catharanthus roseus cells

    Ramani, Shilpa; Patil, Nandadevi; Jayabaskaran, Chelliah

    2010-01-01

    The enzyme 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthase (EC 4.1.2.15) catalyzes the first committed step in the shikimate pathway of tryptophan synthesis, an important precursor for the production of terpenoid indole alkaloids (TIAs). A full-length cDNA encoding nuclear coded chloroplast-specific DAHP synthase transcript was isolated from a Catharanthus roseus cDNA library. This had high sequence similarity with other members of plant DAHP synthase family. This transcript accum...

  12. Characterization of 10-Hydroxygeraniol Dehydrogenase from Catharanthus roseus Reveals Cascaded Enzymatic Activity in Iridoid Biosynthesis

    Ramakrishnan Krithika; Prabhakar Lal Srivastava; Bajaj Rani; Kolet, Swati P.; Manojkumar Chopade; Mantri Soniya; Hirekodathakallu V. Thulasiram

    2015-01-01

    Catharanthus roseus [L.] is a major source of the monoterpene indole alkaloids (MIAs), which are of significant interest due to their therapeutic value. These molecules are formed through an intermediate, cis-trans-nepetalactol, a cyclized product of 10-oxogeranial. One of the key enzymes involved in the biosynthesis of MIAs is an NAD(P)+ dependent oxidoreductase system, 10-hydroxygeraniol dehydrogenase (Cr10HGO), which catalyses the formation of 10-oxogeranial from 10-hydroxygeraniol via 10-...

  13. Analyses of Catharanthus roseus and Arabidopsis thaliana WRKY transcription factors reveal involvement in jasmonate signaling

    Schluttenhofer, Craig; Pattanaik, Sitakanta; Patra, Barunava; Ling YUAN

    2014-01-01

    Background To combat infection to biotic stress plants elicit the biosynthesis of numerous natural products, many of which are valuable pharmaceutical compounds. Jasmonate is a central regulator of defense response to pathogens and accumulation of specialized metabolites. Catharanthus roseus produces a large number of terpenoid indole alkaloids (TIAs) and is an excellent model for understanding the regulation of this class of valuable compounds. Recent work illustrates a possible role for the...

  14. Effect of Glomus species on physiology and biochemistry of Catharanthus roseus

    Ratti, Neelima; H N Verma; Gautam, S. P.

    2010-01-01

    The present study on efficacy of different Glomus species, an arbuscular mycorrhizal (AM) fungus (G. aggregatum, G. fasciculatum, G. mosseae, G. intraradices) on various growth parameters such as biomass, macro and micronutrients, chlorophyll, protein, cytokinin and alkaloid content and phosphatase activity of pink flowered Catharanthus roseus plants showed that all Glomus species except G. intraradices enhanced the chlorophyll, protein, crude alkaloid, phosphorus, sulphur, manganese and copp...

  15. Somatic embryo mediated mass production of Catharanthus roseus in culture vessel (bioreactor) – A comparative study

    Mujib, A.; Ali, Muzamil; Isah, Tasiu; Dipti

    2014-01-01

    The purpose of this study was to evaluate and compare the use of liquid and solid Murashige and Skoog (MS) medium in different culture vessels for mass production of Catharanthus roseus, an important source of anticancerous compounds, vincristine and vinblastine. Three media conditions i.e. agar-solidified medium (S), liquid medium in agitated conical flask (L) and growtek bioreactor (B) were used. Rapid propagation was achieved through in vitro somatic embryogenesis pathway. The process of e...

  16. ATP-binding cassette transporter controls leaf surface secretion of anticancer drug components in Catharanthus roseus

    Yu, Fang; De Luca, Vincenzo

    2013-01-01

    The presence of biologically active monoterpenoid indole alkaloids (MIAs) on the leaf surfaces of medicinally important Catharanthus roseus has led to questions about the secretion processes involved and their prevalence within MIA-producing species of plants. This report shows that a transporter closely related to those involved in cuticle assembly in plants and belonging to the pleiotropic drug resistance family of ATP-binding cassette transporters is specialized for transport of the MIA ca...

  17. Development of a kinetic metabolic model: application to Catharanthus roseus hairy root

    Leduc, M.; Tikhomiroff, C.; Cloutier, M.; Perrier, M.; Jolicoeur, M.

    2006-01-01

    A kinetic metabolic model describing Catharanthus roseus hairy root growth and nutrition was developed. The metabolic network includes glycolysis, pentose-phosphate pathway, TCA cycle and the catabolic reactions leading to cell building blocks such as amino acids, organic acids, organic phosphates, lipids and structural hexoses. The central primary metabolic network was taken at pseudo-steady state and metabolic flux analysis technique allowed reducing from 31 metabolic fluxes to 20 independe...

  18. A reliable protocol for transformation of Catharanthus roseus through Agrobacterium tumefaciens

    Srivastava, Toolika; Das, Sandip; Sopory, Sudhir Kumar; Srivastava, P. S.

    2009-01-01

    Proliferation of axillary shoot buds and multiple shoot formation in Catharanthus roseus was obtained in 96 % explants on MS medium (3 % sucrose) containing NAA + BA. 2,4-D induced callusing in both, the nodal as well as in leaf segments. Leaf-derived callus was used for transformation with Agrobacterium tumefaciens LBA4404/pBI-S1. Bacterial cell concentration, duration of co-cultivation and acetosyringone concentration influenced transformation efficiency. Under optimal co-cultivation condit...

  19. Endophytic filamentous fungi from a Catharanthus roseus: Identification and its hydrolytic enzymes

    Farah Wahida Ayob; Khanom Simarani

    2016-01-01

    This paper reported on the various filamentous fungi strains that were isolated from a wild grown Catharanthus roseus. Based on the morphological characteristics and molecular technique through a Polymerase Chain Reaction and DNA sequencing method using internal transcribed spacer (ITS), these fungi had been identified as a Colletotrichum sp., Macrophomina phaseolina, Nigrospora sphaerica and Fusarium solani. The ultrastructures of spores and hyphae were observed under a Scanning Electron Mic...

  20. Assessment of genetic diversity in a highly valuable medicinal plant Catharanthus roseus using molecular markers

    Ranjan Kumar Shaw; Laxmikanta Acharya; Arup Kumar Mukherjee

    2009-01-01

    Genetic diversity was evaluated among 14 cultivars of Catharanthus roseus using RAPD and ISSR markers.The RAPD primers resulted in the amplification of 56 bands, among which 46 (82%) bands were polymorphic Four ISSRprimers amplified 31 loci out of which 17 were polymorphic and 14 are monomorphic. The Jaccard's similarity derived fromthe combined marker system showed that the varieties First Kiss Coral and Cooler Orchid were the most closely relatedcultivars, with 98% similarity. In the dendro...

  1. Isolation and Characterization of Antineoplastic Alkaloids from Catharanthus Roseus L. Don. Cultivated in Egypt

    Shams, Khaled A; Nazif, Naglaa M; Abdel Azim, Nahla S; Abdel Shafeek, Khaled A; El-Missiry, Mostafa M.; Ismail, Shams I; Seif El Nasr, Medhat M

    2009-01-01

    Vinblastine and vincristine (the antileukemic agents) were isolated, in a pure form, from Catharanthus roseus L. Don., cultivated in Egypt, by several chromatographic techniques. Five modified methods for the preparation of total alkaloids were carried out. All the isolated mixtures were evaluated by HPLC and HPTLC analyses. The antineoplastic alkaloids; vinblastine and vincristine, were isolated by the use of vacuum liquid chromatographic column on silica gel : aluminium oxide (1:1) mixed be...

  2. Induced dwarf mutant in Catharanthus roseus with enhanced antibacterial activity

    Verma A; Singh R

    2010-01-01

    Evaluation of an ethyl methane sulphonate-induced dwarf mutant of Catharanthus roseus (L.) G. Don revealed that the mutant exhibited marked variation in morphometric parameters. The in vitro antibacterial activity of the aqueous and alcoholic leaf extracts of the mutant and control plants was investigated against medically important bacteria. The mutant leaf extracts showed enhanced antibacterial activity against all the tested bacteria except Bacillus subtilis.

  3. [Identification and expression analysis of WRKY transcription factors in medicinal plant Catharanthus roseus].

    Yang, Zhirong; Wang, Xingchun; Xue, Jin'ai; Meng, Lingzhi; Li, Runzhi

    2013-06-01

    WRKY transcription factors, one of the largest families of transcriptional regulators in plants, involve in multiple life activities including plant growth and development as well as stress responses. However, little is known about the types and functions of WRKY transcription factors in Catharanthus roseus, an important medicinal plant. In this study, we identified 47 CrWRKY transcriptional factors from 26 009 proteins in Catharanthus roseus, and classified them into three distinct groups (G1, G2 and G3) according to the structure of WRKY domain and evolution of the protein family. The expression profiling showed that these CrWRKY genes expressed in a tissue/organ specific manner. The 47 CrWRKY genes were clustered into three types of expression patterns. The first type includes the CrWRKYs highly expressed in flowers and the protoplast treated with methy jasmonate (MeJA) or yeast extraction (YE). The second type contains the CrWRKYs highly expressed in stem and hairy root. The third type represents the CrWRKYs highly expressed in root, stem, leaf, seedling and the hairy root treated by MeJA. Real time quantitative PCR was employed to further identify the expression patterns of the 16 selected CrWRKY genes in various organs, the MeJA-treated protoplasts and hairy roots of Catharanthus roseus, and similar results were obtained. Notably, the expresion of more than 1/3 CrWRKY genes were regulated by MeJA or YE, indicating that these CrWRKYs are likely involed in the signalling webs which modulate the biosynthesis of terpenoid indole alkaloid and plant responses to various stresses. The present results provide a framework for functional identification of the CrWRKYs and understanding of the regulation network of terpenoid indole alkaloid biosynthesis in Catharanthus roseus. PMID:24063238

  4. Somatic Embryos in Catharanthus roseus: A Scanning Electron Microscopic Study

    Aslam, Junaid; MUJIB, Abdul; Mahendra Prasad SHARMA

    2014-01-01

    Catharanthus roseus (L.) G. Don is an important medicinal plant as it contains several anti-cancerous compounds, like vinblastine and vincristine. Plant tissue culture technology (organogenesis and embryogenesis) has currently been used in fast mass propagating raw materials for secondary metabolite synthesis. In this present communication, scanning electron microscopic (SEM) study of somatic embryos was conducted and discussed. The embryogenic callus was first induced from hypocotyls of in ...

  5. Effect of Salinity Stress on Seed Germination Catharanthus roseus Don. Cvs. Rosea and Alba

    Zahra Rezaee; Mehrangiz Chehrazi; Norollah Moalemi

    2012-01-01

    This experiment was carried out aiming to determine the Catharanthus roseus Don resistance against salinity, due to the increasing salinity of soil, and the importance of this plant as an ornamental flower, as well as the little information available on its tolerance against salinity during the germination period. In order to an experiment was conducted in randomized completely design factorial. Sodium chloride was used for induce salinity stress. These factors include cultivar and salinity l...

  6. Influence of Some Heavy Metals on Growth, Alkaloid Content and Composition in Catharanthus roseus L.

    Srivastava, N. K.; A.K. Srivastava

    2010-01-01

    Shoot biomass production, alkaloid content and composition as influence by cadmium, manganese, nickel and lead at uniform dose of 5 mM were investigated in Catharanthus roseus plants grown in sand culture. Treatment with Mn, Ni, and Pb significantly enhanced total root alkaloid accumulation. Cd and Ni treatment resulted in two-fold where as Pb treatment resulted in three fold increase in serpentine content of roots. The non-significant affect on biomass suggests that plants can withstand meta...

  7. Soil Salinity Alters Growth, Chlorophyll Content, and Secondary Metabolite Accumulation in Catharanthus roseus

    Jaleel, Cheruth Abdul; SANKAR, Beemarao; SRIDHARAN, Ramalingam

    2008-01-01

    The effect of salinity on growth, photosynthetic pigment content, and alkaloid secondary metabolite accumulation were studied in an economically important medicinal plant, Catharanthus roseus (L.) G. Don., under pot culture conditions. Plants were treated with different concentrations of NaCl, (e.g. 50 and 100 mM) 30, 45, 60, and 75 days after sowing (DAS). The plants were uprooted randomly 90 DAS to analyse growth, and chlorophyll and alkaloid content. Salinity affected all the morphological...

  8. In vitro evaluation of crude extracts of Catharanthus roseus for potential antibacterial activity

    Goyal Pankaj

    2008-01-01

    Full Text Available Context: Catharanthus roseus (periwinkle is an important medicinal plant, mentioned in Ayurveda, an ancient Indian Sanskrit literature. The plant is selected to evaluate the possibility for novel pharmaceuticals since most of the bacterial pathogens are developing resistance against currently available antibiotics. Aims: To determine the antibacterial activity of crude extracts from different parts of Catharanthus roseus against several bacterial species of clinical significance. Materials and Methods: Extraction of each plant part in appropriate solvent followed by evaluation of antibacterial activity by agar well diffusion assay against a total of six bacterial stains. Further, minimum inhibitory concentration(s was evaluated for active crude extracts. Results: Data indicated that the pattern of inhibition depends largely upon the extraction procedure, the plant part used for extraction, state of plant part (fresh or dry, solvent used for extraction and the microorganism tested. Dry powder extracts of all plant parts demonstrated more antibacterial activity than extracts prepared from fresh parts. Furthermore, extracts prepared from leaves were shown to have better efficacy than stem, root, and flower extracts. Organic extracts provided more potent antibacterial activity as compared to aqueous extracts. Among all the extracts, the ethanolic extract was found to be most active against almost all the bacterial species tested. Hot water and cold water extracts were completely inactive. Gram-positive bacteria were found more sensitive than Gram-negative bacteria. Conclusions: The study promises an interesting future for designing potentially active antibacterial agents from Catharanthus roseus.

  9. Comparative studies of elemental composition in leaves and flowers of Catharanthus roseus growing in Bangladesh

    Shahin Aziz; Koushik Saha; Nasim Sultana; Husna Parvin Nur; Md Aminul Ahsan; Shamim Ahmed; Md Kamal Hossain

    2016-01-01

    Objective: To investigate the elemental composition of the leaves and flowers of Catharanthus roseus (C. roseus) due to the plant's wide application in the indigenous medicinal system and its chemical constituents' importance. Methods: The atomic absorption spectrophotometer was used for quantitative analysis of various elements. Results: Total 13 important elements were analyzed in leaves and flowers of C. roseus. Results indicated the presence of Na, K, Ca, Mg, Cr, Fe, Zn, Al, Cu, Ni, Pb, Cd and Mn in both leaves and flowers. The most important finding of the work was that, leaves of C. roseus showed high concentration of all elements except K and Zn while flowers of C. roseus showed higher concentration of K and Zn. Conclusions: The elemental composition in both leaves and flowers of C. roseus were found to be different. Therefore, different parts of this medicinal plant are enriched in some micro and macro nutrients like Fe, Ca, Na, K, Zn, which are very important for biological metabolic system as well as human health.

  10. Transcriptome analysis of Catharanthus roseus for gene discovery and expression profiling.

    Verma, Mohit; Ghangal, Rajesh; Sharma, Raghvendra; Sinha, Alok K; Jain, Mukesh

    2014-01-01

    The medicinal plant, Catharanthus roseus, accumulates wide range of terpenoid indole alkaloids, which are well documented therapeutic agents. In this study, deep transcriptome sequencing of C. roseus was carried out to identify the pathways and enzymes (genes) involved in biosynthesis of these compounds. About 343 million reads were generated from different tissues (leaf, flower and root) of C. roseus using Illumina platform. Optimization of de novo assembly involving a two-step process resulted in a total of 59,220 unique transcripts with an average length of 1284 bp. Comprehensive functional annotation and gene ontology (GO) analysis revealed the representation of many genes involved in different biological processes and molecular functions. In total, 65% of C. roseus transcripts showed homology with sequences available in various public repositories, while remaining 35% unigenes may be considered as C. roseus specific. In silico analysis revealed presence of 11,620 genic simple sequence repeats (excluding mono-nucleotide repeats) and 1820 transcription factor encoding genes in C. roseus transcriptome. Expression analysis showed roots and leaves to be actively participating in bisindole alkaloid production with clear indication that enzymes involved in pathway of vindoline and vinblastine biosynthesis are restricted to aerial tissues. Such large-scale transcriptome study provides a rich source for understanding plant-specialized metabolism, and is expected to promote research towards production of plant-derived pharmaceuticals. PMID:25072156

  11. Transcriptome analysis of Catharanthus roseus for gene discovery and expression profiling.

    Mohit Verma

    Full Text Available The medicinal plant, Catharanthus roseus, accumulates wide range of terpenoid indole alkaloids, which are well documented therapeutic agents. In this study, deep transcriptome sequencing of C. roseus was carried out to identify the pathways and enzymes (genes involved in biosynthesis of these compounds. About 343 million reads were generated from different tissues (leaf, flower and root of C. roseus using Illumina platform. Optimization of de novo assembly involving a two-step process resulted in a total of 59,220 unique transcripts with an average length of 1284 bp. Comprehensive functional annotation and gene ontology (GO analysis revealed the representation of many genes involved in different biological processes and molecular functions. In total, 65% of C. roseus transcripts showed homology with sequences available in various public repositories, while remaining 35% unigenes may be considered as C. roseus specific. In silico analysis revealed presence of 11,620 genic simple sequence repeats (excluding mono-nucleotide repeats and 1820 transcription factor encoding genes in C. roseus transcriptome. Expression analysis showed roots and leaves to be actively participating in bisindole alkaloid production with clear indication that enzymes involved in pathway of vindoline and vinblastine biosynthesis are restricted to aerial tissues. Such large-scale transcriptome study provides a rich source for understanding plant-specialized metabolism, and is expected to promote research towards production of plant-derived pharmaceuticals.

  12. Catharanthus roseus: a natural source for the synthesis of silver nanoparticles

    Mukunthan, KS; Elumalai, EK; Patel, Trupti N; Murty, V Ramachandra

    2011-01-01

    Objective To develop a simple rapid procedure for bioreduction of silver nanoparticles (AgNPs) using aqueous leaves extracts of Catharanthus roseus (C. roseus). Methods Characterization were determined by using UV-Vis spectrophotometry, scanning electron microscopy (SEM), energy dispersive X-ray and X-ray diffraction. Results SEM showed the formation of silver nanoparticles with an average size of 67 nm to 48 nm. X-ray diffraction analysis showed that the particles were crystalline in nature with face centered cubic geometry. Conclusions C. roseus demonstrates strong potential for synthesis of silver nanoparticles by rapid reduction of silver ions (Ag+ to Ag0). This study provides evidence for developing large scale commercial production of value-added products for biomedical/nanotechnology-based industries. PMID:23569773

  13. Analysis of Karyotype of Catharanthus roseus(Apocynaceae)%长春花核型的研究

    贾彩红; 代正福; 徐碧玉; 金志强; 张蕾; 陈业渊; 王家保

    2008-01-01

    对长春花属的长春花(Catharanthus roseus(L.)G.Don)、白长春花(C. roseus(L.)G.Don'Albus')和黄长春花(C.roseus(L.)G.Don'Flavus')的染色体数目和核型进行了研究.结果表明,它们的核型公式均为2n=2x=16=2m+12sm+2T,均属于"3A"核型,染色体数目均为2n=16,但它们的端部和中部着丝点染色体在核型分析中的排列次序不同.

  14. Overexpression of ORCA3 and G10H in Catharanthus roseus Plants Regulated Alkaloid Biosynthesis and Metabolism Revealed by NMR-Metabolomics

    Qifang Pan; Quan Wang; Fang Yuan; Shihai Xing; Jingya Zhao; Young Hae Choi; Robert Verpoorte; Yuesheng Tian; Guofeng Wang; Kexuan Tang

    2012-01-01

    In order to improve the production of the anticancer dimeric indole alkaloids in Catharanthuse roseus, much research has been dedicated to culturing cell lines, hairy roots, and efforts to elucidate the regulation of the monoterpenoid indole alkaloid (MIA) biosynthesis. In this study, the ORCA3 (Octadecanoid-derivative Responsive Catharanthus AP2-domain) gene alone or integrated with the G10H (geraniol 10-hydroxylase) gene were first introduced into C. roseus plants. Transgenic C. roseus plan...

  15. Antihyperglycemic activity of Catharanthus roseus leaf powder in streptozotocin-induced diabetic rats

    Karuna Rasineni

    2010-01-01

    Full Text Available Catharanthus roseus Linn (Apocynaceae, is a traditional medicinal plant used to control diabetes, in various regions of the world. In this study we evaluated the possible antidiabetic and hypolipidemic effect of C. roseus (Catharanthus roseus leaf powder in diabetic rats. Diabetes was induced by intraperitoneal injection of streptozotocin (STZ, 55 mg/kg body wt to male Wistar rats. The animals were divided into four groups: Control, control-treated, diabetic, and diabetic-treated group. Diabetic-treated and control-treated rats were treated with C. roseus leaf powder suspension in 2 ml distilled water, orally (100 mg/kg body weight/day/60 days. In diabetic rats (D-group the plasma glucose was increased and the plasma insulin was decreased gradually. In the diabetic-treated group lowering of plasma glucose and an increase in plasma insulin were observed after 15 days and by the end of the experimental period the plasma glucose had almost reached the normal level, but insulin had not. The significant enhancement in plasma total cholesterol, triglycerides, LDL and VLDL-cholesterol, and the atherogenic index of diabetic rats were normalized in diabetic-treated rats. Decreased hepatic and muscle glycogen content and alterations in the activities of enzymes of glucose metabolism (glycogen phosphorylase, hexokinase, phosphofructokinase, pyruvate kinase, and glucose-6-phosphate dehydrogenase, as observed in the diabetic control rats, were prevented with C. roseus administration. Our results demonstrated that C. roseus with its antidiabetic and hypolipidemic properties could be a potential herbal medicine in treating diabetes.

  16. Antihyperglycemic activity of Catharanthus roseus leaf powder in streptozotocin-induced diabetic rats

    Rasineni, Karuna; Bellamkonda, Ramesh; Singareddy, Sreenivasa Reddy; Desireddy, Saralakumari

    2010-01-01

    Catharanthus roseus Linn (Apocynaceae), is a traditional medicinal plant used to control diabetes, in various regions of the world. In this study we evaluated the possible antidiabetic and hypolipidemic effect of C. roseus (Catharanthus roseus) leaf powder in diabetic rats. Diabetes was induced by intraperitoneal injection of streptozotocin (STZ, 55 mg/kg body wt) to male Wistar rats. The animals were divided into four groups: Control, control-treated, diabetic, and diabetic-treated group. Diabetic-treated and control-treated rats were treated with C. roseus leaf powder suspension in 2 ml distilled water, orally (100 mg/kg body weight/day/60 days). In diabetic rats (D-group) the plasma glucose was increased and the plasma insulin was decreased gradually. In the diabetic-treated group lowering of plasma glucose and an increase in plasma insulin were observed after 15 days and by the end of the experimental period the plasma glucose had almost reached the normal level, but insulin had not. The significant enhancement in plasma total cholesterol, triglycerides, LDL and VLDL-cholesterol, and the atherogenic index of diabetic rats were normalized in diabetic-treated rats. Decreased hepatic and muscle glycogen content and alterations in the activities of enzymes of glucose metabolism (glycogen phosphorylase, hexokinase, phosphofructokinase, pyruvate kinase, and glucose-6-phosphate dehydrogenase), as observed in the diabetic control rats, were prevented with C. roseus administration. Our results demonstrated that C. roseus with its antidiabetic and hypolipidemic properties could be a potential herbal medicine in treating diabetes. PMID:21808566

  17. Binary stress induces an increase in indole alkaloid biosynthesis in Catharanthus roseus.

    Zhu, Wei; Yang, Bingxian; Komatsu, Setsuko; Lu, Xiaoping; Li, Ximin; Tian, Jingkui

    2015-01-01

    Catharanthus roseus is an important medicinal plant, which produces a variety of indole alkaloids of significant pharmaceutical relevance. In the present study, we aimed to investigate the potential stress-induced increase of indole alkaloid biosynthesis in C. roseus using proteomic technique. The contents of the detectable alkaloids ajmalicine, vindoline, catharanthine, and strictosidine in C. roseus were significantly increased under binary stress. Proteomic analysis revealed that the abundance of proteins related to tricarboxylic acid cycle and cell wall was largely increased; while, that of proteins related to tetrapyrrole synthesis and photosynthesis was decreased. Of note, 10-hydroxygeraniol oxidoreductase, which is involved in the biosynthesis of indole alkaloid was two-fold more abundant in treated group compared to the control. In addition, mRNA expression levels of genes involved in the indole alkaloid biosynthetic pathway indicated an up-regulation in their transcription in C. roseus under UV-B irradiation. These results suggest that binary stress might negatively affect the process of photosynthesis in C. roseus. In addition, the induction of alkaloid biosynthesis appears to be responsive to binary stress. PMID:26284098

  18. Radiation Processed Carrageenan Improves Plant Growth, Physiological Activities, and Alkaloids Production in Catharanthus roseus L.

    M. Naeem

    2015-01-01

    Full Text Available Catharanthus roseus (L. G. Don (Apocynaceae is a medicinal plant that produces indole alkaloids used in cancer chemotherapy. Commercially important antineoplastic alkaloids, namely, vinblastine and vincristine, are mainly present in the leaves of C. roseus. Gamma-rays irradiated carrageenan (ICR has been proven as plant growth promoting substance for a number of medicinal and agricultural plants. Considering the importance of ICR as a promoter of plant growth and alkaloids production in C. roseus, a pot experiment was carried out to explore the effect of ICR on the plant growth, physiological activities, and production of anticancer alkaloids in C. roseus at 120 and 150 days after planting (DAP. Foliar application of ICR (at 0, 20, 40, 60, 80, and 100 mg L−1 significantly improved the performance of C. roseus. 80 mg L−1 of ICR enhanced the leaf yield by 29.2 and 35.4% and the herbage yield by 32.5 and 37.4% at 120 and 150 DAP, respectively, over the control. The spray of ICR at 80 mg L−1 increased the yield of vinblastine by 64.3 and 65.0% and of vincristine by 75.5 and 77.0% at 120 and 150 DAP, respectively, as compared to the control.

  19. Binary Stress Induces an Increase in Indole Alkaloid Biosynthesis in Catharanthus roseus

    Wei eZhu

    2015-07-01

    Full Text Available Catharanthus roseus is an important medicinal plant, which produces a variety of indole alkaloids of significant pharmaceutical relevance. In the present study, we aimed to investigate the potential stress-induced increase of indole alkaloid biosynthesis in C. roseus using proteomic technique. The contents of the detectable alkaloids ajmalicine, vindoline, catharanthine, and strictosidine in C. roseus were significantly increased under binary stress. Proteomic analysis revealed that the abundance of proteins related to tricarboxylic acid cycle and cell wall was largely increased; while, that of proteins related to tetrapyrrole synthesis and photosynthesis was decreased. Of note, 10-hydroxygeraniol oxidoreductase, which is involved in the biosynthesis of indole alkaloid was two-fold more abundant in treated group compared to that in control. In addition, mRNA expression levels of genes involved in the indole alkaloid biosynthetic pathway indicated an up-regulation in their transcription in C. roseus under UV-B irradiation. These results suggest that binary stress might negatively affect the process of photosynthesis in C. roseus. In addition, the induction of alkaloid biosynthesis appears to be responsive to binary stress.

  20. A virus-induced gene silencing approach to understanding alkaloid metabolism in Catharanthus roseus

    Liscombe, David K.; O’Connor, Sarah E.

    2011-01-01

    The anticancer agents vinblastine and vincristine are bisindole alkaloids derived from coupling vindoline and catharanthine, monoterpenoid indole alkaloids produced exclusively by Madagascar periwinkle (Catharanthus roseus) plants. Industrial production of vinblastine and vincristine currently relies on isolation from C. roseus leaves, a process that affords these compounds in 0.0003–0.01% yields. Metabolic engineering efforts to improve alkaloid content or provide alternative sources of the bisindole alkaloids ultimately rely on the isolation and characterization of the genes involved. Several vindoline biosynthetic genes have been isolated, and the cellular and subcellular organization of the corresponding enzymes has been well studied. However, due to the leaf-specific localization of vindoline biosynthesis, and the lack of production of this precursor in cell suspension and hairy root cultures of C. roseus, further elucidation of this pathway demands the development of reverse genetics approaches to assay gene function in planta. The bipartite pTRV vector system is a Tobacco Rattle Virus-based virus-induced gene silencing (VIGS) platform that has provided efficient and effective means to assay gene function in diverse plant systems. We have developed a VIGS method to investigate gene function in C. roseus plants using the pTRV vector system. The utility of this approach in understanding gene function in C. roseus leaves is demonstrated by silencing known vindoline biosynthetic genes previously characterized in vitro. PMID:21802100

  1. Production Pattern of Ajmalicine in Catharanthus roseus (L.) G. Don. Cell Aggregates Culture in the Airlift Bioreactor

    RIZKITA RACHMI ESYANTI; AIDA MUSPIAH

    2006-01-01

    A research has been conducted to optimize the rate of aeration and initial weight of cell aggregates in the production of ajmalicine in Catharanthus roseus cell culture in airlift bioreactor. Catharanthus roseus culture were grown in Zenk medium with the addition of 2.50 x 10-6 M naphthalene acetic acid (NAA) and 10-5 M benzyl amino purine (BAP). Cell aggregates were sub-cultured two times before transferring 20 and 30 g/fw of cell aggregates into bioreactor, respectively, and aerated with th...

  2. Interaction Effects of Arbuscular Mycorrhizal Fungi and Different Phosphate Levels on Growth Performance of Catharanthus roseus Linn.

    Mohd AYOOB

    2011-08-01

    Full Text Available Catharanthus roseus L. (Apocynaceae, a valuable medicinal plant with potential therapeutic value was inoculated with AM fungi Glomus fasciculatum under three different phosphate conditions. Catharanthus roseus plants raised in presence of the AM fungi showed increased growth in terms of (shoot length, root length, leaf number, fresh weight and dry weight. Total chlorophyll content and phosphate content of the shoot was found to be significantly higher in AM inoculated plants as compared to non AM Catharanthus plants. The activities of phosphatase enzymes were found to be increased in AM inoculated plants as compared to non AM plants. Root colonization percent was significantly higher in AM inoculated plants at zero and at all three phosphate levels after 60, 90 and 120 days of AM inoculation, but decreased at third phosphate level after 120 days of AM inoculation. The study suggests that Catharanthus roseus is dependent on the mycorrhizal fungi to a large extent for its growth and survival and also shows the potential of AM fungi Glomus fasciculatum in increasing growth and biomass of Catharanthus roseus L.

  3. Selection and validation of reference genes for transcript normalization in gene expression studies in Catharanthus roseus.

    Pollier, Jacob; Vanden Bossche, Robin; Rischer, Heiko; Goossens, Alain

    2014-10-01

    Quantitative Real-Time PCR (qPCR), a sensitive and commonly used technique for gene expression analysis, requires stably expressed reference genes for normalization of gene expression. Up to now, only one reference gene for qPCR analysis, corresponding to 40S Ribosomal protein S9 (RPS9), was available for the medicinal plant Catharanthus roseus, the only source of the commercial anticancer drugs vinblastine and vincristine. Here, we screened for additional reference genes for this plant species by mining C. roseus RNA-Seq data for orthologs of 22 genes known to be stably expressed in Arabidopsis thaliana and qualified as superior reference genes for this model plant species. Based on this, eight candidate C. roseus reference genes were identified and, together with RPS9, evaluated by performing qPCR on a series of different C. roseus explants and tissue cultures. NormFinder, geNorm and BestKeeper analyses of the resulting qPCR data revealed that the orthologs of At2g28390 (SAND family protein, SAND), At2g32170 (N2227-like family protein, N2227) and At4g26410 (Expressed protein, EXP) had the highest expression stability across the different C. roseus samples and are superior as reference genes as compared to the traditionally used RPS9. Analysis of publicly available C. roseus RNA-Seq data confirmed the expression stability of SAND and N2227, underscoring their value as reference genes for C. roseus qPCR analysis. PMID:25058454

  4. Hypoglycaemic Effects of Methanolic Leaf Extract of Catharanthus Roseus (LINN.) G. DON (Apocynaceae) in Normal and Diabetic Mice

    Ojewole, John A O; Adewunmi, Clement O

    2000-01-01

    In Southern Africa, as in many other parts of Africa, Catharanthus roseus (Linn.) G. Don is traditionally used in forklore medicine to treat diabetes and a number of other human ailments. Consequently, the hypoglycaemic effects of methanolic leaf extract of C. roseus have been investigated in streptozotocin (STZ)-induced diabetic (hyperglycaemic) mice as well as in normal (normoglycaemic), STZ-untreated mice. Tolbutamide has been used as reference hypoglycaemic agent. While tolbutamide produc...

  5. Molecular And Radiation Studies On Improving The Ajmalicine Production In Catharanthus roseus

    Elicitations are considered to be an important strategy towards improve in vitro production of secondary metabolites. In seedling cultures, biotic and abiotic elicitors have effectively stimulated the production of plant secondary metabolites. However, molecular basis of elicitor signaling cascades leading to increased production of secondary metabolites of plant cell is largely unknown. Exposure of Catharanthus roseus cultures to low dose of Gamma irradiation was found to increase the amount of catharanthine and transcription of genes encoding tryptophan decarboxylase (TDC) and strictosidine synthase (STR). In the present study, the signaling pathway mediating Gamma irradiation -induced catharanthine accumulation in C. roseus seedling cultures were investigated. Catharanthus roseus seedling cultures were exposed to different low dose of Gamma irradiation in order to induce alkaloid metabolism. The exposure to Gamma irradiation elicitors resulted in the transcriptional activation of tryptophan decarboxylase and in the accumulation of the monoterpenoid indole alkaloids ajmalicine and catharanthine but not of vindoline. The inability of the seedling cultures to produce vindoline was related to a lack of expression of the tryptophan decarboxylase (TDC) and strictosidine synthase (STR) genes.

  6. Structural identification of putative USPs in Catharanthus roseus.

    Bahieldin, Ahmed; Atef, Ahmed; Shokry, Ahmed M; Al-Karim, Saleh; Al Attas, Sanaa G; Gadallah, Nour O; Edris, Sherif; Al-Kordy, Magdy A; Omer, Abdulkader M Shaikh; Sabir, Jamal S M; Ramadan, Ahmed M; Al-Hajar, Abdulrahman S M; Makki, Rania M; Hassan, Sabah M; El-Domyati, Fotouh M

    2015-10-01

    Nucleotide sequences of the C. roseus SRA database were assembled and translated in order to detect putative universal stress proteins (USPs). Based on the known conserved USPA domain, 24 Pfam putative USPA proteins in C. roseus were detected and arranged in six architectures. The USPA-like domain was detected in all architectures, while the protein kinase-like (or PK-like), (tyr)PK-like and/or U-box domains are shown downstream it. Three other domains were also shown to coexist with the USPA domain in C. roseus putative USPA sequences. These domains are tetratricopeptide repeat (or TPR), apolipophorin III (or apoLp-III) and Hsp90 co-chaperone Cdc37. Subsequent analysis divided USPA-like domains based on the ability to bind ATP. The multiple sequence alignment indicated the occurrence of eight C. roseus residues of known features of the bacterial 1MJH secondary structure. The data of the phylogenetic tree indicated several distinct groups of USPA-like domains confirming the presence of high level of sequence conservation between the plant and bacterial USPA-like sequences. PMID:26318047

  7. Subcellular Localization of Enzymes Involved in Indole Alkaloid Biosynthesis in Catharanthus roseus1

    De Luca, Vincenzo; Cutler, Adrian J.

    1987-01-01

    The subcellular localization of enzymes involved in indole alkaloid biosynthesis in leaves of Catharanthus roseus has been investigated. Tryptophan decarboxylase and strictosidine synthase which together produce strictosidine, the first indole alkaloid of this pathway, are both cytoplasmic enzymes. S-Adenosyl-l-methionine: 16-methoxy-2,3-dihydro-3-hydroxytabersonine-N-methyltransferase which catalyses the third to last step in vindoline biosynthesis could be localized in the chloroplasts of Catharanthus leaves and is specifically associated with thylakoids. Acetyl-coenzyme-A-deacetylvindoline-O-acetyltransferase which catalyses the last step in vindoline biosynthesis could also be localized in the cytoplasm. The participation of the chloroplast in this pathway suggests that indole alkaloid intermediates enter and exit this compartment during the biosynthesis of vindoline. PMID:16665811

  8. Development of a kinetic metabolic model: application to Catharanthus roseus hairy root

    Leduc, M.; Tikhomiroff, C.; Cloutier, M.; Perrier, M.

    2006-01-01

    A kinetic metabolic model describing Catharanthus roseus hairy root growth and nutrition was developed. The metabolic network includes glycolysis, pentose-phosphate pathway, TCA cycle and the catabolic reactions leading to cell building blocks such as amino acids, organic acids, organic phosphates, lipids and structural hexoses. The central primary metabolic network was taken at pseudo-steady state and metabolic flux analysis technique allowed reducing from 31 metabolic fluxes to 20 independent pathways. Hairy root specific growth rate was described as a function of intracellular concentration in cell building blocks. Intracellular transport and accumulation kinetics for major nutrients were included. The model uses intracellular nutrients as well as energy shuttles to describe metabolic regulation. Model calibration was performed using experimental data obtained from batch and medium exchange liquid cultures of C. roseus hairy root using a minimal medium in Petri dish. The model is efficient in estimating the growth rate. PMID:16453114

  9. Influence of Some Heavy Metals on Growth, Alkaloid Content and Composition in Catharanthus roseus L.

    Srivastava, N. K.; Srivastava, A. K.

    2010-01-01

    Shoot biomass production, alkaloid content and composition as influence by cadmium, manganese, nickel and lead at uniform dose of 5 mM were investigated in Catharanthus roseus plants grown in sand culture. Treatment with Mn, Ni, and Pb significantly enhanced total root alkaloid accumulation. Cd and Ni treatment resulted in two-fold where as Pb treatment resulted in three fold increase in serpentine content of roots. The non-significant affect on biomass suggests that plants can withstand metal stress at the level tested with positive affect on root alkaloid content. PMID:21969751

  10. Metabolism of Aromatic Amino Acids during the Growth Cycle of Batch Suspension Cultures of Catharanthus roseus

    Nagaoka, Noriko; ASHIHARA, Hiroshi

    1988-01-01

    Profiles of the levels and metabolism of aromatic compounds in suspension-cultured cells of Catharanthus roseus during the growth cycle were determined. The level of total protein-amino acids, i.e., sum of the amounts of amino acids in hydrolyzates of proteins, and the level of total phenolic acids increased after transfer of the cells in the stationary phase to fresh Murashige-Skoog medium. The maximum levels of the proteinamino acids and those of the phenolic acids were observed on days 3-5...

  11. Sub-acute oral toxicity study of methanol leaves extract of Catharanthus roseus in rats

    LYW Kevin; AH Hussin; I Zhari; JH Chin

    2012-01-01

    Objective: To examine the sub-acute (14 d) oral toxic effects of methanol leaves extract ofCatharanthus roseus (C. roseus) (Family: Apocynaceae) on liver and kidney functions in Sprague Dawley (SD) rats. Methods: Twenty four female SD rats were used throughout the experiment. The first group was orally treated with distilled water and served as control, whereas the remaining three groups were orally treated with single dose daily of 0.1 g/kg, 0.5 g/kg, 1 g/kg of C. roseus extract, respectively for 14 d. Cage-side observations were done daily. Any animal died during the experiment was dissected for gross organ examination. Body weight changed, food consumption and water intake were recorded weekly. Blood was collected via cardiac puncture on day-15 and used for determination of serum levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, creatinine and urea. The relative organ weights were also measured. All results were expressed as mean ± S.E.M and analysed using Dunnett’s test. The level of significance was set at P<0.05 when compared to the control group. Results: Repeated oral administration of 0.5 g/kg and 1 g/kg of methanol leaves extract of C. roseus caused mortality and diarrhoea in rats after few days of treatment. There were no significant changes observed in serum biochemical markers, body weight changed, water and food intake and relative organ weight in rats treated with a single dose daily of 0.1 g/kg of C. roseus extract treatment for 14 d when compared to control group. Conclusionds: Fourteen days repeated oral administration of 0.1 g/kg of methanol leaves extract of C. roseus was safe in female SD rats without causing any significant damages to liver and kidney.

  12. Synthesis of silver nanoparticles using leaves of Catharanthus roseus Linn. G. Don and their antiplasmodial activities

    Ponarulselvam, S; Panneerselvam, C; Murugan, K; Aarthi, N; Kalimuthu, K; Thangamani, S

    2012-01-01

    Objective To develop a novel approach for the green synthesis of silver nanoparticles using aqueous leaves extracts of Catharanthus roseus (C. roseus) Linn. G. Don which has been proven active against malaria parasite Plasmodium falciparum (P. falciparum). Methods Characterizations were determined by using ultraviolet-visible (UV-Vis) spectrophotometry, scanning electron microscopy (SEM), energy dispersive X-ray and X-ray diffraction. Results SEM showed the formation of silver nanoparticles with an average size of 35–55 nm. X-ray diffraction analysis showed that the particles were crystalline in nature with face centred cubic structure of the bulk silver with the broad peaks at 32.4, 46.4 and 28.0. Conclusions It can be concluded that the leaves of C. roseus can be good source for synthesis of silver nanoparticle which shows antiplasmodial activity against P. falciparum. The important outcome of the study will be the development of value added products from medicinal plants C. roseus for biomedical and nanotechnology based industries. PMID:23569974

  13. Synthesis of silver nanoparticles using leaves of Catharanthus roseus Linn. G. Don and their antiplasmodial activities

    Ponarulselvam S; Panneerselvam C; Murugan K; Aarthi N; Kalimuthu K; Thangamani S

    2012-01-01

    Objective: To develop a novel approach for the green synthesis of silver nanoparticles using aqueous leaves extracts of Catharanthus roseus (C. roseus) Linn. G. Don which has been proven active against malaria parasite Plasmodium falciparum (P. falciparum). Methods:Characterizations were determined by using ultraviolet-visible (UV-Vis) spectrophotometry, scanning electron microscopy (SEM), energy dispersive X-ray and X-ray diffraction. Results:SEM showed the formation of silver nanoparticles with an average size of 35-55 nm. X-ray diffraction analysis showed that the particles were crystalline in nature with face centred cubic structure of the bulk silver with the broad peaks at 32.4, 46.4 and 28.0. Conclusions: It can be concluded that the leaves of C. roseus can be good source for synthesis of silver nanoparticle which shows antiplasmodial activity against P. falciparum. The important outcome of the study will be the development of value added products from medicinal plants C. roseus for biomedical and nanotechnology based industries.

  14. Computational identification of microRNAs and their targets in Catharanthus roseus expressed sequence tags.

    Pani, Alok; Mahapatra, Rajani Kanta

    2013-12-01

    No study has been performed on identifying microRNAs (miRNAs) and their targets in the medicinal plant, Catharanthus roseus. In the present study, using the comparative genomics approach, we have predicted two potential C. roseus miRNAs. Furthermore, twelve potential mRNA targets were identified in C. roseus genome based on the characteristics that miRNAs exhibit perfect or nearly perfect complementarity with their targeted mRNA sequences. Among them many of the targets were predicted to encode enzymes that regulate the biosynthesis of terpenoid indole alkaloids (TIA). In addition, most of the predicted targets were the gene coding for transcription factors which are mainly involved in cell growth and development, signaling and metabolism. This is the first in silico study to indicate that miRNA target gene encoding enzymes involved in vinblastine and vincristine biosynthesis, which may help to understand the miRNA-mediated regulation of TIA alkaloid biosynthesis in C. roseus. PMID:26484050

  15. PHOTOCHEMICAL INVESTIGATION OF VINBLASTINE IN 43 CULTIVARS OF CATHARANTHUS ROSEUS L.

    S.E AJADI

    2000-12-01

    Full Text Available Background. Vinblastine is one of the alkaloids extracted from Catharanthus roseus L. which is used for the treatment of Hodgkin's disease, lymphoma, and leukemia in children. In spite of the progresses in the synthesis of many drugs, synthesis of vinblastine is not feasible and it has been extracted from plant so far. In this study we used a screening method to facilitate selection for the best cultivar with a higher yield. Methods. The leaves of the 43 cultivars of C. roseus L was freeze dried and vinblastine was extracted with trifluroacetic acid 0.06% in water. The extraction was analysed by HPLC, using a gradient solvent system. Results and Discussion. The results of this study indicates that the amount of vinblastine is variable in different cultivars. The concentration of vinblastine in cultivar NO. 41 [C. roseus L (G. DON] is 5.1 times mare than the means of vinblastine in all cultivars. The cultivar No. 23 [C. roseus L (Pacifica Punch] has the minimum concentration of this alkaloid.

  16. UV-B induced transcript accumulation of DAHP synthase in suspension-cultured Catharanthus roseus cells

    2010-01-01

    The enzyme 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthase (EC 4.1.2.15) catalyzes the first committed step in the shikimate pathway of tryptophan synthesis, an important precursor for the production of terpenoid indole alkaloids (TIAs). A full-length cDNA encoding nuclear coded chloroplast-specific DAHP synthase transcript was isolated from a Catharanthus roseus cDNA library. This had high sequence similarity with other members of plant DAHP synthase family. This transcript accumulated in suspension cultured C. roseus cells on ultraviolet (UV-B) irradiation. Pretreatment of C.roseus cells with variety of agents such as suramin, N-acetyl cysteine, and inhibitors of calcium fluxes and protein kinases and MAP kinase prevented this effect of UV-B irriadiation. These data further show that the essential components of the signaling pathway involved in accumulation DAHP synthase transcript in C. roseus cells include suramin-sensitive cell surface receptor, staurosporine-sensitive protein kinase and MAP kinase. PMID:20704760

  17. ‘Candidatus Phytoplasma hispanicum’, a novel taxon associated with Mexican periwinkle virescence disease of Catharanthus roseus

    Mexican periwinkle virescence (MPV) phytoplasma was originally discovered in diseased plants of Madagascar periwinkle (Catharanthus roseus) in Yucatán, Mexico. On the basis of results from RFLP analysis of PCR-amplified 16S rRNA gene sequences, strain MPV was previously classified as the first know...

  18. 长春花组织培养研究进展%Progress on tissue culture of Catharanthus roseus

    曾智发; 柳润辉

    2012-01-01

    长春花含有100多种吲哚类生物碱,具有抗肿瘤、降血压等多种生物活性,有较高的药用价值,但是含量偏低.长春花组织培养可从提高繁殖系数、调控次生代谢产物的累积等来提高长春花生物碱类成分含量.本文就外植体和培养基的选择、药用成分累积的影响因素、悬浮细胞培养应用、影响毛状根因素和基因工程技术应用等方面,综述了长春花组织培养的主要研究进展,为进一步开发利用长春花药物资源提供参考.%Catharanthus roseus ( L. ) G. Don contained more than 100 known indole alkaloids, some of which had anli-cancer, lowering blood pressure and other biological activity. Catharanthus roseus had a high medicinal value, but the content of alkaloids was very low. In order to improve the content of alkaloid, the technology of tissue culture had been applied in Catharanthus roseus, such as improving the propagation coefficient and controlling the accumulation of secondary metabolites. To provide reference in further development and utilization of Catharanthus roseus drug resources, the main research progress of tissue culture of Catharanthus roseus including the impact factors of medicinal components accumulation, suspension cell culture applications, the impact factors of hairy roots and application of genetic engineering technology were reviewed in this paper.

  19. Development of efficient catharanthus roseus regeneration and transformation system using agrobacterium tumefaciens and hypocotyls as explants

    2012-01-01

    Background As a valuable medicinal plant, Madagascar periwinkle (Catharanthus roseus) produces many terpenoid indole alkaloids (TIAs), such as vindoline, ajamlicine, serpentine, catharanthine, vinblastine and vincristine et al. Some of them are important components of drugs treating cancer and hypertension. However, the yields of these TIAs are low in wild-type plants, and the total chemical synthesis is impractical in large scale due to high-cost and their complicated structures. The recent development of metabolic engineering strategy offers a promising solution. In order to improve the production of TIAs in C. roseus, the establishment of an efficient genetic transformation method is required. Results To develop a genetic transformation method for C. roseus, Agrobacterium tumefaciens strain EHA105 was employed which harbors a binary vector pCAMBIA2301 containing a report β-glucuronidase (GUS) gene and a selectable marker neomycin phosphotransferase II gene (NTPII). The influential factors were investigated systematically and the optimal transformation condition was achieved using hypocotyls as explants, including the sonication treatment of 10 min with 80 W, A. tumefaciens infection of 30 min and co-cultivation of 2 d in 1/2 MS medium containing 100 μM acetosyringone. With a series of selection in callus, shoot and root inducing kanamycin-containing resistance media, we successfully obtained stable transgenic regeneration plants. The expression of GUS gene was confirmed by histochemistry, polymerase chain reaction, and genomic southern blot analysis. To prove the efficiency of the established genetic transformation system, the rate-limiting gene in TIAs biosynthetic pathway, DAT, which encodes deacetylvindoline-4-O-acetyltransferase, was transferred into C. roseus using this established system and 9 independent transgenic plants were obtained. The results of metabolite analysis using high performance liquid chromatography (HPLC) showed that

  20. Development of efficient catharanthus roseus regeneration and transformation system using agrobacterium tumefaciens and hypocotyls as explants

    Wang Quan

    2012-06-01

    Full Text Available Abstract Background As a valuable medicinal plant, Madagascar periwinkle (Catharanthus roseus produces many terpenoid indole alkaloids (TIAs, such as vindoline, ajamlicine, serpentine, catharanthine, vinblastine and vincristine et al. Some of them are important components of drugs treating cancer and hypertension. However, the yields of these TIAs are low in wild-type plants, and the total chemical synthesis is impractical in large scale due to high-cost and their complicated structures. The recent development of metabolic engineering strategy offers a promising solution. In order to improve the production of TIAs in C. roseus, the establishment of an efficient genetic transformation method is required. Results To develop a genetic transformation method for C. roseus, Agrobacterium tumefaciens strain EHA105 was employed which harbors a binary vector pCAMBIA2301 containing a report β-glucuronidase (GUS gene and a selectable marker neomycin phosphotransferase II gene (NTPII. The influential factors were investigated systematically and the optimal transformation condition was achieved using hypocotyls as explants, including the sonication treatment of 10 min with 80 W, A. tumefaciens infection of 30 min and co-cultivation of 2 d in 1/2 MS medium containing 100 μM acetosyringone. With a series of selection in callus, shoot and root inducing kanamycin-containing resistance media, we successfully obtained stable transgenic regeneration plants. The expression of GUS gene was confirmed by histochemistry, polymerase chain reaction, and genomic southern blot analysis. To prove the efficiency of the established genetic transformation system, the rate-limiting gene in TIAs biosynthetic pathway, DAT, which encodes deacetylvindoline-4-O-acetyltransferase, was transferred into C. roseus using this established system and 9 independent transgenic plants were obtained. The results of metabolite analysis using high performance liquid chromatography (HPLC

  1. Antidiabetic and antioxidant properties of alkaloids from Catharanthus roseus (L.) G. Don.

    Tiong, Soon Huat; Looi, Chung Yeng; Hazni, Hazrina; Arya, Aditya; Paydar, Mohammadjavad; Wong, Won Fen; Cheah, Shiau-Chuen; Mustafa, Mohd Rais; Awang, Khalijah

    2013-01-01

    Catharanthus roseus (L.) G. Don is a herbal plant traditionally used by local populations in India, South Africa, China and Malaysia to treat diabetes. The present study reports the in vitro antioxidant and antidiabetic activities of the major alkaloids isolated from Catharanthus roseus (L.) G. Don leaves extract. Four alkaloids--vindoline I, vindolidine II, vindolicine III and vindolinine IV--were isolated and identified from the dichloromethane extract (DE) of this plant's leaves. DE and compounds I-III were not cytotoxic towards pancreatic β-TC6 cells at the highest dosage tested (25.0 µg/mL). All four alkaloids induced relatively high glucose uptake in pancreatic β-TC6 or myoblast C2C12 cells, with III showing the highest activity. In addition, compounds II-IV demonstrated good protein tyrosine phosphatase-1B (PTP-1B) inhibition activity, implying their therapeutic potential against type 2 diabetes. III showed the highest antioxidant potential in ORAC and DPPH assays and it also alleviated H₂O₂-induced oxidative damage in β-TC6 cells at 12.5 µg/mL and 25.0 µg/mL. PMID:23955322

  2. Immunological Detection and Quantitation of Tryptophan Decarboxylase in Developing Catharanthus roseus Seedlings 1

    Fernandez, Jesus Alvarez; Owen, Terence G.; Kurz, Wolfgang G. W.; De Luca, Vincenzo

    1989-01-01

    l-Tryptophan decarboxylase (TDC) (EC 4.2.1.27) enzyme activity was induced in cell suspension cultures of Catharanthus roseus after treatment with a Pythium aphanidermatum elicitor preparation. The enzyme was extracted from lyophilized cells containing high levels of TDC and the protein was purified to homogeneity. The pure protein was used to produce highly specific polyclonal antibodies, and an enzyme-linked immunosorbent assay (ELISA) was developed to quantitate the level of TDC antigen during seedling development and in leaves of the mature plant. Western immunoblotting of proteins after SDS-PAGE with anti-TDC antibodies detected several immunoreactive proteins (40, 44, 54.8, 55, and 67 kilodaltons) which appeared at different stages during seedling development and in leaves of the mature plant. The major 54.8 and 55 kilodalton antigenic proteins in immunoblots appeared transiently between days 1 to 5 and 5 to 8 of seedling development, respectively. The 54.8 kilodalton protein was devoid of TDC enzyme activity, whereas the appearance of the 55 kilodalton protein coincided with the appearance of this decarboxylase activity. The minor immunoreactive proteins (40, 44, and 67 kilodaltons) appeared after day 5 of seedling development and in older leaves of the mature plant, and their relationship, if any, to TDC is presently unknown. Results suggest that the synthesis and degradation of TDC protein is highly regulated in Catharanthus roseus and that this regulation follows a preset developmental program. Images Figure 3 Figure 5 PMID:16667047

  3. Antidiabetic and Antioxidant Properties of Alkaloids from Catharanthus roseus (L. G. Don

    Won Fen Wong

    2013-08-01

    Full Text Available Catharanthus roseus (L. G. Don is a herbal plant traditionally used by local populations in India, South Africa, China and Malaysia to treat diabetes. The present study reports the in vitro antioxidant and antidiabetic activities of the major alkaloids isolated from Catharanthus roseus (L. G. Don leaves extract. Four alkaloids—vindoline I, vindolidine II, vindolicine III and vindolinine IV—were isolated and identified from the dichloromethane extract (DE of this plant’s leaves. DE and compounds I–III were not cytotoxic towards pancreatic β-TC6 cells at the highest dosage tested (25.0 µg/mL. All four alkaloids induced relatively high glucose uptake in pancreatic β-TC6 or myoblast C2C12 cells, with III showing the highest activity. In addition, compounds II–IV demonstrated good protein tyrosine phosphatase-1B (PTP-1B inhibition activity, implying their therapeutic potential against type 2 diabetes. III showed the highest antioxidant potential in ORAC and DPPH assays and it also alleviated H2O2-induced oxidative damage in β-TC6 cells at 12.5 µg/mL and 25.0 µg/mL.

  4. Characterization of an endophytic whorl-forming Streptomyces from Catharanthus roseus stems producing polyene macrolide antibiotic.

    Rakotoniriana, Erick Francisco; Chataigné, Gabrielle; Raoelison, Guy; Rabemanantsoa, Christian; Munaut, Françoise; El Jaziri, Mondher; Urveg-Ratsimamanga, Suzanne; Marchand-Brynaert, Jacqueline; Corbisier, Anne-Marie; Declerck, Stéphane; Quetin-Leclercq, Joëlle

    2012-05-01

    An endophytic whorl-forming Streptomyces sp. designated as TS3RO having antifungal activity against a large number of fungal pathogens, including Sclerotinia sclerotiorum, Rhizoctonia solani, Colletotrichum gloeosporioides, Cryphonectria parasitica, Fusarium oxysporum, Pyrenophora tritici-repentis, Epidermophyton floccosum, and Trichophyton rubrum, was isolated from surface-sterilized Catharanthus roseus stems. Preliminary identification showed that Streptomyces cinnamoneus subsp. sparsus was its closest related species. However, strain TS3RO could readily be distinguished from this species using a combination of phenotypic properties, 16S rDNA sequence similarity, and phylogenetic analyses. Thus, the whorl-forming Streptomyces sp. strain TS3RO is likely a new subspecies within the Streptomyces cinnamoneus group. Direct bioautography on a thin-layer chromatography plate with Cladosporium cucumerinum was conducted throughout the purification steps for bioassay-guided isolation of the active antifungal compounds from the crude extract. Structural elucidation of the isolated bioactive compound was obtained via LC-MS spectrometry, UV-visible spectra, and nuclear magnetic resonance data. It revealed that fungichromin, a known methylpentaene macrolide antibiotic, was the main antifungal component of TS3RO strain, as shown by thin-layer chromatography bioautography. This is the first report of an endophytic whorl-forming Streptomyces isolated from the medically important plant Catharanthus roseus. PMID:22524528

  5. The Leaf Epidermome of Catharanthus roseus Reveals Its Biochemical Specialization[W][OA

    Murata, Jun; Roepke, Jonathon; Gordon, Heather; De Luca, Vincenzo

    2008-01-01

    Catharanthus roseus is the sole commercial source of the monoterpenoid indole alkaloids (MIAs), vindoline and catharanthine, components of the commercially important anticancer dimers, vinblastine and vincristine. Carborundum abrasion technique was used to extract leaf epidermis–enriched mRNA, thus sampling the epidermome, or complement, of proteins expressed in the leaf epidermis. Random sequencing of the derived cDNA library established 3655 unique ESTs, composed of 1142 clusters and 2513 singletons. Virtually all known MIA pathway genes were found in this remarkable set of ESTs, while only four known genes were found in the publicly available Catharanthus EST data set. Several novel MIA pathway candidate genes were identified, as demonstrated by the cloning and functional characterization of loganic acid O-methyltransferase involved in secologanin biosynthesis. The pathways for triterpene biosynthesis were also identified, and metabolite analysis showed that oleanane-type triterpenes were localized exclusively to the cuticular wax layer. The pathways for flavonoid and very-long-chain fatty acid biosynthesis were also located in this cell type. The results illuminate the biochemical specialization of Catharanthus leaf epidermis for the production of multiple classes of metabolites. The value and versatility of this EST data set for biochemical and biological analysis of leaf epidermal cells is also discussed. PMID:18326827

  6. Purification and cDNA Cloning of Isochorismate Synthase from Elicited Cell Cultures of Catharanthus roseus

    van Tegelen, Léon J.P.; Moreno, Paolo R.H.; Croes, Anton F.; Verpoorte, Robert; Wullems, George J.

    1999-01-01

    Isochorismate is an important metabolite formed at the end of the shikimate pathway, which is involved in the synthesis of both primary and secondary metabolites. It is synthesized from chorismate in a reaction catalyzed by the enzyme isochorismate synthase (ICS; EC 5.4.99.6). We have purified ICS to homogeneity from elicited Catharanthus roseus cell cultures. Two isoforms with an apparent molecular mass of 64 kD were purified and characterized. The Km values for chorismate were 558 and 319 μm for isoforms I and II, respectively. The isoforms were not inhibited by aromatic amino acids and required Mg2+ for enzyme activity. Polymerase chain reaction on a cDNA library from elicited C. roseus cells with a degenerated primer based on the sequence of an internal peptide from isoform II resulted in an amplification product that was used to screen the cDNA library. This led to the first isolation, to our knowledge, of a plant ICS cDNA. The cDNA encodes a protein of 64 kD with an N-terminal chloroplast-targeting signal. The deduced amino acid sequence shares homology with bacterial ICS and also with anthranilate synthases from plants. Southern analysis indicates the existence of only one ICS gene in C. roseus. PMID:9952467

  7. Virus-induced gene silencing in Catharanthus roseus by biolistic inoculation of tobacco rattle virus vectors.

    Carqueijeiro, I; Masini, E; Foureau, E; Sepúlveda, L J; Marais, E; Lanoue, A; Besseau, S; Papon, N; Clastre, M; Dugé de Bernonville, T; Glévarec, G; Atehortùa, L; Oudin, A; Courdavault, V

    2015-11-01

    Catharanthus roseus constitutes the unique source of several valuable monoterpenoid indole alkaloids, including the antineoplastics vinblastine and vincristine. These alkaloids result from a complex biosynthetic pathway encompassing between 30 and 50 enzymatic steps whose characterisation is still underway. The most recent identifications of genes from this pathway relied on a tobacco rattle virus-based virus-induced gene silencing (VIGS) approach, involving an Agrobacterium-mediated inoculation of plasmids encoding the two genomic components of the virus. As an alternative, we developed a biolistic-mediated approach of inoculation of virus-encoding plasmids that can be easily performed by a simple bombardment of young C. roseus plants. After optimisation of the transformation conditions, we showed that this approach efficiently silenced the phytoene desaturase gene, leading to strong and reproducible photobleaching of leaves. This biolistic transformation was also used to silence a previously characterised gene from the alkaloid biosynthetic pathway, encoding iridoid oxidase. Plant bombardment caused down-regulation of the targeted gene (70%), accompanied by a correlated decreased in MIA biosynthesis (45-90%), similar to results obtained via agro-transformation. Thus, the biolistic-based VIGS approach developed for C. roseus appears suitable for gene function elucidation and can readily be used instead of the Agrobacterium-based approach, e.g. when difficulties arise with agro-inoculations or when Agrobacterium-free procedures are required to avoid plant defence responses. PMID:26284695

  8. Catharanthus roseus mitogen-activated protein kinase 3 confers UV and heat tolerance to Saccharomyces cerevisiae

    Raina, Susheel Kumar; Wankhede, Dhammaprakash Pandhari; Sinha, Alok Krishna

    2013-01-01

    Catharanthus roseus is an important source of pharmaceutically important Monoterpenoid Indole Alkaloids (MIAs). Accumulation of many of the MIAs is induced in response to abiotic stresses such as wound, ultra violet (UV) irradiations, etc. Recently, we have demonstrated a possible role of CrMPK3, a C. roseus mitogen-activated protein kinase in stress-induced accumulation of a few MIAs. Here, we extend our findings using Saccharomyces cerevisiae to investigate the role of CrMPK3 in giving tolerance to abiotic stresses. Yeast cells transformed with CrMPK3 was found to show enhanced tolerance to UV and heat stress. Comparison of CrMPK3 and SLT2, a MAPK from yeast shows high-sequence identity particularly at conserved domains. Additionally, heat stress is also shown to activate a 43 kDa MAP kinase, possibly CrMPK3 in C. roseus leaves. These findings indicate the role of CrMPK3 in stress-induced MIA accumulation as well as in stress tolerance. PMID:23221751

  9. Effects of mercury (II) species on cell suspension cultures of catharanthus roseus

    Zhu, L. (Hangzhou Univ. (China)); Cullen, W.R. (Univ. of British Columbia, Vancouver, British Columbia (Canada))

    1994-11-01

    Mercury has received considerable attention because of its high toxicity. Widespread contamination with mercury poses severe environmental problems despite our extensive knowledge of its toxicity in living systems. It is generally accepted that the toxicity of mercury is related to its oxidation states and species, the organic forms being more toxic than the inorganic forms. In the aquatic environment, the toxicity of mercury depends on the aqueous speciation of the mercuric ion (Hg[sup 2+]). Because of the complex coordination chemistry of mercury in aqueous systems, the nature of the Hg[sup 2+] species present in aquatic environments is influenced greatly by water chemistry (e. g, pH, inorganic ion composition, and dissolved organics). Consequently, the influence of environmental factors on the aqueous speciation of mercury has been the focus of much attention. However, there is very little information available regarding the effects of the species and speciation on Hg (II) toxicity in plant-tissue cultures. Catharanthus roseus (C. roseus), commonly called the Madagascar Periwinkle, is a member of the alkaloid rich family Apocynaceae. The present investigation was concerned with the toxicity of mercury on the growth of C. roseus cell suspension cultures as influenced by mercury (II) species and speciation. The specific objectives of the study were to (a) study the effects of mercury species on the growth of C. roseus cultures from the point of view of environmental biology and toxicology; (b) evaluate the effects of selenate, selenite and selected ligands such as chloride, 1-cysteine in the media on the acute toxicity of mercuric oxide; (c) determine the impact of the initial pH of the culture media on the toxicities of mercuric compounds; (d) discuss the dependence of the toxicity on the chemical species and speciation of Hg (II). 11 refs., 7 figs., 2 tabs.

  10. Radiolytically degraded sodium alginate enhances plant growth, physiological activities and alkaloids production in Catharanthus roseus L.

    M. Naeem

    2015-10-01

    Full Text Available Catharanthus roseus (L. G. Don (Family Apocynaceae is a medicinal plant that produces indole alkaloids used in cancer chemotherapy. The anticancerous alkaloids, viz. vinblastine and vincristine, are mainly present in the leaves of C. roseus. High demand and low yield of these alkaloids in the plant has led to explore the alternative means for their production. Gamma irradiated sodium alginate (ISA has proved as a plant growth promoting substance for various medicinal and agricultural crops. A pot culture experiment was carried out to explore the effect of ISA on plant growth, physiological activities and production of anticancer alkaloids (vinblastine and vincristine in C. roseus at 120 and 150 days after planting (DAP. Foliar application of ISA (0, 20, 40, 60, 80 and 100 mg L−1 significantly improved the performance of C. roseus. 80 mg L−1 of ISA enhanced the leaf-yield by 25.3 and 30.2% and the herbage-yield by 29.4 and 34.4% at 120 and 150 DAP, respectively, as compared to the control. The spray treatment of ISA at 80 mg L−1 improved the yield of vinblastine by 66.7 and 71.4% and of vincristine by 67.6 and 75.6% at 120 and 150 DAP, respectively, in comparison to the control. As compared to control, the application of ISA at 80 mg L−1 resulted in the maximum swell in the content and yield of vindoline, increasing them by 18.9 and 20.8% and by 81.8 and 87.2% at 120 and 150 DAP, respectively.

  11. Phytoremediation of 137Cs from low level nuclear waste using Catharanthus roseus

    Remediation of radionuclides has been carried out using the phytoremediation technology. The green plants have been screened for the uptake potential of radionuclide and found that Catharanthus roseus has the high potential for radionuclides in particular 137Cs. Low level nuclear waste (LLNW) collected from effluent treatment plant, BARC has been characterized for physico-chemical and the presence of traces of radionuclides. LLNW was spiked with 3.7 x 104 kBqL-1 activity level of 137Cs. The plants of C. roseus were grown in (i) LLNW, (ii) 137Cs spiked LLNW and (iii) with the control. The radio activities were measured in the solution at the intervals of 0, 1, 2, 3, 6, 8 and 15 days in triplicate set of the experiment. The plants were harvested after the growth. The depletion of 137Cs in LLNW was found to be 19, 21, 24, 38, 60 and 76% at intervals of 1, 2, 3, 6, 8 and 15 days, respectively. The bio-accumulation of 137Cs has been measured in the roots and shoots of the harvested plants. The activity of 137Cs was found higher in shoots (998 kBq g-1 dw) as compared to the roots (735 kBq g-1 dw). The uptake of radionuclide-137Cs, bio-accumulation in the shoot via the active transport from the root, shows the high efficiency and potentiality of C. roseus for the remediation of radionuclide. The bio-accumulation of 137Cs in the shoot will remediate the radionuclide contamination from LLNW. C. roseus can also be made applicable for effective remediation of radionuclides present in the LLNW. (author)

  12. Antimicrobial potentials of Catharanthus roseus by disc diffusion assay.

    Bakht, Jehan; Syed, Fatema; Shafi, Mohammad

    2015-05-01

    The present research work investigates the in vitro antimicrobial activity of different solvent extracted samples from the aerial parts (stem, leaf, fruit and flower) of C. roseus against different microbial species using disc diffusion assay at two different concentrations of 1 and 2 mg disc-1. Hexane extracted samples inhibited the growth of all tested microbial strains except S. typhi. Similarly, ethyl acetate extracted samples was effective to control the activity of all the tested microbial strains. E. coli and S. typhi showed resistance to chloroform extracted samples and the remaining eight microbial strains were susceptible to the same extract. Butanol extracted samples did not inhibit the growth of K. pneumonia and S. typhi at low concentration, however, at higher concentration the same extract reduced the growth of different microbes. Methanol extracted samples effectively controlled the growth of all tested microbes at both concentrations except for S. typhi. Water extracted samples did not inhibit the growth at low concentration except E. coli, K. pneumonia and S. aureus and were ineffective against P. aeroginosa at both concentration. C. albicans, showed resistance against chloroform and water extracted samples at low concentration and susceptible to other solvent extracted samples at both concentration. All fractions were effective against plant pathogens i.e. E. carotovora and A. tumefaciens. PMID:26004715

  13. The juice of fresh leaves of Catharanthus roseus Linn. reduces blood glucose in normal and alloxan diabetic rabbits

    Lodagala Srinivas D; Boini Murthy K; Nammi Srinivas; Behara Ravindra Babu S

    2003-01-01

    Abstract Background The leaf juice or water decoction of Catharanthus roseus L. (Apocyanaceae) is used as a folk medicine for the treatment of diabetes all over the world. In the present investigation, the leaf juice of C. roseus has been evaluated for its hypoglycemic activity in normal and alloxan-induced diabetic rabbits. Methods The blood glucose lowering activity of the leaf juice was studied in normal and alloxan-induced (100 mg/kg, i.v.) diabetic rabbits, after oral administration at d...

  14. Synthesis and characterization of palladium nanoparticles using Catharanthus roseus leaf extract and its application in the photo-catalytic degradation

    Kalaiselvi, Aasaithambi; Roopan, Selvaraj Mohana; Madhumitha, Gunabalan; Ramalingam, C.; Elango, Ganesh

    2015-01-01

    The potential effect of Catharanthus roseus leaf extract for the formation of palladium nanoparticles and its application on dye degradation was discussed. The efficiency of C.roseus leaves are used as a bio-material for the first time as reducing agent. Synthesized palladium nanoparticles were supported by UV-vis spectrometry, XRD, FT-IR and TEM analysis. The secondary metabolites which are responsible for the formation of nanoparticles were identified by GC-MS. The results showed that effect of time was directly related to synthesized nanoparticles and functional groups has a critical role in reducing the metal ions and stabilizing the palladium nanoparticles in an eco-friendly process.

  15. Strictosidine synthase from Catharanthus roseus: purification and characterization of multiple forms.

    de Waal, A; Meijer, A H; Verpoorte, R

    1995-01-01

    Multiple (six) forms of strictosidine synthase from Catharanthus roseus cell suspension cultures were purified and characterized. A purification protocol is presented composed of hydrophobic-interaction, gel-permeation and ion-exchange chromatography and chromatofocusing. Four of six isoforms were purified to apparent homogeneity, whereas two others were nearly homogeneous. All strictosidine synthase isoforms were found to be glycoproteins. The isoforms were also found in leaves and roots of the plant, in seedlings and in hairy root cultures. The ratio of the different isoforms differed slightly between these sources. The kinetic parameters of the isoforms showed no significant differences. The maximal velocity (300-400 nkat/mg of protein) is the highest reported so far. It was demonstrated that the apparent Michaelis constant for tryptamine (approx. 9 microM) is much lower than values reported previously. The presence of weak product inhibition (Kp approx. 35 times Km) was established, whereas substrate inhibition was not detected. PMID:7887913

  16. Endophytic filamentous fungi from a Catharanthus roseus: Identification and its hydrolytic enzymes

    Farah Wahida Ayob

    2016-05-01

    Full Text Available This paper reported on the various filamentous fungi strains that were isolated from a wild grown Catharanthus roseus. Based on the morphological characteristics and molecular technique through a Polymerase Chain Reaction and DNA sequencing method using internal transcribed spacer (ITS, these fungi had been identified as a Colletotrichum sp., Macrophomina phaseolina, Nigrospora sphaerica and Fusarium solani. The ultrastructures of spores and hyphae were observed under a Scanning Electron Microscope. The hydrolytic enzyme test showed that all strains were positive in secreting cellulase. Colletotrichum sp. and F. solani strains also gave a positive result for amylase while only F. solani was capable to secrete protease. These fungi were putatively classified as endophytic fungi since they produced extracellular enzymes that allow them to penetrate plant cell walls and colonize with symbiotic properties.

  17. Uptake and metabolism of sugars by suspension-cultured catharanthus roseus cells

    Ashihara, Hiroshi; Sagishima, Kyoko; Kubota, Kaoru (Ochanomizu Univ., Tokyo (Japan))

    1989-04-01

    The Uptake and metabolism of sugars by suspension-cultured Catharanthus roseus cells were investigated. Substantially all the sucrose in the culture medium was hydrolyzed to glucose and fructose before being taken up by the cells. The activity of invertase bound to cell walls, determined in situ, was high at the early stage of culture. Glucose was more easily taken up by the cells than was fructose. Tracer experiments using (U-{sup 14}C)glucose and (U-{sup 14}C)fructose indicated that glucose is a better precursor for respiration than fructose, while fructose is preferentially utilized for the synthesis of sucrose, especially in the early phase of cell growth. These results suggest that fructose is utilized for the synthesis of sucrose via the reaction catalyzed by sucrose synthase, prior to the phosphorylation by hexokinase or fructokinase.

  18. Morphogenetic and chemical stability of long-term maintained Agrobacterium-mediated transgenic Catharanthus roseus plants.

    Verma, Priyanka; Sharma, Abhishek; Khan, Shamshad Ahmad; Mathur, Ajay Kumar; Shanker, Karuna

    2015-01-01

    Transgenic Catharanthus roseus plants (transgenic Dhawal [DT] and transgenic Nirmal [NT]) obtained from the Agrobacterium tumefaciens and Agrobacterium rhizognenes-mediated transformations, respectively, have been maintained in vitro for 5 years. Plants were studied at regular intervals for various parameters such as plant height, leaf size, multiplication rate, alkaloid profile and presence of marker genes. DT plant gradually lost the GUS gene expression and it was not detected in the fifth year while NT plant demonstrated the presence of genes rolA, rolB and rolC even in the fifth year, indicating the more stable nature of Ri transgene. Vindoline content in the DT was two times more than in non-transformed control plants. Alkaloid and tryptophan profiles were almost constant during the 5 years. The cluster analysis revealed that the DT plant is more close to the control Nirmal plant followed by NT plant. PMID:25102992

  19. Developmental Regulation of Enzymes of Indole Alkaloid Biosynthesis in Catharanthus roseus1

    De Luca, Vincenzo; Fernandez, Jesus Alvarez; Campbell, Douglas; Kurz, Wolfgang G. W.

    1988-01-01

    Developing seedlings of Catharanthus roseus were analyzed for appearance of tryptophan decarboxylase (TDC), strictosidine synthase (SS), N-methyltransferase (NMT) and O-acetyltransferase (DAT) enzyme activities. SS enzyme activity appeared early after germination and was present throughout most of the developmental study. TDC activity was highly regulated and peaked over a 48 hour period achieving a maximum by day of 5 of seedling development. Both TDC and SS were present in all tissues of the seedling. NMT and DAT enzyme activities were induced after TDC and SS had peaked and these activities could only be found in hypocotyls and cotyledons. TDC, SS, and NMT did not require light for induction whereas DAT enzyme activity was increased approximately 10-fold after light treatment of dark grown seedlings. PMID:16665928

  20. Effects of tip-pruning treatment on source-sink regulation of Catharanthus roseus seedlings

    GAO Yang; ZHANG Xue-ke; GUO Xiao-rui; SUN Yan-fei; ZU Yuang-gang

    2006-01-01

    Fifty cultivated Catharanthus roseus seedlings were selected for tip-pruning treatment and the effects of tip-pruning on seedling growth and source-sink regulation were investigated for revealing physiological mechanisms of plants. The results showed that tip-pruning treatment resulted in obvious inhibition of apical dominance and enhancement of branching numbers. The contents of soluble sugars, acid sucrose invertase activity (AI) had a great change in differently positional leaves of the seedling. The sink strength in tip leaves of seedlings dramatically declined after tip-pruning treatment, while that in the leaves at the middle and bottom of seedlings had no obvious changes. The inhibition of apical dominance of tip leaves of seedlings was caused by the diminished sink strength due to tip-pruning treatment,

  1. Endophytic filamentous fungi from a Catharanthus roseus: Identification and its hydrolytic enzymes.

    Ayob, Farah Wahida; Simarani, Khanom

    2016-05-01

    This paper reported on the various filamentous fungi strains that were isolated from a wild grown Catharanthus roseus. Based on the morphological characteristics and molecular technique through a Polymerase Chain Reaction and DNA sequencing method using internal transcribed spacer (ITS), these fungi had been identified as a Colletotrichum sp., Macrophomina phaseolina, Nigrospora sphaerica and Fusarium solani. The ultrastructures of spores and hyphae were observed under a Scanning Electron Microscope. The hydrolytic enzyme test showed that all strains were positive in secreting cellulase. Colletotrichum sp. and F. solani strains also gave a positive result for amylase while only F. solani was capable to secrete protease. These fungi were putatively classified as endophytic fungi since they produced extracellular enzymes that allow them to penetrate plant cell walls and colonize with symbiotic properties. PMID:27275114

  2. Two new vinblastine-type N-oxide alkaloids from Catharanthus roseus.

    Zhang, Wei-Ku; Xu, Jie-Kun; Tian, Hai-Yan; Wang, Lei; Zhang, Xiao-Qi; Xiao, Xu-Zhi; Li, Ping; Ye, Wen-Cai

    2013-10-01

    Two new vinblastine-type N-oxide alkaloids, 17-desacetoxyvinblastine N'b-oxide (1) and 20'-deoxyvinblastine N'b-oxide (2), were isolated from the leaves of Catharanthus roseus. The structures of 1 and 2 were established by the analysis of their nuclear magnetic resonance and HR-ESI-MS spectroscopic data. All alkaloids were evaluated for their cytotoxic activities against the human hepatocellular carcinoma (HepG2) cell line, human colorectal carcinoma (Lovo) cell line and human breast carcinoma (MCF-7) cell line by the MTT method in vitro, respectively. The results showed that cytotoxic activities of alkaloids 1 and 2 exhibited moderate inhibitory activity on the proliferation of three cancer cells. PMID:23621523

  3. Uptake and metabolism of sugars by suspension-cultured catharanthus roseus cells

    The Uptake and metabolism of sugars by suspension-cultured Catharanthus roseus cells were investigated. Substantially all the sucrose in the culture medium was hydrolyzed to glucose and fructose before being taken up by the cells. The activity of invertase bound to cell walls, determined in situ, was high at the early stage of culture. Glucose was more easily taken up by the cells than was fructose. Tracer experiments using [U-14C]glucose and [U-14C]fructose indicated that glucose is a better precursor for respiration than fructose, while fructose is preferentially utilized for the synthesis of sucrose, especially in the early phase of cell growth. These results suggest that fructose is utilized for the synthesis of sucrose via the reaction catalyzed by sucrose synthase, prior to the phosphorylation by hexokinase or fructokinase

  4. Synthesis of silver nanoparticles using Catharanthus roseus root extract and its larvicidal effects.

    Rajagopal, Thangavel; Jemimah, Irudayaraj Anto Amal; Ponmanickam, Ponnirul; Ayyanar, Muniappan

    2015-11-01

    Phytosynthesis of silver nanoparticles has attracted considerable attention due to their biocompatibility, low toxicity, cost-effectiveness and being a novel method has an eco-friendly approach. Biological activity of root extracts as well as synthesized silver nanoparticles of Catharanthus roseus were evaluated against larvae of Aedes aegyptiand Culex quinquefasciatus. The structure and proportion of the synthesized nanoparticles was defined by exploitation ultraviolet spectrophotometry, X-ray diffraction, fourier transform infrared spectroscopy, energy dispersive X-ray spectroscopy and scanning electron microscopy methods. Reduction of silver ions occurred when silver nitrate solution was treated with aqueous root extract at 60°C. Synthesized silver nanoparticles (AgNPs) were confirmed by analyzing the excitation of surface plasmon resonance (SPR) using UV-vis spectrophotometer at 423 nm. FTIR showed aliphatic amines and alkanes corresponding peaks to be presence of responsible compounds to produced nanoparticles in the reaction mixture. Spherical shaped and crystalline nature of particles was recorded under XRD analysis. Presence of silver metal and 35-55nm sized particles were recorded using EDAX and SEM respectively. Larvicidal activitywas observed after24 hrs of exposure to root extracts and synthesized silver nanoparticles. The highest larval mortality was observed in synthesized silver nanopartiucles against Aedes aegypti (LC50= 2.01 ± 0.34; LC90= 5.29 ± 0.07 at 5.0 mg(-1) concentration) and Culex quinquefasciatus (LC50= 1.18 ± 0.15; LC90= 2.55 ± 0.76 at 3.5 to 5.0 mgl(-1) concentration) respectively. The present study provides evidence that synthesized silver nanoparticles of Catharanthus roseus offer potential source for larvicidal activity againstthe larvae of both dengue and filariasis vectors. PMID:26688962

  5. Fungal endophytes of Catharanthus roseus enhance vindoline content by modulating structural and regulatory genes related to terpenoid indole alkaloid biosynthesis

    Pandey, Shiv S.; Sucheta Singh; C. S. Vivek Babu; Karuna Shanker; Srivastava, N. K.; Ashutosh K Shukla; Alok Kalra

    2016-01-01

    Not much is known about the mechanism of endophyte-mediated induction of secondary metabolite production in Catharanthus roseus. In the present study two fungal endophytes, Curvularia sp. CATDLF5 and Choanephora infundibulifera CATDLF6 were isolated from the leaves of the plant that were found to enhance vindoline content by 229–403%. The isolated endophytes did not affect the primary metabolism of the plant as the maximum quantum efficiency of PSII, net CO2 assimilation, plant biomass and st...

  6. Analysis of Several Popular Cultivars of Madagascar Periwinkle (Catharanthus roseus (L.) G. Don.) using Biochemical Markers

    Owk ANIEL KUMAR; Sape S. TATA; Kancharla PAVAN KUMAR

    2013-01-01

    Band designs of esterase (EST), peroxidase (PO) and polyphenol oxidase (PPO) isozymes in several selected cultivars of Catharanthus roseus by using native polyacrylamide gel electrophoresis (PAGE) were investigated in this study. It was confirmed that cultivar differences in isozyme polymorphism can be revealed by applied electrophoretic patterns. Three isozyme systems produced a total of 16 bands with polymorphism ranged from 66.6-100%. Considering the patterns of isozyme variations in the f...

  7. Ornamental Exterior versus Therapeutic Interior of Madagascar Periwinkle (Catharanthus roseus): The Two Faces of a Versatile Herb

    Naghmeh Nejat; Alireza Valdiani; David Cahill; Yee-How Tan; Mahmood Maziah; Rambod Abiri

    2015-01-01

    Catharanthus roseus (L.) known as Madagascar periwinkle (MP) is a legendary medicinal plant mostly because of possessing two invaluable antitumor terpenoid indole alkaloids (TIAs), vincristine and vinblastine. The plant has also high aesthetic value as an evergreen ornamental that yields prolific blooms of splendid colors. The plant possesses yet another unique characteristic as an amiable experimental host for the maintenance of the smallest bacteria found on earth, the phytoplasmas and spir...

  8. Alterations in seedling vigour and antioxidant enzyme activities in Catharanthus roseus under seed priming with native diazotrophs

    B.Karthikeyan; Jaleel, C.A.; Gopi, R.; Deiveekasundaram, M.

    2007-01-01

    An experiment was conducted on Catharanthus roseus to study the effect of seed treatments with native diazotrophs on its seedling growth and antioxidant enzyme activities. The treatments had significant influence on various seedling parameters. There is no significant influence on dry matter production with the diazotrophs, Azospirillum and Azotobacter. However, the vital seedling parameters such as germination percentage and vigour index were improved. Azotobacter treatment influenced maximu...

  9. Isolasi dan Uji Ekstrak Metanol Bakteri Endofit Tapak Dara (Catharanthus roseus) dalam Menghambat Pertumbuhan Beberapa Mikroba Patogen

    Pandiangan, Febrin Setiani

    2014-01-01

    A study of isolation of endophytic bacteria from tapak dara (Catharanthus roseus) and examination of methanol extracts in inhibited the growth of several pathogenic microbes such as Aspergillus flavus, Streptococcus mutans, Salmonella typhii and Escherichia coli has been conducted. Examination of methanol extract of endophytic bacteria was done with concentrations of 40, 60, 80, and 100%. Methanol extract was prepared by maceration method and centrifugated at 5000 rpm. Examination of isolates...

  10. Effect of Chromium on Antioxidant Potential of Catharanthus roseus Varieties and Production of Their Anticancer Alkaloids: Vincristine and Vinblastine

    Vartika Rai; Pramod Kumar Tandon; Sayyada Khatoon

    2014-01-01

    Catharanthus roseus (L.) G. Don, a medicinal plant, has a very important place in the traditional as well as modern pharmaceutical industry. Two common varieties of this plant rosea and alba are named so because of pink and white coloured flowers, respectively. This plant comprises of about 130 terpenoid indole alkaloids and two of them, vincristine and vinblastine, are common anticancer drugs. The effect of chromium (Cr) on enzymatic and non-enzymatic antioxidant components and on secondary ...

  11. A comparative study of release profiles of Coccinia cordifolia and Catharanthus roseus with standard antidiabetic agent using rat intestine

    Most. Afia Ahktar

    2014-01-01

    Matured leaves of Coccinia cordifolia and Catharanthus roseus were collected, dried and extracted with 95% ethanol. Solvents were evaporated and suspensions at a concentration of 40 mg/ml were prepared from the residues using phosphate buffer. The aim of the study is to elucidate the release pattern of these extracts in acidic and basic environment. 1 ml suspension of each plant was poured into the intestine fragment prepared from sacrificed rats. The filled pieces of intestine were bound ver...

  12. Direct regeneration of Periwinkle (Catharanthus roseus) via node explants culture and different combinations of plant growth regulators

    M. Talebi; F Etesam; B.E. Sayed-Tabatabaei; Gh. Khaksar

    2012-01-01

    Periwinkle (Catharanthus roseus L., Apocynaceae) contains more than 130 different terpenoid indole alkaloids (TIAs), of which two dimeric alkaloids, Vinblastine and Vincristine, have antineoplastic activity and are useful in treatment of various cancers. Specific production of some alkaloids in differentiated tissues such as leaf and stem led to use direct regeneration of explants in order to increase the production of these important alkaloids in the plant. In this research, 30 combinations ...

  13. Biological Synthesis of Zinc oxide Nanoparticles from Catharanthus roseus (l.) G. Don. Leaf extract and validation for antibacterial activity

    G. Bhumi; N. SAVITHRAMMA

    2014-01-01

    Biologically synthesized nanoparticles have been widely using in the field of medicine. Research in nanotechnology highlights the possibility of green chemistry pathways to produce technologically important nanomaterials. Present study focuses on the Biological synthesis of Zinc oxide nanoparticles (ZnO-NPs) by Zinc acetate and sodium hydroxide utilizing the biocomponents of leaves of Catharanthus roseus. The samples were characterized by x-ray diffraction (XRD), Scanning Electron Microscopy ...

  14. Exploiting EST databases for the mining and characterization of short sequence repeat (SSR) markers in Catharanthus roseus L.

    Joshi, Raj Kumar; Kar, Basudeba; Nayak, Sanghamitra

    2011-01-01

    Periwinkle (Catharanthus roseus L.) (Family: Apocyanaceae) is a ornamental plants with great medicinal properties. Although it is represented by seven species, little work has been carried out on its genetic characterization due to non-availability of reliable molecular markers. Simple sequence repeats (SSRs) have been widely applied as molecular markers in genetic studies. With the rapid increase in the deposition of nucleotide sequences in the public databases and advent of bioinformatics t...

  15. PERKEMBANGAN LATISIFER PADA KULTUR KALUS CATHARANTHUS ROSEUS (L) G. DON YANG DIINDUKSI DENGAN KOMBINASI ZAT PENGATUR TUMBUH KINETIN + NAA

    NI NYOMAN DARSINI

    2011-01-01

    The development of laticifer on callus culture of Catharanthus roseus (L) G Don in Zenk medium supplemented with combination of plant growth regulator kinetin + NAA was studied. The explants were taken from the second folium from shoot apex. Development of laticifer was observed using descriptive analysis method for callus anatomy and percentage of laticifer was observed during 4–14 weeks of callus development. The percentage of laticifer was determined by counting the average number of the l...

  16. Alkaloid Accumulation in Catharanthus roseus Increases with Addition of Seawater Salts to the Nutrient Solution

    WANG Jing-Yan; LIU Zhao-Pu

    2010-01-01

    A sand culture experiment was conducted to determine the effects of different seawater (5% and 10%) treatments on plant growth,inorganic ions,indole alkaloid concentrations and yields of Catharanthus roseus,in an effort to increase the alkaloid yield by artificial cultivation.The total fresh and dry weights and tissue K+ concentrations decreased,but Na+ concentrations increased in the plant roots,stems and leaves of C.roseus under seawater stress as compared to the control.The concentrations and yields of vindoline,catharanthine,vinblastine and vincristine increased under seawater stress.The concentrations and yields of these alkaloids were higher in 5% seawater-treated plants than those in the 10% seawater-treated plants.Considering the industrial production,5% seawater treatments could reduce the cost of producing alkaloid.In the control plants,the highest alkaloid concentrations reached a peak at 100 days after planting,suggesting that plant harvest must be optimized in terms of growth duration.

  17. Indole alkaloids from Catharanthus roseus: bioproduction and their effect on human health.

    Almagro, Lorena; Fernández-Pérez, Francisco; Pedreño, Maria Angeles

    2015-01-01

    Catharanthus roseus is a medicinal plant belonging to the family Apocynaceae which produces terpenoid indole alkaloids (TIAs) of high medicinal importance. Indeed, a number of activities like antidiabetic, bactericide and antihypertensive are linked to C. roseus. Nevertheless, the high added value of this plant is based on its enormous pharmaceutical interest, producing more than 130 TIAs, some of which exhibit strong pharmacological activities. The most striking biological activity investigated has been the antitumour effect of dimeric alkaloids such as anhydrovinblastine, vinblastine and vincristine which are already in pre-, clinical or in use. The great pharmacological importance of these indole alkaloids, contrasts with the small amounts of them found in this plant, making their extraction a very expensive process. To overcome this problem, researches have looked for alternative sources and strategies to produce them in higher amounts. In this sense, intensive research on the biosynthesis of TIAs and the regulation of their pathways has been developed with the aim to increase by biotechnological approaches, the production of these high added value compounds. This review is focused on the different strategies which improve TIA production, and in the analysis of the beneficial effects that these compounds exert on human health. PMID:25685907

  18. Study of the effect of nickel heavy metals on some physiological parameters of Catharanthus roseus.

    Arefifard, Matin; Mahdieh, Majid; Amirjani, Mohammadreza

    2014-01-01

    Plants, in their life cycle, are usually exposed to various kinds of non-biological stresses including heavy metals. One of these heavy metals is nickel which affects many physiological processes of plants. Studies have shown that the changes in planting conditions can affect the qualitative and quantitative features of Catharanthus roseus; therefore, creating stressful conditions (e.g. NiCl2) can be an effective way to investigate the changes. In this research, we investigated the effect of 0, 2.5, 5, 10, 25 and 50 mM concentrations of NiCl2 on the degree of catalase enzyme activity, amount of proline aggregation and photosynthetic parameters on seeds of pink variety of C. roseus. The results indicated that the degree of catalase enzyme activity and the amount of proline aggregation increased in plants which were exposed to NiCl2 treatments, especially in high concentrations, while the total protein decreased. The stress of Ni also affected photosynthetic parameters, and decreased the amount of pigments, as well as the efficiency of photosystem II. PMID:24870880

  19. A Stereoselective Hydroxylation Step of Alkaloid Biosynthesis by a Unique Cytochrome P450 in Catharanthus roseus*

    Giddings, Lesley-Ann; Liscombe, David K.; Hamilton, John P.; Childs, Kevin L.; DellaPenna, Dean; Buell, C. Robin; O'Connor, Sarah E.

    2011-01-01

    Plant cytochrome P450s are involved in the production of over a hundred thousand metabolites such as alkaloids, terpenoids, and phenylpropanoids. Although cytochrome P450 genes constitute one of the largest superfamilies in plants, many of the catalytic functions of the enzymes they encode remain unknown. Here, we report the identification and functional characterization of a cytochrome P450 gene in a new subfamily of CYP71, CYP71BJ1, involved in alkaloid biosynthesis. Co-expression analysis of putative cytochrome P450 genes in the Catharanthus roseus transcriptome identified candidate genes with expression profiles similar to known terpene indole alkaloid biosynthetic genes. Screening of these candidate genes by functional expression in Saccharomyces cerevisiae yielded a unique P450-dependent enzyme that stereoselectively hydroxylates the alkaloids tabersonine and lochnericine at the 19-position of the aspidosperma-type alkaloid scaffold. Tabersonine, which can be converted to either vindoline or 19-O-acetylhörhammericine, represents a branch point in alkaloid biosynthesis. The discovery of CYP71BJ1, which forms part of the pathway leading to 19-O-acetylhörhammericine, will help illuminate how this branch point is controlled in C. roseus. PMID:21454651

  20. Functional characterization of amyrin synthase involved in ursolic acid biosynthesis in Catharanthus roseus leaf epidermis.

    Yu, Fang; Thamm, Antje M K; Reed, Darwin; Villa-Ruano, Nemesio; Quesada, Alfonso Lara; Gloria, Edmundo Lozoya; Covello, Patrick; De Luca, Vincenzo

    2013-07-01

    Catharanthus roseus accumulates high levels of the pentacyclic triterpene, ursolic acid, as a component of its wax exudate on the leaf surface. Bioinformatic analyses of transcripts derived from the leaf epidermis provide evidence for the specialized role of this tissue in the biosynthesis of ursolic acid. Cloning and functional expression in yeast of a triterpene synthase derived from this tissue showed it to be predominantly an α-amyrin synthase (CrAS), since the α-amyrin to β-amyrin reaction products accumulated in a 5:1 ratio. Expression analysis of CrAS showed that triterpene biosynthesis occurs predominantly in the youngest leaf tissues and in the earliest stages of seedling development. Further studies using laser capture microdissection to harvest RNA from epidermis, mesophyll, idioblasts, laticifers and vasculature of leaves showed the leaf epidermis to be the preferred sites of CrAS expression and provide conclusive evidence for the involvement of this tissue in the biosynthesis of ursolic acid in C. roseus. PMID:22652241

  1. Gene-to-metabolite networks for terpenoid indole alkaloid biosynthesis in Catharanthus roseus cells

    Rischer, Heiko; Orešič, Matej; Seppänen-Laakso, Tuulikki; Katajamaa, Mikko; Lammertyn, Freya; Ardiles-Diaz, Wilson; Van Montagu, Marc C. E.; Inzé, Dirk; Oksman-Caldentey, Kirsi-Marja; Goossens, Alain

    2006-01-01

    Rational engineering of complicated metabolic networks involved in the production of biologically active plant compounds has been greatly impeded by our poor understanding of the regulatory and metabolic pathways underlying the biosynthesis of these compounds. Whereas comprehensive genome-wide functional genomics approaches can be successfully applied to analyze a select number of model plants, these holistic approaches are not yet available for the study of nonmodel plants that include most, if not all, medicinal plants. We report here a comprehensive profiling analysis of the Madagascar periwinkle (Catharanthus roseus), a source of the anticancer drugs vinblastine and vincristine. Genome-wide transcript profiling by cDNA-amplified fragment-length polymorphism combined with metabolic profiling of elicited C. roseus cell cultures yielded a collection of known and previously undescribed transcript tags and metabolites associated with terpenoid indole alkaloids. Previously undescribed gene-to-gene and gene-to-metabolite networks were drawn up by searching for correlations between the expression profiles of 417 gene tags and the accumulation profiles of 178 metabolite peaks. These networks revealed that the different branches of terpenoid indole alkaloid biosynthesis and various other metabolic pathways are subject to differing hormonal regulation. These networks also served to identify a select number of genes and metabolites likely to be involved in the biosynthesis of terpenoid indole alkaloids. This study provides the basis for a better understanding of periwinkle secondary metabolism and increases the practical potential of metabolic engineering of this important medicinal plant. PMID:16565214

  2. Characterization of 10-hydroxygeraniol dehydrogenase from Catharanthus roseus reveals cascaded enzymatic activity in iridoid biosynthesis.

    Krithika, Ramakrishnan; Srivastava, Prabhakar Lal; Rani, Bajaj; Kolet, Swati P; Chopade, Manojkumar; Soniya, Mantri; Thulasiram, Hirekodathakallu V

    2015-01-01

    Catharanthus roseus [L.] is a major source of the monoterpene indole alkaloids (MIAs), which are of significant interest due to their therapeutic value. These molecules are formed through an intermediate, cis-trans-nepetalactol, a cyclized product of 10-oxogeranial. One of the key enzymes involved in the biosynthesis of MIAs is an NAD(P)(+) dependent oxidoreductase system, 10-hydroxygeraniol dehydrogenase (Cr10HGO), which catalyses the formation of 10-oxogeranial from 10-hydroxygeraniol via 10-oxogeraniol or 10-hydroxygeranial. This work describes the cloning and functional characterization of Cr10HGO from C. roseus and its role in the iridoid biosynthesis. Substrate specificity studies indicated that, Cr10HGO has good activity on substrates such as 10-hydroxygeraniol, 10-oxogeraniol or 10-hydroxygeranial over monohydroxy linear terpene derivatives. Further it was observed that incubation of 10-hydroxygeraniol with Cr10HGO and iridoid synthase (CrIDS) in the presence of NADP(+) yielded a major metabolite, which was characterized as (1R, 4aS, 7S, 7aR)-nepetalactol by comparing its retention time, mass fragmentation pattern, and co-injection studies with that of the synthesized compound. These results indicate that there is concerted activity of Cr10HGO with iridoid synthase in the formation of (1R, 4aS, 7S, 7aR)-nepetalactol, an important intermediate in iridoid biosynthesis. PMID:25651761

  3. Indole Alkaloids from Catharanthus roseus: Bioproduction and Their Effect on Human Health

    Lorena Almagro

    2015-02-01

    Full Text Available Catharanthus roseus is a medicinal plant belonging to the family Apocynaceae which produces terpenoid indole alkaloids (TIAs of high medicinal importance. Indeed, a number of activities like antidiabetic, bactericide and antihypertensive are linked to C. roseus. Nevertheless, the high added value of this plant is based on its enormous pharmaceutical interest, producing more than 130 TIAs, some of which exhibit strong pharmacological activities. The most striking biological activity investigated has been the antitumour effect of dimeric alkaloids such as anhydrovinblastine, vinblastine and vincristine which are already in pre-, clinical or in use. The great pharmacological importance of these indole alkaloids, contrasts with the small amounts of them found in this plant, making their extraction a very expensive process. To overcome this problem, researches have looked for alternative sources and strategies to produce them in higher amounts. In this sense, intensive research on the biosynthesis of TIAs and the regulation of their pathways has been developed with the aim to increase by biotechnological approaches, the production of these high added value compounds. This review is focused on the different strategies which improve TIA production, and in the analysis of the beneficial effects that these compounds exert on human health.

  4. A vacuolar class III peroxidase and the metabolism of anticancer indole alkaloids in Catharanthus roseus

    Duarte, Patrícia; Figueiredo, Raquel; Ros Barceló, Alfonso

    2008-01-01

    Plants possess a unique metabolic diversity commonly designated as secondary metabolism, of which the anticancer alkaloids from Catharanthus roseus are among the most studied. Recently, in a classical function-to-protein-to-gene approach, we have characterized the main class III peroxidase (Prx) expressed in C. roseus leaves, CrPrx1, implicated in a key biosynthetic step of the anticancer alkaloids. We have shown the vacuolar sorting determination of CrPrx1 using GFP fusions and we have obtained further evidence supporting the role of this enzyme in alkaloid biosynthesis, indicating the potential of CrPrx1 as a molecular tool for the manipulation of alkaloid metabolism. Here, we discuss how plant cells may regulate Prx reactions. In fact, Prxs form a large multigenic family whose members accept a broad range of substrates and, in their two subcellular localizations, the cell wall and the vacuole, Prxs co-locate with a large variety of secondary metabolites which can be accepted as substrates. How then, are Prx reactions regulated? Localization data obtained in our lab suggest that arabinogalactan proteins (AGPs) and Prxs may be associated in membrane microdomains, evocative of lipid rafts. Whether plasma membrane and/or tonoplast microcompartmentation involve AGPs and Prxs and whether this enables metabolic channeling determining Prx substrate selection are challenging questions ahead. PMID:19704535

  5. Construction of genetic linkage map of the medicinal and ornamental plant Catharanthus roseus

    Sarika Gupta; Sashi Pandey-Rai; Suchi Srivastava; Subhas Chandra Naithani; Manoj Prasad; Sushil Kumar

    2007-12-01

    An integrated genetic linkage map of the medicinal and ornamental plant Catharanthus roseus, based on different types of molecular and morphological markers was constructed, using a F2 population of 144 plants. The map defines 14 linkage groups (LGs) and consists of 131 marker loci, including 125 molecular DNA markers (76 RAPD, 3 RAPD combinations; 7 ISSR; 2 EST-SSR from Medicago truncatula and 37 other PCR based DNA markers), selected from a total of 472 primers or primer pairs, and six morphological markers (stem pigmentation, leaf lamina pigmentation and shape, leaf petiole and pod size, and petal colour). The total map length is 1131.9 cM (centiMorgans), giving an average map length and distance between two markers equal to 80.9 cM and 8.6 cM, respectively. The morphological markers/genes were found linked with nearest molecular or morphological markers at distances varying from 0.7 to 11.4 cM. Linkage was observed between the morphological markers concerned with lamina shape and petiole size of leaf on LG1 and leaf, stem and petiole pigmentation and pod size on LG8. This is the first genetic linkage map of C. roseus.

  6. CathaCyc, a metabolic pathway database built from Catharanthus roseus RNA-Seq data.

    Van Moerkercke, Alex; Fabris, Michele; Pollier, Jacob; Baart, Gino J E; Rombauts, Stephane; Hasnain, Ghulam; Rischer, Heiko; Memelink, Johan; Oksman-Caldentey, Kirsi-Marja; Goossens, Alain

    2013-05-01

    The medicinal plant Madagascar periwinkle (Catharanthus roseus) synthesizes numerous terpenoid indole alkaloids (TIAs), such as the anticancer drugs vinblastine and vincristine. The TIA pathway operates in a complex metabolic network that steers plant growth and survival. Pathway databases and metabolic networks reconstructed from 'omics' sequence data can help to discover missing enzymes, study metabolic pathway evolution and, ultimately, engineer metabolic pathways. To date, such databases have mainly been built for model plant species with sequenced genomes. Although genome sequence data are not available for most medicinal plant species, next-generation sequencing is now extensively employed to create comprehensive medicinal plant transcriptome sequence resources. Here we report on the construction of CathaCyc, a detailed metabolic pathway database, from C. roseus RNA-Seq data sets. CathaCyc (version 1.0) contains 390 pathways with 1,347 assigned enzymes and spans primary and secondary metabolism. Curation of the pathways linked with the synthesis of TIAs and triterpenoids, their primary metabolic precursors, and their elicitors, the jasmonate hormones, demonstrated that RNA-Seq resources are suitable for the construction of pathway databases. CathaCyc is accessible online (http://www.cathacyc.org) and offers a range of tools for the visualization and analysis of metabolic networks and 'omics' data. Overlay with expression data from publicly available RNA-Seq resources demonstrated that two well-characterized C. roseus terpenoid pathways, those of TIAs and triterpenoids, are subject to distinct regulation by both developmental and environmental cues. We anticipate that databases such as CathaCyc will become key to the study and exploitation of the metabolism of medicinal plants. PMID:23493402

  7. Alterations in seedling vigour and antioxidant enzyme activities in Catharanthus roseus under seed priming with native diazotrophs

    KARTHIKEYAN B.; JALEEL C.A.; GOPI R.; DEIVEEKASUNDARAM M.

    2007-01-01

    An experiment was conducted on Catharanthus roseus to study the effect of seed treatments with native diazotrophs on its seedling growth and antioxidant enzyme activities. The treatments had significant influence on various seedling parameters.There is no significant influence on dry matter production with the diazotrophs, Azospirillum and Azotobacter. However, the vital seedling parameters such as germination percentage and vigour index were improved. Azotobacter treatment influenced maximum of 50% germination, whereas Azospirillum and Azotobacter were on par with C. roseus with respect to their vigour index. There was significant difference in the population of total diazotrophs. Azospirillum and Azotobacter between rhizosphere and non-rhizosphere soils of C. roseus had the same trend and were observed at various locations of the study. The activities of antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POX) and catalase (CAT) were increased to a significant extent due to the treatment with diazotrophs.

  8. Alterations in seedling vigour and antioxidant enzyme activities in Catharanthus roseus under seed priming with native diazotrophs

    Karthikeyan, B.; Jaleel, C.A.; Gopi, R.; Deiveekasundaram, M.

    2007-01-01

    An experiment was conducted on Catharanthus roseus to study the effect of seed treatments with native diazotrophs on its seedling growth and antioxidant enzyme activities. The treatments had significant influence on various seedling parameters. There is no significant influence on dry matter production with the diazotrophs, Azospirillum and Azotobacter. However, the vital seedling parameters such as germination percentage and vigour index were improved. Azotobacter treatment influenced maximum of 50% germination, whereas Azospirillum and Azotobacter were on par with C. roseus with respect to their vigour index. There was significant difference in the population of total diazotrophs. Azospirillum and Azotobacter between rhizosphere and non-rhizosphere soils of C. roseus had the same trend and were observed at various locations of the study. The activities of antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POX) and catalase (CAT) were increased to a significant extent due to the treatment with diazotrophs. PMID:17610323

  9. Enhancement of vindoline and vinblastine production in suspension-cultured cells of Catharanthus roseus by artemisinic acid elicitation.

    Liu, Jinwei; Zhu, Jianhua; Tang, Le; Wen, Wei; Lv, Shuangshuang; Yu, Rongmin

    2014-01-01

    Elicitation is an important strategy to improve production of secondary metabolites in vitro. Artemisinic acid was studied as a novel elicitor to enhance the yield of terpenoid indole alkaloids in the present paper. Our results demonstrated that the concentrations of vindoline and vinblastine were increased by sixfold and twofold, respectively, compared to those of the control group after treatment with artemisinic acid. To elucidate the underlying mechanism, we investigated the gene expression of four enzymes involved in the biosynthetic pathway of vinblastine in the suspension-cultured cells of Catharanthu sroseus. RT-PCR experiment showed that artemisinic acid was able to up-regulate the transcriptions of tryptophan decarboxylase, geraniol 10-hydroxylase, tabersonine 16-hydroxylase and deacetoxyvindoline 4-hydroxylase. PMID:23864440

  10. The juice of fresh leaves of Catharanthus roseus Linn. reduces blood glucose in normal and alloxan diabetic rabbits

    Lodagala Srinivas D

    2003-09-01

    Full Text Available Abstract Background The leaf juice or water decoction of Catharanthus roseus L. (Apocyanaceae is used as a folk medicine for the treatment of diabetes all over the world. In the present investigation, the leaf juice of C. roseus has been evaluated for its hypoglycemic activity in normal and alloxan-induced diabetic rabbits. Methods The blood glucose lowering activity of the leaf juice was studied in normal and alloxan-induced (100 mg/kg, i.v. diabetic rabbits, after oral administration at doses of 0.5, 0.75 and 1.0 ml/kg body weight. Blood samples were collected from the marginal ear vein before and also at 4, 6, 8, 10, 12, 16, 18, 20 & 24 h after drug administration and blood glucose was analyzed by Nelson-Somogyi's method using a visible spectrophotometer. The data was compared statistically by using Student's t-test. Results The leaf juice of C. roseus produced dose-dependent reduction in blood glucose of both normal and diabetic rabbits and comparable with that of the standard drug, glibenclamide. The results indicate a prolonged action in reduction of blood glucose by C. roseus and the mode of action of the active compound(s of C. roseus is probably mediated through enhance secretion of insulin from the β-cells of Langerhans or through extrapancreatic mechanism. Conclusions The present study clearly indicated a significant antidiabetic activity with the leaf juice of Catharanthus roseus and supports the traditional usage of the fresh leaves by Ayurvedic physicians for the control of diabetes.

  11. The juice of fresh leaves of Catharanthus roseus Linn. reduces blood glucose in normal and alloxan diabetic rabbits

    Nammi, Srinivas; Boini, Murthy K; Lodagala, Srinivas D; Behara, Ravindra Babu S

    2003-01-01

    Background The leaf juice or water decoction of Catharanthus roseus L. (Apocyanaceae) is used as a folk medicine for the treatment of diabetes all over the world. In the present investigation, the leaf juice of C. roseus has been evaluated for its hypoglycemic activity in normal and alloxan-induced diabetic rabbits. Methods The blood glucose lowering activity of the leaf juice was studied in normal and alloxan-induced (100 mg/kg, i.v.) diabetic rabbits, after oral administration at doses of 0.5, 0.75 and 1.0 ml/kg body weight. Blood samples were collected from the marginal ear vein before and also at 4, 6, 8, 10, 12, 16, 18, 20 & 24 h after drug administration and blood glucose was analyzed by Nelson-Somogyi's method using a visible spectrophotometer. The data was compared statistically by using Student's t-test. Results The leaf juice of C. roseus produced dose-dependent reduction in blood glucose of both normal and diabetic rabbits and comparable with that of the standard drug, glibenclamide. The results indicate a prolonged action in reduction of blood glucose by C. roseus and the mode of action of the active compound(s) of C. roseus is probably mediated through enhance secretion of insulin from the β-cells of Langerhans or through extrapancreatic mechanism. Conclusions The present study clearly indicated a significant antidiabetic activity with the leaf juice of Catharanthus roseus and supports the traditional usage of the fresh leaves by Ayurvedic physicians for the control of diabetes. PMID:12950994

  12. Cell-specific localization of alkaloids in Catharanthus roseus stem tissue measured with Imaging MS and Single-cell MS.

    Yamamoto, Kotaro; Takahashi, Katsutoshi; Mizuno, Hajime; Anegawa, Aya; Ishizaki, Kimitsune; Fukaki, Hidehiro; Ohnishi, Miwa; Yamazaki, Mami; Masujima, Tsutomu; Mimura, Tetsuro

    2016-04-01

    Catharanthus roseus (L.) G. Don is a medicinal plant well known for producing antitumor drugs such as vinblastine and vincristine, which are classified as terpenoid indole alkaloids (TIAs). The TIA metabolic pathway in C. roseus has been extensively studied. However, the localization of TIA intermediates at the cellular level has not been demonstrated directly. In the present study, the metabolic pathway of TIA in C. roseus was studied with two forefront metabolomic techniques, that is, Imaging mass spectrometry (MS) and live Single-cell MS, to elucidate cell-specific TIA localization in the stem tissue. Imaging MS indicated that most TIAs localize in the idioblast and laticifer cells, which emit blue fluorescence under UV excitation. Single-cell MS was applied to four different kinds of cells [idioblast (specialized parenchyma cell), laticifer, parenchyma, and epidermal cells] in the stem longitudinal section. Principal component analysis of Imaging MS and Single-cell MS spectra of these cells showed that similar alkaloids accumulate in both idioblast cell and laticifer cell. From MS/MS analysis of Single-cell MS spectra, catharanthine, ajmalicine, and strictosidine were found in both cell types in C. roseus stem tissue, where serpentine was also accumulated. Based on these data, we discuss the significance of TIA synthesis and accumulation in the idioblast and laticifer cells of C. roseus stem tissue. PMID:27001858

  13. Metabolic Discrimination of Catharanthus roseus Leaves Infected by Phytoplasma Using 1H-NMR Spectroscopy and Multivariate Data Analysis1

    Choi, Young Hae; Tapias, Elisabet Casas; Kim, Hye Kyong; Lefeber, Alfons W.M.; Erkelens, Cornelis; Verhoeven, Jacobus Th.J.; Brzin, Jernej; Zel, Jana; Verpoorte, Robert

    2004-01-01

    A comprehensive metabolomic profiling of Catharanthus roseus L. G. Don infected by 10 types of phytoplasmas was carried out using one-dimensional and two-dimensional NMR spectroscopy followed by principal component analysis (PCA), an unsupervised clustering method requiring no knowledge of the data set and used to reduce the dimensionality of multivariate data while preserving most of the variance within it. With a combination of these techniques, we were able to identify those metabolites that were present in different levels in phytoplasma-infected C. roseus leaves than in healthy ones. The infection by phytoplasma in C. roseus leaves causes an increase of metabolites related to the biosynthetic pathways of phenylpropanoids or terpenoid indole alkaloids: chlorogenic acid, loganic acid, secologanin, and vindoline. Furthermore, higher abundance of Glc, Glu, polyphenols, succinic acid, and Suc were detected in the phytoplasma-infected leaves. The PCA of the 1H-NMR signals of healthy and phytoplasma-infected C. roseus leaves shows that these metabolites are major discriminating factors to characterize the phytoplasma-infected C. roseus leaves from healthy ones. Based on the NMR and PCA analysis, it might be suggested that the biosynthetic pathway of terpenoid indole alkaloids, together with that of phenylpropanoids, is stimulated by the infection of phytoplasma. PMID:15286294

  14. Adaptation of lettuce mosaic virus to Catharanthus roseus involves mutations in the central domain of the VPg.

    Svanella-Dumas, Laurence; Verdin, Eric; Faure, Chantal; German-Retana, Sylvie; Gognalons, Patrick; Danet, Jean Luc; Marais, Armelle; Candresse, Thierry

    2014-05-01

    An isolate of Lettuce mosaic virus (LMV, a Potyvirus) infecting Madagascar periwinckle (Catharanthus roseus) was identified and characterized by Illumina deep sequencing. LMV-Cr has no close affinities to previously sequenced LMV isolates and represents a novel, divergent LMV clade. Inoculation experiments with other representative LMV isolates showed that they are unable to infect C. roseus, which was not known to be a host for LMV. However, three C. roseus variants of one of these isolates, LMV-AF199, could be selected and partially or completely sequenced. These variants are characterized by the accumulation of mutations affecting the C-terminal part of the cylindrical inclusion (CI) helicase and the central part of the VPg. In particular, a serine to proline mutation at amino acid 143 of the VPg was observed in all three independently selected variants and is also present in the LMV-Cr isolate, making it a prime candidate as a host-range determinant. Other mutations at VPg positions 65 and 144 could also contribute to the ability to infect C. roseus. Inoculation experiments involving a recombinant LMV expressing a permissive lettuce eukaryotic translation initiation factor 4E (eIF4E) suggest that eIF4E does not contribute to the interaction of most LMV isolates with C. roseus. PMID:24400938

  15. Characterization of Alkaloid Uptake by Catharanthus roseus (L.) G. Don Protoplasts 1

    McCaskill, David G.; Martin, DeAndra L.; Scott, A. Ian

    1988-01-01

    The accumulation of alkaloids by protoplasts of Catharanthus roseus (L.) G. Don var. Little Bright Eye was studied to determine the specificity of uptake and the role of ion trapping in the storage of alkaloids. Accumulation of the indole alkaloids vindoline, ajmalicine, tabersonine, and vinblastine was found to be biphasic, with an initial burst of uptake followed by a slow, prolonged phase of accumulation. The concentration and pH dependence of the initial burst of uptake for vindoline suggested that uptake occurred by simple diffusion. Uptake of nicotine was monophasic, with a half life of 5.2 minutes. The accumulation ratio (Ci/Ce) for nicotine at steady state and for the initial burst of uptake for vindoline and ajmalicine suggested that accumulation was driven by the pH gradient between the vacuole and the external assay medium. The second, sustained phase of uptake of vindoline was sensitive to inhibition by either 20 millimolar NaN3 or 0.5 millimolar Cu2+. In azide-treated protoplasts, the uptake for vindoline conformed to the kinetics of simple diffusion, with a half life of 4 minutes. The second phase of uptake for ajmalicine, although sensitive to inhibition by Cu2+, was insensitive to inhibition by NaN3. The biphasic uptake of the indole alkaloids was not due to any significant metabolism. It is concluded that accumulation and storage of the indole alkaloids is due only partly to ion trapping of the alkaloids by the low pH of the vacuole lumen. In the case of vindoline, there appears to be a specific energy-requiring uptake that is not seen with nicotine (which is not endogenous to Catharanthus). Accumulation of ajmalicine appears to involve both ion trapping and an azide-insensitive component, which may be due to complexation with organic counterions and phenolics. PMID:16666154

  16. Influence of native arbuscular mycorrhizal fungi on growth, nutrition and phytochemical constituents of Catharanthus roseus (L.) G. Don

    Rajendran Srinivasan; Chinnavenkataraman Govindasamy

    2014-01-01

    Objective: To study the isolation, identification, mass production and the effect of native arbuscular mycorrhizal fungi (AM fungi) on growth parameters of the Catharanthus roseus (C. roseus).Methods:Acaulospora marrowae, Glomus aggregatum (G. aggregatum), Glomus fasciculatum, Glomusgeosporum, Gigaspora margarita, Gigaspora nigra, Scutellospora heterogama and Scutellospora pellucida were isolated and identified from the root zone soil of C. roseus.Results:A total of nine different AM fungi species such as Acaulospora scrobiculata, The phytochemical analyses showed high concentration of chlorophyll a (0.152±0.0140 µg/g), chlorophyll b (0.081±0.006 µg/g), total chlorophyll (0.233±0.020 µg/g), soluble sugar (0.051±0.004 µg/g), reducing sugar (0.060±0,007 µg/g), phenols (0.293±0.032 µg/g), ortho-dihydroxy phenols (0.275±0.022 µg/g), lipids (0.300±0.025 µg/g), proteins (0.063±0.003 µg/g) and amino acids (1.042±0.056 µg/g) in G. aggregatum inoculated C. roseus. G. aggregatum was found to perform better on growth when compared to others and phytochemical constituents of C. roseus.Conclusions:fasciculatum can be used as a potential growth promoters for the C. roseus for better yielding in the agricultural sectors. It is concluded from the present findings that the G. aggregatum and Glomus fasciculatum can be used as a potential growth promoters for the C. roseus for better yielding in the agricultural sectors.

  17. Development of SSR and gene-targeted markers for construction of a framework linkage map of Catharanthus roseus

    Shokeen, Bhumika; Choudhary, Shalu; Sethy, Niroj Kumar; Bhatia, Sabhyata

    2011-01-01

    Background and Aims Catharanthus roseus is a plant of great medicinal importance, yet inadequate knowledge of its genome structure and the unavailability of genomic resources have been major impediments in the development of improved varieties. The aims of this study were to develop co-dominant sequence-tagged microsatellite sites (STMS) and gene-targeted markers (GTMs) and utilize them for the construction of a framework intraspecific linkage map of C. roseus. Methods For simple sequence repeat (SSR) isolation, a genomic library enriched for (GA)n repeats was constructed from C. roseus ‘Nirmal’ (CrN1). In addition, GTMs were also designed from 12 genes of the TIA (terpenoid indole alkaloid) pathway – the medicinally most significant pathway in C. roseus. An F2 mapping population was also generated by crossing two diverse accessions of C. roseus CrN1 (Nirmal)×CrN82 (Kew). Key Results A new set of 314 STMS markers and 64 GTMs were developed in this study. A segregating F2 mapping population consisting of 111 F2 individuals was generated. For generating the linkage map, a set of 423 co-dominant markers (378 newly developed and 45 published earlier) were screened for polymorphism between the parental genotypes, of which 134 were identified to be polymorphic. A total of 114 markers were mapped on eight linkage groups that spanned a 632·7 cM region of the genome with an average marker distance of 5·55 cM. Further, the mechanism of hypervariability at the gene-targeted loci was investigated at the sequence level. Conclusions For the first time, a large array of STMS markers and GTMs was generated in the model medicinal plant C. roseus. Moreover, the first microsatellite marker-based linkage map was described in this study. Together, these will serve as a foundation for future genomics studies related to quantitative trait loci analysis and molecular breeding in C. roseus. PMID:21788377

  18. Abnormalities in carbohydrate and lipid metabolisms in high-fructose dietfed insulin-resistant rats: amelioration by Catharanthus roseus treatments.

    Rasineni, Karuna; Bellamkonda, Ramesh; Singareddy, Sreenivasa Reddy; Desireddy, Saralakumari

    2013-09-01

    High intake of dietary fructose has been shown to exert a number of adverse metabolic effects in humans and experimental animals. The present study was proposed to elucidate the effect of Catharanthus roseus (C. roseus) leaf powder treatment on alterations in carbohydrate and lipid metabolisms in rats fed with high-fructose diet. Male Wistar rats of body weight around 180 g were divided into four groups, two of these groups (groups C and C+CR) were fed with standard pellet diet and the other two groups (groups F and F+CR) were fed with high-fructose (66 %) diet. C. roseus leaf powder suspension in water (100 mg/kg body weight/day) was administered orally to group C+CR and group F+CR. At the end of a 60-day experimental period, biochemical parameters related to carbohydrate and lipid metabolisms were assayed. C. roseus treatment completely prevented the fructose-induced increased body weight, hyperglycemia, and hypertriglyceridemia. Hyperinsulinemia and insulin resistance observed in group F was significantly decreased with C. roseus treatment in group F+CR. The alterations observed in the activities of enzymes of carbohydrate and lipid metabolisms and contents of hepatic tissue lipids in group F rats were significantly restored to near normal values by C. roseus treatment in group F+CR. In conclusion, our study demonstrates that C. roseus treatment is effective in preventing fructose-induced insulin resistance and hypertriglyceridemia while attenuating the fructose-induced alterations in carbohydrate and lipid metabolisms. This study suggests that the plant can be used as an adjuvant for the prevention and/or management of insulin resistance and disorders related to it. PMID:23334857

  19. Effect of Gloriosa superba and Catharanthus roseus Extracts on IFN-γ-Induced Keratin 17 Expression in HaCaT Human Keratinocytes

    Nattaporn Pattarachotanant; Varaporn Rakkhitawatthana; Tewin Tencomnao

    2014-01-01

    Gloriosa superba and Catharanthus roseus are useful in traditional medicine for treatment of various skin diseases and cancer. However, their molecular effect on psoriasis has not been investigated. In this study, the effect of ethanol extracts derived from G. superba leaves and C. roseus stems on the expression of psoriatic marker, keratin 17 (K17), was investigated in human keratinocytes using biochemical and molecular experimental approaches. Both extracts could reduce the expression of K1...

  20. Induction and Flow Cytometry Identification of Tetraploids from Seed-Derived Explants through Colchicine Treatments in Catharanthus roseus (L.) G. Don

    Shi-Hai Xing; Xin-Bo Guo; Quan Wang; Qi-Fang Pan; Yue-Sheng Tian; Pin Liu; Jing-Ya Zhao; Guo-Feng Wang; Xiao-Fen Sun; Ke-Xuan Tang

    2011-01-01

    The tetraploid plants of Catharanthus roseus (L.) G. Don was obtained by colchicine induction from seeds explants, and the ploidy of the plants was identified by flow cytometry. The optimal treatment is 0.2% colchicine solution treated for 24 hours, and the induction rate reaches up to 30%. Comparing with morphological characteristics and growth habits between tetraploids and the control, we found that tetraploids of C. roseus had larger stoma and more branches and leaves. HPLC analysis showe...

  1. Polyamines and the Cell Cycle of Catharanthus roseus Cells in Culture 1

    Maki, Hisae; Ando, Satoshi; Kodama, Hiroaki; Komamine, Atsushi

    1991-01-01

    Investigation was made on the effect of partial depletion of polyamines (PAs), induced by treatment with inhibitors of the biosynthesis of PAs, on the distribution of cells at each phase of the cell cycle in Catharanthus roseus (L.) G. Don. cells in suspension cultures, using flow cytometry. More cells treated with inhibitors of arginine decarboxylase (ADC) and ornithine decarboxylase (ODC) were accumulated in the G1 phase than those in the control, while the treatment with an inhibitor of spermidine (SPD) synthase showed no effect on the distribution of cells. The endogenous levels of the PAs, putrescine (PUT), SPD, and spermine (SPM), were determined during the cell cycle in synchronous cultures of C. roseus. Two peaks of endogenous level of PAs, in particular, of PUT and SPD, were observed during the cell cycle. Levels of PAs increased markedly prior to synthesis of DNA in the S phase and prior to cytokinesis. Activities of ADC and ODC were also assayed during the cell cycle. Activities of ADC was much higher than that of ODC throughout the cell cycle, but both activities of ODC and ADC changed in concert with changes in levels of PAs. Therefore, it is suggested that these enzymes may regulate PA levels during the cell cycle. These results indicate that inhibitors of PUT biosynthesis caused the suppression of cell proliferation by prevention of the progression of the cell cycle, probably from the G1 to the S phase, and PUT may play more important roles in the progression of the cell cycle than other PAs. PMID:16668290

  2. Sombreamento de plantas de Catharanthus roseus (L.) G. Don 'Pacifica White' por malhas coloridas: desenvolvimento vegetativo Shading of 'Pacifica White' Catharanthus roseus (L.) G. Don plants with colored nets: vegetative development

    Anderson Adriano Martins Melo; Amauri Alves de Alvarenga

    2009-01-01

    As malhas coloridas têm sido utilizadas para manipular o desenvolvimento vegetativo, melhorando a utilização da radiação solar por plantas ornamentais. Objetivou-se, neste trabalho estudar o efeito da redução de 50% da radiação fotossinteticamente ativa sobre o crescimento vegetativo de plantas de Catharanthus roseus (L.) G. Don, por meio de malhas azul e vermelha e malha preta, em comparação com plantas crescidas na ausência de sombreamento (pleno sol). As plantas foram obtidas a partir de s...

  3. Multicellular compartmentation of catharanthus roseus alkaloid biosynthesis predicts intercellular translocation of a pathway intermediate

    St-Pierre, B; Vazquez-Flota, FA; De Luca V

    1999-01-01

    In situ RNA hybridization and immunocytochemistry were used to establish the cellular distribution of monoterpenoid indole alkaloid biosynthesis in Madagascar periwinkle (Catharanthus roseus). Tryptophan decarboxylase (TDC) and strictosidine synthase (STR1), which are involved in the biosynthesis of the central intermediate strictosidine, and desacetoxyvindoline 4-hydroxylase (D4H) and deacetylvindoline 4-O-acetyltransferase (DAT), which are involved in the terminal steps of vindoline biosynthesis, were localized. tdc and str1 mRNAs were present in the epidermis of stems, leaves, and flower buds, whereas they appeared in most protoderm and cortical cells around the apical meristem of root tips. In marked contrast, d4h and dat mRNAs were associated with the laticifer and idioblast cells of leaves, stems, and flower buds. Immunocytochemical localization for TDC, D4H, and DAT proteins confirmed the differential localization of early and late stages of vindoline biosynthesis. Therefore, we concluded that the elaboration of the major leaf alkaloids involves the participation of at least two cell types and requires the intercellular translocation of a pathway intermediate. A basipetal gradient of expression in maturing leaves also was shown for all four genes by in situ RNA hybridization studies and by complementary studies with dissected leaves, suggesting that expression of the vindoline pathway occurs transiently during early leaf development. These results partially explain why attempts to produce vindoline by cell culture technology have failed. PMID:10330473

  4. Colonization of Madagascar periwinkle (Catharanthus roseus), by endophytes encoding gfp marker.

    Torres, Adalgisa Ribeiro; Araújo, Welington Luiz; Cursino, Luciana; de Barros Rossetto, Priscilla; Mondin, Mateus; Hungria, Mariangela; Azevedo, João Lúcio

    2013-07-01

    This study reports the introduction of gfp marker in two endophytic bacterial strains (Pantoea agglomerans C33.1, isolated from cocoa, and Enterobacter cloacae PR2/7, isolated from citrus) to monitor the colonization in Madagascar perinwinkle (Catharanthus roseus). Stability of the plasmid encoding gfp was confirmed in vitro for at least 72 h of bacterial growth and after the colonization of tissues, under non-selective conditions. The colonization was observed using fluorescence microscopy and enumeration of culturable endophytes in inoculated perinwinkle plants that grew for 10 and 20 days. Gfp-expressing strains were re-isolated from the inner tissues of surface-sterilized roots and stems of inoculated plants, and the survival of the P. agglomerans C33:1gfp in plants 20 days after inoculation, even in the absence of selective pressure, suggests that is good colonizer. These results indicated that both gfp-tagged strains, especially P. agglomerans C33.1, may be useful tools to deliver enzymes or other proteins in plant. PMID:23695435

  5. Effect of Salinity Stress on Seed Germination Catharanthus roseus Don. Cvs. Rosea and Alba

    Zahra Rezaee

    2012-03-01

    Full Text Available This experiment was carried out aiming to determine the Catharanthus roseus Don resistance against salinity, due to the increasing salinity of soil, and the importance of this plant as an ornamental flower, as well as the little information available on its tolerance against salinity during the germination period. In order to an experiment was conducted in randomized completely design factorial. Sodium chloride was used for induce salinity stress. These factors include cultivar and salinity levels with Electrical Conductivity (EC of 1, 2, 4, 6, 8, and 10 dS/m. The results of variance analysis showed that salinity effect at level 1% on the germination percentage, germination rate, radicle and plumule length, seedling wet and dry weight and ratio of plumule length to radicle length were significant and they were reduced. While the cultivar and interaction of cultivar and salinity treatments were non significant. According to the results, response of two cultivars to the salinity was the same and there was no difference between them.

  6. Kinetic Analysis of Phospholipase C from Catharanthus roseus Transformed Roots Using Different Assays1

    Hernández-Sotomayor, S.M. Teresa; De Los Santos-Briones, César; Muñoz-Sánchez, J. Armando; Loyola-Vargas, Victor M.

    1999-01-01

    The properties of phospholipase C (PLC) partially purified from Catharanthus roseus transformed roots were analyzed using substrate lipids dispersed in phospholipid vesicles, phospholipid-detergent mixed micelles, and phospholipid monolayers spread at an air-water interface. Using [33P]phosphatidylinositol 4,5-bisphosphate (PIP2) of high specific radioactivity, PLC activity was monitored directly by measuring the loss of radioactivity from monolayers as a result of the release of inositol phosphate and its subsequent dissolution on quenching in the subphase. PLC activity was markedly affected by the surface pressure of the monolayer, with reduced activity at extremes of initial pressure. The optimum surface pressure for PIP2 hydrolysis was 20 mN/m. Depletion of PLC from solution by incubation with sucrose-loaded PIP2 vesicles followed by ultracentrifugation demonstrated stable attachment of PLC to the vesicles. A mixed micellar system was established to assay PLC activity using deoxycholate. Kinetic analyses were performed to determine whether PLC activity was dependent on both bulk PIP2 and PIP2 surface concentrations in the micelles. The interfacial Michaelis constant was calculated to be 0.0518 mol fraction, and the equilibrium dissociation constant of PLC for the lipid was 45.5 μm. These findings will add to our understanding of the mechanisms of regulation of plant PLC. PMID:10444091

  7. Biochemical and Ultrastructural Changes in Sida cordifolia L. and Catharanthus roseus L. to Auto Pollution.

    Verma, Vijeta; Chandra, Neelam

    2014-01-01

    Auto pollution is the by-product of our mechanized mobility, which adversely affects both plant and human life. However, plants growing in the urban locations provide a great respite to us from the brunt of auto pollution by absorbing the pollutants at their foliar surface. Foliar surface configuration and biochemical changes in plant species, namely, Sida cordifolia L. and Catharanthus roseus L. grown at roadside (polluted site 1, Talkatora; polluted site 2, Charbagh) in Lucknow city and in the garden of the university campus, which has been taken as reference site, were investigated. It was observed that air pollution caused by auto exhaust showed marked alterations in photosynthetic pigments (chlorophyll, carotenoid, and phaeophytin), and relative water content was reduced while antioxidative enzymes like catalase and peroxidase were found to be enhanced. The changes in the foliar configuration reveal marked alteration in epidermal traits, with decreased number of stomata, stomatal indices, and epidermal cells per unit area, while length and breadth of stomata and epidermal cells were found to be increased in leaves samples wich can be used as biomarkers of auto pollution. PMID:27355010

  8. Influence of Precursor Availability on Alkaloid Accumulation by Transgenic Cell Line of Catharanthus roseus1

    Whitmer, Serap; Canel, Camilo; Hallard, Didier; Gonçalves, Cecilia; Verpoorte, Robert

    1998-01-01

    We have used a transgenic cell line of Catharanthus roseus (L.) G. Don to study the relative importance of the supply of biosynthetic precursors for the synthesis of terpenoid indole alkaloids. Line S10 carries a recombinant, constitutively overexpressed version of the endogenous strictosidine synthase (Str) gene. Various concentrations and combinations of the substrate tryptamine and of loganin, the immediate precursor of secologanin, were added to suspension cultures of S10. Our results indicate that high rates of tryptamine synthesis can take place under conditions of low tryptophan decarboxylase activity, and that high rates of strictosidine synthesis are possible in the presence of a small tryptamine pool. It appears that the utilization of tryptamine for alkaloid biosynthesis enhances metabolic flux through the indole pathway. However, a deficiency in the supply of either the iridoid or the indole precursor can limit flux through the step catalyzed by strictosidine synthase. Precursor utilization for the synthesis of strictosidine depends on the availability of the cosubstrate; the relative abundance of these precursors is a cell-line-specific trait that reflects the metabolic status of the cultures. PMID:9490777

  9. ATP-binding cassette transporter controls leaf surface secretion of anticancer drug components in Catharanthus roseus.

    Yu, Fang; De Luca, Vincenzo

    2013-09-24

    The Madagascar periwinkle (Catharanthus roseus) is highly specialized for the biosynthesis of many different monoterpenoid indole alkaloids (MIAs), many of which have powerful biological activities. Such MIAs include the commercially important chemotherapy drugs vinblastine, vincristine, and other synthetic derivatives that are derived from the coupling of catharanthine and vindoline. However, previous studies have shown that biosynthesis of these MIAs involves extensive movement of metabolites between specialized internal leaf cells and the leaf epidermis that require the involvement of unknown secretory processes for mobilizing catharanthine to the leaf surface and vindoline to internal leaf cells. Spatial separation of vindoline and catharanthine provides a clear explanation for the low levels of dimers that accumulate in intact plants. The present work describes the molecular cloning and functional identification of a unique catharanthine transporter (CrTPT2) that is expressed predominantly in the epidermis of young leaves. CrTPT2 gene expression is activated by treatment with catharanthine, and its in planta silencing redistributes catharanthine to increase the levels of catharanthine-vindoline drug dimers in the leaves. Phylogenetic analysis shows that CrTPT2 is closely related to a key transporter involved in cuticle assembly in plants and that may be unique to MIA-producing plant species, where it mediates secretion of alkaloids to the plant surface. PMID:24019465

  10. Cytoplasmic Acidification Induced by Inorganic Phosphate Uptake in Suspension Cultured Catharanthus roseus Cells

    Sakano, Katsuhiro; Yazaki, Yoshiaki; Mimura, Tetsuro

    1992-01-01

    Cytoplasmic acidification during inorganic phosphate (Pi) absorption by Catharanthus roseus cells were studied by means of a fluorescent pH indicator, 2′,7′-bis-(2-carboxyethyl)-5 carboxyfluorescein (acetomethylester) (BCECF), and 31P-nuclear magnetic resonance spectroscopy. Cytoplasmic acidification measured by decrease in the fluorescence intensity started immediately after Pi application. Within a minute or so, a stable state was attained and no further acidification occurred, whereas Pi absorption was still proceeding. As soon as Pi in the medium was exhausted, cytoplasmic pH started to recover. Coincidentally, the medium pH started to recover toward the original acidic pH. The Pi-induced changes in the cytoplasmic pH were confirmed by 31P-nuclear magnetic resonance study. Maximum acidification of the cytoplasm induced by 1.7 millimolar Pi was 0.2 pH units. Vacuolar pH was also affected by Pi. In some experiments, but not all, pH decreased reversibly by 0.2 to 0.3 pH units during Pi absorption. Results suggest that the cytoplasmic pH is regulated by proton pumps in the plasma membrane and in the tonoplast. In addition, other mechanisms that could consume extra protons in the cytoplasm are suggested. ImagesFigure 1 PMID:16668939

  11. Vindogentianine, a hypoglycemic alkaloid from Catharanthus roseus (L.) G. Don (Apocynaceae).

    Tiong, Soon Huat; Looi, Chung Yeng; Arya, Aditya; Wong, Won Fen; Hazni, Hazrina; Mustafa, Mohd Rais; Awang, Khalijah

    2015-04-01

    Vindogentianine, a new indole alkaloid together with six known alkaloids, vindoline, vindolidine, vindolicine, vindolinine, perivine and serpentine were isolated from leaf extract (DA) of Catharanthus roseus (L.) G. Don. Their structures were elucidated by spectroscopic methods; NMR, MS, UV and IR. Vindogentianine is a dimer containing a vindoline moiety coupled to a gentianine moiety. After 24h incubation, vindogentianine exhibited no cytotoxic effect in C2C12 mouse myoblast and β-TC6 mouse pancreatic cells (IC50>50μg/mL). Real-time cell proliferation monitoring also indicated vindogentianine had little or no effect on C2C12 mouse myoblast cell growth at the highest dose tested (200μg/mL), without inducing cell death. Vindogentianine exhibited potential hypoglycemic activity in β-TC6 and C2C12 cells by inducing higher glucose uptake and significant in vitro PTP-1B inhibition. However, in vitro α-amylase and α-glucosidase inhibition assay showed low inhibition under treatment of vindogentianine. This suggests that hypoglycemic activity of vindogentianine may be due to the enhancement of glucose uptake and PTP-1B inhibition, implying its therapeutic potential against type 2 diabetes. PMID:25665941

  12. Novel drug target identification on UDP-Glucose 4-epimerase enzyme in Catharanthus roseus by insilico model

    Ramachandran M; Elumalai EK

    2012-01-01

    Objective: To investigate the definite crystal structure of UDP-glucose 4-epimerase enzymes (EC 5.1.3.2) from Catharanthus roseus (C. roseus) for further research activities. Methods: The structure was modeled using homologous templates. The model validated under PROCHECK and WHAT-IF. Results: The model constructed using Modeller9v7 was validated. Moreover, 89% of residues lie in the most favored region. The model was checked for its grand average of hydropathicity and three binding sites were predicted using Molsoft ICM Pro v3.5. Conclusions:The model was suggested to be the good model. The constructed model can be used for further pharmacological studies and it can act as potential target against novel inhibitors.

  13. Studies on induced mutation frequency in Catharanthus roseus (L.) G. Don by gamma rays and EMS individually and in combination

    Seeds of pink flowered (PF) and white flowered (WF) Catharanthus roseus were soaked in distilled water for 24 h and treated with gamma rays and 0.1% EMS separately and in combination. Six types of chlorophyll mutations, viz., xantha, albina, chlorina, viridis, maculata and tigrina were recovered to M2 generation of both forms. The frequency of chlorophyll mutations was found to be dependent on the dose, of gamma rays and duration of treatment with EMS. Higher frequency of chlorophyll mutations was noticed in PF, which is mutagenically more sensitive than WF. It was also noticed that the combination treatments of gamma rays and EMS enhanced the frequency of chlorophyll mutations

  14. Antioxidative Potentials as a Protective Mechanism in Catharanthus roseus (L.) G.Don. Plants under Salinity Stress

    Jaleel, Cheruth Abdul; Gopi, Ragupathi; MANIVANNAN, Paramasivam; Panneerselvam, Rajaram

    2007-01-01

    Antioxidant responses were analysed in Catharanthus roseus (L.) G.Don. under 0, 50 and 100 mM NaCl in order to investigate the plant´s protective mechanisms against long-term salt-induced oxidative stress. The NaCl treatments were repeated in 4 different stages of growth, i.e. 30, 45, 60 and 75 days after sowing (DAS). The plants were uprooted randomly 90 DAS and the non-enzymatic and enzymatic antioxidant potentials were analysed. High salinity caused a decrease in reduced glutathione (GSH) ...

  15. A novel cytochrome P450 gene from Catharanthus roseus cell line C20hi: cloning and characterization of expression

    Lihong He; Shujuan Zhao; Zhibi Hu

    2012-01-01

    An expressed sequence tag (EST) obtained from a subtractive-suppression hybridization cDNA library constructed using Catharanthus roseus cell line C20hi and its parental cell line C20D was used to clone a full-length cytochrome P450 cDNA of cyp71d1. The encoded polypeptide contained 507 amino acids with 39–56% identity to other CYP71D subfamily members at the amino acid level. Expression characteristics of cyp71d1 were determined using semi-quantitative RT-PCR. The cyp71d1 transcript was expr...

  16. Influence of native arbuscular mycorrhizal fungi on growth, nutrition and phytochemical constituents of Catharanthus roseus (L. G. Don.

    Rajendran Srinivasan

    2014-01-01

    Full Text Available Objective: To study the isolation, identification, mass production and the effect of native arbuscular mycorrhizal fungi (AM fungi on growth parameters of the Catharanthus roseus (C. roseus. Methods: A total of nine different AM fungi species such as Acaulospora scrobiculata, Acaulospora marrowae, Glomus aggregatum (G. aggregatum, Glomus fasciculatum, Glomus geosporum, Gigaspora margarita, Gigaspora nigra, Scutellospora heterogama and Scutellospora pellucida were isolated and identified from the root zone soil of C. roseus. Results: The phytochemical analyses showed high concentration of chlorophyll a (0.152±0.0140 µg/g, chlorophyll b (0.081±0.006 µg/g, total chlorophyll (0.233±0.020 µg/g, soluble sugar (0.051±0.004 µg/g, reducing sugar (0.060±0,007 µg/g, phenols (0.293±0.032 µg/g, ortho-dihydroxy phenols (0.275±0.022 µg/g, lipids (0.300±0.025 µg/g, proteins (0.063±0.003 µg/g and amino acids (1.042±0.056 µg/g in G. aggregatum inoculated C. roseus. G. aggregatum was found to perform better on growth when compared to others and phytochemical constituents of C. roseus. Conclusions: It is concluded from the present findings that the G. aggregatum and Glomus fasciculatum can be used as a potential growth promoters for the C. roseus for better yielding in the agricultural sectors.

  17. Evaluation of the nutritive and organoleptic values of food products developed by incorporated Catharanthus roseus (Sadabahar) fresh leaves explore their hypoglycemic potential.

    Bisla, Gita; Choudhary, Shailza; Chaudhary, Vijeta

    2014-01-01

    Diabetes becomes a real problem of public health in developing countries, where its prevalence is increasing steadily. Diabetes mellitus can be found in almost every population in the world. Since the Ayurvedic practice started in India, plants are being used in the cure of diseases. Although the Catharanthus roseus have been used for their alleged health benefits and avail their hypoglycemic effect, used as medicine by diabetics. Medicinal plants have rarely been incorporated in food preparations. To fill these lacunae, food products were prepared by using Catharanthus roseus (Sadabahar) fresh leaves with hypoglycemic properties. Commonly consumed recipes in India are prepared for diabetic patients and were developed at different levels at 3 g, 4 g, and 6 g per serving. Food product development and their acceptability appraisal through organoleptic evaluation were carried out by semitrained panel comprising 15 trained panelists from the department of Food Science and Nutrition, Banasthali University. Seven products were developed by incorporating Catharanthus roseus fresh leaves. Nine point hedonic scale was used as a medium to know about the product acceptability at various variances. All products are moderately acceptable at different concentrations except product fare "6 g" which was more acceptable than the standard. Among the three variations of incorporating the Catharanthus roseus (Sadabahar) Leaves, 3 g variation is more acceptable than other variations. PMID:24790561

  18. Evaluation of the Nutritive and Organoleptic Values of Food Products Developed by Incorporated Catharanthus roseus (Sadabahar Fresh Leaves Explore Their Hypoglycemic Potential

    Gita Bisla

    2014-01-01

    Full Text Available Diabetes becomes a real problem of public health in developing countries, where its prevalence is increasing steadily. Diabetes mellitus can be found in almost every population in the world. Since the Ayurvedic practice started in India, plants are being used in the cure of diseases. Although the Catharanthus roseus have been used for their alleged health benefits and avail their hypoglycemic effect, used as medicine by diabetics. Medicinal plants have rarely been incorporated in food preparations. To fill these lacunae, food products were prepared by using Catharanthus roseus (Sadabahar fresh leaves with hypoglycemic properties. Commonly consumed recipes in India are prepared for diabetic patients and were developed at different levels at 3 g, 4 g, and 6 g per serving. Food product development and their acceptability appraisal through organoleptic evaluation were carried out by semitrained panel comprising 15 trained panelists from the department of Food Science and Nutrition, Banasthali University. Seven products were developed by incorporating Catharanthus roseus fresh leaves. Nine point hedonic scale was used as a medium to know about the product acceptability at various variances. All products are moderately acceptable at different concentrations except product fare “6 g” which was more acceptable than the standard. Among the three variations of incorporating the Catharanthus roseus (Sadabahar Leaves, 3 g variation is more acceptable than other variations.

  19. Study on the Photosynthesis Character of Catharanthus roseus and Catharanthus roseus cv. Albus in Glasshouse%温室内两种长春花光合特性的研究

    王非; 李雷鸿; 雷声武

    2007-01-01

    以温室内生长的红长春花[Catharanthus roseus(L.G.Don]和白长春花[Catharanthus roseus(L.)G.Don cv.Albus]为研究对象,通过对长春花的净光合速率日变化、叶绿素含量、光响应曲线及几项重要的光合参数等的研究,分析比较红长春花和白长春花光合生理的特性.结果表明,红长春花光合能力及利用光能的能力均强于白长春花.具体表现在:红长春花的叶绿素含量高于白长春花;红长春花的净光合速率高于白长春花;长春花在对强光的利用上表现出喜光植物的生理特性;红长春花的利用强光的能力高于白长春花;两种长春花净光合速率Pn日变化曲线趋势相同,均为单峰型.

  20. Somatic embryo mediated mass production of Catharanthus roseus in culture vessel (bioreactor) – A comparative study

    Mujib, A.; Ali, Muzamil; Isah, Tasiu; Dipti

    2014-01-01

    The purpose of this study was to evaluate and compare the use of liquid and solid Murashige and Skoog (MS) medium in different culture vessels for mass production of Catharanthus roseus, an important source of anticancerous compounds, vincristine and vinblastine. Three media conditions i.e. agar-solidified medium (S), liquid medium in agitated conical flask (L) and growtek bioreactor (B) were used. Rapid propagation was achieved through in vitro somatic embryogenesis pathway. The process of embryogenesis has been categorized into induction, proliferation, maturation and germination stages. All in vitro embryogenesis stages were conducted by withdrawing spent liquid medium and by adding fresh MS medium. In optimized 4.52 μM 2,4-D added MS, the callus biomass growth was low in solid (1.65 g) compared to liquid medium in agitated conical flask (1.95 g) and in bioreactor (2.11 g). The number of normal somatic embryos was more in solid medium (99.75/50 mg of callus mass) compared to liquid medium used in conical flask (83.25/callus mass) and growtek bioreactor (84.88/callus mass). The in vitro raised embryos maturated in GA3 (2.60 μM) added medium; and in bioreactor the embryo growth was high, a maximum length of 9.82 mm was observed at the end of four weeks. These embryos germinated into seedlings in BAP (2.22 μM) added medium and the embryo germination ability was more (59.41%) in bioreactor compared to liquid medium in conical flask (55.5%). Shoot length (11.25 mm) was also high in bioreactor compared to agitated conical flask. The liquid medium used in agitated conical flask and bioreactor increased seedling production efficiency, at the same time it also reduced plant recovery time. The embryo generated plants grew normally in outdoor conditions. The exploitation of medium to large culture vessel or bioreactor may make the process more efficient in getting large number of Catharanthus plant as it is the only source of anti-cancerous alkaloids

  1. Somatic embryo mediated mass production of Catharanthus roseus in culture vessel (bioreactor) - A comparative study.

    Mujib, A; Ali, Muzamil; Isah, Tasiu; Dipti

    2014-11-01

    The purpose of this study was to evaluate and compare the use of liquid and solid Murashige and Skoog (MS) medium in different culture vessels for mass production of Catharanthus roseus, an important source of anticancerous compounds, vincristine and vinblastine. Three media conditions i.e. agar-solidified medium (S), liquid medium in agitated conical flask (L) and growtek bioreactor (B) were used. Rapid propagation was achieved through in vitro somatic embryogenesis pathway. The process of embryogenesis has been categorized into induction, proliferation, maturation and germination stages. All in vitro embryogenesis stages were conducted by withdrawing spent liquid medium and by adding fresh MS medium. In optimized 4.52 μM 2,4-D added MS, the callus biomass growth was low in solid (1.65 g) compared to liquid medium in agitated conical flask (1.95 g) and in bioreactor (2.11 g). The number of normal somatic embryos was more in solid medium (99.75/50 mg of callus mass) compared to liquid medium used in conical flask (83.25/callus mass) and growtek bioreactor (84.88/callus mass). The in vitro raised embryos maturated in GA3 (2.60 μM) added medium; and in bioreactor the embryo growth was high, a maximum length of 9.82 mm was observed at the end of four weeks. These embryos germinated into seedlings in BAP (2.22 μM) added medium and the embryo germination ability was more (59.41%) in bioreactor compared to liquid medium in conical flask (55.5%). Shoot length (11.25 mm) was also high in bioreactor compared to agitated conical flask. The liquid medium used in agitated conical flask and bioreactor increased seedling production efficiency, at the same time it also reduced plant recovery time. The embryo generated plants grew normally in outdoor conditions. The exploitation of medium to large culture vessel or bioreactor may make the process more efficient in getting large number of Catharanthus plant as it is the only source of anti-cancerous alkaloids

  2. Production Pattern of Ajmalicine in Catharanthus roseus (L. G. Don. Cell Aggregates Culture in the Airlift Bioreactor

    RIZKITA RACHMI ESYANTI

    2006-12-01

    Full Text Available A research has been conducted to optimize the rate of aeration and initial weight of cell aggregates in the production of ajmalicine in Catharanthus roseus cell culture in airlift bioreactor. Catharanthus roseus culture were grown in Zenk medium with the addition of 2.50 x 10-6 M naphthalene acetic acid (NAA and 10-5 M benzyl amino purine (BAP. Cell aggregates were sub-cultured two times before transferring 20 and 30 g/fw of cell aggregates into bioreactor, respectively, and aerated with the rate of 0.25 l min-1 and 0.34 l min-1, respectively. The pattern of ajmalicine production in bioreactor were observed in every three days within 24 days. Qualitative and quantitative analysis were conducted using HPLC connected to Cromatopac CL-7A Plus. The results showed that the cell aggregates and medium contain ajmalicine. The highest concentration was obtained in combination of 30 g/fw and 0.34 l min-1 aeration compare to 20 g/fw - 0.25 l min-1, 20 g/fw - 0.34 l min-1, as well as 30 g/fw – 0.25 l min-1. The highest ajmalicine content in cell aggregates was obtained on the 12 days (79.23 µg g-1 whilst in medium was obtained in the 18th days (981.15 µg l-1.

  3. A comparative study of release profiles of Coccinia cordifolia and Catharanthus roseus with standard antidiabetic agent using rat intestine

    Most. Afia Ahktar

    2014-09-01

    Full Text Available Matured leaves of Coccinia cordifolia and Catharanthus roseus were collected, dried and extracted with 95% ethanol. Solvents were evaporated and suspensions at a concentration of 40 mg/ml were prepared from the residues using phosphate buffer. The aim of the study is to elucidate the release pattern of these extracts in acidic and basic environment. 1 ml suspension of each plant was poured into the intestine fragment prepared from sacrificed rats. The filled pieces of intestine were bound vertically to the paddle of  modified dissolution apparatus and rotated at 50 rpm in phosphate buffer incubation medium at pH 3 and pH 7.4. Metformin hydrochloride was used as the standard. Percent releases of Coccinia cordifolia, Catharanthus roseus and metformin hydrochloride were determined by analyzing the UV absorbance data at different time interval and the result obtained at pH 3 was compared with that of pH 7.4. Experimental result with these extracts and drug showed burst release initially followed by gradual release at pH 3.0 and the value of R2 (correlation coefficient of percent release versus time indicates that the release pattern was better maintained at pH 3.0 for coccinia and catharanths while that was better maintained at pH 7.4 for metformin hydrochloride.

  4. Exploiting EST databases for the mining and characterization of short sequence repeat (SSR) markers in Catharanthus roseus L.

    Joshi, Raj Kumar; Kar, Basudeba; Nayak, Sanghamitra

    2011-01-01

    Periwinkle (Catharanthus roseus L.) (Family: Apocyanaceae) is a ornamental plants with great medicinal properties. Although it is represented by seven species, little work has been carried out on its genetic characterization due to non-availability of reliable molecular markers. Simple sequence repeats (SSRs) have been widely applied as molecular markers in genetic studies. With the rapid increase in the deposition of nucleotide sequences in the public databases and advent of bioinformatics tools, it has become a cost effective and fast approach to scan for microsatellite repeats and exploit the possibility of converting it into potential genetic markers. Expressed sequence tags (EST's) from Catharanthus roseus were used for the screening of Class I (hyper variable) simple sequence repeats (SSR's). A total of 502 microsatellite repeats were detected from 21730 EST sequences of turmeric after redundancy elimination. The average density of Class I SSRs account to 1 SSR per 10.21 kb of EST. Mononucleotides was the most abundant class of microsatellite motifs. It accounted for 44.02% of the total, followed by the trinucleotide (26.09%) and dinucleotide repeats (14.34%). Among all the repeat motifs, (A/T)n accounted for the highest Proportion (36.25%) followed by (AAG)n. These detected SSRs can be used to design primers that have functional importance and should also facilitate the analysis of genetic diversity, variability, linkage mapping and evolutionary relationships in plants especially medicinal plants. PMID:21383904

  5. UV-B-induced signaling events leading to enhanced-production of catharanthine in Catharanthus roseus cell suspension cultures

    Ramani, Shilpa; Chelliah, Jayabaskaran

    2007-01-01

    Background Elicitations are considered to be an important strategy towards improved in vitro production of secondary metabolites. In cell cultures, biotic and abiotic elicitors have effectively stimulated the production of plant secondary metabolites. However, molecular basis of elicitor-signaling cascades leading to increased production of secondary metabolites of plant cell is largely unknown. Exposure of Catharanthus roseus cell suspension culture to low dose of UV-B irradiation was found to increase the amount of catharanthine and transcription of genes encoding tryptophan decarboxylase (Tdc) and strictosidine synthase (Str). In the present study, the signaling pathway mediating UV-B-induced catharanthine accumulation in C. roseus suspension cultures were investigated. Results Here, we investigate whether cell surface receptors, medium alkalinization, Ca2+ influx, H2O2, CDPK and MAPK play required roles in UV-B signaling leading to enhanced production of catharanthine in C. roseus cell suspension cultures. C. roseus cells were pretreated with various agonists and inhibitors of known signaling components and their effects on the accumulation of Tdc and Str transcripts as well as amount of catharanthine production were investigated by various molecular biology techniques. It has been found that the catharanthine accumulation and transcription of Tdc and Str were inhibited by 3–4 fold upon pretreatment of various inhibitors like suramin, N-acetyl cysteine, inhibitors of calcium fluxes, staurosporine etc. Conclusion Our results demonstrate that cell surface receptor(s), Ca2+ influx, medium alkalinization, CDPK, H2O2 and MAPK play significant roles in UV-B signaling leading to stimulation of Tdc and Str genes and the accumulation of catharanthine in C. roseus cell suspension cultures. Based on these findings, a model for signal transduction cascade has been proposed. PMID:17988378

  6. UV-B-induced signaling events leading to enhanced-production of catharanthine in Catharanthus roseus cell suspension cultures

    Chelliah Jayabaskaran

    2007-11-01

    Full Text Available Abstract Background Elicitations are considered to be an important strategy towards improved in vitro production of secondary metabolites. In cell cultures, biotic and abiotic elicitors have effectively stimulated the production of plant secondary metabolites. However, molecular basis of elicitor-signaling cascades leading to increased production of secondary metabolites of plant cell is largely unknown. Exposure of Catharanthus roseus cell suspension culture to low dose of UV-B irradiation was found to increase the amount of catharanthine and transcription of genes encoding tryptophan decarboxylase (Tdc and strictosidine synthase (Str. In the present study, the signaling pathway mediating UV-B-induced catharanthine accumulation in C. roseus suspension cultures were investigated. Results Here, we investigate whether cell surface receptors, medium alkalinization, Ca2+ influx, H2O2, CDPK and MAPK play required roles in UV-B signaling leading to enhanced production of catharanthine in C. roseus cell suspension cultures. C. roseus cells were pretreated with various agonists and inhibitors of known signaling components and their effects on the accumulation of Tdc and Str transcripts as well as amount of catharanthine production were investigated by various molecular biology techniques. It has been found that the catharanthine accumulation and transcription of Tdc and Str were inhibited by 3–4 fold upon pretreatment of various inhibitors like suramin, N-acetyl cysteine, inhibitors of calcium fluxes, staurosporine etc. Conclusion Our results demonstrate that cell surface receptor(s, Ca2+ influx, medium alkalinization, CDPK, H2O2 and MAPK play significant roles in UV-B signaling leading to stimulation of Tdc and Str genes and the accumulation of catharanthine in C. roseus cell suspension cultures. Based on these findings, a model for signal transduction cascade has been proposed.

  7. Involvement of nitric oxide signaling in mammalian Bax-induced terpenoid indole alkaloid production of Catharanthus roseus cells

    2007-01-01

    Bax, a mammalian pro-apoptotic member of the Bcl-2 family, has been demonstrated to be a potential regulatory factor for plant secondary metabolite biosynthesis recently. To investigate the molecular mechanism of Bax-induced secondary metabolite biosynthesis, we determined the contents of nitric oxide (NO) of the transgenic Catharanthus roseus cells overexpressing a mouse Bax protein and checked the effects of NO specific scavenger 2,4-carboxyphenyl-4,4,5,5-tetramethylimidazoline-1- oxyl-3-oxide (cPITO) on Bax-induced terpenoid indole alkaloid (TIA) production of the cells. The data showed that overexpression of the mouse Bax in C. roseus cells triggered NO generation of the cells. Treatment of cPITO not only inhibited the Bax-triggered NO burst but also suppressed the Bax-induced TIA production. The results indicated that the mouse Bax might activate the NO signaling in C. roseus cells and induce TIA production through the NO-dependent signal pathway in the cells. Furthermore, the activities of nitric oxide synthase (NOS) were significantly increased in the transgenic Bax cells as compared to those in the control cells, showing that the mouse Bax may induce NOS of C. roseus cells. Treatment of the transgenic Bax cells with NOS inhibitor PBITU blocked both Bax-induced NO generation and TIA production, which suggested that the mouse Bax might trigger NO generation and TIA production through NOS. However, the NOS-like activities and NO generation in the transgenic Bax cells did not match kinetically and the Bax-induced NOS-like activity was much later and lower than NO production. Moreover, the Bax-induced NO generation and TIA production were only partially inhibited by PBITU. Thus, our results suggested that the Bax-induced NO production and secondary metabolite biosynthesis in C. roseus cells was not entirely dependent on NOS or NOS-like enzymes.

  8. Developmental and Light Regulation of Desacetoxyvindoline 4-Hydroxylase in Catharanthus roseus (L.) G. Don.1

    Vazquez-Flota, Felipe A.; De Luca, Vincenzo

    1998-01-01

    The expression of desacetoxyvindoline 4-hydroxylase (D4H), which catalyzes the second to the last reaction in vindoline biosynthesis in Catharanthus roseus, appears to be under complex, multilevel developmental and light regulation. Developmental studies with etiolated and light-treated seedlings suggested that although light had variable effects on the levels of d4h transcripts, those of D4H protein and enzyme activity could be increased, depending on seedling development, up to 9- and 8-fold, respectively, compared with etiolated seedlings. However, light treatment of etiolated seedlings could stop and reverse the decline of d4h transcripts at later stages of seedling development. Repeated exposure of seedlings to light was also required to maintain the full spectrum of enzyme activity observed during seedling development. Further studies showed that a photoreversible phytochrome appeared to be involved in the activation of D4H, since red-light treatment of etiolated seedlings increased the detectable levels of d4h transcripts, D4H protein, and D4H enzyme activity, whereas far-red-light treatment completely reversed this process. Additional studies also confirmed that different major isoforms of D4H protein exist in etiolated (isoelectric point, 4.7) and light-grown (isoelectric point, 4.6) seedlings, suggesting that a component of the light-mediated activation of D4H may involve an undetermined posttranslational modification. The biological reasons for this complex control of vindoline biosynthesis may be related to the need to produce structures that could sequester away from cellular activities the cytotoxic vinblastine and vincristine dimers that are derived partially from vindoline. PMID:9701591

  9. Interaction between abscisic acid and nitric oxide in PB90-induced catharanthine biosynthesis of catharanthus roseus cell suspension cultures.

    Chen, Qian; Chen, Zunwei; Lu, Li; Jin, Haihong; Sun, Lina; Yu, Qin; Xu, Hongke; Yang, Fengxia; Fu, Mengna; Li, Shengchao; Wang, Huizhong; Xu, Maojun

    2013-01-01

    Elicitations are considered to be an important strategy to improve production of secondary metabolites of plant cell cultures. However, mechanisms responsible for the elicitor-induced production of secondary metabolites of plant cells have not yet been fully elucidated. Here, we report that treatment of Catharanthus roseus cell suspension cultures with PB90, a protein elicitor from Phytophthora boehmeriae, induced rapid increases of abscisic acid (ABA) and nitric oxide (NO), subsequently followed by the enhancement of catharanthine production and up-regulation of Str and Tdc, two important genes in catharanthine biosynthesis. PB90-induced catharanthine production and the gene expression were suppressed by the ABA inhibitor and NO scavenger respectively, showing that ABA and NO are essential for the elicitor-induced catharanthine biosynthesis. The relationship between ABA and NO in mediating catharanthine biosynthesis was further investigated. Treatment of the cells with ABA triggered NO accumulation and induced catharanthine production and up-regulation of Str and Tdc. ABA-induced catharanthine production and gene expressions were suppressed by the NO scavenger. Conversely, exogenous application of NO did not stimulate ABA generation and treatment with ABA inhibitor did not suppress NO-induced catharanthine production and gene expressions. Together, the results showed that both NO and ABA were involved in PB90-induced catharanthine biosynthesis of C. roseus cells. Furthermore, our data demonstrated that ABA acted upstream of NO in the signaling cascade leading to PB90-induced catharanthine biosynthesis of C. roseus cells. PMID:23554409

  10. Vacuolar Transport of the Medicinal Alkaloids from Catharanthus roseus Is Mediated by a Proton-Driven Antiport1[W

    Carqueijeiro, Inês; Noronha, Henrique; Duarte, Patrícia; Gerós, Hernâni; Sottomayor, Mariana

    2013-01-01

    Catharanthus roseus is one of the most studied medicinal plants due to the interest in their dimeric terpenoid indole alkaloids (TIAs) vinblastine and vincristine, which are used in cancer chemotherapy. These TIAs are produced in very low levels in the leaves of the plant from the monomeric precursors vindoline and catharanthine and, although TIA biosynthesis is reasonably well understood, much less is known about TIA membrane transport mechanisms. However, such knowledge is extremely important to understand TIA metabolic fluxes and to develop strategies aimed at increasing TIA production. In this study, the vacuolar transport mechanism of the main TIAs accumulated in C. roseus leaves, vindoline, catharanthine, and α-3′,4′-anhydrovinblastine, was characterized using a tonoplast vesicle system. Vindoline uptake was ATP dependent, and this transport activity was strongly inhibited by NH4+ and carbonyl cyanide m-chlorophenyl hydrazine and was insensitive to the ATP-binding cassette (ABC) transporter inhibitor vanadate. Spectrofluorimetry assays with a pH-sensitive fluorescent probe showed that vindoline and other TIAs indeed were able to dissipate an H+ gradient preestablished across the tonoplast by either vacuolar H+-ATPase or vacuolar H+-pyrophosphatase. The initial rates of H+ gradient dissipation followed Michaelis-Menten kinetics, suggesting the involvement of mediated transport, and this activity was species and alkaloid specific. Altogether, our results strongly support that TIAs are actively taken up by C. roseus mesophyll vacuoles through a specific H+ antiport system and not by an ion-trap mechanism or ABC transporters. PMID:23686419

  11. Vacuolar transport of the medicinal alkaloids from Catharanthus roseus is mediated by a proton-driven antiport.

    Carqueijeiro, Inês; Noronha, Henrique; Duarte, Patrícia; Gerós, Hernâni; Sottomayor, Mariana

    2013-07-01

    Catharanthus roseus is one of the most studied medicinal plants due to the interest in their dimeric terpenoid indole alkaloids (TIAs) vinblastine and vincristine, which are used in cancer chemotherapy. These TIAs are produced in very low levels in the leaves of the plant from the monomeric precursors vindoline and catharanthine and, although TIA biosynthesis is reasonably well understood, much less is known about TIA membrane transport mechanisms. However, such knowledge is extremely important to understand TIA metabolic fluxes and to develop strategies aimed at increasing TIA production. In this study, the vacuolar transport mechanism of the main TIAs accumulated in C. roseus leaves, vindoline, catharanthine, and α-3',4'-anhydrovinblastine, was characterized using a tonoplast vesicle system. Vindoline uptake was ATP dependent, and this transport activity was strongly inhibited by NH4(+) and carbonyl cyanide m-chlorophenyl hydrazine and was insensitive to the ATP-binding cassette (ABC) transporter inhibitor vanadate. Spectrofluorimetry assays with a pH-sensitive fluorescent probe showed that vindoline and other TIAs indeed were able to dissipate an H(+) gradient preestablished across the tonoplast by either vacuolar H(+)-ATPase or vacuolar H(+)-pyrophosphatase. The initial rates of H(+) gradient dissipation followed Michaelis-Menten kinetics, suggesting the involvement of mediated transport, and this activity was species and alkaloid specific. Altogether, our results strongly support that TIAs are actively taken up by C. roseus mesophyll vacuoles through a specific H(+) antiport system and not by an ion-trap mechanism or ABC transporters. PMID:23686419

  12. A Cytochrome P-450 Monooxygenase Catalyzes the First Step in the Conversion of Tabersonine to Vindoline in Catharanthus roseus.

    St-Pierre, B.; De Luca, V.

    1995-01-01

    Hydroxylation at the C-16 position of the indole alkaloid tabersonine has been suggested as the first step toward vindoline biosynthesis in Catharanthus roseus. Tabersonine 16-hydroxylase (16-OH) activity was detected in total protein extracts from young leaves of C. roseus using a novel coupled assay system. Enzyme activity was dependent on NADPH and molecular oxygen and was inhibited by CO, clotrimazole, miconazole, and cytochrome c. 16-OH was localized to the endoplasmic reticulum by linear sucrose density gradient centrifugation. These data suggest that 16-OH is a cytochrome P-450-dependent monooxygenase. The activity of 16-OH reached a maximum in seedlings 9 d postimbibition and was induced by light. The leaf-specific distribution of 16-OH in the mature plant is consistent with the localization of other enzymes in the tabersonine to vindoline pathway. However, in contrast to enzymes that catalyze the last four steps of vindoline biosynthesis, enzymes responsible for the first two steps from tabersonine (16-OH and 16-O-methyltransfersase) were detected in C. roseus cell-suspension cultures. These data complement the complex model of vindoline biosynthesis that has evolved with respect to enzyme compartmentalization, metabolic transport, and control mechanisms. PMID:12228585

  13. Induction and Flow Cytometry Identification of Tetraploids from Seed-Derived Explants through Colchicine Treatments in Catharanthus roseus (L.) G. Don

    Xing, Shi-Hai; Guo, Xin-Bo; Wang, Quan; Pan, Qi-Fang; Tian, Yue-Sheng; Liu, Pin; Zhao, Jing-Ya; Wang, Guo-Feng; Sun, Xiao-Fen; Tang, Ke-Xuan

    2011-01-01

    The tetraploid plants of Catharanthus roseus (L.) G. Don was obtained by colchicine induction from seeds explants, and the ploidy of the plants was identified by flow cytometry. The optimal treatment is 0.2% colchicine solution treated for 24 hours, and the induction rate reaches up to 30%. Comparing with morphological characteristics and growth habits between tetraploids and the control, we found that tetraploids of C. roseus had larger stoma and more branches and leaves. HPLC analysis showed tetraploidization could increase the contents of terpenoid indole alkaloids in C. roseus. Thus, tetraploidization could be used to produce higher alkaloids lines for commercial use. QRT-PCR results showed that the expression of enzymes involved in terpenoid indole alkaloids biosynthesis pathway had increased in the tetraploid plants. To our knowledge, this was the first paper to explore the secondary metabolism in autotetraploid C. roseus induced by colchicine. PMID:21660143

  14. Induction and Flow Cytometry Identification of Tetraploids from Seed-Derived Explants through Colchicine Treatments in Catharanthus roseus (L. G. Don

    Shi-Hai Xing

    2011-01-01

    Full Text Available The tetraploid plants of Catharanthus roseus (L. G. Don was obtained by colchicine induction from seeds explants, and the ploidy of the plants was identified by flow cytometry. The optimal treatment is 0.2% colchicine solution treated for 24 hours, and the induction rate reaches up to 30%. Comparing with morphological characteristics and growth habits between tetraploids and the control, we found that tetraploids of C. roseus had larger stoma and more branches and leaves. HPLC analysis showed tetraploidization could increase the contents of terpenoid indole alkaloids in C. roseus. Thus, tetraploidization could be used to produce higher alkaloids lines for commercial use. QRT-PCR results showed that the expression of enzymes involved in terpenoid indole alkaloids biosynthesis pathway had increased in the tetraploid plants. To our knowledge, this was the first paper to explore the secondary metabolism in autotetraploid C. roseus induced by colchicine.

  15. Effect of Gloriosa superba and Catharanthus roseus Extracts on IFN-γ-Induced Keratin 17 Expression in HaCaT Human Keratinocytes

    Nattaporn Pattarachotanant

    2014-01-01

    Full Text Available Gloriosa superba and Catharanthus roseus are useful in traditional medicine for treatment of various skin diseases and cancer. However, their molecular effect on psoriasis has not been investigated. In this study, the effect of ethanol extracts derived from G. superba leaves and C. roseus stems on the expression of psoriatic marker, keratin 17 (K17, was investigated in human keratinocytes using biochemical and molecular experimental approaches. Both extracts could reduce the expression of K17 in a dose-dependent manner through JAK/STAT pathway as demonstrated by an observation of reduced phosphorylation of STAT3 (p-STAT3. The inhibitory activity of G. superba extract was more potent than that of C. roseus. The Pearson's correlation between K17 and cell viability was shown positive. Taken together, the extracts of G. superba and C. roseus may be developed as alternative therapies for psoriasis.

  16. Effect of Gloriosa superba and Catharanthus roseus Extracts on IFN-γ-Induced Keratin 17 Expression in HaCaT Human Keratinocytes.

    Pattarachotanant, Nattaporn; Rakkhitawatthana, Varaporn; Tencomnao, Tewin

    2014-01-01

    Gloriosa superba and Catharanthus roseus are useful in traditional medicine for treatment of various skin diseases and cancer. However, their molecular effect on psoriasis has not been investigated. In this study, the effect of ethanol extracts derived from G. superba leaves and C. roseus stems on the expression of psoriatic marker, keratin 17 (K17), was investigated in human keratinocytes using biochemical and molecular experimental approaches. Both extracts could reduce the expression of K17 in a dose-dependent manner through JAK/STAT pathway as demonstrated by an observation of reduced phosphorylation of STAT3 (p-STAT3). The inhibitory activity of G. superba extract was more potent than that of C. roseus. The Pearson's correlation between K17 and cell viability was shown positive. Taken together, the extracts of G. superba and C. roseus may be developed as alternative therapies for psoriasis. PMID:25435888

  17. Molecular Analysis and Heterologous Expression of an Inducible Cytochrome P-450 Protein from Periwinkle (Catharanthus roseus L.) 1

    Vetter, Hans-Peter; Mangold, Ursula; Schröder, Gudrun; Marner, Franz-Josef; Werck-Reichhart, Danielle; Schröder, Joachim

    1992-01-01

    We screened cDNA libraries from periwinkle (Catharanthus roseus) cell cultures induced for indole alkaloid synthesis and selected clones for induced cytochrome P-450 (P-450) proteins by differential hybridization, size of the hybridizing mRNA, and presence of amino acid motifs conserved in many P-450 families. Four cDNAs satisfying these criteria were analyzed in detail. They were grouped in two classes (pCros1, pCros2) that represented two closely related genes of a new P-450 family designated CYP72. Antiserum against a cDNA fusion protein overexpressed in Escherichia coli recognized in C. roseus a protein band of 56 kD. Quantification of western blots showed that it represented 1.5 ± 0.5 and 6 ± 1 μg/mg of protein in the membranes from noninduced and induced cells, respectively, and analysis of the total P-450 content suggested that the cDNA-encoded protein was one of the dominant P-450 proteins. The pathway to indole alkaloids contains two known P-450 enzymes, geraniol-10-hydroxylase (GE10H) and nerol-10-hydroxylase (NE10H). The induction kinetics of the cloned P-450 protein and of GE10H activity were similar, but those of NE10H were different. Western blots with membranes from other plants suggested that P-450 CYP72 is specific for C. roseus and other plants with GE10H activity. A tentative assignment of CYP72 as GE10H is discussed. The cDNA was recloned for expression in Saccharomyces cerevisiae, and the presence of the protein was demonstrated by western blots. Assays for GE10H failed to detect enzyme activity, and the same negative result was obtained for NE10H and other P-450 enzymes that are present in C. roseus. Images Figure 5 Figure 7 PMID:16653087

  18. A pair of tabersonine 16-hydroxylases initiates the synthesis of vindoline in an organ-dependent manner in Catharanthus roseus.

    Besseau, Sébastien; Kellner, Franziska; Lanoue, Arnaud; Thamm, Antje M K; Salim, Vonny; Schneider, Bernd; Geu-Flores, Fernando; Höfer, René; Guirimand, Grégory; Guihur, Anthony; Oudin, Audrey; Glevarec, Gaëlle; Foureau, Emilien; Papon, Nicolas; Clastre, Marc; Giglioli-Guivarc'h, Nathalie; St-Pierre, Benoit; Werck-Reichhart, Danièle; Burlat, Vincent; De Luca, Vincenzo; O'Connor, Sarah E; Courdavault, Vincent

    2013-12-01

    Hydroxylation of tabersonine at the C-16 position, catalyzed by tabersonine 16-hydroxylase (T16H), initiates the synthesis of vindoline that constitutes the main alkaloid accumulated in leaves of Catharanthus roseus. Over the last decade, this reaction has been associated with CYP71D12 cloned from undifferentiated C. roseus cells. In this study, we isolated a second cytochrome P450 (CYP71D351) displaying T16H activity. Biochemical characterization demonstrated that CYP71D12 and CYP71D351 both exhibit high affinity for tabersonine and narrow substrate specificity, making of T16H, to our knowledge, the first alkaloid biosynthetic enzyme displaying two isoforms encoded by distinct genes characterized to date in C. roseus. However, both genes dramatically diverge in transcript distribution in planta. While CYP71D12 (T16H1) expression is restricted to flowers and undifferentiated cells, the CYP71D351 (T16H2) expression profile is similar to the other vindoline biosynthetic genes reaching a maximum in young leaves. Moreover, transcript localization by carborundum abrasion and RNA in situ hybridization demonstrated that CYP71D351 messenger RNAs are specifically located to leaf epidermis, which also hosts the next step of vindoline biosynthesis. Comparison of high- and low-vindoline-accumulating C. roseus cultivars also highlights the direct correlation between CYP71D351 transcript and vindoline levels. In addition, CYP71D351 down-regulation mediated by virus-induced gene silencing reduces vindoline accumulation in leaves and redirects the biosynthetic flux toward the production of unmodified alkaloids at the C-16 position. All these data demonstrate that tabersonine 16-hydroxylation is orchestrated in an organ-dependent manner by two genes including CYP71D351, which encodes the specific T16H isoform acting in the foliar vindoline biosynthesis. PMID:24108213

  19. CrMPK3, a mitogen activated protein kinase from Catharanthus roseus and its possible role in stress induced biosynthesis of monoterpenoid indole alkaloids

    Raina Susheel

    2012-08-01

    Full Text Available Abstract Background Mitogen activated protein kinase (MAPK cascade is an important signaling cascade that operates in stress signal transduction in plants. The biologically active monoterpenoid indole alkaloids (MIA produced in Catharanthus roseus are known to be induced under several abiotic stress conditions such as wounding, UV-B etc. However involvement of any signaling component in the accumulation of MIAs remains poorly investigated so far. Here we report isolation of a novel abiotic stress inducible Catharanthus roseus MAPK, CrMPK3 that may have role in accumulation of MIAs in response to abiotic stress. Results CrMPK3 expressed in bacterial system is an active kinase as it showed auto-phosphorylation and phosphorylation of Myelin Basic Protein. CrMPK3 though localized in cytoplasm, moves to nucleus upon wounding. Wounding, UV treatment and MeJA application on C. roseus leaves resulted in the transcript accumulation of CrMPK3 as well as activation of MAPK in C. roseus leaves. Immuno-precipitation followed by immunoblot analysis revealed that wounding, UV treatment and methyl jasmonate (MeJA activate CrMPK3. Transient over-expression of CrMPK3 in C. roseus leaf tissue showed enhanced expression of key MIA biosynthesis pathway genes and also accumulation of specific MIAs. Conclusion Results from our study suggest a possible involvement of CrMPK3 in abiotic stress signal transduction towards regulation of transcripts of key MIA biosynthetic pathway genes, regulators and accumulation of major MIAs.

  20. CrMPK3, a mitogen activated protein kinase from Catharanthus roseus and its possible role in stress induced biosynthesis of monoterpenoid indole alkaloids

    2012-01-01

    Background Mitogen activated protein kinase (MAPK) cascade is an important signaling cascade that operates in stress signal transduction in plants. The biologically active monoterpenoid indole alkaloids (MIA) produced in Catharanthus roseus are known to be induced under several abiotic stress conditions such as wounding, UV-B etc. However involvement of any signaling component in the accumulation of MIAs remains poorly investigated so far. Here we report isolation of a novel abiotic stress inducible Catharanthus roseus MAPK, CrMPK3 that may have role in accumulation of MIAs in response to abiotic stress. Results CrMPK3 expressed in bacterial system is an active kinase as it showed auto-phosphorylation and phosphorylation of Myelin Basic Protein. CrMPK3 though localized in cytoplasm, moves to nucleus upon wounding. Wounding, UV treatment and MeJA application on C. roseus leaves resulted in the transcript accumulation of CrMPK3 as well as activation of MAPK in C. roseus leaves. Immuno-precipitation followed by immunoblot analysis revealed that wounding, UV treatment and methyl jasmonate (MeJA) activate CrMPK3. Transient over-expression of CrMPK3 in C. roseus leaf tissue showed enhanced expression of key MIA biosynthesis pathway genes and also accumulation of specific MIAs. Conclusion Results from our study suggest a possible involvement of CrMPK3 in abiotic stress signal transduction towards regulation of transcripts of key MIA biosynthetic pathway genes, regulators and accumulation of major MIAs. PMID:22871174

  1. Efekt inokulacije korijena korijenovim bakterijama za poticanje rasta (PGPR) na rast biljke, sadržaj alkaloida i nutrijenata kod biljke Catharanthus roseus (L.) G. Don.

    Karthikeyan, Balathandayutham; Joe, Manoharan Melvin; Jaleel, Cheruth Abdul; Deiveekasundaram, Muthukumar

    2010-01-01

    Testirani su učinci korijenovih bakterija za poticanje rasta, kao što su Azotobacter, Bacillus i Pseudomonas, zasebno ili u kombinaciji, na biljci Catharanthus roseus i to tijekom dviju godina (2005 i 2006). Kombinacije gorespomenutih PGPR sojeva značajno su povećale visinu biljaka, duljinu korijena, debljinu korijena i sadržaj alkaloida kod C. roseus u usporedbi s kontrolom. Uz to sadržaj svih nutrijenata (N, P, K, Ca i Mg) bio je značajno povišen u usporedbi s kontrolom. Maksimalni sadržaj ...

  2. 7-deoxyloganetic acid synthase catalyzes a key 3 step oxidation to form 7-deoxyloganetic acid in Catharanthus roseus iridoid biosynthesis.

    Salim, Vonny; Wiens, Brent; Masada-Atsumi, Sayaka; Yu, Fang; De Luca, Vincenzo

    2014-05-01

    Iridoids are key intermediates required for the biosynthesis of monoterpenoid indole alkaloids (MIAs), as well as quinoline alkaloids. Although most iridoid biosynthetic genes have been identified, one remaining three step oxidation required to form the carboxyl group of 7-deoxyloganetic acid has yet to be characterized. Here, it is reported that virus-induced gene silencing of 7-deoxyloganetic acid synthase (7DLS, CYP76A26) in Catharanthus roseus greatly decreased levels of secologanin and the major MIAs, catharanthine and vindoline in silenced leaves. Functional expression of this gene in Saccharomyces cerevisiae confirmed its function as an authentic 7DLS that catalyzes the 3 step oxidation of iridodial-nepetalactol to form 7-deoxyloganetic acid. The identification of CYP76A26 removes a key bottleneck for expression of iridoid and related MIA pathways in various biological backgrounds. PMID:24594312

  3. CrMPK3, a mitogen activated protein kinase from Catharanthus roseus and its possible role in stress induced biosynthesis of monoterpenoid indole alkaloids

    Raina Susheel; Wankhede Dhammaprakash; Jaggi Monika; Singh Pallavi; Jalmi Siddhi; Raghuram Badmi; Sheikh Arsheed; Sinha Alok

    2012-01-01

    Abstract Background Mitogen activated protein kinase (MAPK) cascade is an important signaling cascade that operates in stress signal transduction in plants. The biologically active monoterpenoid indole alkaloids (MIA) produced in Catharanthus roseus are known to be induced under several abiotic stress conditions such as wounding, UV-B etc. However involvement of any signaling component in the accumulation of MIAs remains poorly investigated so far. Here we report isolation of a novel abiotic ...

  4. Isolation of Catharanthus roseus (L.) G. Don Nuclei and Measurement of Rate of Tryptophan decarboxylase Gene Transcription Using Nuclear Run-On Transcription Assay

    Santosh Kumar; Sabhyata Bhatia

    2015-01-01

    Background An accurate assessment of transcription ‘rate’ is often desired to describe the promoter activity. In plants, isolation of transcriptionally active nuclei and their subsequent use in nuclear run-on assays has been challenging and therefore limit an accurate measurement of gene transcription ‘rate’. Catharanthus roseus has emerged as a model medicinal plant as it exhibits an unsurpassed spectrum of chemodiversity, producing over 130 alkaloids through the terpenoid indole alkaloid (T...

  5. Evaluation of the Nutritive and Organoleptic Values of Food Products Developed by Incorporated Catharanthus roseus (Sadabahar) Fresh Leaves Explore Their Hypoglycemic Potential

    Gita Bisla; Shailza Choudhary; Vijeta Chaudhary

    2014-01-01

    Diabetes becomes a real problem of public health in developing countries, where its prevalence is increasing steadily. Diabetes mellitus can be found in almost every population in the world. Since the Ayurvedic practice started in India, plants are being used in the cure of diseases. Although the Catharanthus roseus have been used for their alleged health benefits and avail their hypoglycemic effect, used as medicine by diabetics. Medicinal plants have rarely been incorporated in food prepara...

  6. The Complete Plastid Genome Sequence of Madagascar Periwinkle Catharanthus roseus (L.) G. Don: Plastid Genome Evolution, Molecular Marker Identification, and Phylogenetic Implications in Asterids

    Chuan Ku; Wan-Chia Chung; Ling-Ling Chen; Chih-Horng Kuo

    2013-01-01

    The Madagascar periwinkle ( Catharanthus roseus in the family Apocynaceae) is an important medicinal plant and is the source of several widely marketed chemotherapeutic drugs. It is also commonly grown for its ornamental values and, due to ease of infection and distinctiveness of symptoms, is often used as the host for studies on phytoplasmas, an important group of uncultivated plant pathogens. To gain insights into the characteristics of apocynaceous plastid genomes (plastomes), we used a re...

  7. Progress of Research on Chemical and Active Constituents of Catharanthus Roseus%长春花化学成分研究进展

    杨莹莹; 张广晶; 徐雅娟; 徐暾海; 刘铜华; 张舒媛

    2014-01-01

    对长春花的化学成分研究进展进行文献整理,为其进一步开发利用提供有价值的参考。%This paper sorted the literatures on progress of researches on chemical contents of Catharanthus Roseus,so as to provide val-uable reference for its further development and utilization.

  8. Indirect regeneration from in vitro leaf tissue of periwinkle (Catharanthus roseus L.) in response to different treatments of plant growth regulators

    B.E. Sayed-Tabatabaei; F. Eatesam; M. Talebi

    2012-01-01

    Periwinkle (Catharanthus roseus L.) belongs to the Apocynaceae family and accumulates more than 130 terpenoid indole alkaloids (TIAs), of which two dimeric alkaloids Vinblastine and Vincristine have antineoplastic activity and are useful for treatment of various cancers. Therefore, the production of these drugs has been emphasized in plant tissue culture. In this research, 25 treatments of plant growth regulators to produce callus from leaf explants and seven treatments for regeneration of ca...

  9. Fungal endophytes of Catharanthus roseus enhance vindoline content by modulating structural and regulatory genes related to terpenoid indole alkaloid biosynthesis.

    Pandey, Shiv S; Singh, Sucheta; Babu, C S Vivek; Shanker, Karuna; Srivastava, N K; Shukla, Ashutosh K; Kalra, Alok

    2016-01-01

    Not much is known about the mechanism of endophyte-mediated induction of secondary metabolite production in Catharanthus roseus. In the present study two fungal endophytes, Curvularia sp. CATDLF5 and Choanephora infundibulifera CATDLF6 were isolated from the leaves of the plant that were found to enhance vindoline content by 229-403%. The isolated endophytes did not affect the primary metabolism of the plant as the maximum quantum efficiency of PSII, net CO2 assimilation, plant biomass and starch content of endophyte-inoculated plants was similar to endophyte-free control plants. Expression of terpenoid indole alkaloid (TIA) pathway genes, geraniol 10-hydroxylase (G10H), tryptophan decarboxylase (TDC), strictosidine synthase (STR), 16-hydoxytabersonine-O-methyltransferase (16OMT), desacetoxyvindoline-4-hydroxylase (D4H), deacetylvindoline-4-O-acetyltransferase (DAT) were upregulated in endophyte-inoculated plants. Endophyte inoculation upregulated the expression of the gene for transcriptional activator octadecanoid-responsive Catharanthus AP2-domain protein (ORCA3) and downregulated the expression of Cys2/His2-type zinc finger protein family transcriptional repressors (ZCTs). The gene for the vacuolar class III peroxidase (PRX1), responsible for coupling vindoline and catharanthine, was upregulated in endophyte-inoculated plants. These endophytes may enhance vindoline production by modulating the expression of key structural and regulatory genes of vindoline biosynthesis without affecting the primary metabolism of the host plant. PMID:27220774

  10. 土壤条件对长春花生物碱含量的影响%Effects of Different Soil on Alkaloid Content of Catharanthus roseus

    何际婵; 董志超; 王建荣; 晏小霞

    2012-01-01

    Objective:To studied three kinds of Alkaloid contents of Catharanthus roseus ( L.) G. Don ( bright - eyes, rose periwinkle, Madagascar periwinkle) in different soil. Methods: Rp - Hplc method was used to determine the contents of vinblastine, catharanthus and vindohne from C. roseus. Results: Contents were the highest under sandy soil, with the values being 0. 0058,0. 0317 ,0. 0823mg/mL respectively. Conclusion: C. roseus growing in coastal saline soil can improve the contents of vinblastine, catharanthus and vindoline.%目的:研究不同土壤条件对长春花体内3种生物碱含量的影响.方法:采用反相高效液相色谱(RP-HPLC)法测定不同土壤下长春花中长春碱、长春质碱、文多灵含量.结果:沙质壤土中长春碱、长春质碱、文多灵含量最高,分别为0.0058、0.0317、0.0823mg/mL.结论:以沙质壤土种植长春花可提高其植株体内长春碱、长春质碱、文多灵的含量.

  11. Investigation of a substrate-specifying residue within Papaver somniferum and Catharanthus roseus aromatic amino acid decarboxylases.

    Torrens-Spence, Michael P; Lazear, Michael; von Guggenberg, Renee; Ding, Haizhen; Li, Jianyong

    2014-10-01

    Plant aromatic amino acid decarboxylases (AAADs) catalyze the decarboxylation of aromatic amino acids with either benzene or indole rings. Because the substrate selectivity of AAADs is intimately related to their physiological functions, primary sequence data and their differentiation could provide significant physiological insights. However, due to general high sequence identity, plant AAAD substrate specificities have been difficult to identify through primary sequence comparison. In this study, bioinformatic approaches were utilized to identify several active site residues within plant AAAD enzymes that may impact substrate specificity. Next a Papaver somniferum tyrosine decarboxylase (TyDC) was selected as a model to verify our putative substrate-dictating residues through mutation. Results indicated that mutagenesis of serine 372 to glycine enables the P. somniferum TyDC to use 5-hydroxytryptophan as a substrate, and reduces the enzyme activity toward 3,4-dihydroxy-L-phenylalanine (dopa). Additionally, the reverse mutation in a Catharanthus roseus tryptophan decarboxylase (TDC) enables the mutant enzyme to utilize tyrosine and dopa as substrates with a reduced affinity toward tryptophan. Molecular modeling and molecular docking of the P. somniferum TyDC and the C. roseus TDC enzymes provided a structural basis to explain alterations in substrate specificity. Identification of an active site residue that impacts substrate selectivity produces a primary sequence identifier that may help differentiate the indolic and phenolic substrate specificities of individual plant AAADs. PMID:25107664

  12. Silencing the Transcriptional Repressor, ZCT1, Illustrates the Tight Regulation of Terpenoid Indole Alkaloid Biosynthesis in Catharanthus roseus Hairy Roots.

    Rizvi, Noreen F; Weaver, Jessica D; Cram, Erin J; Lee-Parsons, Carolyn W T

    2016-01-01

    The Catharanthus roseus plant is the source of many valuable terpenoid indole alkaloids (TIAs), including the anticancer compounds vinblastine and vincristine. Transcription factors (TFs) are promising metabolic engineering targets due to their ability to regulate multiple biosynthetic pathway genes. To increase TIA biosynthesis, we elicited the TIA transcriptional activators (ORCAs and other unidentified TFs) with the plant hormone, methyl jasmonate (MJ), while simultaneously silencing the expression of the transcriptional repressor ZCT1. To silence ZCT1, we developed transgenic hairy root cultures of C. roseus that expressed an estrogen-inducible Zct1 hairpin for activating RNA interference. The presence of 17β-estradiol (5μM) effectively depleted Zct1 in hairy root cultures elicited with MJ dosages that either optimize or inhibit TIA production (250 or 1000μM). However, silencing Zct1 was not sufficient to increase TIA production or the expression of the TIA biosynthetic genes (G10h, Tdc, and Str), illustrating the tight regulation of TIA biosynthesis. The repression of the TIA biosynthetic genes at the inhibitory MJ dosage does not appear to be solely regulated by ZCT1. For instance, while Zct1 and Zct2 levels decreased through activating the Zct1 hairpin, Zct3 levels remained elevated. Since ZCT repressors have redundant yet distinct functions, silencing all three ZCTs may be necessary to relieve their repression of alkaloid biosynthesis. PMID:27467510

  13. Correspondence between flowers and leaves in terpenoid indole alkaloid metabolism of the phytoplasma-infected Catharanthus roseus plants.

    Srivastava, Suchi; Pandey, Richa; Kumar, Sushil; Nautiyal, Chandra Shekhar

    2014-11-01

    Several plants of Catharanthus roseus cv 'leafless inflorescence (lli)' showing phenotype of phytoplasma infection were observed for symptoms of early flowering, virescence, phyllody, and apical clustering of branches. Symptomatic plants were studied for the presence/absence and identity of phytoplasma in flowers. Transcription levels of several genes involved in plants' metabolism and development, accumulation of pharmaceutically important terpenoid indole alkaloids in flowers and leaves and variation in the root-associated microbial flora were examined. The expression profile of 12 genes studied was semi-quantitatively similar in control leaves and phytoplasma-infected leaves and flowers, in agreement with the symptoms of virescence and phyllody in phytoplasma-infected plants. The flowers of phytoplasma-infected plants possessed the TIA profile of leaves and accumulated catharanthine, vindoline, and vincristine and vinblastine in higher concentrations than leaves. The roots of the infected plants displayed lower microbial diversity than those of normal plants. In conclusion, phytoplasma affected the biology of C. roseus lli plants multifariously, it reduced the differences between the metabolite accumulates of the leaves and flowers and restrict the microbial diversity of rhizosphere. PMID:24658891

  14. Effect of chromium on antioxidant potential of Catharanthus roseus varieties and production of their anticancer alkaloids: vincristine and vinblastine.

    Rai, Vartika; Tandon, Pramod Kumar; Khatoon, Sayyada

    2014-01-01

    Catharanthus roseus (L.) G. Don, a medicinal plant, has a very important place in the traditional as well as modern pharmaceutical industry. Two common varieties of this plant rosea and alba are named so because of pink and white coloured flowers, respectively. This plant comprises of about 130 terpenoid indole alkaloids and two of them, vincristine and vinblastine, are common anticancer drugs. The effect of chromium (Cr) on enzymatic and non-enzymatic antioxidant components and on secondary metabolites vincristine and vinblastine was studied under pot culture conditions of both varieties of C. roseus. Antioxidant responses of these varieties were analyzed under 0, 10, 50, and 100  μM chromium (Cr) level in order to investigate the plant's protective mechanisms against Cr induced oxidative stress. The results indicated that Cr affects all the studied parameters and decreases growth performance. However, vincristine and vinblastine contents were increased under Cr stress. Results are quite encouraging, as this plant shows good antioxidant potential and increased the level of active constituents under Cr stress. PMID:24734252

  15. PERKEMBANGAN LATISIFER PADA KULTUR KALUS CATHARANTHUS ROSEUS (L G. DON YANG DIINDUKSI DENGAN KOMBINASI ZAT PENGATUR TUMBUH KINETIN + NAA

    NI NYOMAN DARSINI

    2011-12-01

    Full Text Available The development of laticifer on callus culture of Catharanthus roseus (L G Don in Zenk medium supplemented with combination of plant growth regulator kinetin + NAA was studied. The explants were taken from the second folium from shoot apex. Development of laticifer was observed using descriptive analysis method for callus anatomy and percentage of laticifer was observed during 4–14 weeks of callus development. The percentage of laticifer was determined by counting the average number of the laticifer and the average number of surrounding cells in every optical field of few under light microscope. The results showed that early development of laticifer which was induced with plant regulator growth kinetin + NAA was found in the 9 weeks old callus. The laticifer has specific characteristics i.e. thicker cell wall and longer cell than sorounding cell. Elongated laticifer was observed at 12 weeks old callus. The highest percentage of laticifer on callus C. roseus induced with combination of kinetin and NAA was found in 12 weeks old callus i.e. 0,12%. At 13 and 14 weeks old callus, the anatomy of laticifer was similar to that at 12 weeks old callus, but the percentage was lower.

  16. Identification and quantification of active alkaloids in Catharanthus roseus by liquid chromatography-ion trap mass spectrometry.

    Chen, Qinhua; Zhang, Wenpeng; Zhang, Yulin; Chen, Jing; Chen, Zilin

    2013-08-15

    Catharanthus roseus is an important dicotyledonous medicinal plant that produces anticancer compounds. The active alkaloids vinblastine, vindoline, ajmalicine, catharanthine, and vinleurosine were identified by direct-injection ion trap-mass spectrometry (IT-MS) for collecting MS(1-2) spectra. The determinations of five alkaloids were accomplished by liquid chromatography (LC) with UV and MS detections. The analytes provided good signals corresponding to the protonated molecular ions [M+H](+) and product ions. The precursor ions and product ions for quantification of vinblastine, vindoline, ajmalicine, catharanthine, and vinleurosine were m/z 825→807, 457→397, 353→144, 337→144 and 809→748 by LC-IT-MS, respectively. Two methods were used to evaluate a number of validation characteristics (repeatability, LOD, calibration range, and recovery). MS provided a high selectivity and sensitivity for determination of five alkaloids in positive mode. After optimisation of the methods, separation, identification and quantification of the five components in C. roseus were comprehensively accomplished by HPLC with UV and MS detection. PMID:23561180

  17. A novel cytochrome P450 gene from Catharanthus roseus cell line C20hi: cloning and characterization of expression

    Lihong He

    2012-06-01

    Full Text Available An expressed sequence tag (EST obtained from a subtractive-suppression hybridization cDNA library constructed using Catharanthus roseus cell line C20hi and its parental cell line C20D was used to clone a full-length cytochrome P450 cDNA of cyp71d1. The encoded polypeptide contained 507 amino acids with 39–56% identity to other CYP71D subfamily members at the amino acid level. Expression characteristics of cyp71d1 were determined using semi-quantitative RT-PCR. The cyp71d1 transcript was expressed in all three cell lines with the highest level in the cell line C20hi. In the mature C. roseus plant, the cyp71d1 cDNA was highly expressed in petals, roots and stems, but very weakly expressed in young leaves. Its transcription level increased with the development of flowers. 2,4-D could down-regulate the transcription of cyp71d1, as did KT, but only to a minor degree. Neither light nor yeast elicitor could induce the transcription of cyp71d1.

  18. Effect of Chromium on Antioxidant Potential of Catharanthus roseus Varieties and Production of Their Anticancer Alkaloids: Vincristine and Vinblastine

    Vartika Rai

    2014-01-01

    Full Text Available Catharanthus roseus (L. G. Don, a medicinal plant, has a very important place in the traditional as well as modern pharmaceutical industry. Two common varieties of this plant rosea and alba are named so because of pink and white coloured flowers, respectively. This plant comprises of about 130 terpenoid indole alkaloids and two of them, vincristine and vinblastine, are common anticancer drugs. The effect of chromium (Cr on enzymatic and non-enzymatic antioxidant components and on secondary metabolites vincristine and vinblastine was studied under pot culture conditions of both varieties of C. roseus. Antioxidant responses of these varieties were analyzed under 0, 10, 50, and 100 μM chromium (Cr level in order to investigate the plant’s protective mechanisms against Cr induced oxidative stress. The results indicated that Cr affects all the studied parameters and decreases growth performance. However, vincristine and vinblastine contents were increased under Cr stress. Results are quite encouraging, as this plant shows good antioxidant potential and increased the level of active constituents under Cr stress.

  19. Heteromeric and homomeric geranyl diphosphate synthases from Catharanthus roseus and their role in monoterpene indole alkaloid biosynthesis.

    Rai, Avanish; Smita, Shachi S; Singh, Anup Kumar; Shanker, Karuna; Nagegowda, Dinesh A

    2013-09-01

    Catharanthus roseus is the sole source of two most important monoterpene indole alkaloid (MIA) anti-cancer agents: vinblastine and vincristine. MIAs possess a terpene and an indole moiety derived from terpenoid and shikimate pathways, respectively. Geranyl diphosphate (GPP), the entry point to the formation of terpene moiety, is a product of the condensation of isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) by GPP synthase (GPPS). Here, we report three genes encoding proteins with sequence similarity to large subunit (CrGPPS.LSU) and small subunit (CrGPPS.SSU) of heteromeric GPPSs, and a homomeric GPPSs. CrGPPS.LSU is a bifunctional enzyme producing both GPP and geranyl geranyl diphosphate (GGPP), CrGPPS.SSU is inactive, whereas CrGPPS is a homomeric enzyme forming GPP. Co-expression of both subunits in Escherichia coli resulted in heteromeric enzyme with enhanced activity producing only GPP. While CrGPPS.LSU and CrGPPS showed higher expression in older and younger leaves, respectively, CrGPPS.SSU showed an increasing trend and decreased gradually. Methyl jasmonate (MeJA) treatment of leaves significantly induced the expression of only CrGPPS.SSU. GFP localization indicated that CrGPPS.SSU is plastidial whereas CrGPPS is mitochondrial. Transient overexpression of AmGPPS.SSU in C. roseus leaves resulted in increased vindoline, immediate monomeric precursor of vinblastine and vincristine. Although C. roseus has both heteromeric and homomeric GPPS enzymes, our results implicate the involvement of only heteromeric GPPS with CrGPPS.SSU regulating the GPP flux for MIA biosynthesis. PMID:23543438

  20. Cytosine hypomethylation at CHG and CHH sites in the pleiotropic mutants of Mendelian inheritance in Catharanthus roseus.

    Kumari, Renu; Yadav, Gitanjali; Sharma, Vishakha; Sharma, Vinay; Kumar, Sushil

    2013-12-01

    The 5S and 18S rDNA sequences of Catharanthus roseus cv 'Nirmal' (wild type) and its leafless inflorescence (lli), evergreen dwarf (egd) and irregular leaf lamina (ill) single mutants and lli egd, lli ill and egd ill double mutants were characterized. The lli, egd and ill mutants of Mendelian inheritance bore the names after their most conspicuous morphological feature(s). They had been chemically induced and isolated for their salt tolerance. The double mutants were isolated as morphological segregants from crosses between single mutants. The morphological features of the two parents accompanied salt tolerance in the double mutants. All the six mutants were hypomethylated at repeat sequences, upregulated and downregulated for many genes and carried pleiotropic alterations for several traits. Here the 5S and 18S rDNAs of C. roseus were found to be relatively low in cytosine content. Cytosines were preponderantly in CG context (53%) and almost all of them were methylated (97%). The cytosines in CHH and CHG (where H = A, T or C) contexts were largely demethylated (92%) in mutants. The demethylation was attributable to reduced expression of RDR2 and DRM2 led RNA dependant DNA methylation and CMT3 led maintenance methylation pathways. Mutants had gained some cytosines by substitution of C at T sites. These perhaps arose on account of errors in DNA replication, mediated by widespread cytosine demethylation at CHG and CHH sites. It was concluded that the regulation of cytosine ethylation mechanisms was disturbed in the mutants. ILL, EGD and LLI genes were identified as the positive regulators of other genes mediating the RdDM and CMT3 pathways, for establishment and maintenance of cytosine methylation in C. roseus. PMID:24371171

  1. Cytosine hypomethylation at CHG and CHH sites in the pleiotropic mutants of Mendelian inheritance in Catharanthus roseus

    Renu Kumari; Gitanjali Yadav; Vishakha Sharma; Vinay Sharma; Sushil Kumar

    2013-12-01

    The 5S and 18S rDNA sequences of Catharanthus roseus cv ‘Nirmal’ (wild type) and its leafless inflorescence (lli), evergreen dwarf (egd) and irregular leaf lamina (ill) single mutants and lli egd, lli ill and egd ill double mutants were characterized. The lli, egd and ill mutants of Mendelian inheritance bore the names after their most conspicuous morphological feature(s). They had been chemically induced and isolated for their salt tolerance. The double mutants were isolated as morphological segregants from crosses between single mutants. The morphological features of the two parents accompanied salt tolerance in the double mutants. All the six mutants were hypomethylated at repeat sequences, upregulated and downregulated for many genes and carried pleiotropic alterations for several traits. Here the 5S and 18S rDNAs of C. roseus were found to be relatively low in cytosine content. Cytosines were preponderantly in CG context (53%) and almost all of them were methylated (97%). The cytosines in CHH and CHG (where H = A, T or C) contexts were largely demethylated (92%) in mutants. The demethylation was attributable to reduced expression of RDR2 and DRM2 led RNA dependant DNA methylation and CMT3 led maintenance methylation pathways. Mutants had gained some cytosines by substitution of C at T sites. These perhaps arose on account of errors in DNA replication, mediated by widespread cytosine demethylation at CHG and CHH sites. It was concluded that the regulation of cytosine methylation mechanisms was disturbed in the mutants. ILL, EGD and LLI genes were identified as the positive regulators of other genes mediating the RdDM and CMT3 pathways, for establishment and maintenance of cytosine methylation in C. roseus.

  2. Overexpression of ORCA3 and G10H in Catharanthus roseus Plants Regulated Alkaloid Biosynthesis and Metabolism Revealed by NMR-Metabolomics

    Pan, Qifang; Wang, Quan; Yuan, Fang; Xing, Shihai; Zhao, Jingya; Choi, Young Hae; Verpoorte, Robert; Tian, Yuesheng; Wang, Guofeng; Tang, Kexuan

    2012-01-01

    In order to improve the production of the anticancer dimeric indole alkaloids in Catharanthuse roseus, much research has been dedicated to culturing cell lines, hairy roots, and efforts to elucidate the regulation of the monoterpenoid indole alkaloid (MIA) biosynthesis. In this study, the ORCA3 (Octadecanoid-derivative Responsive Catharanthus AP2-domain) gene alone or integrated with the G10H (geraniol 10-hydroxylase) gene were first introduced into C. roseus plants. Transgenic C. roseus plants overexpressing ORCA3 alone (OR lines), or co-overexpressing G10H and ORCA3 (GO lines) were obtained by genetic modification. ORCA3 overexpression induced an increase of AS, TDC, STR and D4H transcripts but did not affect CRMYC2 and G10H transcription. G10H transcripts showed a significant increase under G10H and ORCA3 co-overexpression. ORCA3 and G10H overexpression significantly increased the accumulation of strictosidine, vindoline, catharanthine and ajmalicine but had limited effects on anhydrovinblastine and vinblastine levels. NMR-based metabolomics confirmed the higher accumulation of monomeric indole alkaloids in OR and GO lines. Multivariate data analysis of 1H NMR spectra showed change of amino acid, organic acid, sugar and phenylpropanoid levels in both OR and GO lines compared to the controls. The result indicated that enhancement of MIA biosynthesis by ORCA3 and G10H overexpression might affect other metabolic pathways in the plant metabolism of C. roseus. PMID:22916202

  3. Overexpression of ORCA3 and G10H in Catharanthus roseus plants regulated alkaloid biosynthesis and metabolism revealed by NMR-metabolomics.

    Qifang Pan

    Full Text Available In order to improve the production of the anticancer dimeric indole alkaloids in Catharanthuse roseus, much research has been dedicated to culturing cell lines, hairy roots, and efforts to elucidate the regulation of the monoterpenoid indole alkaloid (MIA biosynthesis. In this study, the ORCA3 (Octadecanoid-derivative Responsive Catharanthus AP2-domain gene alone or integrated with the G10H (geraniol 10-hydroxylase gene were first introduced into C. roseus plants. Transgenic C. roseus plants overexpressing ORCA3 alone (OR lines, or co-overexpressing G10H and ORCA3 (GO lines were obtained by genetic modification. ORCA3 overexpression induced an increase of AS, TDC, STR and D4H transcripts but did not affect CRMYC2 and G10H transcription. G10H transcripts showed a significant increase under G10H and ORCA3 co-overexpression. ORCA3 and G10H overexpression significantly increased the accumulation of strictosidine, vindoline, catharanthine and ajmalicine but had limited effects on anhydrovinblastine and vinblastine levels. NMR-based metabolomics confirmed the higher accumulation of monomeric indole alkaloids in OR and GO lines. Multivariate data analysis of (1H NMR spectra showed change of amino acid, organic acid, sugar and phenylpropanoid levels in both OR and GO lines compared to the controls. The result indicated that enhancement of MIA biosynthesis by ORCA3 and G10H overexpression might affect other metabolic pathways in the plant metabolism of C. roseus.

  4. 长春花(Catharanthus roseus)中吲哚类生物碱含量的比较%Comparative Study on the Contents of Indole Alkaloids in Catharanthus roseus

    张琳; 祖元刚; 牛卉颖; 张颜滨; 孙志强

    2008-01-01

    建立了反相高效液相法测定长春花中吲哚类生物碱文多灵、长春质碱和阿玛碱含量的方法,色谱柱为HiQ sil C18色谱柱(250 mm×4.6 mm,5 μm);流动相为1%二乙胺水溶液(磷酸调pH=7.2)-甲醇-乙腈/2:1:1(V/V);流速为1 mL·min-1;检测波长为215 nm;柱温为40℃.并采用此方法对长春花根、茎、叶、花和种子以及不同产地的长春花中的这3种生物碱进行了检测,结果表明文多灵和长春质碱主要存在于在根、茎、叶、花中,阿玛碱主要存在于种子中;并且随着地理位置的北移,长春花中的文多灵和长春质碱的含量逐渐降低,温室中人为控制长春花中的栽培条件能提高二者的含量.%A RP-HPLC quantification method was established to determine the contents of three indole alkaloids-vindoline, catharanthine and ajmalicine in Catharanthus roseus. HiQ sil C18 column(250 mm×4.6 mm,5 μm) was used. The mobile phase was 1% diethylamine(pH=7.2)-methanol-acetonitrile/2:1:1(V/V). The detection wavelength was 215 nm. The flow rate was 1 mL·min-1. The column temperature was 40℃. Different parts of C. roseus were determined and the results showed that vindoline and catharanthine mainly existed in roots, stems, leaves and flowers, while ajmalicine mainly existed in seeds. C. roseus from different habitats were also determined by this method and the results showed that the contents of vindoline and catharanthine in south habitats samples were higher than those in north habitats. For the cultured C. roseus in greenhouse, both vindoline and catharanthine contents were enhanced greatly.

  5. Preliminary results of indole alkaloids production in different roots of Catharanthus roseus cultured in vitro

    Agnieszka Pietrosiuk; Mirosława Furmanowa

    2014-01-01

    Six groups of untransformed and hairy root cultures of Catharunthus roseus (L.) G. Don were established. Agrobacterium rhizogenes strains: ATCC 15834, LBA 9403, and TR 105 were used for infection of the 3-week old rooted plantlets of C. roseus. The highest contents of examined indole alkaloids were found in: roots of intact plants - yohimbine and serpentine; in hairy roots - catharanthine. Vinblastine and ajmalicine were detected in untransformed roots of plants regenerated in vitro, and tran...

  6. Large scale in-silico identification and characterization of simple sequence repeats (SSRs) from de novo assembled transcriptome of Catharanthus roseus (L.) G. Don.

    Kumar, Santosh; Shah, Niraj; Garg, Vanika; Bhatia, Sabhyata

    2014-06-01

    Transcriptomic data of C. roseus offering ample sequence resources for providing better insights into gene diversity: large resource of genic SSR markers to accelerate genomic studies and breeding in Catharanthus . Next-generation sequencing is an efficient system for generating high-throughput complete transcripts/genes and developing molecular markers. We present here the transcriptome sequencing of a 26-day-old Catharanthus roseus seedling tissue using Illumina GAIIX platform that resulted in a total of 3.37 Gb of nucleotide sequence data comprising 29,964,104 reads which were de novo assembled into 26,581 unigenes. Based on similarity searches 58 % of the unigenes were annotated of which 13,580 unique transcripts were assigned 5016 gene ontology terms. Further, 7,687 of the unigenes were found to have Cluster of Orthologous Group classifications, and 4,006 were assigned to 289 Kyoto Encyclopedia of Genes and Genome pathways. Also, 5,221 (19.64 %) of transcripts were distributed to 81 known transcription factor (TF) families. In-silico analysis of the transcriptome resulted in identification of 11,004 SSRs in 26.62 % transcripts from which 2,520 SSR markers were designed which exhibited a non-random pattern of distribution. The most abundant was the trinucleotide repeats (AAG/CTT) followed by the dinucleotide repeats (AG/CT). Location specific analysis of SSRs revealed that SSRs were preferentially associated with the 5'-UTRs with a predicted role in regulation of gene expression. A PCR validation of a set of 48 primers revealed 97.9 % successful amplification, and 76.6 % of them showed polymorphism across different Catharanthus species as well as accessions of C. roseus. In summary, this study will provide an insight into understanding the seedling development and resources for novel gene discovery and SSR development for utilization in marker-assisted selective breeding in C. roseus. PMID:24482265

  7. Precursor feeding studies and molecular characterization of geraniol synthase establish the limiting role of geraniol in monoterpene indole alkaloid biosynthesis in Catharanthus roseus leaves.

    Kumar, Krishna; Kumar, Sarma Rajeev; Dwivedi, Varun; Rai, Avanish; Shukla, Ashutosh K; Shanker, Karuna; Nagegowda, Dinesh A

    2015-10-01

    The monoterpene indole alkaloids (MIAs) are generally derived from strictosidine, which is formed by condensation of the terpene moiety secologanin and the indole moiety tryptamine. There are conflicting reports on the limitation of either terpene or indole moiety in the production of MIAs in Catharanthus roseus cell cultures. Formation of geraniol by geraniol synthase (GES) is the first step in secologanin biosynthesis. In this study, feeding of C. roseus leaves with geraniol, but not tryptophan (precursor for tryptamine), increased the accumulation of the MIAs catharanthine and vindoline, indicating the limitation of geraniol in MIA biosynthesis. This was further validated by molecular and in planta characterization of C. roseus GES (CrGES). CrGES transcripts exhibited leaf and shoot specific expression and were induced by methyl jasmonate. Virus-induced gene silencing (VIGS) of CrGES significantly reduced the MIA content, which was restored to near-WT levels upon geraniol feeding. Moreover, over-expression of CrGES in C. roseus leaves increased MIA content. Further, CrGES exhibited correlation with MIA levels in leaves of different C. roseus cultivars and has significantly lower expression relative to other pathway genes. These results demonstrated that the transcriptional regulation of CrGES and thus, the in planta geraniol availability plays crucial role in MIA biosynthesis. PMID:26398791

  8. A simple and rapid HPLC-DAD method for simultaneously monitoring the accumulation of alkaloids and precursors in different parts and different developmental stages of Catharanthus roseus plants.

    Pan, Qifang; Saiman, Mohd Zuwairi; Mustafa, Natali Rianika; Verpoorte, Robert; Tang, Kexuan

    2016-03-01

    A rapid and simple reversed phase liquid chromatographic system has been developed for simultaneous analysis of terpenoid indole alkaloids (TIAs) and their precursors. This method allowed separation of 11 compounds consisting of eight TIAs (ajmalicine, serpentine, catharanthine, vindoline, vindolinine, vincristine, vinblastine, and anhydrovinblastine) and three related precursors i.e., tryptophan, tryptamine and loganin. The system has been applied for screening the TIAs and precursors in Catharanthus roseus plant extracts. In this study, different organs i.e., flowers, leaves, stems, and roots of C. roseus were investigated. The results indicate that TIAs and precursor accumulation varies qualitatively and quantitatively in different organs of C. roseus. The precursors showed much lower levels than TIAs in all organs. Leaves and flowers accumulate higher level of vindoline, catharanthine and anhydrovinblastine while roots have higher level of ajmalicine, vindolinine and serpentine. Moreover, the alkaloid profiles of leaves harvested at different ages and different growth stages were studied. The results show that the levels of monoindole alkaloids decreased while bisindole alkaloids increased with leaf aging and upon plant growth. The HPLC method has been successfully applied to detect TIAs and precursors in different types of C. roseus samples to facilitate further study of the TIA pathway and its regulation in C. roseus plants. PMID:26854826

  9. Larvicidal efficacy of Catharanthus roseus Linn. (Family:Apocynaceae) leaf extract and bacterial insecticideBacillus thuringiensis againstAnopheles stephensi Liston

    Chellasamy Panneerselvam; Kadarkarai Murugan; Kalimuthu Kovendan; Palanisamy Mahesh Kumar; Sekar Ponarulselvam; Duraisamy Amerasan; Jayapal Subramaniam; Jiang-Shiou Hwang

    2013-01-01

    Objective:To explore the larvicidal activity ofCatharanthus roseus(C. roseus) leaf extract and Bacillus thuringiensis(B. thuringiensis) against the malarial vectorAnopheles stephensi(An. stephensi), when being used alone or together.Methods:The larvicidal activity was assayed at various concentrations under the laboratory and field conditions.TheLC50 andLC90 values of theC. roseus leaf extract were determined by probit analysis.Results:The plant extract showed larvicidal effects after24 h of exposure;however, the highest larval mortality was found in the petroleum ether extract ofC. roseus against the first to fourth instars larvae withLC50=3.34,4.48, 5.90 and8.17 g/L, respectively;B. thuringiensis against the first to fourth instars larvae with LC50=1.72,1.93,2.17 and2.42 g/L, respectively; and the combined treatment withLC50=2.18,2.41, 2.76 and3.22 g/L, respectively.No mortality was observed in the control.Conclusions:The petroleum ether extract ofC. roseus extract andB. thuringiensis have potential to be used as ideal eco-friendly agents for the control ofAn. stephensi in vector control programs.The combined treatment with this plant crude extract and bacterial toxin has better larvicidal efficacy against An. stephensi.

  10. Effects of β-cyclodextrin and methyl jasmonate on the production of vindoline, catharanthine, and ajmalicine in Catharanthus roseus cambial meristematic cell cultures.

    Zhou, Pengfei; Yang, Jiazeng; Zhu, Jianhua; He, Shuijie; Zhang, Wenjin; Yu, Rongmin; Zi, Jiachen; Song, Liyan; Huang, Xuesong

    2015-09-01

    Long-term stable cell growth and production of vindoline, catharanthine, and ajmalicine of cambial meristematic cells (CMCs) from Catharanthus roseus were observed after 2 years of culture. C. roseus CMCs were treated with β-cyclodextrin (β-CD) and methyl jasmonate (MeJA) individually or in combination and were cultured both in conventional Erlenmeyer flasks (100, 250, and 500 mL) and in a 5-L stirred hybrid airlift bioreactor. CMCs of C. roseus cultured in the bioreactor showed higher yields of vindoline, catharanthine, and ajmalicine than those cultured in flasks. CMCs of C. roseus cultured in the bioreactor and treated with 10 mM β-CD and 150 μM MeJA gave the highest yields of vindoline (7.45 mg/L), catharanthine (1.76 mg/L), and ajmalicine (58.98 mg/L), concentrations that were 799, 654, and 426 % higher, respectively, than yields of CMCs cultured in 100-mL flasks without elicitors. Quantitative reverse transcription (RT)-PCR showed that β-CD and MeJA upregulated transcription levels of genes related to the biosynthesis of terpenoid indole alkaloids (TIAs). This is the first study to report that β-CD induced the generation of NO, which plays an important role in mediating the production of TIAs in C. roseus CMCs. These results suggest that β-CD and MeJA can enhance the production of TIAs in CMCs of C. roseus, and thus, CMCs of C. roseus have significant potential to be an industrial platform for production of bioactive alkaloids. PMID:25981997

  11. An Endophytic Fungus, Talaromyces radicus, Isolated from Catharanthus roseus, Produces Vincristine and Vinblastine, Which Induce Apoptotic Cell Death.

    Palem, Padmini P C; Kuriakose, Gini C; Jayabaskaran, Chelliah

    2015-01-01

    Endophytic fungi isolated from Catharanthus roseus were screened for the production of vincristine and vinblastine. Twenty-two endophytic fungi isolated from various tissues of C. roseus were characterized taxonomically by sequence analysis of the internal transcribed spacer (ITS) region of rDNA and grouped into 10 genera: Alternaria, Aspergillus, Chaetomium, Colletotrichum, Dothideomycetes, Eutypella, Eutypa, Flavodon, Fusarium and Talaromyces. The antiproliferative activity of these fungi was assayed in HeLa cells using the MTT assay. The fungal isolates Eutypella sp--CrP14, obtained from stem tissues, and Talaromyces radicus--CrP20, obtained from leaf tissues, showed the strongest antiproliferative activity, with IC50 values of 13.5 μg/ml and 20 μg/ml, respectively. All 22 endophytic fungi were screened for the presence of the gene encoding tryptophan decarboxylase (TDC), the key enzyme in the terpenoid indole alkaloid biosynthetic pathway, though this gene could only be amplified from T. radicus--CrP20 (NCBI GenBank accession number KC920846). The production of vincristine and vinblastine by T. radicus--CrP20 was confirmed and optimized in nine different liquid media. Good yields of vincristine (670 μg/l) in modified M2 medium and of vinblastine (70 μg/l) in potato dextrose broth medium were obtained. The cytotoxic activity of partially purified fungal vincristine was evaluated in different human cancer cell lines, with HeLa cells showing maximum susceptibility. The apoptosis-inducing activity of vincristine derived from this fungus was established through cell cycle analysis, loss of mitochondrial membrane potential and DNA fragmentation patterns. PMID:26697875

  12. Enzyme inhibitor studies reveal complex control of methyl-D-erythritol 4-phosphate (MEP pathway enzyme expression in Catharanthus roseus.

    Mei Han

    Full Text Available In Catharanthus roseus, the monoterpene moiety exerts a strong flux control for monoterpene indole alkaloid (MIA formation. Monoterpene synthesis depends on the methyl-D-erythritol 4-phosphate (MEP pathway. Here, we have explored the regulation of this pathway in response to developmental and environmental cues and in response to specific enzyme inhibitors. For the MEP pathway entry enzyme 1-deoxy-D-xylulose 5-phosphate synthase (DXS, a new (type I DXS isoform, CrDXS1, has been cloned, which, in contrast to previous reports on type II CrDXS, was not transcriptionally activated by the transcription factor ORCA3. Regulation of the MEP pathway in response to metabolic perturbations has been explored using the enzyme inhibitors clomazone (precursor of 5-ketochlomazone, inhibitor of DXS and fosmidomycin (inhibitor of deoxyxylulose 5-phosphate reductoisomerase (DXR, respectively. Young leaves of non-flowering plants were exposed to both inhibitors, adopting a non-invasive in vivo technique. Transcripts and proteins of DXS (3 isoforms, DXR, and hydroxymethylbutenyl diphosphate synthase (HDS were monitored, and protein stability was followed in isolated chloroplasts. Transcripts for DXS1 were repressed by both inhibitors, whereas transcripts for DXS2A&B, DXR and HDS increased after clomazone treatment but were barely affected by fosmidomycin treatment. DXS protein accumulated in response to both inhibitors, whereas DXR and HDS proteins were less affected. Fosmidomycin-induced accumulation of DXS protein indicated substantial posttranscriptional regulation. Furthermore, fosmidomycin effectively protected DXR against degradation in planta and in isolated chloroplasts. Thus our results suggest that DXR protein stability may be affected by substrate binding. In summary, the present results provide novel insight into the regulation of DXS expression in C. roseus in response to MEP-pathway perturbation.

  13. Synthesis of molecularly imprinted polymers for the application of selective clean-up vinblastine from Catharanthus roseus extract

    2010-01-01

    Molecularly imprinted polymers (MIPs) are synthetic tailor-made polymers with high selectivity towards a particular substance (template).An MIP using vinblastine (VLB) as the template molecule was synthesized and characterized.The presence of monomer-template complexes in a non-covalent way was confirmed by UV-vis spectrometry analysis.The polymerization was performed using methacrylic acid (MAA) as the functional monomer,ethylene glycol dimethacrylate (EGDMA) as the cross-linking agent,and toluene as the porogenic solvent by a thermo-polymerization method.The characterization of the obtained MIP was evaluated by scanning electron microscopy (SEM) and Brunauer-Emmett-Teller (BET) analysis.It was observed that the morphology of the MIP was more porous and rough,and the surface area had a significant increase compared with that of the non-imprinted polymer (NIP).This MIP was used as the sorbents of solid-phase extraction (SPE) to assess the selectivity of the MIP after optimization of the SPE protocol.VLB was specifically adsorbed on the MIP cartridge,while to vincristine (VCR),the chemical analog of VLB,almost no selective binding appeared.On the basis of the results,Catharanthus roseus extract was applied to the MIP cartridge for investigating its capability to extract VLB from the plant extract,and the capacity of the MIP cartridge was also evaluated.It was shown that the MIP could effectively enrich VLB from C.roseus extract and the recovery amounted to 93.8%.The solvents dissolving the samples had significant influence on the capacity of the MIP cartridge;it was 750 μg/g in toluene,625 μg/g in chloroform,and 250 μg/g in methanol.

  14. Enzyme Inhibitor Studies Reveal Complex Control of Methyl-D-Erythritol 4-Phosphate (MEP) Pathway Enzyme Expression in Catharanthus roseus

    Han, Mei; Heppel, Simon C.; Su, Tao; Bogs, Jochen; Zu, Yuangang; An, Zhigang; Rausch, Thomas

    2013-01-01

    In Catharanthus roseus, the monoterpene moiety exerts a strong flux control for monoterpene indole alkaloid (MIA) formation. Monoterpene synthesis depends on the methyl-D-erythritol 4-phosphate (MEP) pathway. Here, we have explored the regulation of this pathway in response to developmental and environmental cues and in response to specific enzyme inhibitors. For the MEP pathway entry enzyme 1-deoxy-D-xylulose 5-phosphate synthase (DXS), a new (type I) DXS isoform, CrDXS1, has been cloned, which, in contrast to previous reports on type II CrDXS, was not transcriptionally activated by the transcription factor ORCA3. Regulation of the MEP pathway in response to metabolic perturbations has been explored using the enzyme inhibitors clomazone (precursor of 5-ketochlomazone, inhibitor of DXS) and fosmidomycin (inhibitor of deoxyxylulose 5-phosphate reductoisomerase (DXR)), respectively. Young leaves of non-flowering plants were exposed to both inhibitors, adopting a non-invasive in vivo technique. Transcripts and proteins of DXS (3 isoforms), DXR, and hydroxymethylbutenyl diphosphate synthase (HDS) were monitored, and protein stability was followed in isolated chloroplasts. Transcripts for DXS1 were repressed by both inhibitors, whereas transcripts for DXS2A&B, DXR and HDS increased after clomazone treatment but were barely affected by fosmidomycin treatment. DXS protein accumulated in response to both inhibitors, whereas DXR and HDS proteins were less affected. Fosmidomycin-induced accumulation of DXS protein indicated substantial posttranscriptional regulation. Furthermore, fosmidomycin effectively protected DXR against degradation in planta and in isolated chloroplasts. Thus our results suggest that DXR protein stability may be affected by substrate binding. In summary, the present results provide novel insight into the regulation of DXS expression in C. roseus in response to MEP-pathway perturbation. PMID:23650515

  15. An Endophytic Fungus, Talaromyces radicus, Isolated from Catharanthus roseus, Produces Vincristine and Vinblastine, Which Induce Apoptotic Cell Death.

    Padmini P C Palem

    Full Text Available Endophytic fungi isolated from Catharanthus roseus were screened for the production of vincristine and vinblastine. Twenty-two endophytic fungi isolated from various tissues of C. roseus were characterized taxonomically by sequence analysis of the internal transcribed spacer (ITS region of rDNA and grouped into 10 genera: Alternaria, Aspergillus, Chaetomium, Colletotrichum, Dothideomycetes, Eutypella, Eutypa, Flavodon, Fusarium and Talaromyces. The antiproliferative activity of these fungi was assayed in HeLa cells using the MTT assay. The fungal isolates Eutypella sp--CrP14, obtained from stem tissues, and Talaromyces radicus--CrP20, obtained from leaf tissues, showed the strongest antiproliferative activity, with IC50 values of 13.5 μg/ml and 20 μg/ml, respectively. All 22 endophytic fungi were screened for the presence of the gene encoding tryptophan decarboxylase (TDC, the key enzyme in the terpenoid indole alkaloid biosynthetic pathway, though this gene could only be amplified from T. radicus--CrP20 (NCBI GenBank accession number KC920846. The production of vincristine and vinblastine by T. radicus--CrP20 was confirmed and optimized in nine different liquid media. Good yields of vincristine (670 μg/l in modified M2 medium and of vinblastine (70 μg/l in potato dextrose broth medium were obtained. The cytotoxic activity of partially purified fungal vincristine was evaluated in different human cancer cell lines, with HeLa cells showing maximum susceptibility. The apoptosis-inducing activity of vincristine derived from this fungus was established through cell cycle analysis, loss of mitochondrial membrane potential and DNA fragmentation patterns.

  16. Vindoline Formation in Shoot Cultures of Catharanthus roseus is Synchronously Activated with Morphogenesis Through the Last Biosynthetic Step

    Campos-Tamayo, Freddy; Hernández-Domínguez, Elizabeta; Vázquez-Flota, Felipe

    2008-01-01

    Background and Aims The Madagascar periwinkle (Catharanthus roseus) produces the monoterpenoid alkaloid vindoline, which requires a specialized cell organization present only in the aerial tissues. Vindoline content can be affected by photoperiod and this effect seems to be associated with the morphogenetic capacity of branches; this association formed the basis of the study reported here. Methods Vindoline-producing in vitro shoot cultures were exposed either to continuous light or a 16-h photoperiod regime. New plantlet formation and alkaloid biosynthesis were analysed throughout a culture cycle. Key Results In cultures under the photoperiod, the formation of new plantlets occurred in a more synchronized fashion as compared to those under continuous light. The accumulation of vindoline in cultures under the photoperiod occurred in co-ordination with plantlet formation, in constrast to cultures under continuous light, and coincided with a peak of activity of deacetylvindoline acetyl CoA acetyltransferase (DAT), the enzyme that catalyses the last step in vindoline biosynthesis. When new plantlet formation was blocked in cultures under the photoperiod by treatment with phytoregulators, vindoline synthesis was also reduced via an effect on DAT activity. No association between plantlet formation and other biosynthetic enzymes, such as tryptophan decarboxylase (TDC) and deacetoxyvindoline 4-hydroxylase (D4H), was found. Effects of light treatment on vindoline synthesis were not mediated by ORCA-3 proteins (which are involved in the induction of alkaloid synthesis in response to elicitation), suggesting that the presence of a different set of regulatory proteins. Conclusions The data suggest that vindoline biosynthesis is associated with morphogenesis in shoot cultures of C. roseus. PMID:18587132

  17. Preliminary results of indole alkaloids production in different roots of Catharanthus roseus cultured in vitro

    Agnieszka Pietrosiuk

    2014-02-01

    Full Text Available Six groups of untransformed and hairy root cultures of Catharunthus roseus (L. G. Don were established. Agrobacterium rhizogenes strains: ATCC 15834, LBA 9403, and TR 105 were used for infection of the 3-week old rooted plantlets of C. roseus. The highest contents of examined indole alkaloids were found in: roots of intact plants - yohimbine and serpentine; in hairy roots - catharanthine. Vinblastine and ajmalicine were detected in untransformed roots of plants regenerated in vitro, and transferred to the soil for 5 months.

  18. The bHLH transcription factor BIS1 controls the iridoid branch of the monoterpenoid indole alkaloid pathway in Catharanthus roseus.

    Van Moerkercke, Alex; Steensma, Priscille; Schweizer, Fabian; Pollier, Jacob; Gariboldi, Ivo; Payne, Richard; Vanden Bossche, Robin; Miettinen, Karel; Espoz, Javiera; Purnama, Purin Candra; Kellner, Franziska; Seppänen-Laakso, Tuulikki; O'Connor, Sarah E; Rischer, Heiko; Memelink, Johan; Goossens, Alain

    2015-06-30

    Plants make specialized bioactive metabolites to defend themselves against attackers. The conserved control mechanisms are based on transcriptional activation of the respective plant species-specific biosynthetic pathways by the phytohormone jasmonate. Knowledge of the transcription factors involved, particularly in terpenoid biosynthesis, remains fragmentary. By transcriptome analysis and functional screens in the medicinal plant Catharanthus roseus (Madagascar periwinkle), the unique source of the monoterpenoid indole alkaloid (MIA)-type anticancer drugs vincristine and vinblastine, we identified a jasmonate-regulated basic helix-loop-helix (bHLH) transcription factor from clade IVa inducing the monoterpenoid branch of the MIA pathway. The bHLH iridoid synthesis 1 (BIS1) transcription factor transactivated the expression of all of the genes encoding the enzymes that catalyze the sequential conversion of the ubiquitous terpenoid precursor geranyl diphosphate to the iridoid loganic acid. BIS1 acted in a complementary manner to the previously characterized ethylene response factor Octadecanoid derivative-Responsive Catharanthus APETALA2-domain 3 (ORCA3) that transactivates the expression of several genes encoding the enzymes catalyzing the conversion of loganic acid to the downstream MIAs. In contrast to ORCA3, overexpression of BIS1 was sufficient to boost production of high-value iridoids and MIAs in C. roseus suspension cell cultures. Hence, BIS1 might be a metabolic engineering tool to produce sustainably high-value MIAs in C. roseus plants or cultures. PMID:26080427

  19. Comparative studies of elemental composition in leaves and flowers of Catharanthus roseus growing in Bangladesh

    Shahin Aziz

    2016-01-01

    Conclusions: The elemental composition in both leaves and flowers of C. roseus were found to be different. Therefore, different parts of this medicinal plant are enriched in some micro and macro nutrients like Fe, Ca, Na, K, Zn, which are very important for biological metabolic system as well as human health.

  20. Alkaloid variations in Catharanthus roseus seedlings treated by different temperatures in short term and long term

    GUO Xiao-rui; YANG Lei; YU Jing-hua; TANG Zhong-hua; ZU Yuan-gang

    2007-01-01

    A study was conducted to investigate the effects of high temperature on variations of alkaloid metabolism in C. Roseus seedlings in Key Laboratory of Forest Plant Ecology, Northeast Forestry University, Heilongiang, China. 60-day-old C. Roseus seedlings with 3-4 pairs of leaves were incubated in chambers with temperature of 30℃ and 40℃ for short-term heat shock experiment and 20℃, 25℃ and 35℃ for long-term experiment. The contents ofvindoline, catharanthine, vinblastine and vincristine in C. Roseus leaves and root were checked at different temperatures in short term (1-6 h) and long term (1-16 d). Results showed that under short-term heat shock, the contents of vindoline,catharanthine and vinblastine in leaves of the seedlings were higher at 40℃ than at 30℃, but after 6 h treatment, the contents of vindoline and catharanthine under the two temperatures came to the same level. Catharanthine was exclusively distributed in C. Roseus roots and its content was increased by 40% after two hours incubation at 40℃, while increased slowly at 30℃ incubation and reached the highest value at 6 h. In the Long-term experiment, concentrations of monomeric alkaloids catharanthine and vindoline were higher at 20℃ than at 25℃ and had a sharp increase under the condition of 35℃. While for dimeric alkaloid, it showed that the higher the temperature, the earlier the peak value of vinblastine content appears. Vincristine had a continuous enhancement and attained 0.027 mg·g-1 at 16th day under 35℃ condition which was higher than those in the other conditions. It was concluded that high temperature could promote the accumulation of different alkaloids in C roseus and the accumulation characteristic is highly related to treatment time.

  1. Ornamental Exterior versus Therapeutic Interior of Madagascar Periwinkle (Catharanthus roseus: The Two Faces of a Versatile Herb

    Naghmeh Nejat

    2015-01-01

    Full Text Available Catharanthus roseus (L. known as Madagascar periwinkle (MP is a legendary medicinal plant mostly because of possessing two invaluable antitumor terpenoid indole alkaloids (TIAs, vincristine and vinblastine. The plant has also high aesthetic value as an evergreen ornamental that yields prolific blooms of splendid colors. The plant possesses yet another unique characteristic as an amiable experimental host for the maintenance of the smallest bacteria found on earth, the phytoplasmas and spiroplasmas, and serves as a model for their study. Botanical information with respect to synonyms, vernacular names, cultivars, floral morphology, and reproduction adds to understanding of the plant while the geography and ecology of periwinkle illustrate the organism’s ubiquity. Good agronomic practices ensure generous propagation of healthy plants that serve as a source of bioactive compounds and multitudinous horticultural applications. The correlation between genetic diversity, variants, and TIA production exists. MP is afflicted with a whole range of diseases that have to be properly managed. The ethnobotanical significance of MP is exemplified by its international usage as a traditional remedy for abundant ailments and not only for cancer. TIAs are present only in micro quantities in the plant and are highly poisonous per se rendering a challenge for researchers to increase yield and reduce toxicity.

  2. Isolation, Purification and Characterization of Vinblastine and Vincristine from Endophytic Fungus Fusarium oxysporum Isolated from Catharanthus roseus

    Kumar, Ashutosh; Patil, Deepak; Rajamohanan, Pattuparambil Ramanpillai; Ahmad, Absar

    2013-01-01

    Endophytic fungi reside in a symbiotic fashion inside their host plants, mimic their chemistry and interestingly, produce the same natural products as their hosts and are thus being screened for the production of valuable compounds like taxol, camptothecin, podophyllotoxin, etc. Vinblastine and vincristine are excellent anti-cancer drugs but their current production using plants is non-abundant and expensive. In order to make these drugs readily available to the patients at affordable prices, we isolated the endophytic fungi from Catharanthus roseus plant and found a fungus AA-CRL-6 which produces vinblastine and vincristine in appreciable amounts. These drugs were purified by TLC and HPLC and characterized using UV-Vis spectroscopy, ESI-MS, MS/MS and 1H NMR. One liter of culture filtrate yielded 76 µg and 67 µg of vinblastine and vincristine respectively. This endophytic fungal strain was identified as Fusarium oxysporum based upon its cultural and morphological characteristics and internal transcribed spacer (ITS) sequence analysis. PMID:24066024

  3. Salicylic acid restrains nickel toxicity, improves antioxidant defence system and enhances the production of anticancer alkaloids in Catharanthus roseus (L.).

    Idrees, Mohd; Naeem, M; Aftab, Tariq; Khan, M Masroor A; Moinuddin

    2013-05-15

    Salicylic acid (SA) has been reported to ameliorate various stresses in plants. In order to explore the role of SA under nickel (Ni) stress, thirty-days old plants of periwinkle (Catharanthus roseus L.) were supplied with eight treatments comprising basal application of Ni (0, 50, 100 and 150 mg kg(-1)) and foliar application of SA (0 and 10(-5)M) under net house conditions. Ni application significantly reduced the growth attributes including plant height, leaf-area index and fresh and dry weights of shoot and root. Increasing Ni concentration led to a gradual decrease in photosynthetic parameters and activities of nitrate reductase and carbonic anhydrase. The plants, undergoing Ni stress, exhibited a significant increase in the activity of superoxide dismutase, catalase and peroxidase together with an increase in electrolyte leakage and proline content. Total alkaloid content was also declined in Ni-treated plants. Foliar application of SA (10(-5)M) reduced the deleterious effects of Ni on plant growth, accelerating the restoration of growth processes. SA also improved the total alkaloid content under normal as well as adverse conditions. Foliar spray of SA significantly improved the content of anticancer alkaloids vincristine (by 22.2%) and vinblastine (by 50.0%) in plants treated with 150 mg kg(-1) of Ni. PMID:23597961

  4. Phosphatidate Kinase, A Novel Enzyme in Phospholipid Metabolism (Characterization of the Enzyme from Suspension-Cultured Catharanthus roseus Cells).

    Wissing, J. B.; Kornak, B.; Funke, A.; Riedel, B.

    1994-01-01

    Phosphatidate kinase (adenosine 5[prime]-triphosphate:phosphatidic acid phosphotransferase), a novel enzyme of phospholipid metabolism, was detected recently in the plasma membranes of suspension-cultured Catharanthus roseus cells and purified (J.B. Wissing, H. Behrbohm [1993] Plant Physiol 102: 1243-1249). In the present work the properties of phosphatidate kinase are described. The enzyme showed a pH optimum of 6.1 and an isoelectric point of 4.8, and was rather stable in the presence of its substrates. Although the kinase accepted both ATP and GTP, with Km values of about 12 and 18 [mu]M, respectively, the only lipid substrate was phosphatidic acid; neither lysophosphatidic acid nor any other lipid tested was phosphorylated. With 32P- and 14C-labeled diacylglycerol pyrophosphate, the product of the enzyme, it was shown that the kinase catalyzes a reversible reaction. The activity of the extracted enzyme depended on the presence of surfactants such as Triton X-100 or [beta]-octylglucoside, whereas deoxycholate was strongly inhibitory. Kinetic analysis with Triton X-100/phosphatidate mixed micelles performed according to the "surface dilution" kinetic model showed saturation kinetics with respect to both bulk and surface concentration of phosphatidate. The interfacial Michaelis constant for phosphatidate was determined as 0.6 mol %. PMID:12232252

  5. Proton/Phosphate Stoichiometry in Uptake of Inorganic Phosphate by Cultured Cells of Catharanthus roseus (L.) G. Don

    Sakano, Katsuhiro

    1990-01-01

    Upon absorption of phosphate, cultured cells of Catharanthus roseus (L.) G. Don caused a rapid alkalinization of the medium in which they were suspended. The alkalinization continued until the added phosphate was completely exhausted from the medium, at which time the pH of the medium started to drop sharply toward the original pH value. Phosphate exposure caused the pH of the medium to increase from pH 3.5 to values as high as 5.8, while the rate of phosphate uptake was constant throughout (10-17 micromoles per hour per gram fresh weight). This indicates that no apparent pH optimum exists for the phosphate uptake by the cultured cells. The amount of protons cotransported with phosphate was calculated from the observed pH change up to the maximum alkalinization and the titration curve of the cell suspension. Proton/phosphate transport stoichiometry ranged from less than unity to 4 according to the amount of phosphate applied. At low phosphate doses, the stoichiometries were close to 4, while at high phosphate doses, smaller stoichiometries were observed. This suggests that, at high phosphate doses, activation of the proton pump is induced by the longer lasting proton influx acidifying the cytoplasm. The increased H+ efflux due to the proton pump could partially compensate protons taken up via the proton-phosphate cotransport system. Thus, the H+/H2PO4− stoichiometry of the cotransport is most likely to be 4. PMID:16667491

  6. Ornamental exterior versus therapeutic interior of Madagascar periwinkle (Catharanthus roseus): the two faces of a versatile herb.

    Nejat, Naghmeh; Valdiani, Alireza; Cahill, David; Tan, Yee-How; Maziah, Mahmood; Abiri, Rambod

    2015-01-01

    Catharanthus roseus (L.) known as Madagascar periwinkle (MP) is a legendary medicinal plant mostly because of possessing two invaluable antitumor terpenoid indole alkaloids (TIAs), vincristine and vinblastine. The plant has also high aesthetic value as an evergreen ornamental that yields prolific blooms of splendid colors. The plant possesses yet another unique characteristic as an amiable experimental host for the maintenance of the smallest bacteria found on earth, the phytoplasmas and spiroplasmas, and serves as a model for their study. Botanical information with respect to synonyms, vernacular names, cultivars, floral morphology, and reproduction adds to understanding of the plant while the geography and ecology of periwinkle illustrate the organism's ubiquity. Good agronomic practices ensure generous propagation of healthy plants that serve as a source of bioactive compounds and multitudinous horticultural applications. The correlation between genetic diversity, variants, and TIA production exists. MP is afflicted with a whole range of diseases that have to be properly managed. The ethnobotanical significance of MP is exemplified by its international usage as a traditional remedy for abundant ailments and not only for cancer. TIAs are present only in micro quantities in the plant and are highly poisonous per se rendering a challenge for researchers to increase yield and reduce toxicity. PMID:25667940

  7. A Simplified Procedure for Indole Alkaloid Extraction from Catharanthus roseus Combined with a Semi-synthetic Production Process for Vinblastine

    Marja-Liisa Riekkola

    2007-07-01

    Full Text Available Dried leaves of Catharanthus roseus were extracted with aqueous acidic 0.1 M solution of HCl. Alkaloid-embonate complexes were obtained as precipitates by treating the extract with an alkaline (NaOH solution of embonic acid (4,4-methylene-bis-3-hydroxynaphtalenecarboxylic acid. The precipitate mainly consisted of catharanthine and vindoline embonates and it was directly used as the starting material for a semi-synthesis of the anti-cancer bisindole alkaloid vinblastine. The coupling reaction involved oxidation of catharanthine in aqueous acidic medium by singlet oxygen (1O2, continuously produced in situ by the reaction between H2O2 with NaClO. An excess of NaBH4 was used for the reduction step. Analysis of the reaction mixture indicated a maximum yield of 20% for vinblastine at pH 8.3, based on the initial amount of catharanthine concentration. Direct-injection electrospray ionization mass spectrometry in positive ion mode was used for the identification of vinblastine. The mass spectra of vinblastine were dominated by the corresponding protonated molecular ion [M+H]+ at m/z 811 and the characteristic fragment ions matched with those of the standard compound.

  8. Biological Synthesis of Zinc oxide Nanoparticles from Catharanthus roseus (l. G. Don. Leaf extract and validation for antibacterial activity

    G. Bhumi

    2014-03-01

    Full Text Available Biologically synthesized nanoparticles have been widely using in the field of medicine. Research in nanotechnology highlights the possibility of green chemistry pathways to produce technologically important nanomaterials. Present study focuses on the Biological synthesis of Zinc oxide nanoparticles (ZnO-NPs by Zinc acetate and sodium hydroxide utilizing the biocomponents of leaves of Catharanthus roseus. The samples were characterized by x-ray diffraction (XRD, Scanning Electron Microscopy (SEM, Energy Dispersive Spectroscopy (EDAX and FT-Raman Spectroscopy. The synthesized ZnO-NPs were found to be spherical in shape with an average size of 23 to 57 nm. These ZnO-NPs were evaluated for antibacterial activity. The maximum diameter of inhibition zones around the ZnONPs disk used for Bacillus thuringiensis indicates the resistance to ZnO NPs followed by Escherichia coli. Among the four bacterial species tested, the Pseudomonas aeuroginosa is more susceptible when compared with other three species. It is concluded that the biological synthesis of ZnO NPs is very fast, easy, cost effective and eco-friendly and without any side effects and ZnO Nps may be used for the preparation of antibacterial formulations against Pseudomonas aeuroginosa

  9. Direct regeneration of Periwinkle (Catharanthus roseus via node explants culture and different combinations of plant growth regulators

    M. Talebi

    2012-09-01

    Full Text Available Periwinkle (Catharanthus roseus L., Apocynaceae contains more than 130 different terpenoid indole alkaloids (TIAs, of which two dimeric alkaloids, Vinblastine and Vincristine, have antineoplastic activity and are useful in treatment of various cancers. Specific production of some alkaloids in differentiated tissues such as leaf and stem led to use direct regeneration of explants in order to increase the production of these important alkaloids in the plant. In this research, 30 combinations of plant growth regulators and activated charcoal were used in MS media for direct regeneration of node explants. Application of BAP in media containing 1 g/l activated charcoal showed the best direct regeneration of node explants and shoot proliferation. Although application of activated charcoal is necessary for periwinkle growth in media due to many phenolic compounds, but it has negative effects on adsorption of plant growth regulators and consequently reduce shoot proliferation. Therefore, it seems that 1 g/l activated charcoal is an appropriate concentration for preparing shoot proliferation media. In addition, transporting regenerated shoots to culture media containing NAA resulted in increasing shoot length. Proliferated shoots rooted in media without PGR and with 2 g/l activated charcoal and acclimated with environmental conditions after transferring to the soil.

  10. Isolation, purification and characterization of vinblastine and vincristine from endophytic fungus Fusarium oxysporum isolated from Catharanthus roseus.

    Ashutosh Kumar

    Full Text Available Endophytic fungi reside in a symbiotic fashion inside their host plants, mimic their chemistry and interestingly, produce the same natural products as their hosts and are thus being screened for the production of valuable compounds like taxol, camptothecin, podophyllotoxin, etc. Vinblastine and vincristine are excellent anti-cancer drugs but their current production using plants is non-abundant and expensive. In order to make these drugs readily available to the patients at affordable prices, we isolated the endophytic fungi from Catharanthus roseus plant and found a fungus AA-CRL-6 which produces vinblastine and vincristine in appreciable amounts. These drugs were purified by TLC and HPLC and characterized using UV-Vis spectroscopy, ESI-MS, MS/MS and (1H NMR. One liter of culture filtrate yielded 76 µg and 67 µg of vinblastine and vincristine respectively. This endophytic fungal strain was identified as Fusarium oxysporum based upon its cultural and morphological characteristics and internal transcribed spacer (ITS sequence analysis.

  11. In vitro plant propagation of Catharanthus roseus and assessment of genetic fidelity of micropropagated plants by RAPD marker assay.

    Kumar, Ajay; Prakash, Krishna; Sinha, Rajesh Kumar; Kumar, Nitish

    2013-02-01

    An investigation was carried out to develop an efficient micropropagation protocol for Catharanthus roseus. Experiments were conducted to optimize suitable media for in vitro shoot multiplication and root induction. Out of the different media compared for in vitro shoot multiplication, Murashige and Skoog (MS) medium supplemented with 1 mg/l of 6-benzylaminopurine and 0.2 mg/l α-naphthaleneacetic acid showed better response in terms of the emergence of shoots from axillary buds as well as proliferation and multiplication of shoots. The shoots when placed on half strength of MS medium having 1 mg/l indole 3-butyric acid and 0.25 % charcoal showed cent percent root induction with maximum number of roots per shoot (4.2) as well as maximum root length (1.72 cm). Further, clonal fidelity of the in vitro-raised plants was carried out using randomly amplified polymorphic DNA marker and results indicated that all the tissue culture-derived plants are true-to-type and there were no somaclonal variations among these plants. PMID:23292901

  12. A polymorphic (GA/CT)n- SSR influences promoter activity of Tryptophan decarboxylase gene in Catharanthus roseus L. Don.

    Kumar, Santosh; Bhatia, Sabhyata

    2016-01-01

    Simple Sequence Repeats (SSRs) of polypurine-polypyrimidine type motifs occur very frequently in the 5' flanks of genes in plants and have recently been implicated to have a role in regulation of gene expression. In this study, 2 accessions of Catharanthus roseus having (CT)8 and (CT)21 varying motifs in the 5'UTR of Tryptophan decarboxylase (Tdc) gene, were investigated for its role in regulation of gene expression. Extensive Tdc gene expression analysis in the 2 accessions was carried out both at the level of transcription and translation. Transcript abundance was estimated using Northern analysis and qRT-PCR, whereas the rate of Tdc gene transcription was assessed using in-situ nuclear run-on transcription assay. Translation status of Tdc gene was monitored by quantification of polysome associated Tdc mRNA using qRT-PCR. These observations were validated through transient expression analysis using the fusion construct [CaM35S:(CT)8-21:GUS]. Our study demonstrated that not only does the length of (CT)n -SSRs influences the promoter activity, but the presence of SSRs per se in the 5'-UTR significantly enhances the level of gene expression. We termed this phenomenon as "microsatellite mediated enhancement" (MME) of gene expression. Results presented here will provide leads for engineering plants with enhanced amounts of medicinally important alkaloids. PMID:27623355

  13. 长春花生物碱提取技术的研究进展%Research Progress on the Extraction Technology of Alkaloids from Catharanthus roseus

    董晓宁; 赵海福; 刘文博

    2012-01-01

    Alkaloid is a kind of nitrogen-containing organic compounds that widely existed in the nature,which is of special medicinal value. The extraction, separation and content determination methods of alkaloid from natural medicinal herb Catharanthus roseus were summarized, so as to provide certain reference for the industrial production and scientific research of alkaloid in C. roseus.%生物碱广泛存在于自然界的动植物中,是一类含氮有机化合物,具有特殊药用价值。通过对天然药用植物长春花中生物碱的提取、分离和含量测定方法的总结,从而达到为长春花生物碱的工业生产和科学研究提供一定借鉴的目的。

  14. 长春花Crlea基因的克隆及原核表达初步分析%Cloning and prokaryotic expression of Crlea gene from Catharanthus roseus

    聂明珠; 祖元刚; 房思良

    2006-01-01

    晚期胚胎丰富(Late Embryogenesis Abundant, LEA)蛋白是植物在干旱胁迫下响应并被描述为具有潜在的抗旱功能的一类重要的抗旱蛋白.通过建立干旱胁迫下长春花(Catharanthus roseus)的cDNA文库并进行测序筛选分析,首次分离得到Crlea(Crlea for Catharanthus roseus late embryogenesis abundant)全长基因.该基因具有492 bp的开放读码框,编码163个氨基酸,其中偏性氨基酸含量占总蛋白的55.9%.同源性分析表明该假定蛋白与胡萝卜(Daucus carota)LEA DC3 的同源性达69%.亲水性分析表明具有极强的亲水性.为进一步验证CrLEA蛋白的功能,构建了Crlea基因的原核表达载体并在大肠杆菌中对其表达进行了分析.结果表明,原核载体成功的表达了CrLEA蛋白,亲水性实验及热稳定性实验表明CrLEA蛋白具有极强的亲水性和热稳定性.%Late Embryogenesis Abundant (LEA) proteins are one of the important proteins in stress response which have been described as having a potential role in drought tolerance. Crlea(Crlea for Catharanthus roseus late embryogenesis abundant) gene was first cloned from cDNA library of Catharanthus roseus under drought stress through sequencing screening method. The sequence analysis of the Crlea gene showed a complete 492bp ORF encoding a putative 163 amino acids polypeptide with a biased amino acid of 55.9% to total component. Homology analysis indicated a 69% similarity between CrLEA protein and carrot (Daucus carota)LEA DC3 protein. Hydrophobicity analysis found that CrLEA protein was extremely hydrophilic. For further identifying the function of CrLEA protein, prokaryotic recombinant vector of Crlea gene was constructed and the expression in E.coli was studied. Results showed recombinant CrLEA protein was successfully expressed in E.coli. Hydrophilic and heat-stable analysis of recombinant CrLEA protein showed a strong hydrophilic and heat-stable ability.

  15. Micorriza arbuscular y estrés abiótico en el contenido de alcaloides (vinblastima y vincristina) de Catharanthus roseus (L.) G. Don

    Rosa Mera, Claudia Janette de la

    2012-01-01

    Este estudio evaluó el incremento de vinblastina y vincristina en plantas micorrizadas de Catharanthus roseus (L.) G. Don., establecidas en invernadero, sometidas a estrés abiótico causado por aplicaciones de bicarbonato de potasio (2.5 y 7.5 mM) y cloruro de sodio (40 mM). El presente trabajo fue dividido en tres fases experimentales. La primera fase describe la propagación de consorcios e identificación de hongos micorrízicos arbusculares (HMA) obtenidos de la rizósfera ...

  16. Dust loadings on some common plants near Lucknow city. [Acacia melanoxylon, Bauhinia malabarica, Bougain-villea glabra, Calotropis procera, Catharanthus roseus, Eucalyptus globulus, Ipomoea fistulosa and Peltophorum pterocarpum

    Yunus, M.; Dwivedi, A.K.; Kulshreshtha, K.; Ahmad, K.J.

    1985-01-01

    Eight plant species - Acacia melanoxylon, Bauhinia malabarica, Bougain-villea glabra, Calotropis procera, Catharanthus roseus, Eucalyptus globulus, Ipomoea fistulosa and Peltophorum pterocarpum - were collected from a newly established suburb colony of Lucknow city, where the major pollutant is dust, to study the dust cleansing efficiency of the plant canopy and also to establish the correlation between the leaf morphological characteristics and their dust trapping potential. The dust load, in milligrams per square centimeter of leaf surface, was measured and related to foliar epidermal and cuticular characteristics, and morphological features.

  17. The Transcription Factor CrWRKY1 Positively Regulates the Terpenoid Indole Alkaloid Biosynthesis in Catharanthus roseus1[W][OA

    Suttipanta, Nitima; Pattanaik, Sitakanta; Kulshrestha, Manish; Patra, Barunava; Singh, Sanjay K.; Yuan, Ling

    2011-01-01

    Catharanthus roseus produces a large array of terpenoid indole alkaloids (TIAs) that are an important source of natural or semisynthetic anticancer drugs. The biosynthesis of TIAs is tissue specific and induced by certain phytohormones and fungal elicitors, indicating the involvement of a complex transcriptional control network. However, the transcriptional regulation of the TIA pathway is poorly understood. Here, we describe a C. roseus WRKY transcription factor, CrWRKY1, that is preferentially expressed in roots and induced by the phytohormones jasmonate, gibberellic acid, and ethylene. The overexpression of CrWRKY1 in C. roseus hairy roots up-regulated several key TIA pathway genes, especially Tryptophan Decarboxylase (TDC), as well as the transcriptional repressors ZCT1 (for zinc-finger C. roseus transcription factor 1), ZCT2, and ZCT3. However, CrWRKY1 overexpression repressed the transcriptional activators ORCA2, ORCA3, and CrMYC2. Overexpression of a dominant-repressive form of CrWRKY1, created by fusing the SRDX repressor domain to CrWRKY1, resulted in the down-regulation of TDC and ZCTs but the up-regulation of ORCA3 and CrMYC2. CrWRKY1 bound to the W box elements of the TDC promoter in electrophoretic mobility shift, yeast one-hybrid, and C. roseus protoplast assays. Up-regulation of TDC increased TDC activity, tryptamine concentration, and resistance to 4-methyl tryptophan inhibition of CrWRKY1 hairy roots. Compared with control roots, CrWRKY1 hairy roots accumulated up to 3-fold higher levels of serpentine. The preferential expression of CrWRKY1 in roots and its interaction with transcription factors including ORCA3, CrMYC2, and ZCTs may play a key role in determining the root-specific accumulation of serpentine in C. roseus plants. PMID:21988879

  18. Sombreamento de plantas de Catharanthus roseus (L. G. Don 'Pacifica White' por malhas coloridas: desenvolvimento vegetativo Shading of 'Pacifica White' Catharanthus roseus (L. G. Don plants with colored nets: vegetative development

    Anderson Adriano Martins Melo

    2009-04-01

    Full Text Available As malhas coloridas têm sido utilizadas para manipular o desenvolvimento vegetativo, melhorando a utilização da radiação solar por plantas ornamentais. Objetivou-se, neste trabalho estudar o efeito da redução de 50% da radiação fotossinteticamente ativa sobre o crescimento vegetativo de plantas de Catharanthus roseus (L. G. Don, por meio de malhas azul e vermelha e malha preta, em comparação com plantas crescidas na ausência de sombreamento (pleno sol. As plantas foram obtidas a partir de sementes e tratadas por 180 dias. Foram avaliados o ganho de biomassa e a distribuição de matéria seca nas plantas, o conteúdo de pigmentos foliares (clorofilas e carotenóides e de nitrogênio foliar. A malha vermelha provocou um aumento de matéria seca total e de área foliar das plantas em comparação com as malhas azul, preta e ao tratamento a pleno sol, porém, exceto em relação a esse tratamento, a malha vermelha causou menor conteúdo de nitrogênio e pigmentos foliares. A maior relação raiz/parte aérea e relação clorofila a/b, menores razões de área foliar e de massa foliar das plantas crescidas a pleno sol em relação às plantas sombreadas indicam um efeito mais proeminente da irradiância mais alta do que da alteração do espectro de luz. O sombreamento altera significativamente a distribuição de matéria seca e o uso de malhas de diferentes cores modifica o conteúdo de pigmentos fotossintéticos dessa espécie.Colored shade nets have been used to manipulate the vegetative development, improving the utilization of solar radiation by ornamental plants. This work aimed to study the effect of 50% reduction of PAR on vegetative growth of plants of Catharanthus roseus (L. G. Don, using blue and red nets, and black net, in comparison to plants growing under full sunlight (lack of shading. The plants were obtained from seeds and treated for 180 days. Biomass increment and distribution, pigment content (chlorophylls and

  19. 刈割对栽培长春花(Catharanthus roseus)生活史型转变的影响%The effects of harvest on transformation of plant life cycle forms of Catharanthus roseus

    唐中华; 郭晓瑞; 张学科; 高扬; 祖元刚; 杨蕾; 杨逢建; 张衷华

    2006-01-01

    为了提高栽培长春花(Catharanthus roseus)的生物碱产量,应用植物生活史型的理论和方法,研究了刈割对栽培长春花生活史型转变及其生物碱代谢的影响.运用主成分分析法(Principal Component Analysis,PCA)对刈割后的长春花后生活史型变化进行定量和定性划分,发现在对照栽培环境下生长的长春花处于DE生境,定性划分结果为SV生活史型,定量划分结果为V0.3638S0.6174 C0.0187,属于SV型.刈割使长春花的生活史型转变为V0.2847 S0.6684 C0.0469,属于SC型.同时,对两种生活史型的长春花中长春碱及其前体文朵灵和长春质碱的含量进行了检测分析,发现刈割后的SC型长春花不同叶位叶片中的生物碱含量均显著提高(p<0.05),可以为提高栽培长春花生物碱含量提供科学指导,也进一步验证了生活史型理论.

  20. Differential induction of meristematic stem cells of Catharanthus roseus and their characterization.

    Moon, So Hyun; Venkatesh, Jelli; Yu, Jae-Woong; Park, Se Won

    2015-11-01

    Plant cell culture technology has been introduced for the mass production of the many useful components. A variety of plant-derived compounds is being used in various fields, such as pharmaceuticals, foods, and cosmetics. Plant cell cultures are believed to be derived from the dedifferentiation process. In the present study, an undifferentiated cambial meristematic cell (CMCs) of Catharanthus is isolated using histological and genetic methods, and compared with dedifferentiation-derived callus (DDCs) cultures. Furthermore, differential culture conditions for both DDCs- and CMCs-derived cell lines were established. A suitable media for the increased accumulation of terpenoid indole alkaloids (TIAs) was also standardized. Compared with DDCs, CMCs showed marked accumulation of TIAs in cell lines grown on media with 1.5 mg·mL(-1) of NAA and 0.5 mg·mL(-1) of kinetin. CMCs-derived cultures of Catharanthus, as a source of key anticancer drugs (viblastine and vincristine), would overcome the obstacles usually associated with the production of natural metabolites through the use of DDCs. Cell culture systems that are derived from CMCs may also provide a cost-effective and eco-friendly basis for the sustainable production of a number of important plant natural products. PMID:26298518

  1. Efficacy of larvicidal and pupicidal activity of Catharanthus roseus aqueous and solvent extracts against Anopheles stephensi Liston and Culex quinquefasciatus Say (Diptera:Culicidae)

    Selladurai Subarani; Selvi Sabhanayakam; Chinnaperumal Kamaraj; Gandhi Elango; Mohamed Abdul Kadir

    2013-01-01

    Objective:To investigate the larvicidal and pupicidal activities of aqueous, ethyl acetate and methanol extracts of Catharanthus roseus (C. roseus) against malaria and filariasis vectors. Methods:The larvicidal and pupicidal activities of C. roseus leaf extracts were tested against the fourth instar larvae and pupae of Anopheles stephensi (An. stephensi) and Culex quinquefasciatus (Cx. quinquefasciatus). The mortality was observed after 24 and 48 h post the treatment. The data were subjected to probit analysis to determine the lethal concentrations (LC50 and LC90) at which 50%and 90%of the treated larvae or pupae of the tested species were killed. Results:The larval and pupal mortality were observed after 24 and 48 h of exposure of aqueous, ethyl acetate and methanol extracts of C. roseus; no mortality was observed in the control group. The LC50 values against the fourth-instar larvae of An. stephensi were 68.62 and 72.04 mg/mL for the aqueous extract, 82.47 mg/mL for the ethyl acetate extract, and 78.80 and 86.64 mg/mL for the methanol extract, while the aqueous, ethyl acetate and methanol extracts had LC50 values of 85.21, 76.84 and 94.20 mg/mL against the fourth-instar larvae of Cx. quinquefasciatus. The aqueous, ethyl acetate and methanol extracts had LC50 values of 118.08, 182.47 and 143.80 mg/mL against the pupae of An. stephensi and 146.20, 226.84 and 156.62 mg/mL against the pupae of Cx. quinquefasciatus, respectively. Conclusions: The aqueous and methanol extracts of C. roseus leaves had an excellent potential to control the malarial vector An. stephensi and filariasis vector Cx. quinquefasciatus.

  2. The Complete Plastid Genome Sequence of Madagascar Periwinkle Catharanthus roseus (L. G. Don: Plastid Genome Evolution, Molecular Marker Identification, and Phylogenetic Implications in Asterids.

    Chuan Ku

    Full Text Available The Madagascar periwinkle (Catharanthusroseus in the family Apocynaceae is an important medicinal plant and is the source of several widely marketed chemotherapeutic drugs. It is also commonly grown for its ornamental values and, due to ease of infection and distinctiveness of symptoms, is often used as the host for studies on phytoplasmas, an important group of uncultivated plant pathogens. To gain insights into the characteristics of apocynaceous plastid genomes (plastomes, we used a reference-assisted approach to assemble the complete plastome of C. roseus, which could be applied to other C. roseus-related studies. The C. roseus plastome is the second completely sequenced plastome in the asterid order Gentianales. We performed comparative analyses with two other representative sequences in the same order, including the complete plastome of Coffeaarabica (from the basal Gentianales family Rubiaceae and the nearly complete plastome of Asclepiassyriaca (Apocynaceae. The results demonstrated considerable variations in gene content and plastome organization within Apocynaceae, including the presence/absence of three essential genes (i.e., accD, clpP, and ycf1 and large size changes in non-coding regions (e.g., rps2-rpoC2 and IRb-ndhF. To find plastome markers of potential utility for Catharanthus breeding and phylogenetic analyses, we identified 41 C. roseus-specific simple sequence repeats. Furthermore, five intergenic regions with high divergence between C. roseus and three other euasterids I taxa were identified as candidate markers. To resolve the euasterids I interordinal relationships, 82 plastome genes were used for phylogenetic inference. With the addition of representatives from Apocynaceae and sampling of most other asterid orders, a sister relationship between Gentianales and Solanales is supported.

  3. The Complete Plastid Genome Sequence of Madagascar Periwinkle Catharanthus roseus (L.) G. Don: Plastid Genome Evolution, Molecular Marker Identification, and Phylogenetic Implications in Asterids.

    Ku, Chuan; Chung, Wan-Chia; Chen, Ling-Ling; Kuo, Chih-Horng

    2013-01-01

    The Madagascar periwinkle (Catharanthusroseus in the family Apocynaceae) is an important medicinal plant and is the source of several widely marketed chemotherapeutic drugs. It is also commonly grown for its ornamental values and, due to ease of infection and distinctiveness of symptoms, is often used as the host for studies on phytoplasmas, an important group of uncultivated plant pathogens. To gain insights into the characteristics of apocynaceous plastid genomes (plastomes), we used a reference-assisted approach to assemble the complete plastome of C. roseus, which could be applied to other C. roseus-related studies. The C. roseus plastome is the second completely sequenced plastome in the asterid order Gentianales. We performed comparative analyses with two other representative sequences in the same order, including the complete plastome of Coffeaarabica (from the basal Gentianales family Rubiaceae) and the nearly complete plastome of Asclepiassyriaca (Apocynaceae). The results demonstrated considerable variations in gene content and plastome organization within Apocynaceae, including the presence/absence of three essential genes (i.e., accD, clpP, and ycf1) and large size changes in non-coding regions (e.g., rps2-rpoC2 and IRb-ndhF). To find plastome markers of potential utility for Catharanthus breeding and phylogenetic analyses, we identified 41 C. roseus-specific simple sequence repeats. Furthermore, five intergenic regions with high divergence between C. roseus and three other euasterids I taxa were identified as candidate markers. To resolve the euasterids I interordinal relationships, 82 plastome genes were used for phylogenetic inference. With the addition of representatives from Apocynaceae and sampling of most other asterid orders, a sister relationship between Gentianales and Solanales is supported. PMID:23825699

  4. Jasmonic Acid Effect on the Fatty Acid and Terpenoid Indole Alkaloid Accumulation in Cell Suspension Cultures of Catharanthus roseus

    Guitele Dalia Goldhaber-Pasillas

    2014-07-01

    Full Text Available The stress response after jasmonic acid (JA treatment was studied in cell suspension cultures of Catharanthus roseus. The effect of JA on the primary and secondary metabolism was based on changes in profiles of fatty acids (FA and terpenoid indole alkaloids (TIA. According to multivariate data analyses (MVDA, three major time events were observed and characterized according to the variations of specific FA and TIA: after 0–30 min of induction FA such as C18:1, C20:0, C22:0 and C24:0 were highly induced by JA; 90–360 min after treatment was characterized by variations of C14:0 and C15:0; and 1440 min after induction JA had the largest effect on both group of metabolites were C18:1, C18:2, C18:3, C16:0, C20:0, C22:0, C24:0, catharanthine, tabersonine-like 1, serpentine, tabersonine and ajmalicine-like had the most significant variations. These results unambiguously demonstrate the profound effect of JA particularly on the accumulation of its own precursor, C18:3 and the accumulation of TIA, which can be considered as late stress response events to JA since they occurred only after 1440 min. These observations show that the early events in the JA response do not involve the de novo biosynthesis of neither its own precursor nor TIA, but is due to an already present biochemical system.

  5. Carrier-Mediated Uptake of 1-(Malonylamino)cyclopropane-1-Carboxylic Acid in Vacuoles Isolated from Catharanthus roseus Cells 1

    Bouzayen, Mondher; Latché, Alain; Pech, Jean-Claude; Marigo, Gérard

    1989-01-01

    The uptake of 1-(malonylamino)cyclopropane-1-carboxylic acid (MACC), the conjugated form of the ethylene precursor, into vacuoles isolated from Catharanthus roseus cells has been studied by silicone layer floatation filtering. The transport across the tonoplast of MACC is stimulated fourfold by 5 millimolar MgATP, has a Km of about 2 millimolar, an optimum pH around 7, and an optimum temperature at 30°C. Several effectors known to inhibit ATPase (N,N′-dicyclohexylcarbodiimide) and to collapse the transtonoplastic H+ electrochemical gradient (carbonylcyanide m-chlorophenylhydrazone, gramicidin, and benzylamine) all reduced MACC uptake. Abolishing the membrane potential with SCN− and valinomycin also greatly inhibited MACC transport. Our data demonstrate that MACC accumulates in the vacuole against a concentration gradient by means of a proton motive force generated by a tonoplastic ATPase. The involvement of a protein carrier is suggested by the strong inhibition of uptake by compounds known to block SH—, OH—, and NH2— groups. MACC uptake is antagonized competitively by malonyl-d-tryptophan, indicating that the carrier also accepts malonyl-d-amino acids. Neither the moities of these compounds taken separately [1-aminocyclopropane-1-carboxylic acid, malonate, d-tryptophan or d-phenylalanine] nor malate act as inhibitors of MACC transport. The absence of inhibition of malate uptake by MACC suggests that MACC and malate are taken up by two different carriers. We propose that the carrier identified here plays an important physiological role in withdrawing from the cytosol MACC and malonyl-d-amino acids generated under stress conditions. PMID:16667182

  6. The improved resistance to high salinity induced by trehalose is associated with ionic regulation and osmotic adjustment in Catharanthus roseus.

    Chang, Bowen; Yang, Lei; Cong, Weiwei; Zu, Yuangang; Tang, Zhonghua

    2014-04-01

    The effects of exogenous trehalose (Tre) on salt tolerance of pharmaceutical plant Catharanthus roseus and the physiological mechanisms were both investigated in this study. The results showed that the supplement of Tre in saline condition (250 mM NaCl) largely alleviated the inhibitory effects of salinity on plant growth, namely biomass accumulation and total leaf area per plant. In this saline condition, the decreased level of relative water content (RWC) and photosynthetic rate were also greatly rescued by exogenous Tre. This improved performance of plants under high salinity induced by Tre could be partly ascribed to its ability to decrease accumulation of sodium, and increase potassium in leaves. The exogenous Tre led to high levels of fructose, glucose, sucrose and Tre inside the salt-stressed plants during whole the three-week treatment. The major free amino acids such as proline, arginine, threonine and glutamate were also largely elevated in the first two-week course of treatment with Tre in saline solution. It was proposed here that Tre might act as signal to make the salt-stressed plants actively increase internal compatible solutes, including soluble sugars and free amino acids, to control water loss, leaf gas exchange and ionic flow at the onset of salt stress. The application of Tre in saline condition also promoted the accumulation of alkaloids. The regulatory role of Tre in improving salt tolerance was optimal with an exogenous concentration of 10 mM Tre. Larger concentrations of Tre were supra-optimum and adversely affected plant growth. PMID:24589477

  7. Jasmonic acid effect on the fatty acid and terpenoid indole alkaloid accumulation in cell suspension cultures of Catharanthus roseus.

    Goldhaber-Pasillas, Guitele Dalia; Mustafa, Natali Rianika; Verpoorte, Robert

    2014-01-01

    The stress response after jasmonic acid (JA) treatment was studied in cell suspension cultures of Catharanthus roseus. The effect of JA on the primary and secondary metabolism was based on changes in profiles of fatty acids (FA) and terpenoid indole alkaloids (TIA). According to multivariate data analyses (MVDA), three major time events were observed and characterized according to the variations of specific FA and TIA: after 0-30 min of induction FA such as C18:1, C20:0, C22:0 and C24:0 were highly induced by JA; 90-360 min after treatment was characterized by variations of C14:0 and C15:0; and 1440 min after induction JA had the largest effect on both group of metabolites were C18:1, C18:2, C18:3, C16:0, C20:0, C22:0, C24:0, catharanthine, tabersonine-like 1, serpentine, tabersonine and ajmalicine-like had the most significant variations. These results unambiguously demonstrate the profound effect of JA particularly on the accumulation of its own precursor, C18:3 and the accumulation of TIA, which can be considered as late stress response events to JA since they occurred only after 1440 min. These observations show that the early events in the JA response do not involve the de novo biosynthesis of neither its own precursor nor TIA, but is due to an already present biochemical system. PMID:25029072

  8. Isolation of Catharanthus roseus (L. G. Don Nuclei and Measurement of Rate of Tryptophan decarboxylase Gene Transcription Using Nuclear Run-On Transcription Assay.

    Santosh Kumar

    Full Text Available An accurate assessment of transcription 'rate' is often desired to describe the promoter activity. In plants, isolation of transcriptionally active nuclei and their subsequent use in nuclear run-on assays has been challenging and therefore limit an accurate measurement of gene transcription 'rate'. Catharanthus roseus has emerged as a model medicinal plant as it exhibits an unsurpassed spectrum of chemodiversity, producing over 130 alkaloids through the terpenoid indole alkaloid (TIA pathway and therefore serves as a 'molecular hub' to understand gene expression profiles.The protocols presented here streamline, adapt and optimize the existing methods of nuclear run-on assay for use in C. roseus. Here, we fully describe all the steps to isolate transcriptionally active nuclei from C. roseus leaves and utilize them to perform nuclear run-on transcription assay. Nuclei isolated by this method transcribed at a level consistent with their response to external stimuli, as transcription rate of TDC gene was found to be higher in response to external stimuli i.e. when seedlings were subjected to UV-B light or to methyl jasmonate (MeJA. However, the relative transcript abundance measured parallel through qRT-PCR was found to be inconsistent with the synthesis rate indicating that some post transcriptional events might have a role in transcript stability in response to stimuli.Our study provides an optimized, efficient and inexpensive method of isolation of intact nuclei and nuclear 'run-on' transcription assay to carry out in-situ measurement of gene transcription rate in Catharanthus roseus. This would be valuable in investigating the transcriptional and post transcriptional response of other TIA pathway genes in C. roseus. Isolated nuclei may also provide a resource that could be used for performing the chip assay as well as serve as the source of nuclear proteins for in-vitro EMSA studies. Moreover, nascent nuclear run-on transcript could be further

  9. Leaf blight of catharanthus roseus (l). g. don caused by macrophomina phaseolina (tassi) goid and its in vitro control through bio-pesticides

    Catharanthus roseus (L.).G. Don, a highly valued medicinal plant suffers from a serious disease. A survey on the symptom and severity of the leaf blight of C. roseus was performed in the nurseries of the Institute of Forestry and Environmental Sciences (IFESCU), Bangladesh Council of Industrial Research (BCSIR) and Bangladesh Forest Research Institute (BFRI), Bangladesh. An observation was also made in the avenues and garden of King Saud University (KSU), Kingdom of Saudi Arabia. No disease was recorded in KSU, but severe infections were found in all the locations surveyed in Bangladesh. The highest infection percentage and the highest disease index were found in BCSIR nursery followed by BFRI and the lowest was recorded in IFESCU nursery. Macrophomina phaseolina (Tassi) Goid was isolated and it was proved to be pathogenic. This is the first report of leaf blight of C. roseus caused by M. phaseolina. Azadirachta indica, Ocimum sanctum Vitex negundo, Mucuna pruriens, Calotropis procera, Terminalia arjuna, Allium sativum, Zingiber officinalis and Allium cepa were used as biopesticides. Out of the nine plant species screened, T. arjuna showed the highest (58.37%) inhibition percentage which was followed by A. indica (55.72%) and the lowest (27.4%) inhibition percentage were obtained with V. negundo whereas the rest of the plant extracts showed more or less same inhibitory effect. (author)

  10. Aislamiento de consorcios de hongos micorrícicos arbusculares de plantas medicinales y su efecto en el crecimiento de vinca (Catharanthus roseus) Isolation of arbuscular mycorrhizal fungi consortia from medicinal plants and their effectiveness on growth of vinca (Catharanthus roseus)

    CLAUDIA DE LA ROSA-MERA; RONALD FERRERA-CERRATO; ALEJANDRO ALARCÓN; MARÍA DE JESÚS SÁNCHEZ-COLÍN; ALICIA FRANCO-RAMÍREZ

    2012-01-01

    Este trabajo consistió en propagar e identificar hongos micorrícicos arbusculares (HMA) recolectados de plantas medicinales (PM) de áreas naturales de bosques mixtos, y seleccionar consorcios micorrícicos con base en la promoción del crecimiento de vinca Catharanthus roseus (L) G. Don, planta medicinal cuyos alcaloides tienen propiedades antineoplásicas. En la primera fase experimental se recolectaron raíces y suelo rizosférico de 13 PM establecidas en campo para evaluar el porcentaje de colo...

  11. Effects of different water conditions on life cycle forms and physiological metabolisms of Catharanthus roseus%土壤不同水分条件对长春花(Catharanthus roseus)生活史型的影响

    唐中华; 杨蕾; 梁胜楠; 祖元刚

    2007-01-01

    为了研究土壤中不同水分条件对长春花生活史型形成及生理代谢的影响,设置对照、轻度干旱、中度干旱和重度干旱等土壤水分梯度,对长春花(Catharanthus roseus(L.)G.Don)幼苗进行处理.对长春花形态指标进行聚类分析发现选择的20个聚类实体被分为2组,第1组为对照(CK)和轻度干旱(LD)处理的植株,第2组为中度干旱(MD)和重度干旱(HD)处理的植株.运用主成分分析(Principal component analysis,PCA)方法对不同土壤水分条件下长春花营养生长(Vegetative growth,V)、有性生殖(Sexual reproduction,S)和无性繁殖(Clone reproduction,C)等3类15种性状进行统计.结果显示,长春花在对照条件下生活史型为V0.39 S0.54 C0.07,轻度干旱为V0.36 S0.50 C0.14,中度干旱为V0.53 S0.27 C0.20,重度干旱为V0.45 S0.09 C0.46,干旱程度加强显著提高了无性繁殖的比重,降低了有性生殖的比例.同时,对长春花中文朵灵、长春质碱和脱水长春碱等生物碱的含量进行了动态测定,发现重度干旱下的文朵灵、长春质碱和脱水长春碱的含量在16d时分别是对照水平的1.5倍、2.3倍和3.1倍,表明干旱胁迫诱导生物碱积累,为长春花高效栽培提供了理论依据.

  12. A Pair of Tabersonine 16-Hydroxylases Initiates the Synthesis of Vindoline in an Organ-Dependent Manner in Catharanthus roseus1[C][W

    Besseau, Sébastien; Kellner, Franziska; Lanoue, Arnaud; Thamm, Antje M.K.; Salim, Vonny; Schneider, Bernd; Geu-Flores, Fernando; Höfer, René; Guirimand, Grégory; Guihur, Anthony; Oudin, Audrey; Glevarec, Gaëlle; Foureau, Emilien; Papon, Nicolas; Clastre, Marc; Giglioli-Guivarc’h, Nathalie; St-Pierre, Benoit; Werck-Reichhart, Danièle; Burlat, Vincent; De Luca, Vincenzo; O’Connor, Sarah E.; Courdavault, Vincent

    2013-01-01

    Hydroxylation of tabersonine at the C-16 position, catalyzed by tabersonine 16-hydroxylase (T16H), initiates the synthesis of vindoline that constitutes the main alkaloid accumulated in leaves of Catharanthus roseus. Over the last decade, this reaction has been associated with CYP71D12 cloned from undifferentiated C. roseus cells. In this study, we isolated a second cytochrome P450 (CYP71D351) displaying T16H activity. Biochemical characterization demonstrated that CYP71D12 and CYP71D351 both exhibit high affinity for tabersonine and narrow substrate specificity, making of T16H, to our knowledge, the first alkaloid biosynthetic enzyme displaying two isoforms encoded by distinct genes characterized to date in C. roseus. However, both genes dramatically diverge in transcript distribution in planta. While CYP71D12 (T16H1) expression is restricted to flowers and undifferentiated cells, the CYP71D351 (T16H2) expression profile is similar to the other vindoline biosynthetic genes reaching a maximum in young leaves. Moreover, transcript localization by carborundum abrasion and RNA in situ hybridization demonstrated that CYP71D351 messenger RNAs are specifically located to leaf epidermis, which also hosts the next step of vindoline biosynthesis. Comparison of high- and low-vindoline-accumulating C. roseus cultivars also highlights the direct correlation between CYP71D351 transcript and vindoline levels. In addition, CYP71D351 down-regulation mediated by virus-induced gene silencing reduces vindoline accumulation in leaves and redirects the biosynthetic flux toward the production of unmodified alkaloids at the C-16 position. All these data demonstrate that tabersonine 16-hydroxylation is orchestrated in an organ-dependent manner by two genes including CYP71D351, which encodes the specific T16H isoform acting in the foliar vindoline biosynthesis. PMID:24108213

  13. Rapid and simultaneous determination of five vinca alkaloids in Catharanthus roseus and human serum using trilinear component modeling of liquid chromatography-diode array detection data.

    Liu, Zhi; Wu, Hai-Long; Li, Yong; Gu, Hui-Wen; Yin, Xiao-Li; Xie, Li-Xia; Yu, Ru-Qin

    2016-07-15

    A novel chemometrics-assisted high performance liquid chromatography method coupled with diode array detector (HPLC-DAD) was proposed for the simultaneous determination of vincristine (VCR), vinblastine (VLB), vindoline (VDL), catharanthine (CAT) and yohimbine (YHB) in Catharanthus roseus (C. roseus) and human serum samples. With the second-order advantage of the alternating trilinear decomposition (ATLD) method, the resolution and rapid determination of five components of interest in complex matrices were performed, even in the present of heavy overlaps and unknown interferences. Therefore, multi-step purification was omitted and five components could be fast eluted out within 7.5min under simple isocratic elution condition (acetonitrile/0.2% formic acid water, 37:63, v/v). Statistical parameters, such as the linear correlation coefficient (R(2)), root-mean-square error of prediction (RMSEP), limit of detection (LOD) and limit of quantitation (LOQ) had been calculated to investigate the accuracy and reliability of the method. The average recoveries of five vinca alkaloids ranged from 97.1% to 101.9% and 98.8% to 103.0% in C. roseus and human serum samples, respectively. The five vinca alkaloids were adequately determined with limits of detection (LODs) of 29.5-49.3ngmL(-1) in C. roseus and 12.4-27.2ngmL(-1) in human serum samples, respectively. The obtained results demonstrated that the analytical strategy provided a feasible alternative for synchronously monitoring the quality of raw herb and the concentration of blood drugs. PMID:26321366

  14. High-throughput transcriptome analysis of the leafy flower transition of Catharanthus roseus induced by peanut witches'-broom phytoplasma infection.

    Liu, Li-Yu Daisy; Tseng, Hsin-I; Lin, Chan-Pin; Lin, Yen-Yu; Huang, Yuan-Hung; Huang, Chien-Kang; Chang, Tean-Hsu; Lin, Shih-Shun

    2014-05-01

    Peanut witches'-broom (PnWB) phytoplasma are obligate bacteria that cause leafy flower symptoms in Catharanthus roseus. The PnWB-mediated leafy flower transitions were studied to understand the mechanisms underlying the pathogen-host interaction; however, our understanding is limited because of the lack of information on the C. roseus genome. In this study, the whole-transcriptome profiles from healthy flowers (HFs) and stage 4 (S4) PnWB-infected leafy flowers of C. roseus were investigated using next-generation sequencing (NGS). More than 60,000 contigs were generated using a de novo assembly approach, and 34.2% of the contigs (20,711 genes) were annotated as putative genes through name-calling, open reading frame determination and gene ontology analyses. Furthermore, a customized microarray based on this sequence information was designed and used to analyze samples further at various stages of PnWB infection. In the NGS profile, 87.8% of the genes showed expression levels that were consistent with those in the microarray profiles, suggesting that accurate gene expression levels can be detected using NGS. The data revealed that defense-related and flowering gene expression levels were altered in S4 PnWB-infected leafy flowers, indicating that the immunity and reproductive stages of C. roseus were compromised. The network analysis suggested that the expression levels of >1,000 candidate genes were highly associated with CrSVP1/2 and CrFT expression, which might be crucial in the leafy flower transition. In conclusion, this study provides a new perspective for understanding plant pathology and the mechanisms underlying the leafy flowering transition caused by host-pathogen interactions through analyzing bioinformatics data obtained using a powerful, rapid high-throughput technique. PMID:24492256

  15. Patterns of indole alkaloids synthesis in response to heat shock, 5-azacytidine and Na-butyrate treatment of cultured catharanthus roseus mesophyll protoplasts

    Alkaloids of C. roseus are in high demand for therapeutic and other reasons. Cultured Catharanthus cells can produce limited quantities of these alkaloids. The authors have found that cultured mesophyll protoplasts in the presence of 14C-Tryptamine are capable of synthesizing alkaloids. The pattern of alkaloids synthesis changes when protoplasts are subjected to a heat shock at 370C. The heat shocked protoplasts incorporated 33% more 14C-Tryptamine and produced 3 new types of alkaloids. Treatment of protoplasts with 5-azacytidine, a DNA hypomethylating agent and Na-butyrate which induces hyperacetylation of histones produced qualitative and quantitative changes in the alkaloid pattern. Four new alkaloids following the above treatments were detected by TLC and HPLC of the extracts. It is suggested that the alkaloid pattern of the cultured protoplasts can be altered by treatment with compounds known as regulators of gene expression. Work is in progress to isolate and identify these new alkaloids

  16. Effects of Adding Vindoline and MeJA on Production of Vincristine and Vinblastine, and Transcription of their Biosynthetic Genes in the Cultured CMCs of Catharanthus roseus.

    Zhang, Wenjin; Yang, Jiazeng; Zi, Jiachen; Zhu, Jianhua; Song, Liyan; Yu, Rongmin

    2015-12-01

    Vincristine and vinblastine were found by Liquid Chromatography-Mass Spectrometry (LC-MS) in Catharanthus roseuscambial meristem cells (CMCs) jointly treated with 0.25 mM vindoline and methyl jasmonate (MeJA), suggesting that C. roseus CMCs contain a complete set of the enzymes which are in response to convert vindoline into vincristine and vinblastine. Based on the facts that the transcript levels of vindoline-biosynthetic genes (STR, SGD and D4H) were up-regulated instead of being down-regulated by adding itself to the culture, and that the transcriptional factor ORCA3 was up-regulated simultaneously, we further confirmed that the transcription of STR, SGD, D4H was manipulated by ORCA3. PMID:26882673

  17. Patterns of indole alkaloids synthesis in response to heat shock, 5-azacytidine and Na-butyrate treatment of cultured catharanthus roseus mesophyll protoplasts

    Saleem, M.; Cutler, A.J.

    1986-04-01

    Alkaloids of C. roseus are in high demand for therapeutic and other reasons. Cultured Catharanthus cells can produce limited quantities of these alkaloids. The authors have found that cultured mesophyll protoplasts in the presence of /sup 14/C-Tryptamine are capable of synthesizing alkaloids. The pattern of alkaloids synthesis changes when protoplasts are subjected to a heat shock at 37/sup 0/C. The heat shocked protoplasts incorporated 33% more /sup 14/C-Tryptamine and produced 3 new types of alkaloids. Treatment of protoplasts with 5-azacytidine, a DNA hypomethylating agent and Na-butyrate which induces hyperacetylation of histones produced qualitative and quantitative changes in the alkaloid pattern. Four new alkaloids following the above treatments were detected by TLC and HPLC of the extracts. It is suggested that the alkaloid pattern of the cultured protoplasts can be altered by treatment with compounds known as regulators of gene expression. Work is in progress to isolate and identify these new alkaloids.

  18. 长春花悬浮培养细胞对天麻素的生物转化%Biotransformation of Gastrodin by Cell Suspension Cultures of Catharanthus roseus

    戴均贵; 巩卓; 朱丹萌; 郭洪祝; 郑俊华; 果德安

    2002-01-01

    应用长春花(Catharanthus roseus (L.) G. Don)悬浮细胞培养体系对天麻素进行了生物转化反应研究.经过8 d培养形成一个转化产物,应用光谱方法鉴定转化产物的结构为对羟基苯甲醇,为天麻素水解后形成的甙元.

  19. Synthesis and characterization of Gold and Silver nano-particles using different leaf extracts namely Catharanthus roseus, Datura metel and Azadirachta indica and Estimation of antimicrobial activity of silver nano-particles using disc diffusion method

    Sarbjeet Singh Gujral

    2014-01-01

    Objective: synthesis of gold and silver nano-particles using leaf extracts of Catharanthus roseus, Datura metel and Azadirachta indica and Estimation of antimicrobial activity of silver nano-particles using disc diffusion method. Method: Green approach has been utilized for the synthesis of gold and silver nano-particles. Different aqueous plant extracts has been prepared which was then utilized for the biosynthesis of gold and silver nano-particles. Estimation for the synthesis of nano-parti...

  20. Lack of Control in Inorganic Phosphate Uptake by Catharanthus roseus (L.) G. Don Cells (Cytoplasmic Inorganic Phosphate Homeostasis Depends on the Tonoplast Inorganic Phosphate Transport System?).

    Sakano, K.; Yazaki, Y.; Okihara, K.; Mimura, T.; Kiyota, S.

    1995-01-01

    Inorganic phosphate (Pi) uptake by Catharanthus roseus (L.) G. Don cells was studied in relation to its apparent uncontrolled uptake using 31P-nuclear magnetic resonance spectroscopy. Kinetics of Pi uptake by the cells indicated that apparent Km and Vm were about 7 [mu]M and 20 [mu]mol g-1 fresh weight h-1, respectively. Pi uptake in Murashige-Skoog medium under different Pi concentrations and different initial cell densities followed basically the same kinetics. When supplied with abundant Pi, cells absorbed Pi at a constant rate (Vm) for the first hours and accumulated it in the vacuole. As the endogenous pool expanded, the rate of Pi uptake gradually decreased to nil. Maximum Pi accumulation was 100 to 120 [mu]mol g-1 fresh weight if cell swelling during Pi uptake (about 2-fold in cell volume) was not considered. Results indicated that (a) the rate of Pi uptake by Catharanthus cells was independent of initial cell density and was constant over a wide range of Pi concentrations (2 mM to about 10 [mu]M) unless the cells were preloaded with excess Pi, and (b) there was no apparent feedback control over the Pi uptake process in the plasma membrane to avoid Pi toxicity. The importance of the tonoplast Pi transport system in cytoplasmic Pi homeostasis is discussed. PMID:12228474

  1. Involvement of the Octadecanoid Pathway and Protein Phosphorylation in Fungal Elicitor-Induced Expression of Terpenoid Indole Alkaloid Biosynthetic Genes in Catharanthus roseus

    Menke, Frank L.H.; Parchmann, Stefanie; Mueller, Martin J.; Kijne, Jan W.; Memelink, Johan

    1999-01-01

    Two key genes in terpenoid indole alkaloid biosynthesis, Tdc and Str, encoding tryptophan decarboxylase and strictosidine synthase, respectively, are coordinately induced by fungal elicitors in suspension-cultured Catharanthus roseus cells. We have studied the roles of the jasmonate biosynthetic pathway and of protein phosphorylation in signal transduction initiated by a partially purified elicitor from yeast extract. In addition to activating Tdc and Str gene expression, the elicitor also induced the biosynthesis of jasmonic acid. The jasmonate precursor α-linolenic acid or methyl jasmonate (MeJA) itself induced Tdc and Str gene expression when added exogenously . Diethyldithiocarbamic acid, an inhibitor of jasmonate biosynthesis, blocked both the elicitor-induced formation of jasmonic acid and the activation of terpenoid indole alkaloid biosynthetic genes. The protein kinase inhibitor K-252a abolished both elicitor-induced jasmonate biosynthesis and MeJA-induced Tdc and Str gene expression. Analysis of the expression of Str promoter/gusA fusions in transgenic C. roseus cells showed that the elicitor and MeJA act at the transcriptional level. These results demonstrate that the jasmonate biosynthetic pathway is an integral part of the elicitor-triggered signal transduction pathway that results in the coordinate expression of the Tdc and Str genes and that protein kinases act both upstream and downstream of jasmonates. PMID:10198087

  2. Molecular Cloning and Characterization of a Vacuolar Class III Peroxidase Involved in the Metabolism of Anticancer Alkaloids in Catharanthus roseus1[C

    Costa, Maria Manuela R.; Hilliou, Frederique; Duarte, Patrícia; Pereira, Luís Gustavo; Almeida, Iolanda; Leech, Mark; Memelink, Johan; Barceló, Alfonso Ros; Sottomayor, Mariana

    2008-01-01

    Catharanthus roseus produces low levels of two dimeric terpenoid indole alkaloids, vinblastine and vincristine, which are widely used in cancer chemotherapy. The dimerization reaction leading to α-3′,4′-anhydrovinblastine is a key regulatory step for the production of the anticancer alkaloids in planta and has potential application in the industrial production of two semisynthetic derivatives also used as anticancer drugs. In this work, we report the cloning, characterization, and subcellular localization of an enzyme with anhydrovinblastine synthase activity identified as the major class III peroxidase present in C. roseus leaves and named CrPrx1. The deduced amino acid sequence corresponds to a polypeptide of 363 amino acids including an N-terminal signal peptide showing the secretory nature of CrPrx1. CrPrx1 has a two-intron structure and is present as a single gene copy. Phylogenetic analysis indicates that CrPrx1 belongs to an evolutionary branch of vacuolar class III peroxidases whose members seem to have been recruited for different functions during evolution. Expression of a green fluorescent protein-CrPrx1 fusion confirmed the vacuolar localization of this peroxidase, the exact subcellular localization of the alkaloid monomeric precursors and dimeric products. Expression data further supports the role of CrPrx1 in α-3′,4′-anhydrovinblastine biosynthesis, indicating the potential of CrPrx1 as a target to increase alkaloid levels in the plant. PMID:18065566

  3. The Combined Effects of Ethylene and MeJA on Metabolic Profiling of Phenolic Compounds in Catharanthus roseus Revealed by Metabolomics Analysis.

    Liu, Jia; Liu, Yang; Wang, Yu; Zhang, Zhong-Hua; Zu, Yuan-Gang; Efferth, Thomas; Tang, Zhong-Hua

    2016-01-01

    Phenolic compounds belong to a class of secondary metabolites and are implicated in a wide range of responsive mechanisms in plants triggered by both biotic and abiotic elicitors. In this study, we approached the combinational effects of ethylene and MeJA (methyl jasmonate) on phenolic compounds profiles and gene expressions in the medicinal plant Catharanthus roseus. In virtue of a widely non-targeted metabolomics method, we identified a total of 34 kinds of phenolic compounds in the leaves, composed by 7 C6C1-, 11 C6C3-, and 16 C6C3C6 compounds. In addition, 7 kinds of intermediates critical for the biosynthesis of phenolic compounds and alkaloids were identified and discussed with phenolic metabolism. The combinational actions of ethylene and MeJA effectively promoted the total phenolic compounds, especially the C6C1 compounds (such as salicylic acid, benzoic acid) and C6C3 ones (such as cinnamic acid, sinapic acid). In contrast, the C6C3C6 compounds displayed a notably inhibitory trend in this case. Subsequently, the gene-to-metabolite networks were drawn up by searching for correlations between the expression profiles of 5 gene tags and the accumulation profiles of 41 metabolite peaks. Generally, we provide an insight into the controlling mode of ethylene-MeJA combination on phenolic metabolism in C. roseus leaves. PMID:27375495

  4. Promoter analysis reveals cis-regulatory motifs associated with the expression of the WRKY transcription factor CrWRKY1 in Catharanthus roseus.

    Yang, Zhirong; Patra, Barunava; Li, Runzhi; Pattanaik, Sitakanta; Yuan, Ling

    2013-12-01

    WRKY transcription factors (TFs) are emerging as an important group of regulators of plant secondary metabolism. However, the cis-regulatory elements associated with their regulation have not been well characterized. We have previously demonstrated that CrWRKY1, a member of subgroup III of the WRKY TF family, regulates biosynthesis of terpenoid indole alkaloids in the ornamental and medicinal plant, Catharanthus roseus. Here, we report the isolation and functional characterization of the CrWRKY1 promoter. In silico analysis of the promoter sequence reveals the presence of several potential TF binding motifs, indicating the involvement of additional TFs in the regulation of the TIA pathway. The CrWRKY1 promoter can drive the expression of a β-glucuronidase (GUS) reporter gene in native (C. roseus protoplasts and transgenic hairy roots) and heterologous (transgenic tobacco seedlings) systems. Analysis of 5'- or 3'-end deletions indicates that the sequence located between positions -140 to -93 bp and -3 to +113 bp, relative to the transcription start site, is critical for promoter activity. Mutation analysis shows that two overlapping as-1 elements and a CT-rich motif contribute significantly to promoter activity. The CrWRKY1 promoter is induced in response to methyl jasmonate (MJ) treatment and the promoter region between -230 and -93 bp contains a putative MJ-responsive element. The CrWRKY1 promoter can potentially be used as a tool to isolate novel TFs involved in the regulation of the TIA pathway. PMID:23979312

  5. Identification of conserved and novel microRNAs in Catharanthus roseus by deep sequencing and computational prediction of their potential targets.

    Prakash, Pravin; Ghosliya, Dolly; Gupta, Vikrant

    2015-01-10

    MicroRNAs are small endogenous non-coding RNAs of ~19-24 nucleotides and perform regulatory roles in many plant processes. To identify miRNAs involved in regulatory networks controlling diverse biological processes including secondary metabolism in Catharanthus roseus, an important medicinal plant, we employed deep sequencing of small RNA from leaf tissue. A total of 88 potential miRNAs comprising of 81 conserved miRNAs belonging to 35 families and seven novel miRNAs were identified. Precursors for 16 conserved and seven novel cro-miRNAs were identified, and their stem-loop hairpin structures were predicted. Selected cro-miRNAs were analyzed by stem-loop qRT-PCR and differential expression patterns were observed in different vegetative tissues of C. roseus. Targets were predicted for conserved and novel cro-miRNAs, which were found to be involved in diverse biological role(s) including secondary metabolism. Our study enriches available resources and information regarding miRNAs and their potential targets for better understanding of miRNA-mediated gene regulation in plants. PMID:25445288

  6. Catharanthus roseus Aqueous Extract is Cytotoxic to Jurkat Leukaemic T-cells but Induces the Proliferation of Normal Peripheral Blood Mononuclear Cells

    Ahmad, Nor Hazwani; Rahim, Rohanizah Abdul; Mat, Ishak

    2010-01-01

    Research on natural products has been widely used as a strategy to discover new drugs with potential for applications in complementary medicines because they have fewer side effects than conventional drugs. The aim of the present study was to evaluate the in vitro cytotoxic effects of crude aqueous Catharanthus roseus extract on Jurkat cells and normal peripheral blood mononuclear cells (PBMCs). The aqueous extract was standardised to vinblastine by high performance liquid chromatography (HPLC) and was used to determine cytotoxicity by the MTS [3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] assay. DNA fragmentation assay was employed to determine if cell death was due to apoptosis. The results showed that the aqueous extract induced cell death of Jurkat cells at 24, 48 and 72 hours post-treatment in a time- and dose-dependent manner. However, cells treated at 48 and 72 hours produced higher cytotoxic effects with half maximal inhibitory concentration (IC50) values of 2.55 μg/ml and 2.38 μg/ml, respectively. In contrast, the extract induced normal PBMC proliferation, especially after 24 hours treatment with 1000 μg/ml. This result indicates that the C. roseus crude aqueous extract showed differential effects of inhibiting the proliferation of the Jurkat cell line and promoting the growth of PBMCs. These data suggest that the extract may be applicable for modulating the normal and transformed immune cells in leukaemia patients. PMID:24575203

  7. Cytological changes in meristematic cells of Allium cepa L. root tip treated with extracts from callus of Catharanthus roseus (L. G. Don

    Agnieszka Pietrosiuk

    2014-02-01

    Full Text Available The effect of an ethanolic extract from callus of Catharanthus roseus on Allium cepa root cells divisions was investigated. Two lines: white and green callus, were established on solid B5 medium with IAA 1 mg/l and kinetin 0.1 mg/l. The HPLC analysis of callus extracts showed the presence of indole alkaloids, however not known pharmacologically active alkaloids or derivatives used in semisynthesis (vinblastine, ajmalicine, serpentine, yohimbine, vindoline and catharanthine have been found. The ethanolic extract of C. roseus callus inhibited the number of mitoses in Allium cepa root tip cells. Short (1-3 hours treatment resulted in an increase in the index of late prophases, with characteristic light spaces, and the index of metaphases with twisted chromosomes forming an equatorial plate or irregular structures of c-metaphases. At the same time, the percentage of anaphases and telophases decreased significantly. Longer treatment of the root tip inhibited mitotic activity, stopping it completely already after 12 hours. Interphase nuclei became at first denser and homogeneous, eventually their structure became , partitioned into zones and formation of chromatin territories, with distinct large nucleoli has been observed. Electron microscope observations revealed well developed rough endoplasmic reticulum and thick, invaginating cell membrane.

  8. Effects of co-administration of methanol leaf extract of Catharanthus roseus on the hypoglycemic activity of metformin and glibenclamide in rats

    Ohadoma SC; Michael HU

    2011-01-01

    Objective:To investigate the interacting effects of co-administration of methanol leaf extract of Catharanthus roseus (C. roseus) on the hypoglycemic activity of metformin as well as glibenclamide using experimental rats. Methods: Phytochemical analysis as well as acute toxicity and lethality (LD50) test were carried out on its methanol leaf extract. The alloxan model for experimental induction of diabetes in rats was employed. Six groups comprising five rats each were used. GroupsⅡ,ⅢandⅣreceived 250 mg/kg of extract, 100 mg/kg of metformin and 1 mg/kg of glibenclamide respectively, whileⅤandⅥwere administered metformin-extract and glibenclamide-extract combinations respectively at doses as above. GroupⅠserved as negative control and received only distilled water. All administration was done once daily for seven days. Fasting blood glucose was determined at 2, 12, 24, 72 and 168 h using a glucometer. One-way ANOVA with post-hoc tests was used to assess for significant difference due to administration of drug alone and with co-administration of drug and extract. Results:The LD50 was 2 121.32 mg/kg. The phytochemical studies indicated the presence of saponins, tannins, alkaloids, phlotatannins, flavonoids, triterpenoids, reducing sugars, anthraquinones and glycosides. All medicaments significantly reduced blood glucose levels when compared with control alone (P<0.05) with the highest percentage reduction in blood glucose (64.86%) exhibited by metformin-extract combination. Conclusions:The leaf extract of C. roseus significantly increases the hypoglycemic effect of metformin.

  9. Gas Chromatography-Mass Spectrometry (GC-MS Analysis of Indole Alkaloids Isolated from Catharanthus roseus (L. G. Don Cultivated Conventionally and Derived from In vitro Cultures

    Aneta WESOŁOWSKA

    2016-06-01

    Full Text Available Catharanthus roseus (periwinkle is a medicinal plant commonly known for its wide biological activity. In many countries different parts of this plant are used for the treatment of diabetes, hypertension and for menstrual regulation. Due to the ability of production of alkaloids, which can be applied in cancer therapy, is still extensively investigated. Two, the most valuable alkaloids (vincristine and vinblastine are present in C. roseus in very low concentrations. Micropropagation is promising technique used to enhance the level of important secondary metabolites. The main objective of present study was alkaloids extraction from plants cultivated conventionally and derived from in vitro cultures. In this order the aerial parts of periwinkle were extracted with 96% ethanol at room temperature (method I and heated with 96% ethanol at 55 °C for 90 minutes (method II. The obtained mixtures of different indole alkaloids were analyzed by gas chromatography – mass spectrometry (GC-MS. Analysis revealed the presence of 15 alkaloids, among which vindoline, vindorosine, isovindolinine and ajmalicine were the most abundant. The obtained results indicated that the propagation method had a significant effect on the percentage content of alkaloids in C. roseus herb. Plants derived from in vitro cultures were richer in vindorosine and vindoline, while conventionally cultivated – in tetrahydroalstonine and ajmalicine. Moreover, in case of isovindolinine, vindolinine and ajmalicine, extraction at 55 °C was more effective, while for pericyclivine – maceration at room temperature. Interestingly, the pericyclivine was not detected in the mixture of alkaloids obtained from periwinkle herb by the extraction at 55 °C.

  10. Jasmonate-dependent alkaloid biosynthesis in Catharanthus Roseus hairy root cultures is correlated with the relative expression of Orca and Zct transcription factors.

    Goklany, Sheba; Rizvi, Noreen F; Loring, Ralph H; Cram, Erin J; Lee-Parsons, Carolyn W T

    2013-01-01

    The effects of methyl jasmonate (MJ) dosage on terpenoid indole alkaloid (TIA) biosynthesis in Catharanthus roseus are correlated with the relative levels of specific MJ-responsive transcription factors. In this study, the expression of transcription factors (Orca, Zct, Gbf, Myc2, At-hook, and Wrky1), TIA pathway genes (G10h, Tdc, Str, and Sgd), and TIA metabolites (secologanin, strictosidine, and tabersonine) were investigated in C. roseus hairy root cultures elicited with a range of MJ dosages (0-1,000 µM) during mid-exponential growth. The highest production of TIA metabolites occurs at 250 μM MJ, increasing by 150-370% compared with untreated controls. At this MJ dosage, the expression of the transcriptional activators (Orca) is dramatically increased (29-40 fold) while the levels of the transcriptional repressors (Zct) remain low (2-7 fold). Simultaneously, the expression of genes coding for key enzymes involved in TIA biosynthesis increases by 8-15 fold. In contrast, high MJ dosages (1,000 µM) inhibit the production of TIA metabolites. This dosage is correlated with elevated expression levels of Zct (up to 40-fold) relative to Orca (13-19-fold) and minimal induction of the TIA biosynthetic genes (0-6 fold). The significant changes in the expression of Orca and Zct with MJ dosage do not correspond to changes in the expression of the early-response transcription factors (AT-hook, Myc2, and Wrky1) believed to regulate Orca and Zct. In summary, these observations suggest that the dependence of alkaloid production on MJ dosage in C. roseus may be partly mediated through the relative levels of Orca and Zct family transcription factors. PMID:23970483

  11. Suppression of aggressive strains of 'Candidatus phytoplasma mali' by mild strains in Catharanthus roseus and Nicotiana occidentalis and indication of similar action in apple trees.

    Schneider, Bernd; Sule, Sandor; Jelkmann, Wilhelm; Seemüller, Erich

    2014-05-01

    To study antagonistic interactions of 'Candidatus Phytoplasma mali' strains, graft inoculation of Catharanthus roseus and Nicotiana occidentalis was performed with mild strains 1/93Vin and 1/93Tab as suppressors and three aggressive strains as challengers. Inoculation of the suppressors was carried out in either the cross-protection modus prior to grafting of the challengers or by co-inoculating suppressors and challengers. Monitoring using multiplex real-time polymerase chain reaction assays revealed that, in long-term cross-protection trials with C. roseus, suppressor 1/93Vin was present in all root and randomly collected stem samples over the entire observation period. In contrast, the challengers were never detected in such stem samples and rarely in the roots. Following simultaneous inoculation, the suppressor successively colonized all stem and root regions whereas detection of challenger AT steadily decreased. However, this strain remained detectable in up to 13 and 27% of stem and root samples, respectively. The cross-protection trials with N. occidentalis yielded results similar to that of the cross-protection experiments with C. roseus. Comparison of the symptomatology of infected apple trees with the presence of putatively suppressive strains indicated that suppression of severe strains also occurs in apple. Phylogenetic analysis using a variable fragment of AAA+ ATPase gene AP460 of 'Ca. P. mali' revealed that suppressors 1/93Vin and 1/93Tab, together with several other mild strains maintained in apple, cluster distantly from obviously nonsuppressive strains that were predominantly highly virulent. PMID:24724815

  12. 长春花高效快繁技术研究%Rapid Propagation Techniques of Catharanthus roseus (L.) G. Don

    陈泉; 李建国

    2009-01-01

    [目的]建立长春花高效的组织培养与无性快繁体系.[方法]以长春花种子为外植体,分别用1.1%有效氯的次氯酸钠溶液和0.1%HgCl2溶液对长春花种子进行消毒,以MS为基本培养基,通过添加不同浓度的6-BA、IBA和NAA,对脱毒后的长春花试管苗进行增殖和生根培养试验,确定长春花种子的最佳消毒时间,筛选长春花快繁的最佳培养基.[结果]用1.1%有效氯的次氯酸钠消毒长春花种子的最佳消毒时间是30 min,种子发芽率达96.7%;最适宜的增殖培养基为MS+0.40 mg/L 6-BA +0.05 mg/L IBA,外植体的增殖数最多,且长势旺;诱导生根的最佳培养基为1/2MS+0.10 mg/L IBA +0.10 mg/L NAA,试管苗生根率达100%.[结论]次氯酸钠是长春花种子最好的消毒剂,最适宜的增殖培养基和生根培养基分别为MS+0.40 mg/L 6-BA +0.05 mg/L IBA和1/2MS+0.10 mg/L IBA +0.10 mg/L NAA.%[Objective] The study aimed to establish the efficient tissue culture and asexual propagation system of Catharanthus roseus (L.) G. Don. [Method] With seeds of C. roseus as explants, they were disinfected with sodium hypochlorite solution containing 1.1% available chlorine and 0.1% mercuric chloride solution, resp.. MS was used as the basic medium with different concn. of 6-BA, IBA and NAA to configure different media. The tests of proliferation and rooting culture on the virus-free test-tube plantlets of C. roseus were carried out to determine the best disinfection time of C. roseus seeds and screen the optimal medium for rapid propagation of C. roseus. [Result] The best disinfection time by using sodium hypochlorite solution containing 1.1% available chlorine was 30 min and the seed germination rate was 96.7%. The best proliferation medium for C. roseus was MS+0.40 mg/L 6-BA +0.05 mg/L IBA, on which the explants got more proliferated number and strong growth vigor. The best rooting culture medium for C. roseus was 1/2MS+0.10 mg/L IBA +0.10 mg/L NAA, on which the test

  13. Gene transcript profiles of the TIA biosynthetic pathway in response to ethylene and copper reveal their interactive role in modulating TIA biosynthesis in Catharanthus roseus.

    Pan, Ya-Jie; Liu, Jia; Guo, Xiao-Rui; Zu, Yuan-Gang; Tang, Zhong-Hua

    2015-05-01

    Research on transcriptional regulation of terpenoid indole alkaloid (TIA) biosynthesis of the medicinal plant, Catharanthus roseus, has largely been focused on gene function and not clustering analysis of multiple genes at the transcript level. Here, more than ten key genes encoding key enzyme of alkaloid synthesis in TIA biosynthetic pathways were chosen to investigate the integrative responses to exogenous elicitor ethylene and copper (Cu) at both transcriptional and metabolic levels. The ethylene-induced gene transcripts in leaves and roots, respectively, were subjected to principal component analysis (PCA) and the results showed the overall expression of TIA pathway genes indicated as the Q value followed a standard normal distribution after ethylene treatments. Peak gene expression was at 15-30 μM of ethephon, and the pre-mature leaf had a higher Q value than the immature or mature leaf and root. Treatment with elicitor Cu found that Cu up-regulated overall TIA gene expression more in roots than in leaves. The combined effects of Cu and ethephon on TIA gene expression were stronger than their separate effects. It has been documented that TIA gene expression is tightly regulated by the transcriptional factor (TF) ethylene responsive factor (ERF) and mitogen-activated protein kinase (MAPK) cascade. The loading plot combination with correlation analysis for the genes of C. roseus showed that expression of the MPK gene correlated with strictosidine synthase (STR) and strictosidine b-D-glucosidase(SGD). In addition, ERF expression correlated with expression of secologanin synthase (SLS) and tryptophan decarboxylase (TDC), specifically in roots, whereas MPK and myelocytomatosis oncogene (MYC) correlated with STR and SGD genes. In conclusion, the ERF regulates the upstream pathway genes in response to heavy metal Cu mainly in C. roseus roots, while the MPK mainly participates in regulating the STR gene in response to ethylene in pre-mature leaf. Interestingly, the

  14. Pleiotropic phenotypes of the salt-tolerant and cytosine hypomethylated leafless inflorescence, evergreen dwarf and irregular leaf lamina mutants of Catharanthus roseus possessing Mendelian inheritance

    Renu Kumari; Vishakha Sharma; Vinay Sharma; Sushil Kumar

    2013-12-01

    In Catharanthus roseus, three morphological cum salt-tolerant chemically induced mutants ofMendelian inheritance and their wild-type parent cv Nirmal were characterized for overall cytosine methylation at DNA repeats, expression of 119 protein-coding and seven miRNA-coding genes and 50 quantitative traits. The mutants, named after their principal morphological feature(s), were leafless inflorescence (lli), evergreen dwarf (egd) and irregular leaf lamina (ill). The Southern-blot analysis of MspI digested DNAs of mutants probed with centromeric and 5S and 18S rDNA probes indicated that, in comparison to wild type, the mutants were extensively demethylated at cytosine sites. Among the 126 genes investigated for transcriptional expression, 85 were upregulated and 41 were downregulated in mutants. All of the five genes known to be stress responsive had increased expression in mutants. Several miRNA genes showed either increased or decreased expression in mutants. The C. roseus counterparts of CMT3, DRM2 and RDR2 were downregulated in mutants. Among the cell, organ and plant size, photosynthesis and metabolism related traits studied, 28 traits were similarly affected in mutants as compared to wild type. Each of the mutants also expressed some traits distinctively. The egd mutant possessed superior photosynthesis and water retention abilities. Biomass was hyperaccumulated in roots, stems, leaves and seeds of the lli mutant. The ill mutant was richest in the pharmaceutical alkaloids catharanthine, vindoline, vincristine and vinblastine. The nature of mutations, origins of mutant phenotypes and evolutionary importance of these mutants are discussed.

  15. Antagonism of Ca2+ influx via L-type Ca2+ channels mediates the vasorelaxant effect of Catharanthus roseus-derived vindorosine in rat renal artery.

    Wu, Xiao-Lin; Cheang, Wai San; Zhang, Dong-Mei; Li, Yong; Lau, Chi-Wai; Wang, Guo-Cai; Huang, Yu; Ye, Wen-Cai

    2014-12-01

    Catharanthus roseus is a traditional herbal medicine used in Asian and African countries for the treatment of various diseases including hypertension. The present study examined possible cellular mechanisms for the relaxation of rat renal arteries induced by vindorosine extracted from C. roseus. Intrarenal arteries were isolated from 200-300 g male Sprague-Dawley rats and treated with different pharmacological blockers and inhibitors for the measurement of vascular reactivity on a Multi Myograph System. Fluorescence imaging by laser scanning confocal microscopy was utilized to determine the intracellular Ca(2+) level in the vascular smooth muscles of the renal arteries. Vindorosine in micromolar concentrations relaxes renal arteries precontracted by KCl, phenylephrine, 11-dideoxy-9α,11α-epoxymethanoprostaglandin F2α, and serotonin. Vindorosine-induced relaxations were unaffected by endothelium denudation or by treatment with the nitric oxide synthase inhibitor N (G)-nitro-L-arginine methyl ester hydrochloride, the guanylyl cyclase inhibitor 1H-[1, 2, 4]oxadiazolo[4,3-a]quinoxalin-1-one, the cyclooxygenase inhibitor indomethacin, or K(+) channel blockers such as tetraethylammonium ions, glibenclamide, and BaCl2. Vindorosine-induced relaxations were attenuated in the presence of 0.1 µM nifedipine (an L-type Ca(2+) channel blocker). Vindorosine also concentration-dependently suppressed contractions induced by CaCl2 (0.01-5 mM) in Ca-free 60 mM KCl solution. Furthermore, fluorescence imaging using fluo-4 demonstrated that 30 min incubation with 100 µM vindorosine reduced the 60 mM KCl-stimulated Ca(2+) influx in the smooth muscles of rat renal arteries. The present study is probably the first report of blood vessel relaxation by vindorosine and the possible underlying mechanisms involving the inhibition of Ca(2+) entry via L-type Ca(2+) channels in vascular smooth muscles. PMID:25340466

  16. Pleiotropic phenotypes of the salt-tolerant and cytosine hypomethylated leafless inflorescence, evergreen dwarf and irregular leaf lamina mutants of Catharanthus roseus possessing Mendelian inheritance.

    Kumari, Renu; Sharma, Vishakha; Sharma, Vinay; Kumar, Sushil

    2013-12-01

    In Catharanthus roseus, three morphological cum salt-tolerant chemically induced mutants of Mendelian inheritance and their wild-type parent cv Nirmal were characterized for overall cytosine methylation at DNA repeats, expression of 119 protein coding and seven miRNA-coding genes and 50 quantitative traits. The mutants, named after their principal morphological feature(s), were leafless inflorescence (lli), evergreen dwarf (egd) and irregular leaf lamina (ill). The Southern-blot analysis of MspI digested DNAs of mutants probed with centromeric and 5S and 18S rDNA probes indicated that, in comparison to wild type, the mutants were extensively demethylated at cytosine sites. Among the 126 genes investigated for transcriptional expression, 85 were upregulated and 41 were downregulated in mutants. All of the five genes known to be stress responsive had increased expression in mutants. Several miRNA genes showed either increased or decreased expression in mutants. The C. roseus counterparts of CMT3, DRM2 and RDR2 were downregulated in mutants. Among the cell, organ and plant size, photosynthesis and metabolism related traits studied, 28 traits were similarly affected in mutants as compared to wild type. Each of the mutants also expressed some traits distinctively. The egd mutant possessed superior photosynthesis and water retention abilities. Biomass was hyperaccumulated in roots, stems, leaves and seeds of the lli mutant. The ill mutant was richest in the pharmaceutical alkaloids catharanthine, vindoline, vincristine and vinblastine. The nature of mutations, origins of mutant phenotypes and evolutionary importance of these mutants are discussed. PMID:24371160

  17. Aislamiento de consorcios de hongos micorrícicos arbusculares de plantas medicinales y su efecto en el crecimiento de vinca (Catharanthus roseus Isolation of arbuscular mycorrhizal fungi consortia from medicinal plants and their effectiveness on growth of vinca (Catharanthus roseus

    CLAUDIA DE LA ROSA-MERA

    2012-06-01

    Full Text Available Este trabajo consistió en propagar e identificar hongos micorrícicos arbusculares (HMA recolectados de plantas medicinales (PM de áreas naturales de bosques mixtos, y seleccionar consorcios micorrícicos con base en la promoción del crecimiento de vinca Catharanthus roseus (L G. Don, planta medicinal cuyos alcaloides tienen propiedades antineoplásicas. En la primera fase experimental se recolectaron raíces y suelo rizosférico de 13 PM establecidas en campo para evaluar el porcentaje de colonización total (PCT y cuantificar el número de esporas; además, se tomó una parte del suelo para establecer plantas trampa en invernadero durante 10 meses, y posteriormente evaluar el PCT e identificar los principales géneros de HMA. Todas las PM en su condición natural presentaron colonización micorrícica, observándose cuatro géneros de HMA (Glomus, Acaulospora, Gigaspora y Scutellospora, de los cuales Acaulospora y Glomus fueron los predominantes. En la segunda fase experimental se seleccionaron ocho consorcios con base en el PCT (> 40 % obtenido en las plantas trampa, que correspondieron a las muestras recolectadas de Adiantum capillus-veneris L., Castilleja tenuiflora Benth., Erigeron karvinskianus DC., Pimpinella anisum L., Plantago major L., Ricinus communis L., Rubus fruticosus L. y Rumex mexicanus Meisn. Estos consorcios fueron inoculados en plántulas de C. roseus para evaluar su capacidad de estimular el crecimiento de esta especie en condiciones de invernadero. Después de 70 días, a pesar de presentar un solo género predominante (Glomus, el consorcio aislado de R. mexicanus promovió de manera más consistente el crecimiento de C. roseus (número de hojas, área foliar y peso seco foliar en comparación con el resto de los consorcios micorrícicos.This study consisted on propagating and identifying arbuscular mycorrhizal fungi (AMF collected from medicinal plants (MP of natural areas of mixed forest (Estado de Mexico, and

  18. Indirect regeneration from in vitro leaf tissue of periwinkle (Catharanthus roseus L. in response to different treatments of plant growth regulators

    B.E. Sayed-Tabatabaei

    2012-03-01

    Full Text Available Periwinkle (Catharanthus roseus L. belongs to the Apocynaceae family and accumulates more than 130 terpenoid indole alkaloids (TIAs, of which two dimeric alkaloids Vinblastine and Vincristine have antineoplastic activity and are useful for treatment of various cancers. Therefore, the production of these drugs has been emphasized in plant tissue culture. In this research, 25 treatments of plant growth regulators to produce callus from leaf explants and seven treatments for regeneration of calli were considered. Analysis of variance showed that the effect of different hormonal treatments in the production of callus from leaf explants is significant (P<0.01. Application of 0.1 mg/L BAP and 5 mg/L NAA in nutrient media produced a lot of calli and roots. In addition, nutrient media containing activated charcoal and without it were found to be suitable for production of callus and regeneration, respectively. In conclusion, the results showed that indirect regeneration of leaf explants is not a suitable method for micropropagation of periwinkle due to difficult regeneration of callus, probability of mutation and lower number of produced branches. But this method can be utilized for production of secondary metabolites.

  19. Identification of a human ABCC10 orthologue in Catharanthus roseus reveals a U12-type intron determinant for the N-terminal domain feature

    Taissir El-Guizani; Clotilde Guibert; Saïda Triki; Benoit St-Pierre; Eric Ducos

    2014-04-01

    ABC (ATP-binding cassette) transporters are members of a large superfamily of proteins that utilize ATP hydrolysis to translocate a wide range of substrates across biological membranes. In general, members of C subfamily (ABCC) are structurally characterized by an additional (N-terminal) transmembrane domain (TMD0). Phylogenetic analysis of plant ABCCs separates their protein sequences into three distinct clusters: I and II are plant specific whereas cluster III contains both human and plant ABCCs. Screening of the Plant Medicinal Genomics Resource database allowed us to identify 16 ABCCs partial sequences in Catharanthus roseus; two of which belong to the unique CrABCC1 transcript that we identified in cluster III. Genomic organization of CrABCC1 TMD0 coding sequence displays an AT-AC U12-type intron that is conserved in higher plant orthologues. We showed that CrABCC1, like its human orthologue ABCC10, produces alternative transcripts that encode protein sequences with a truncated form of TMD0 without the first transmembrane span (TM1). Subcellular localization of CrABCC1 TMD0 variants using yellow fluorescent protein fusions reveals that the TM1 is required for a correct routing of the TMD0 to the tonoplast. Finally, the specific repartition of CrABCC1 orthologues in some species suggests that this gene was lost several times during evolution and that its physiological function may, rely on a common feature of multicellular eukaryotes.

  20. Identification of a human ABCC10 orthologue in Catharanthus roseus reveals a U12-type intron determinant for the N-terminal domain feature.

    El-Guizani, Taissir; Guibert, Clotilde; Triki, Saida; St-Pierre, Benoit; Ducos, Eric

    2014-04-01

    ABC (ATP-binding cassette) transporters are members of a large superfamily of proteins that utilize ATP hydrolysis to translocate a wide range of substrates across biological membranes. In general, members of C subfamily (ABCC) are structurally characterized by an additional (N-terminal) transmembrane domain (TMD0). Phylogenetic analysis of plant ABCCs separates their protein sequences into three distinct clusters: I and II are plant specific whereas cluster III contains both human and plant ABCCs. Screening of the Plant Medicinal Genomics Resource database allowed us to identify 16 ABCCs partial sequences in Catharanthus roseus; two of which belong to the unique CrABCC1 transcript that we identified in cluster III. Genomic organization of CrABCC1 TMD0 coding sequence displays an AT-AC U12-type intron that is conserved in higher plant orthologues. We showed that CrABCC1, like its human orthologue ABCC10, produces alternative transcripts that encode protein sequences with a truncated form of TMD0 without the first transmembrane span (TM1). Subcellular localization of CrABCC1 TMD0 variants using yellow fluorescent protein fusions reveals that the TM1 is required for a correct routing of the TMD0 to the tonoplast. Finally, the specific repartition of CrABCC1 orthologues in some species suggests that this gene was lost several times during evolution and that its physiological function may, rely on a common feature of multicellular eukaryotes. PMID:24840820

  1. 长春花愈伤组织的诱导和增殖%Effects on induction and proliferation of Catharanthus roseus (L.)G.Don callus

    郭胜娟; 杨春燕; 冯玲玲; 张国彬; 范美华; 张泽宏; 刘树楠; 周吉源

    2004-01-01

    以长春花(Catharanthus roseus (L.)G.Don)幼嫩叶片作为外植体,接种于附加不同浓度的NAA,2,4-D、6-BA、KT及其组合的MS固体培养基上,结果发现除单独使用KT不能产生愈伤组织外,其它3种单独使用时,在一定的浓度范围均有愈伤组织产生;最佳的组合培养基为MS+NAA 0.5 mg/L+2,4-D 0.5 mg/L+6-BA 2.0 mg/L.进一步研究发现,在黑暗和光照培养条件下,愈伤组织增殖呈 "S"型,且生长周期均为27 d.

  2. Phase-specific polypeptides and poly(A) sup + RNAs during the cell cycle in synchronous cultures of Catharanthus roseus cells

    Kodama, Hiroaki; Komamine, Atsushi (Tohoku Univ., Sendai (Japan)); Kawakami, Naoto; Watanabe, Akira (Nagoya Univ. (Japan))

    1989-03-01

    This study shows an overall analysis of gene expression during the cell cycle in synchronous suspension cultures of Catharanthus roseus cells. First, the cellular cytoplasmic proteins were fractionated by two-dimensional gel electrophoresis and visualized by staining with silver. Seventeen polypeptides showed qualitative or quantitative changes during the cell cycle. Second, the rates of synthesis of cytoplasmic proteins were also investigated by autoradiography by labeling cells with ({sup 35}S)methionine at each phase of the cell cycle. The rates of synthesis of 13 polypeptides were found to vary during the cell cycle. The silver-stained electrophoretic pattern of proteins in the G{sub 2} phase in particular showed characteristic changes in levels of polypeptides, while the rates of synthesis of polypeptides synthesized during the G{sub 2} phase did not show such phase-specific changes. This result suggest that posttranslational processing of polypeptides occurs during or prior to the G{sub 2} phase. In the G{sub 1} and S phases and during cytokinesis, several other polypeptides were specifically synthesized. Finally, the variation of mRNAs was analyzed from the autoradiograms of in vitro translation products of poly(A){sup +} RNA isolated at each phase. Three poly(A){sup +} RNAs increased in amount from the G{sub 1} to the S phase and one poly(A){sup +} RNA increased preferentially from the G{sub 2} phase to cytokinesis.

  3. Application of natural deep eutectic solvents to the extraction of anthocyanins from Catharanthus roseus with high extractability and stability replacing conventional organic solvents.

    Dai, Yuntao; Rozema, Evelien; Verpoorte, Robert; Choi, Young Hae

    2016-02-19

    Natural deep eutectic solvents (NADES) have attracted a great deal of attention in recent times as promising green media. They are generally composed of neutral, acidic or basic compounds that form liquids of high viscosity when mixed in certain molar ratio. Despite their potential, viscosity and acid or basic nature of some ingredients may affect the extraction capacity and stabilizing ability of the target compounds. To investigate these effects, extraction with a series of NADES was employed for the analysis of anthocyanins in flower petals of Catharanthus roseus in combination with HPLC-DAD-based metabolic profiling. Along with the extraction yields of anthocyanins their stability in NADES was also studied. Multivariate data analysis indicates that the lactic acid-glucose (LGH), and 1,2-propanediol-choline chloride (PCH) NADES present a similar extraction power for anthocyanins as conventional organic solvents. Furthermore, among the NADES employed, LGH exhibits an at least three times higher stabilizing capacity for cyanidins than acidified ethanol, which facilitates their extraction and analysis process. Comparing NADES to the conventional organic solvents, in addition to their reduced environmental impact, they proved to provide higher stability for anthocyanins, and therefore have a great potential as possible alternatives to those organic solvents in health related areas such as food, pharmaceuticals and cosmetics. PMID:26822320

  4. Virus-induced gene silencing identifies Catharanthus roseus 7-deoxyloganic acid-7-hydroxylase, a step in iridoid and monoterpene indole alkaloid biosynthesis.

    Salim, Vonny; Yu, Fang; Altarejos, Joaquín; De Luca, Vincenzo

    2013-12-01

    Iridoids are a major group of biologically active molecules that are present in thousands of plant species, and one versatile iridoid, secologanin, is a precursor for the assembly of thousands of monoterpenoid indole alkaloids (MIAs) as well as a number of quinoline alkaloids. This study uses bioinformatics to screen large databases of annotated transcripts from various MIA-producing plant species to select candidate genes that may be involved in iridoid biosynthesis. Virus-induced gene silencing of the selected genes combined with metabolite analyses of silenced plants was then used to identify the 7-deoxyloganic acid 7-hydroxylase (CrDL7H) that is involved in the 3rd to last step in secologanin biosynthesis. Silencing of CrDL7H reduced secologanin levels by at least 70%, and increased the levels of 7-deoxyloganic acid to over 4 mg g(-1) fresh leaf weight compared to control plants in which this iridoid is not detected. Functional expression of this CrDL7H in yeast confirmed its biochemical activity, and substrate specificity studies showed its preference for 7-deoxyloganic acid over other closely related substrates. Together, these results suggest that hydroxylation precedes carboxy-O-methylation in the secologanin pathway in Catharanthus roseus. PMID:24103035

  5. Processing of forecast by ARIMA model on the weak electrical signals in Catharanthus roseus%长春花微弱电信号的ARIMA模型

    王兰州; 李峤; 李东升

    2007-01-01

    依据长春花(Catharanthus roseus)植物电信号小波软阈值消噪后的数据进行其电信号时间序列的求和自回归移动平均(ARIMA)模型分析.长春花植物微弱低频电信号的模型是Xt=2Xt-1+X-2+εt-0.119 04εt-1-0.361 83εt-2-0.155 47εt-3-0.363 66εt-4.长春花电信号幅值后向10个点的真实值与预测值的相对误差小于15%,表明利用自回归移动平均(ARIMA)模型对植物微弱电信号特性进行预测是可行的.预测数据可作为以节能为目标依据植物自适应电信号特性建立温室和/或塑料大棚智能自动化控制系统的重要参数.

  6. Molecular characterization of recombinant T1, a non-allergenic periwinkle (Catharanthus roseus) protein, with sequence similarity to the Bet v 1 plant allergen family.

    Laffer, Sylvia; Hamdi, Said; Lupinek, Christian; Sperr, Wolfgang R; Valent, Peter; Verdino, Petra; Keller, Walter; Grote, Monika; Hoffmann-Sommergruber, Karin; Scheiner, Otto; Kraft, Dietrich; Rideau, Marc; Valenta, Rudolf

    2003-01-01

    More than 25% of the population suffer from Type I allergy, an IgE-mediated hypersensitivity disease. Allergens with homology to the major birch ( Betula verrucosa ) pollen allergen, Bet v 1, belong to the most potent elicitors of IgE-mediated allergies. T1, a cytokinin-inducible cytoplasmic periwinkle ( Catharanthus roseus ) protein, with significant sequence similarity to members of the Bet v 1 plant allergen family, was expressed in Escherichia coli. Recombinant T1 (rT1) did not react with IgE antibodies from allergic patients, and failed to induce basophil histamine release and immediate-type skin reactions in Bet v 1-allergic patients. Antibodies raised against purified rT1 could be used for in situ localization of natural T1 by immunogold electron microscopy, but did not cross-react with most of the Bet v 1-related allergens. CD analysis showed significant differences regarding secondary structure and thermal denaturation behaviour between rT1 and recombinant Bet v 1, suggesting that these structural differences are responsible for the different allergenicity of the proteins. T1 represents a non-allergenic member of the Bet v 1 family that may be used to study structural requirements of allergenicity and to engineer hypo-allergenic plants by replacing Bet v 1-related allergens for primary prevention of allergy. PMID:12656672

  7. Phase-Specific Polypeptides and Poly(A)+ RNAs during the Cell Cycle in Synchronous Cultures of Catharanthus roseus Cells 1

    Kodama, Hiroaki; Kawakami, Naoto; Watanabe, Akira; Komamine, Atsushi

    1989-01-01

    This study shows an overall analysis of gene expression during the cell cycle in synchronous suspension cultures of Catharanthus roseus cells. First, the cellular cytoplasmic proteins were fractionated by two-dimensional gel electrophoresis and visualized by staining with silver. Seventeen polypeptides showed qualitative or quantitative changes during the cell cycle. Second, the rates of synthesis of cytoplasmic proteins were also investigated by autoradiography by labeling cells with [35S]methionine at each phase of the cell cycle. The rates of synthesis of 13 polypeptides were found to vary during the cell cycle. The silverstained electrophoretic pattern of proteins in the G2 phase in particular showed characteristic changes in levels of polypeptides, while the rates of synthesis of polypeptides synthesized during the G2 phase did not show such phase-specific changes. This result suggests that posttranslational processing of polypeptides occurs during or prior to the G2 phase. In the G1 and S phases and during cytokinesis, several other polypeptides were specifically synthesized. Finally, the variation of mRNAs was analyzed from the autoradiograms of in vitro translation products of poly(A)+ RNA isolated at each phase. Three poly(A)+ RNAs increased in amount from the G1 to the S phase and one poly (A)+ RNA increased preferentially from the G2 phase to cytokinesis. Images Figure 1 Figure 3 Figure 4 Figure 6 Figure 7 Figure 8 Figure 10 Figure 11 Figure 12 PMID:16666641

  8. 4种外源激素对长春花生物碱积累的影响%Effects of exogenous hormones on the accumulation of alkaloids in Catharanthus roseus (L.)

    邹冬梅; 冯信平; 王建荣; 庞玉新; 蒋昌顺

    2012-01-01

    分别用茉莉酸、乙烯利、乙酰水杨酸、色氨酸等4种试剂的不同质量浓度的溶液,处理温室盆栽的长春花成苗和幼苗。在处理2d后,测定长春花叶片中的吲哚生物总碱、长春碱、长春质碱和文多灵碱的含量。结果表明,4种试剂对长春花植物中吲哚生物碱的积累有明显的促进作用。%Four chemical reagents, jasmonic acid (JA) , ethrel, Aspirin and tryptophan, were used as exogenous hormones to treat pot plants of Catharanthus roseus grown in green house to determine the contents of medicinal components, alkaloids, in C. roseus leaves. The pot plants at the age of 2 and 5 months were treated with the reagents at different concentrations. Two days after treatment, the contents of catharanthine, vinblastine and vindoline in leaves were determined by using HPLC. All the reagents under trial obviously improved the accumulation of indole alkaloids including catharanthine and vinblastine in the leaves of C. roseus, and the optimum treatment concentrations of these reagents were determined. These reagents apparently promote the accumulation of total indole alkoloids in the leaves of C. roseus.

  9. Ethylene-Induced Vinblastine Accumulation Is Related to Activated Expression of Downstream TIA Pathway Genes in Catharanthus roseus

    Xi Wang; Ya-Jie Pan; Bo-Wen Chang; Yan-Bo Hu; Xiao-Rui Guo; Zhong-Hua Tang

    2016-01-01

    We selected different concentrations of ethephon, to stress C. roseus. We used qRT-PCR and HPLC followed by PCA to obtain comprehensive profiling of the vinblastine biosynthesis in response to ethephon. Based on our findings, the results showed that the high concentration of ethephon had a positive effect at both transcriptional and metabolite level. Meanwhile, there was a remarkable decrease of hydrogen peroxide content and a promoted peroxidase activity in leaves. The loading plot combinati...

  10. 有机溶剂对长春花Catharanthus roseus细胞悬浮生长的影响%The Effects of Organic Solvents on Growth of Catharanthus Roseus Cell Suspension Cultures

    朱利中; W. R. Cullen

    1993-01-01

    @@植物细胞悬浮培养已广泛被用于生物碱及有机物的合成中[1-4]。长春花Catharanthus roseus(C.roseus)细胞悬浮培养是常用于生物合成的体系之一。长春花C.roseus是Catha-ranthus G.Don(长春花属,夹竹桃科)的一种,植株含有长春碱,有降低血压之功能,亦能抗癌。早期人们直接从植株中提取生物碱,由于含量太低等原因、成本太高。近年来有人利用长春花C.roseus细胞悬浮培养来制备所需的生物碱(细胞培养生长过程中的代谢物),并在培养介质中加入代谢诱导物(如硫酸钒)来刺激生物碱的产生,以提高产量[5-6]。显然,培养介质对C.roseus细胞生长乃至生物碱的产量起着决定性的作用。此外,在C.roseus细胞悬浮培养体系中,加入一些反应物可达到生物有机合成之目的,以解决人工合成中的一些难题[3]。但有些代谢诱导物或反应物不溶于水,常要将它们先溶于少量乙醇、丙酮等有机溶剂中,然后加到植物细胞培养体系中。那么,这些有机溶剂对C.roseus细胞悬浮生长有何影响,我们还未曾见过有关这方面的报道。本文研究了甲醇、乙醇、丙酮对C.roseus细胞悬浮生长的影响,可为生物合成中有机溶剂的选择和使用提供参考。

  11. Characterization of variation and quantitative trait loci related to terpenoid indole alkaloid yield in a recombinant inbred line mapping population of Catharanthus roseus

    Vishakha Sharma; Swati Chaudhary; Suchi Srivastava; Richa Pandey; Sushil Kumar

    2011-04-01

    Improved Catharanthus roseus cultivars are required for high yields of vinblastine, vindoline and catharanthine and/or serpentine and ajmalicine, the pharmaceutical terpenoid indole alkaloids. An approach to derive them is to map QTL for terpenoid indole alkaloids yields, identify DNA markers tightly linked to the QTL and apply marker assisted selection. Towards the end, 197 recombinant inbred lines from a cross were grown over two seasons to characterize variability for seven biomass and 23 terpenoid indole alkaloids content-traits and yield-traits. The recombinant inbred lines were genotyped for 178 DNA markers which formed a framework genetic map of eight linkage groups (LG), spanning 1786.5 cM, with 10.0 cM average intermarker distance. Estimates of correlations between traits allowed selection of seven relatively more important traits for terpenoid indole alkaloids yields. QTL analysis was performed on them using single marker (regression) analysis, simple interval mapping and composite interval mapping procedures. A total of 20 QTL were detected on five of eight LG, 10 for five traits on LG1, five for four traits on LG2, three for one trait on LG3 and one each for different traits on LG three and four. QTL for the same or different traits were found clustered on three LG. Co-location of two QTL for biomass traits was in accord of correlation between them. The QTL were validated for use in marker assisted selection by the recombinant inbred line which transgressively expressed 16 traits contributory to the yield vinblastine, vindoline and catharanthine from leaves and roots that possessed favourable alleles of 13 relevant QTL.

  12. Three non-autonomous signals collaborate for nuclear targeting of CrMYC2, a Catharanthus roseus bHLH transcription factor

    2010-01-01

    Background CrMYC2 is an early jasmonate-responsive bHLH transcription factor involved in the regulation of the expression of the genes of the terpenic indole alkaloid biosynthesis pathway in Catharanthus roseus. In this paper, we identified the amino acid domains necessary for the nuclear targeting of CrMYC2. Findings We examined the intracellular localization of whole CrMYC2 and of various deletion mutants, all fused with GFP, using a transient expression assay in onion epidermal cells. Sequence analysis of this protein revealed the presence of four putative basic nuclear localization signals (NLS). Assays showed that none of the predicted NLS is active alone. Further functional dissection of CrMYC2 showed that the nuclear targeting of this transcription factor involves the cooperation of three domains located in the C-terminal region of the protein. The first two domains are located at amino acid residues 454-510 and 510-562 and contain basic classical monopartite NLSs; these regions are referred to as NLS3 (KRPRKR) and NLS4 (EAERQRREK), respectively. The third domain, between residues 617 and 652, is rich in basic amino acids that are well conserved in other phylogenetically related bHLH transcription factors. Our data revealed that these three domains are inactive when isolated but act cooperatively to target CrMYC2 to the nucleus. Conclusions This study identified three amino acid domains that act in cooperation to target the CrMYC2 transcription factor to the nucleus. Further fine structure/function analysis of these amino acid domains will allow the identification of new NLS domains and will allow the investigation of the related molecular mechanisms involved in the nuclear targeting of the CrMYC2 bHLH transcription factor. PMID:21073696

  13. Isolation of Genes that Are Preferentially Expressed at the G1/S Boundary during the Cell Cycle in Synchronized Cultures of Catharanthus roseus Cells 1

    Kodama, Hiroaki; Ito, Masaki; Hattori, Tsukaho; Nakamura, Kenzo; Komamine, Atsushi

    1991-01-01

    A cDNA library was screened for genes that may be involved in the progression of the cell cycle of cells of higher plants. The Catharanthus roseus L. (G) Don. cells were synchronized by the double phosphate starvation method, and a λgt11 cDNA library was prepared using poly(A)+ RNA from cells in the S phase of the cell cycle. Two independent sequences, cyc02 and cyc07, were identified by differential screening. The levels of cyc02 and cyc07 mRNAs increased dramatically, but transiently, at the G1/S boundary of the cell cycle. High levels of cyc02 mRNA, but not of cyc07 mRNA, were also present in cells arrested at the G1 phase by phosphate starvation. In an asynchronous batch culture, cyc02 and cyc07 mRNAs accumulated transiently at different stages of the growth cycle, cyc02 mRNA early in the stationary phase, and cyc07 mRNA in the midlogarithmic phase. When the proliferation of cells was arrested by nutrient starvation, i.e. by sucrose or nitrogen starvation, the relative amounts of the cyc02 and cyc07 mRNAs decreased. These results indicate that cyc02 and cyc07 contain nucleotide sequences from growth-related genes. The analysis of nucleotide sequence of cyc02 shows that the predicted product of this gene is basic and is composed of 101 amino acids. No significant homology to other known proteins was detected. Images Figure 1 Figure 4 Figure 5 PMID:16667998

  14. Over-expression of Catharanthus roseus tryptophan decarboxylase and strictosidine synthase in rol gene integrated transgenic cell suspensions of Vinca minor.

    Verma, Priyanka; Sharma, Abhishek; Khan, Shamshad Ahmad; Shanker, Karuna; Mathur, Ajay K

    2015-01-01

    Tryptophan decarboxylase (TDC) and strictosidine synthase (STR) genes from Catharanthus roseus have been successfully over-expressed in the rol gene integrated cell suspensions of V. minor. Thirty seconds SAAT (sonication-assisted Agrobacterium transformation) treatment of plant cell suspension with LBA1119 having construct () generated three stable TDC + STR over-expressing cell lines--PVG1, PVG2, and PVG3. The transgenes were confirmed by β-glucuronidase GUS histochemical assay and PCR amplification of rol genes/GUS gene. All the three cell suspension lines were found to be slow growing. In comparison to the control cell suspensions (GI = 241.0 ± 5.8), PVG3 cell line registered a growth index (GI) of 208.0 ± 10.0 followed by PVG1 (GI = 140.0 ± 14.2) and PVG2 (GI = 85.0 ± 9.6). The PVG3 cell line was also up-scaled in the 5-l stirred tank bioreactor with GI of 745.6 ± 35.3 under optimized parameters. Only PVG3 line registered a twofold increase in total alkaloid content (2.1 ± 0.1% dry wt.) and showed vincamine presence (0.003 ± 0.001% dry wt.) which was further enhanced at the bioreactor level (2.7 ± 0.3 and 0.005 ± 0.001% dry wt., respectively). Real-time (RT) qPCR analysis of PVG3 showed more than sevenfold to eightfold increase in TDC and STR expression [relative quantity value (RQ) = 7.6 ± 0.8 (TDC); RQ = 8.5 ± 0.9 (STR)]. PMID:25106473

  15. Three non-autonomous signals collaborate for nuclear targeting of CrMYC2, a Catharanthus roseus bHLH transcription factor

    Gantet Pascal

    2010-11-01

    Full Text Available Abstract Background CrMYC2 is an early jasmonate-responsive bHLH transcription factor involved in the regulation of the expression of the genes of the terpenic indole alkaloid biosynthesis pathway in Catharanthus roseus. In this paper, we identified the amino acid domains necessary for the nuclear targeting of CrMYC2. Findings We examined the intracellular localization of whole CrMYC2 and of various deletion mutants, all fused with GFP, using a transient expression assay in onion epidermal cells. Sequence analysis of this protein revealed the presence of four putative basic nuclear localization signals (NLS. Assays showed that none of the predicted NLS is active alone. Further functional dissection of CrMYC2 showed that the nuclear targeting of this transcription factor involves the cooperation of three domains located in the C-terminal region of the protein. The first two domains are located at amino acid residues 454-510 and 510-562 and contain basic classical monopartite NLSs; these regions are referred to as NLS3 (KRPRKR and NLS4 (EAERQRREK, respectively. The third domain, between residues 617 and 652, is rich in basic amino acids that are well conserved in other phylogenetically related bHLH transcription factors. Our data revealed that these three domains are inactive when isolated but act cooperatively to target CrMYC2 to the nucleus. Conclusions This study identified three amino acid domains that act in cooperation to target the CrMYC2 transcription factor to the nucleus. Further fine structure/function analysis of these amino acid domains will allow the identification of new NLS domains and will allow the investigation of the related molecular mechanisms involved in the nuclear targeting of the CrMYC2 bHLH transcription factor.

  16. Effects of aluminum on DNA synthesis, cellular polyamines, polyamine biosynthetic enzymes and inorganic ions in cell suspension cultures of a woody plant, Catharanthus roseus

    Minocha, R.; Shortle, W.C. (USDA Forest Service, Durham (US)); Minocha, S.C.; Long, S.L. (Dept. of Plant Biology, Univ. of New Hamshire, Durham (US))

    1992-01-01

    Increased aluminium (Al) solubility in soil waters due to acid precipitation has aroused considerable interest in the problem of Al toxicity in plants. In the present study, an in vitro suspension culture system of Catharanthus roseus (L.) G. Don was used to analyze the effects of aluminum on several biochemical processes in these cells. The aliphatic polyamines, spermine and spermidine, and their precusor, putrescine, have been implicated in a number of stress responses of plants. Addition of 0.2, 0.5 or 1.0 mM AlCl{sub 3} to cells cultured for 3 days caused a small but significant increase in cellular levels of putrescine at 4 h followed by a sharp decline by 16 h. There was no further decline in levels of putrescine during the next 32 h. Spermidine levels did not change appreciably compared to those in the control cultures. However, spermine levels increased by 2-3-fold at 24 and 48 h. Cellular activities of arginine decarboxylase (ADC; EC 4.1.1.19) and S-adenosylmethionine decarboxylase (SAMDC; EC 4.1.1.50) were both inhibited by 20-25% at 4 and 7 h. Ornithine decarboxylase (ODC; EC 4.1.1.17) was less than 10% of ADC activity at all times. Whereas all concentrations of Al caused a slight decrease in total cell number, cell viability was affected only by 1.0 mM Al. There was a decrease in the cellular levels of Ca, Mg, Na, K, Mn, P and Fe in the cells treated with Al at 4 h, but a significant increase by 16 and 24 h. The results presented here suggest that both the absolute amounts of Al and the length of exposure to it are important for cell toxicity. (au).

  17. Ethylene-Induced Vinblastine Accumulation Is Related to Activated Expression of Downstream TIA Pathway Genes in Catharanthus roseus

    Xi Wang

    2016-01-01

    Full Text Available We selected different concentrations of ethephon, to stress C. roseus. We used qRT-PCR and HPLC followed by PCA to obtain comprehensive profiling of the vinblastine biosynthesis in response to ethephon. Based on our findings, the results showed that the high concentration of ethephon had a positive effect at both transcriptional and metabolite level. Meanwhile, there was a remarkable decrease of hydrogen peroxide content and a promoted peroxidase activity in leaves. The loading plot combination with correlation analysis suggested that CrPrx1 could be regarded as a positive regulator and interacts with ethylene response factor (ERF to play a key role in vinblastine content and peroxidase (POD activity. This study provides the foundation for a better understanding of the regulation and accumulation of vinblastine in response to ethephon.

  18. Ethylene-Induced Vinblastine Accumulation Is Related to Activated Expression of Downstream TIA Pathway Genes in Catharanthus roseus.

    Wang, Xi; Pan, Ya-Jie; Chang, Bo-Wen; Hu, Yan-Bo; Guo, Xiao-Rui; Tang, Zhong-Hua

    2016-01-01

    We selected different concentrations of ethephon, to stress C. roseus. We used qRT-PCR and HPLC followed by PCA to obtain comprehensive profiling of the vinblastine biosynthesis in response to ethephon. Based on our findings, the results showed that the high concentration of ethephon had a positive effect at both transcriptional and metabolite level. Meanwhile, there was a remarkable decrease of hydrogen peroxide content and a promoted peroxidase activity in leaves. The loading plot combination with correlation analysis suggested that CrPrx1 could be regarded as a positive regulator and interacts with ethylene response factor (ERF) to play a key role in vinblastine content and peroxidase (POD) activity. This study provides the foundation for a better understanding of the regulation and accumulation of vinblastine in response to ethephon. PMID:27314017

  19. Análisis de iridoides y expresión de genes que codifican enzimas tempranas en la síntesis de alcaloides indol terpenoicos en Catharanthus roseus Analysis of iridoids content and expression studies of genes encoding early enzymes in the indol terpenoid biosynthesis pathway in Catharanthus roseus

    Palacios-Rojas Natalia

    2004-07-01

    Full Text Available Los alcaloides indol terpenoicos (TIA son metabolitos secundarios de importancia medicinal por sus propiedades como agentes anticancerígenos, entre otras. Sin embargo, su explotación en la industria farmacéutica se ha visto limitada, ya que la acumulación de estos compuestos en las plantas que los producen es mínima. Dichos alcaloides son biosintetizados por la vía del shikimato y de los terpenoides, los cuales proveen los precursores: secologanina y triptamina, respectivamente. La secologanina es sintetizada vía terpenoides, y estudios preliminares sugieren que es sintetizada vía del 2-C-metil-D-eritrol-4-fosfato (MEP y no vía acetato/ácido mevalónico. Se cree que la secologanina es una de las moléculas limitantes en la biosíntesis de TIA. En el presente estudio se cuantif ican y evalúan por HPLC los niveles de este compuesto y sus predecesores iridoides, ácido logánico y secologanina, en diferentes partes de la planta medi­cinal Catharanthus roseus. La distribución de estos iridoides varía según el tejido analizado, encontrándose mayor acumulación de secologanina en el segundo par de hojas de la planta. Los niveles de transcriptos de las enzimas involucradas en los pasos tempranos de la vía de los iridoides fueron monitoreados también en diferentes tejidos usando la técnica transferencia de ARN, Northern blots. Adicionalmente, las plantas se elicitaron con jasmonato de metilo (MeJA, con el fin de estudiar el efecto de esta molécula en la trans­cripción de genes que codifican enzimas involucradas en los pasos tempranos de la vía de los iridoides. Los resultados obtenidos en este estudio sugieren que la vía del MEP puede estar más activa que la vía del acetato/ácido mevalónico en tejidos aéreos jóvenes de la planta. Adicionalmente, es evidente que el MeJA afecta la biosíntesis de TIA estimulando la transcripción de diferentes genes. Palabras clave: metabolismo secundario, alcaloides indol terpenoicos

  20. 光照强度对长春花生活史形成和光合作用的影响%Effect of Different Light Intensity on Life Cycle forms and Photosynthesis of Catharanthus roseus

    高扬; 王永勖; 樊梓鸾; 王振宇; 唐中华

    2015-01-01

    长春花是重要的药用植物,广泛栽植于热带和亚热带地区,其次生代谢产物文多灵、长春碱,长春质碱等可治疗多种癌症。如果可以通过调节光照来达到不同的生产目的,将对生产实践具有重要意义。实验分3个强度光照培养长春花,分别为100μmol/m2· s、500μmol/m2· s和900μmol/m2· s ,运用主成分分析法(PCA ),对营养生长、有性生长和无性繁殖3类15种性状进行统计,结果表明:不同光照强度对长春花生活史型的调控作用十分明显,其生活史型随着光照强度的减弱,由VS转变为VC ,体现了植物能量的内部循环。光合数据结果表明,嫩叶主要利用强光,老叶主要利用弱光,使植物充分利用不同生境下的光照,以保证生存。%Catharanthus roseus can be used as medicine ,Its secondary metabolites such as vindoline ,vinblastine can be used to cure various cancers .It would be important for production if we can be by adjusting the illumination to achive diffrent purposes of production .The experiment was divided into three intensity of light culture Catharanthus roseus ,respectively 100μmol/m2 · s ,500μmol/m2 · s and 900μmol/m2 · s ,the results show that the control effects of different light intensity on the life history of Catharanthus roseus type is obvious ,its life cycle forms with the reduction of light intensity ,changes from VS for the VC ,embodies the plant energy internal circulation .The photosynthetic data results show that young leaves use of bright light ,old leaves use of weak light ,make full use of light plants under different habitats in order to ensure survival .

  1. Comparative expression analysis of Crlea gene in Catharanthus roseus under drought stress by real-time quantitative PCR%荧光定量PCR检测干旱胁迫下长春花Crlea基因的相对表达

    祖元刚; 聂明珠; 房思良; 于景华; 郭晓瑞

    2006-01-01

    A full-length Crlea(Crlea for Catharanthus roseus late embryogenesis abundant) gene was first isolated from Catharanthus roseus. Gene expression profiles of Crlea gene in leaves and roots under drought stress were monitored by real-time quantitative PCR. The results showed that a similar accumulation pattern of Crlea gene in leaves and roots over the observation period of 0.5 to 8 hours. The expression of Crlea mRNA was strengthened with the prolongation of stress time. In leaves, expression amounts of Crlea gene were 9.984 and 20.431 times higher than that of control respectively at 6 and 8 h. Similarly, the expression amounts of Crlea gene in root obviously increased (2.831 times higher than that of control) at 8 h. Primary results show the expression of Crlea gene is non-tissue-specific and up-regulated under drought stress.%首次从长春花中克隆了Crlea(Crlea for Catharanthus roseus late embryogenesis abundant)的全长基因,采用荧光定量PCR方法对干旱胁迫下长春花叶片和根部Crlea基因的表达模式进行监测,结果表明,在0.5~8h的胁迫时间中,叶片和根部的Crlea基因表现出相似的积累模式.长春花Crlea基因的表达随着胁迫时间的延长而表达增强.在叶片中,在6 h和8 h的干旱处理后,Crlea基因表达显著提高,分别是未处理材料的9.984和20.431倍.在根部, 在8 h的处理后,Crlea基因的表达量显著提高(2.831倍于对照).初步结果表明Crlea基因的表达没有组织特异性,并且为干旱胁迫正调控.

  2. Effects of Different Calcium Reagents on Photosynthesis Characteristics of Catharanthus roseus%不同钙效应剂对长春花光合特性的影响

    孙金春; 张扬欢; 温泉; 张楠; 唐娟; 吴能表

    2011-01-01

    The effects of LaCl3 (a plasmalemma calcium channel blocker), herpain (an intracellular IP3 channel blocker) and trifluoperazine (an intracellular CaM activity inhibitor) on the photosynthesis of Ca-tharanthus roseus were researched. All the calcium channel blockers studied decreased Pn (net photosynthesis rate), maximal PSII quantum (Fv/Fm), PSII quantum yield (Yield), relative electron transport rate (ETR) and photochemical quenching (qP) of C. Roseus and increased its non-photochemical quenching (qN), chlorophyll a concentration, chlorophyll b concentration and chlorophyll a+b. It is, therefore, concluded that the calcium channel greatly influenced the photosynthesis of C. Roseus by a special way other than promoting the degradation of photosynthetic pigments or the inhibition of their synthesis to inhibit the photosynthesis of C. Roseus. Herpain affected photosynthesis of C. Roseus most significantly, which means the calcium released by intracellular calcium pool through the IP3 channel plays an important role in the photosynthesis of C. Roseus.%研究了细胞质膜钙通道阻断剂氯化镧(LaCl3)、胞内IP3通道阻断剂肝素(Heparin)、胞内CaM活性抑制剂三氟啦嗪(TFP)对长春花光合作用的影响,结果发现:Ca2+通道阻断剂处理后长春花叶片净光合速率(Pn)、PSII最大量子产量(Fv/Fm)、PSⅡ实际量子产量(Yield)、电子传递速率(ETR)、光化学淬灭系数(qP)均下降,非光化学淬灭系数(qN)及Chla、Chlb、Chla+Chlb含量上升,表明Ca2+通道阻断剂对长春花的光合作用有较大影响,且不是通过加速叶片光合色素降解或抑制其合成来实现抑制叶片的光合能力;其中Heparin处理的长春花叶片相关参数变化幅度最大,表明胞内钙库通过IP3通道释放的Ca2+在长春花叶片光合作用过程中发挥了更积极的作用.

  3. Effects of Tryptophan on Alkoloids Contents of Catharanthus roseus Seedlings under Seawater Stress%色氨酸对海水处理下长春花生物碱含量的影响

    刘冲; 刘玲; 刘兆普; 王景艳

    2009-01-01

    以长春花为(Catharanthus roseus(L.)G.Don)材料进行温室实验,研究了不同浓度色氨酸对20%海水处理下长春花幼苗生物碱含量的影响,发现20%海水中加入色氨酸,长春花中色氨酸脱羧酶(TDC)活性提高,吲哚生物总碱以及长春碱、长春质碱、文多灵、长春新碱含量显著增加.其中加入500 mg/L的色氨酸最有利于生物碱含量的增加.

  4. 海南长春花生境土壤环境及其生长规律%Suitable Soil Conditions for Catharanthus roseus in Longlou of Hainan and Its Growth Regularity

    于晓梅; 王非

    2010-01-01

    阐述了长春花(Catharanthus roseus)在海南龙楼地区的分布状况.不同的土壤环境下长春花生长规律有差异,对土壤因子分析,结果表明:长春花适宜生长在沙质土壤;长春花适宜生长在pH为7左右的土壤中;长春花的根系主要分布在土壤的0~20cm深度之间;人为扰动因素是影响长春花生长的一个重要因素.

  5. Monomerie indole alkaloids from the aerial parts of Catharanthus roseus%长春花地上部分单吲哚类生物碱成分研究

    钟祥章; 王国才; 王英; 张晓琦; 叶文才

    2010-01-01

    长春花Catharanthus roseus(L.)G Don为夹竹桃科(Apocynaceae)长春花属(Catharanthus)植物,具有解毒抗癌、清热平肝等功效.为进一步研究其生物碱类活性成分,对其地上部分95%乙醇提取物采用硅胶正相柱色谱及制备高效液相色谱进行了分离纯化,共得到6个单吲哚类生物碱.根据理化性质和波谱数据鉴定了它们的化学结构,分别为vindolinine B(1)、洛柯碱(2)、荷哈默辛碱(3)、文多尼定碱(4)、文多灵(5)和狗牙花定碱(6).其中化合物1为新化合物,命名为vindolinine B.

  6. Biotransformation of artemisinin by Catharanthus roseus and Ginkgo biloba cell suspension cultures%长春花及银杏植物细胞悬浮培养对青蒿素的生物转化研究

    韩健; 戴均贵; 崔亚君; 占纪勋; 郭洪祝; 果德安

    2003-01-01

    Object To investigate the biotransformation of the antimalarial compound artemisinin( Ⅰ ) by Catharanthus roseus and Ginkgo biloba cell suspension cultures. Methods Plant tissue culture technology was employed. The product was isolated on silica gel column chromatography and its structure was elucidated by spectroscopic evidence. Results One product was obtained and its structure was characterized as 3α- hydroxydeoxyartemisinin ( Ⅱ ). Conclusion Both of C. roseus and G. biloba cell suspension cultures can bioconvert artemisinin.%目的对抗疟药物青蒿素(Ⅰ)进行了生物转化研究.方法利用长春花及银杏植物细胞悬浮培养细胞进行生物转化.用硅胶柱色谱进行产物的分离,波谱方法鉴定产物的结构.结果此两种植物悬浮细胞体系均能将青蒿素转化成3α-羟基去氧青蒿素(Ⅱ).结论此两种植物悬浮细胞体系均能有效转化青蒿素.

  7. Research Progress of Terpenoid Indole Alkaloids Metabolism Pathway in Catharanthus Roseus%长春花吲哚萜类生物碱代谢途径研究进展

    章津; 赵淑娟; 胡之璧

    2012-01-01

    长春花(Catharanthus roseus(L.)G.Don.)含有130余种生物碱,既是抗癌药物长春碱和长春新碱的唯一植物来源,也是研究吲哚萜类生物碱代谢途径的模式植物.生成长春花生物碱的前体环烯醚萜主要来源于2-C-甲基-D-赤藓醇4-磷酸途径(MEP),甲羟戊酸途径(MVA)可能在此过程中起辅助作用.吲哚生物碱代谢途径受茉莉酸甲酯正调控.文多灵途径过去被认为在长春花悬浮细胞中没有文多灵产生,而目前的研究发现文多灵在长春花C20hi细胞中可以微量合成.文章对近年来长春花吲哚生物碱代谢途径及部分关键酶研究进展进行了综述.%Caiharanthus roseus ( L. ) can produce more than 130 alkaloids. It is the only natural resource of vinblastine and vincris-tine and the model plant to investigete the biosynthesis of terpenoid indole alkaloids (TIAs). The MEP pathway is the major way to produce iridoid precursors and the MVA pathway maybe plays a minor role in the alkaloids metabolism. The indole pathway could be up-regulated with methyl jasmonate. It was believed that vindoline could not be yielded in cultured cells. But the recent research revealed that C. Roseus C20hi cells could produce vindoline at a minor level. This paper reviews the research progress of TIAs metabolism and some important enzymes in the pathway in C. Roseus.

  8. The effect of extracts from Catharanthus roseus on cholangiocarcinoina in vitro%长春花对胆管癌细胞增殖和细胞周期的影响

    韩鹏; 陈良华; 沈东炎; 童庆宣; 李文岗; 陈清西

    2010-01-01

    Objective To investigate the potential inhibitory effects of herbal Catharanthus roseus on cholangiocarcinoma in vitro. Methods The whole herbal of Catharanthus roseus was used as the material, and total alkaloid extract and water extract were obtained. The effects of the two extracts on human cholangiocarcinoma QBC939 cells were investigated. The inhibition rate of QBC939 cells was measured by methyl thiazolyl tetrazolium (MTT) method. The morphological change of cells was examined by inverted microscopy. The effect on cell cycles was observed by flow cytometry. Results The two extracts could both inhibit the growth of QBC939 cells, but the total alkaloid extract showed stronger proliferation inhibitory effect comparing to the water extract. IC50 value of total alkaloid and water extract was 0. 16 and 49. 02 mg/L, respectively. Obvious morphological changes were both observed after adding the two extracts. Both the two extracts could change the cell cycle of cholangiocarcinoma cells significantly. The ratio of G0/G1 phase cells was decreased greatly from 53. 10% to 5. 87% and that of G2/M phase cells was increased greatly from 26. 4% to 71. 8% , respectively. Compared to the cells treated with the total alkaloid extract, the ratio of Sub-G1 phase cells treated with the water extract changed greatly. The water extract could induce the apoptosis of the cells. Conclusion The extracts from Catharanthus roseus showed the inhibitory effects on proliferation of cholangiocarcinoma cells. They also could induce the morphological change and cell arrest.%目的 观察长春花草提取物对体外培养的人胆管癌QBC939细胞的抑制作用.方法 以长春花全草为原料进行分离提取,获得总生物碱层和水层两种提取物,分别采用噻唑蓝(MTT)法、普通倒置光学显微镜观察法和流式细胞术研究两种提取物对QBC939细胞增殖、细胞形态以及细胞周期的影响.结果 长春花两种提取物均能够显著抑制胆管癌细

  9. Effects of Low Light Stress on the Biomass Distribution and Secondary Metabolism of Catharanthus roseus%弱光胁迫对大田长春花生物量分配及次生代谢的影响

    佟璐; 张宝友; 唐中华; 郭晓瑞; 贾雪莹; 于景华

    2011-01-01

    以长春花(Catharanthus rosetts(L.)Don.)为材料,研究了田间栽培长春花在全光和20%透光率下,经过-个生长季后生物量配置、抗氧化次生代谢产物和文朵灵、长春质碱、长春碱等目的活性物质含量的变化.研究结果表明,弱光条件显著抑制了长春花植株总生物量增长,特别是抑制了有性生殖的投入;弱光组叶片总酚和总黄酮含量显著降低,干重含量分别为对照组的62.50%和50.00%,原花青素含量则略有升高,但与全光组的差异不显著;弱光组叶片文朵灵和长春质碱含量显著高于对照组,长春碱含量略有上升但差异不显著,受生物量降低影响,3种生物碱的产量均显著下降.上述结果表明,长春花能够调控生理代谢以适应低光强环境,特别是文朵灵和长春质碱含量提升显著,林下低光强环境种植长春花可以满足土地资源充分利用和文朵灵、长春质碱优质原料的需求.%Under full exposure and 20% exposure to light, the field-cultivated Catharanthus roseus ( L. ) Don. was used to investigate the changes of biomass distribution, antioxidative secondary metabolites and contents of objective active products such as vindohn, catharanthine and vinblastine after a growing season. The results indicated that the total biomass of C. roseus was extremely restrained by the low light intensity, especially the investment of generative propagation; the contents of total phenolics and total flavonoids in leaves under low light intensity decreased distinctly, and the contents of dry weight were 62.50% and 50.00% of the control, respectively, while the content of proanthocyanidins increased slightly, but not remarkably as compared to the full exposure group; vindolin and catharanthine contents under low light intensity were much higher than that of control while vinblastine content increased slightly; the contents of three alkaloids all reduced, which was affected by the reduction of biomass

  10. Aktivitas Hipolipidemik dan Indeks Aterogenik yang Rendah Ekstrak Air Daun Tapak Dara pada Tikus Hiperkolesterolemia (HYPOLIPIDEMIC ACTIVITY AND LOW ATHEROGENIC INDEX OF AQUEOUS LEAF EXTRACTS OF CATHARANTHUS ROSEUS IN HYPERCHOLESTEROLEMIC RATS

    I Nyoman Suarsana

    2016-01-01

    Full Text Available Catharanthus roseus is one plant recognized as medical potential which can decrease cholesterol. Thepresent study was carried out to evaluate the effect of aqueous leaf extracts of C. roseus on plasma lipidprofil levels in rats cholesterol rich diet and atherogenic index. This study was carried out on 15 SpraqueDawley male rats randomly distributed into five groups (n=3. Rats hypercholesterolemic wereadministrated cholesterol rich diet containing 1% (w/w. Normal control group with normal diet whitoutextracts (K1, positive control hypercholesterolemic group (K2 with cholesterol rich diet whitout extracts,and others three groups (K3-K5 were feed high cholesterol and 1 mL of aqueous leaf extracts of C. roseuswith a dose of 20% (w/v, 40% (w/v and 80% (w/v respectively, administered twice daily 1 mL orally.Treatment was given for 28 days. After treatment, the plasma lipids profile such cholesterol total, highdensity lipoprotein (HDL, low density lipoprotein (LDL, triglyceride (TGA level were measured. Theresults showed that during the period of treatment with cholesterol rich diet have produced under condition hypercholesterolemic of rats with average cholesterol levels of 124.33 ± 4.04 mg/dL (K2 group. Treatmentwith extract dose of 80% showed the best results among the other doses in reducing levels of total cholesterol,triglycerides, LDL and raise HDL levels. Extract treatment dose of 80% up to four weeks resulted incholesterol total (79.33±3.51 mg/dL, TGA (72.33±6.65 mg/dL, HDL (55.00±3.60 mg/dL, and LDL(9.87±5.34mg/dL. In addition, value ratio of cholesterol:HDL was 1.4: 1 and atherogenic index value was0.44. These results suggest that extracts of leaf C. roseus optimum doses 80% (w/v has hipolipidemicactivity in hypercholesterolemic rats and it has low atherogenic index value.

  11. Research Progress of Terpenoid Indole Alkaloids (TIAs) Biosynthetic Pathway of Catharanthus roseus%长春花萜类吲哚生物碱的生物合成途径

    邢世海; 王荃; 潘琪芳; 赵静雅; 唐克轩

    2012-01-01

    There are more than 130 types of terpenoid indole alkaloids (TIAs) in medicinal plant Catharanthus roseus (L. ) G. Don. In this review, the upstream and downstream of the biosynthesis of these alkaloids in C. roseus, and related studies were summarized. During upstream pathway, tryptamine which came from indole biosynthetic pathway and secologamn which came from monoterpenoid pathway were synthesized into 3a(S)-Strictosidine which is the common precurors of different TIAs by the corresponding enzymatic catalysis. Then various TIAs were found from 3α(S)-Strictosidine by enzymes during downstream process. The TIAs biosynthetic pathway in C. roseus were outlined in the article in order to provide useful information for the researchers who are interested in terpenoid indole alkaloids biosynthetic pathway and metabolic regulation.%药用植物长春花含有130余种萜类吲哚生物碱,该文对近年来国内外有关长春花生物碱合成的上游和下游阶段及其相关研究进行详细的归纳总结.长春花上游合成途径中在相应的酶促作用下由吲哚途径产生的色胺和由类萜途径产生的裂环马钱子苷在异胡豆苷合成酶的催化作用下形成了所有长春花TIAs的共同前体物质3α-异胡豆苷,3α-异胡豆苷再由下游途径的各种酶促作用下生成种类各异的长春花TIAs.通过对长春花TIAs合成途径的阐述,为萜类吲哚生物碱合成及其代谢调控的相关研究提供参考.

  12. Identification and expression analysis of WRKY transcription factors in medicinal plant Catharanthus roseus%药用植物长春花WRKY转录因子的鉴定及表达谱分析

    杨致荣; 王兴春; 薛金爱; 孟令芝; 李润植

    2013-01-01

    WRKY是调控植物生长发育和逆境胁迫反应等生命活动的一个转录因子大家族.然而,有关药用植物长春花CrWRKY转录因子的种类和功能却知之甚少.从26 009个长春花蛋白中鉴定出47个CrWRKY转录因子.依据WRKY结构域和系统进化,将CrWRKY分为G1、G2和G3三大类群.表达谱数据分析表明,长春花CrWRKY基因的表达具有器官特异性.47个CrWRKY基因的表达谱可分为3种表达模式:第1类型主要在花、甲基茉莉酸甲酯(MeJA)或酵母提取物(YE)处理的原生质体中高表达;第2类型主要在茎和毛状根中高表达;第3类型在根、茎、叶、幼苗和MeJA处理的毛状根中高表达.进一步用实时定量PCR检测了16个代表性CrWRKY基因在不同器官、MeJA处理原生质体和毛状根中的表达模式,检测结果与上述数字基因表达谱数据基本一致.约1/3以上CrWRKY基因的表达受MeJA和YE的调控,暗示它们可能参与萜类吲哚生物碱的合成和逆境胁迫反应.为进一步解析长春花WRKY转录因子的功能和萜类吲哚生物碱合成调控的网络奠定了基础.%WRKY transcription factors,one of the largest families of transcriptional regulators in plants,involve in multiple life activities including plant growth and development as well as stress responses.However,little is known about the types and functions of WRKY transcription factors in Catharanthus roseus,an important medicinal plant.In this study,we identified 47 CrWRKY transcriptional factors from 26 009 proteins in Catharanthus roseus,and classified them into three distinct groups (G1,G2 and G3) according to the structure of WRKY domain and evolution of the protein family.The expression profiling showed that these CrWRKY genes expressed in a tissue/organ specific manner.The 47 CrWRKY genes were clustered into three types of expression patterns.The first type includes the CrWRKYs highly expressed in flowers and the protoplast treated with methy jasmonate

  13. Identification of a Bipartite Jasmonate-Responsive Promoter Element in the Catharanthus roseus ORCA3 Transcription Factor Gene That Interacts Specifically with AT-Hook DNA-Binding Proteins1[W

    Vom Endt, Débora; Soares e Silva, Marina; Kijne, Jan W.; Pasquali, Giancarlo; Memelink, Johan

    2007-01-01

    Jasmonates are plant signaling molecules that play key roles in defense against certain pathogens and insects, among others, by controlling the biosynthesis of protective secondary metabolites. In Catharanthus roseus, the APETALA2-domain transcription factor ORCA3 is involved in the jasmonate-responsive activation of terpenoid indole alkaloid biosynthetic genes. ORCA3 gene expression is itself induced by jasmonate. By loss- and gain-of-function experiments, we located a 74-bp region within the ORCA3 promoter, which contains an autonomous jasmonate-responsive element (JRE). The ORCA3 JRE is composed of two important sequences: a quantitative sequence responsible for a high level of expression and a qualitative sequence that appears to act as an on/off switch in response to methyl jasmonate. We isolated 12 different DNA-binding proteins having one of four different types of DNA-binding domains, using the ORCA3 JRE as bait in a yeast (Saccharomyces cerevisiae) one-hybrid transcription factor screening. The binding of one class of proteins bearing a single AT-hook DNA-binding motif was affected by mutations in the quantitative sequence within the JRE. Two of the AT-hook proteins tested had a weak activating effect on JRE-mediated reporter gene expression, suggesting that AT-hook family members may be involved in determining the level of expression of ORCA3 in response to jasmonate. PMID:17496112

  14. 长春花钙调素基因克隆及其假基因的识别%Molecular Cloning of A Calmodulin Gene and Identification of Its Pseudogene in Catharanthus roseus

    苏火生; 杨云; 刘宽灿; 张治礼

    2007-01-01

    以长春花[Catharanthus roseus(L.)G.Don]叶片cDNA和基因组DNA为模板,利用PCR技术扩增得到了长春花钙调素基因447 bp的全长编码cDNA序列和2个大小不同的DNA片段.序列分析表明,DNA长片段全长1 551 bp,由2个外显子和1个内含子构成,为长春花钙调素基因编码区DNA片段;DNA小片段全长447 bp,与447 bp的长春花钙调素基因cDNA核苷酸一致性高达87%,有56个碱基的差异,其中位于226 bp处的碱基A突变为T,即由AAG突变为终止密码子TAG使翻译提前终止.推测此447 bp的DNA小片段可能为长春花钙调素基因的假基因,命名为CCaMP1.

  15. 长春花内生真菌的分离及其发酵产生药用成分的初步研究%Preliminary Study on the Isolation of Endophytic Fungus of Catharanthus roseus and Its Fermentation to Produce Products of Therapeutic Value

    张玲琪; 郭波; 李海燕; 曾松荣; 邵华; 谷苏; 魏蓉城

    2000-01-01

    首次报道从长春花Catharanthus roseus (L.) G.Don 茎的韧皮部中分离出尖孢镰刀菌Fusarium oxysporu m,为该植物的一种内生真菌,并用TLC和HPLC对该菌的97CG3菌株培养物进行了分析,初步结果表明该真菌能产生抗癌药长春新碱成分.

  16. 弱光对长春花(Catharanthus roseus)幼苗中可溶性糖、生物碱及激素含量的影响%Effects of low light intensity on changes of soluble sugars, alkaloids and phytohormones in Catharanthus roseus seedlings

    唐中华; 郭晓瑞; 于景华; 杨蕾; 孙艳斐; 祖元刚

    2007-01-01

    阐述了低光照强度和正常光照条件下长春花(Catharanthus roseus)幼苗叶片中可溶性糖、生物碱及内源激素等几种生理活性物质含量的动态变化,以揭示弱光胁迫对长春花生理代谢的影响及上述生理代谢对植物适应弱光环境的生态学意义.结果显示,弱光培养提高了长春花在自身支撑结构的投入,如叶柄长和节间长都显著增加,同时抑制了有性生殖.在弱光培养的第1周,长春花叶片中果糖(fructose, Fru)、葡萄糖(glucose, Glc)和蔗糖(sucrose, Suc)的含量分别由处理前的(0.62±0.01), (1.86±0.12), (0.24±0.01) mg/g FW下降为(0.38±0.02), (0.60±0.03), (0.17±0.02) mg/g FW,均显著低于对照(P<0.01),表明碳同化水平下降.长春花叶片中文朵灵(vindoline, VIN)和长春质碱(catharanthine, CAT)含量在弱光条件下呈显著增加趋势,处理结束时是对照的3倍左右,而它们的耦合产物长春碱(vinblastine, VLB)合成和积累受到抑制,其含量在第3周时仅为对照的50%.弱光条件处理前2周促进了长春花叶片中脱落酸(abscisic acid, ABA)、赤霉素(gibberelline, GA3)和吲哚乙酸(indole-3-acetic acid, IAA)含量水平的积累.这些结果表明,在弱光条件下长春花幼苗的上述生理代谢都发生着显著的变化,可能在植物适应弱光胁迫过程中发挥着积极的调控作用.

  17. Regulación transcripcional de genes involucrados en la producción de alcaloides indol-terpenoides en plántulas de Catharanthus roseus

    Rocha Pedro J.

    2002-12-01

    Full Text Available Cathmnthus roseus es una planta medicinal productora de varios alcaloides indol terpenoides (AIT con actividad farmacológica. Empleando plantas y cultivos celulares de C. roseus la ruta biosintética de AIT ha sido determinada, además, un considerable número de sus enzimas ha sido caracterizado y sus respectivos genes clonados. La producción de AIT en plantas y cultivos celulares de C. roseus está altamente regulada a nivel transcripcional, es dependiente de los estados de desarrollo y puede ser alterada, entre otros factores, por sometimiento a estrés luminoso y tratamiento con elicitores bióticos y abióticos. Estudios sobre la regulación de los genes involucrados en la biosíntesis de AIT han sido llevados a cabo en cultivos celulares, sin embargo poco se conoce acerca de la regulación de los mismos en plantas. En los experimentos reportados aqui, se presenta un examen cualitativo de los niveles de expresión de los genes idcr strl, d4h y dat que codifican enzimas importantes en la biosíntesis de AIT en plántulas de C. roseus en una serie de desarrollo que comprende O a 18 días. Además, se analiza el efecto del estrés hídrico, luminoso y de la elicitación con metil jasmonato (Meja y ácido acetil salicílico (ASA. La comparación entre la evidencia experimental obtenida con plántulas en el presente artículo y los datos reportados en estudios con suspensiones celulares sugiere que la regulación transcripcional de algunos genes difiere de manera considerable. 

  18. 长春花中长春质碱含量与农艺性状的分析%Analysis of catharanthine content and agronomic traits in Catharanthus roseus

    孙小芬; 陈雨; 潘俊松; 王玉亮; 孙克兴; 蔡润; 唐克轩

    2009-01-01

    目的:对国内外长春花主栽品种的长春质碱含量和农艺性状进行了分析.方法:记录长春花盛花期的农艺性状,反相高效认相色谱仪测定长春质碱含量.结果:发现不同品种的长春质碱含量和农艺性状均存在很大差异,太平洋系列(Pacifica)品种Polka Dot(PPD)中长春质碱质量分数最高,达到叶片干重的3.79 mg·g~(-1);清凉系列(cooler)品种Pink(CP)中长春质碱质量分数最低,仅为叶片干重的0.9 mg·g~(-1).长春花中长春质碱含量与农艺性状之间存在一定程度的相关性.通径分析表明,长春花主要农艺性状中,节间距对长春质碱含量的正影响达到了显著水平(P<0.05),通径系数为1.473.结论:本研究对高含量长春质碱长春花引种和育种具有参考价值.%Catharanthine content and agronomic traits in major Catharanthus resells varieties were analyzed.It was found that there existed great difference in catharanthine content and agronomic traits among the varieties.Catharanthine content was the highest in variety Pacifica Polka Dot(PPD),reaching 3.79 mg·g~(-1) dry leaf weight,and the lowest in variety Cooler Pink(CP)with only 0.9 mg·g~(-1) dry leaf weight.Correlation existed in certain extent between catharanthine content and agronomic traits in C. roseus.Path analysis showed that among all the agronomic traits analyzed,intemodal distance positively affected catharanthine content at significant level(P<0.05),with the path coefficient being 1.473.This study provides useful information for high-catharanthine content C.roseus introduction and breeding.

  19. Effects of Nitrate Nitrogen Content on Physiological indicators of Catharanthus roseus%硝态氮浓度对长春花体内生理代谢指标的影响

    朱虹; 郭晓瑞

    2013-01-01

    以药用植物长春花为研究对象,通过施加不同浓度硝态氮,揭示其体内长春碱积累受氮水平的调控特征及其与内源碳/氮( C/N)代谢的联系。结果表明,中等浓度的氮水平能显著诱导根系和叶片中硝酸还原酶活性的提高,并且该效应依赖于处理时间,处理12 h后其诱导效果最为显著。对比氮代谢,测定了叶片中蔗糖的质量分数,在外施18 mmol· L-1 NO-3溶液后6 h内蔗糖质量分数低于对照水平,之后又升高至对照水平。 HPLC检测叶片中长春碱的结果显示,经不同浓度硝态氮处理2h后显著诱导长春碱质量分数增加。%Catharanthus roseus were investigated for the characteristics of the alkaloids accumulation and the metabolism of en-dogenous carbon/nitrogen (C/N) of plants by applying different nitrate contents.The medium concentration of nitrogen can significantly induce NR activity in roots and leaves of C.roseus.In different processing time (20 min, 2 hours, 6 hours, 12 hours and 24 hours), the content of nitrate reductase (NR) activity (μg· h-1· g-1) is the most significant in roots and leaves after 12 hours.Compared with the contents of nitrogen metabolism, the content of sucrose in leaves is less than that of control in the concentration of nutrient solution (NO-3 ) with 18 mmol· L-1 for 6 hours, which is increased to the levels of control after 6 hours.The alkaloids content is increased in Medium content of nitrogen for 2 hours according to the HPLC detection.The alkaloids accumulation is closely related to endogenous carbon and nitrogen metabolism of plants in different exogenous nitrate levels.

  20. Synthesis and characterization of Gold and Silver nano-particles using different leaf extracts namely Catharanthus roseus, Datura metel and Azadirachta indica and Estimation of antimicrobial activity of silver nano-particles using disc diffusion method

    Sarbjeet Singh Gujral

    2014-12-01

    Full Text Available Objective: synthesis of gold and silver nano-particles using leaf extracts of Catharanthus roseus, Datura metel and Azadirachta indica and Estimation of antimicrobial activity of silver nano-particles using disc diffusion method. Method: Green approach has been utilized for the synthesis of gold and silver nano-particles. Different aqueous plant extracts has been prepared which was then utilized for the biosynthesis of gold and silver nano-particles. Estimation for the synthesis of nano-particles were done using UV-Visible spectroscopy and Fourier- Transform infrared spectroscopy. Antimicrobial activity of silver nano-particles prepared using aqueous neem extract was investigated using disc diffusion method. Result: UV- Vis spectroscopy of prepared nano-particles was done which gave a peak at about 550 nm for gold nano-particles and around 430 nm for silver nano-particles. FTIR of collected nano-particles gave an idea about the type of bio-molecules which helped in the reduction of auric and silver salts into corresponding nano-particles. Anti-microbial activity of silver nano-particles showed that the nano-particles have better anti-microbial activity than 2% silver nitrate solution (kept as standard when experiments were performed under similar conditions. Conclusion: Gold and silver nano-particles were successfully synthesized using greener approach and anti-microbial activity of silver nano-particles prepared using aqueous neem extract was estimated against 2% AgNO3 solution. Nano-particles gave better anti-microbial activity than Silver nitrate solution.

  1. Effects of ambient and elevated CO2 on growth, chlorophyll fluorescence, photosynthetic pigments, antioxidants, and secondary metabolites of Catharanthus roseus (L.) G Don. grown under three different soil N levels.

    Singh, Aradhana; Agrawal, Madhoolika

    2015-03-01

    Catharanthus roseus L. plants were grown under ambient (375 ± 30 ppm) and elevated (560 ± 25 ppm) concentrations of atmospheric CO2 at different rates of N supply (without supplemental N, 0 kg N ha(-1); recommended N, 50 kg N ha(-1); and double recommended N, 100 kg N ha(-1)) in open top chambers under field condition. Elevated CO2 significantly increased photosynthetic pigments, photosynthetic efficiency, and organic carbon content in leaves at recommended (RN) and double recommended N (DRN), while significantly decreased total nitrogen content in without supplemental N (WSN). Activities of superoxide dismutase, catalase, and ascorbate peroxidase were declined, while glutathione reductase, peroxidase, and phenylalanine-ammonia lyase were stimulated under elevated CO2. However, the responses of the above enzymes were modified with different rates of N supply. Elevated CO2 significantly reduced superoxide production rate, hydrogen peroxide, and malondialdehyde contents in RN and DRN. Compared with ambient, total alkaloids content increased maximally at recommended level of N, while total phenolics in WSN under elevated CO2. Elevated CO2 stimulated growth of plants by increasing plant height and numbers of branches and leaves, and the magnitude of increment were maximum in DRN. The study suggests that elevated CO2 has positively affected plants by increasing growth and alkaloids production and reducing the level of oxidative stress. However, the positive effects of elevated CO2 were comparatively lesser in plants grown under limited N availability than in moderate and higher N availability. Furthermore, the excess N supply in DRN has stimulated the growth but not the alkaloids production under elevated CO2. PMID:25304238

  2. 不同长春花中3种生物碱的含量测定%Determination of three kinds of important alkaloids in Catharanthus roseus from ten different habitats

    詹小宁

    2014-01-01

    目的:建立快速准确的长春花内生物碱含量测定方法,为寻找高含量品种提供方法和依据。方法:采用反相高效液相色谱法(RP-HPLC),对收集到的10个长春花品种中的文多灵、长春质碱、长春碱含量进行了测定,色谱柱:Dikma Diamonsil -C18(4.6mm ×250mm,5μm);流动相为A∶0.7%二乙胺(磷酸调整pH至7.2);B:乙腈:甲醇=25:40。A∶B=35∶65;流速1ml/min。柱温35℃。检测波长:280nm。结果:文多灵:Y=2.00×106X+5.46×104,r=0.9999,线性范围:0.2~4μg;长春质碱:Y=2.28×106X+8.31×104,r=0.9999,线性范围:0.2~4μg;长春碱:Y=1.76×106X-7.17×104,r=0.9999,线性范围:0.2~4μg。文多灵含量最高的是1号品种,长春质碱含量最高的是3号品种,长春碱含量最高的是2号品种,三种生物碱总和最高的是3号品种。结论:该方法简便、快速,分析准确,适合对长春花内主要生物碱做高通量的筛选。综合考虑总碱的含量和最重要的长春碱的含量,1号品种适合作为代谢调控的本体。%Objective To establish a fast and accurate method of determination of alkaloids in Catharanthus roseus for finding high-content varieties.Method To determinate the contents of Vinblastine,Catharanthine and Vindoline in the 10 varieties of Catharan-thus roseus by RP -HPLC,HPLC method is performed on Dikma Diamonsil -C18 (4. 6mm ×250mm,5μm) column,mobile phase:A:0. 7% diethylamine (pH 7. 2 adjusted by phosphoric acid );B:acetonitrile∶methanol =25∶40.A∶B =35∶65;flow rate is 1ml/min.Column temperature is 35 ℃.Detected at 280nm.Results Vindoline:Y=2. 00 ×106X+5. 46 ×104,r=0. 9999,linear range:0. 2 ~4μg;Catharanthine:Y=2. 28 ×106X+8. 31 ×104,r=0. 9999,linear:0. 2 ~4μg;Vinblastine:Y=1. 76 ×106X-7. 17 ×104,r=0. 9999,linear range:0. 2~4μg.The species of the highest content of vindoline is

  3. EFFECT OF NON – MONETARY INPUTS ON THE YIEDLING PERFORMANCE OF PERIWINKLE (Catharanthus sp.)

    Dahatonde, B. N.

    1985-01-01

    An experiment to study the effect of inter and intra row spacing on the yielding performance of periwinkle species was conducted at Nagarjun Medicinal and Aromatic Plants Research Scheme, P. K. V., Akola (M. S.) during 1980 – 81 to 82 – 83. Three species viz., Catharanthus roseus, Catharanthus alba and Catharanthus oscillatus did not differ significantly in respect of dried foliage and root yields. Inter and intra row spacing of 30 and 20 cm. respectively produced the highest foliage and root...

  4. Research on Purification Process of Vindoline and Catharanthine from Catharanthus roseus by Means of Cation Exchange Resin%阳离子交换树脂纯化长春花中文多灵和长春质碱的工艺优选

    柏道鸣; 周广涛; 代龙

    2012-01-01

    Objective: To select the optimum purifying process of Vindoline and Catharanthine from Catharanthus roseus by means of cation exchange resin. Method: The purifying process of Vindoline and Catharanthine from Catharanthus roseus was performed by cation exchange resin of LSD-001 type, with the solvent of ammonia water, the mixture of ammonia water and ethanol, acid water, the mixture of acid water and ethanol, the mixture of sodium chloride and ethanol, and the content of two alkaloids were determined to choose the optimal technological parameters. Result: The ideal concentration of solvent was: with 80% ethanol ( containing 1% sodium chloride ) eluent elution, two alkaloids eluting attained the highest rate. Conclusion : The purification process has well effect.It can be used for purifying Vindoline and Catharanthine from Catharanthus roseus by means of cation exchange resin.%目的:考察阳离子交换树脂不同洗脱溶剂纯化长春花中文多灵和长春质碱的效果,优选最佳工艺条件.方法:采用LSD-001大孔型阳离子交换树脂对长春花中文多灵和长春质碱进行纯化,以两种生物碱含量为指标,考察以氨水、氨性乙醇、酸水、酸性乙醇、盐醇为洗脱剂纯化两种生物碱的效果.结果:经过优选,离子交换树脂的最佳洗脱条件为:以80%乙醇(含1%氯化钠)洗脱剂洗脱时,两种生物碱的洗脱率最高.结论:优选得到的工艺纯化长春花中文多灵和长春质碱效果好,节省溶剂,工艺简单,可用于长春花中两种生物碱的分离纯化.

  5. Cloning and Expression of Allene Oxide Synthase Gene from Catharanthus roseus%长春花丙二烯氧化物合酶基因的克隆与表达

    王名雪; 陆平; 许菲; 潘琪芳; 唐克轩; 赵静雅

    2012-01-01

    In this study, a full-length cDNA of allene oxide synthase (AOS) gene (named as CrAOS, JQ364955) was cloned from Catharanthus roseus. The gene was 2 118 bp in size containing an open reading frame (1638 bp) encoding 545 amino acids. Comparative and bioinformatic analysis revealed that the deduced protein of CrAOS was highly homologous to AOSs from other plant species. Southern blot analysis revealed that it was a low-copy gene. Real-time Quantitative PCR (qRT-PCR) analysis showed that CrAOS mRNA accumulated most abundantly in old leaves and least in young alabastrums. The qRT-PCR analysis revealed that wound, low temperature, methyl jasmonic acid, ethylene treatments significantly enhanced CrAOS transcript expression,and salicylic acid had no influence.%利用RT-PCR和RACE相结合的方法,从长春花中克隆了丙二烯氧化物合酶(AOS)基因.结果显示:长春花AOS基因(CrAOS) cDNA全长为2 118 bp,包括5 '和3'非翻译区,polyA尾和一个长1 638 bp的开放阅读框,其基因组中不含内含子;CrAOS基因编码的蛋白含545个氨基酸.多重比对表明CrAOS蛋白与其他的AOS蛋白具有较高的相似性,CrAOS蛋白序列中含有AOS家族应有的保守氨基酸残基.Southern杂交表明:CrAOS基因在长春花中为低拷贝.qRT-PCR结果显示:CrAOS在各个组织均有表达但表达量存在差异,在老叶中最高,在幼花中表达最低.对长春花幼苗进行不同处理,结果表明:伤害、低温、甲基茉莉酸、乙烯利处理等可使CrAOS基因表达量显著提高,水杨酸处理对基因表达影响不大.

  6. 黑暗和光照对长春花培养细胞生长和生理生化特性的影响%The Effects of Dark and Light on Growth and Physiological and Biochemical Characteristics in Cell Culture of Catharanthus roseus(L.)G.Don

    郭胜娟; 刘树楠; 冯玲玲; 周吉源

    2004-01-01

    以长春花(Catharanthus roseus (L.)G.Don)幼嫩叶片为外植体,在MS+NAA 0.5 mg/L+2,4-D 0.5 mg/L+6 BA 2.0 mg/L培养基上诱导形成愈伤组织,愈伤组织置于不同条件下培养.结果表明,黑暗和光照下,培养细胞的生长周期为27 d;过氧化物酶(POD)活性和超氧化物歧化酶(SOD)活性均出现2个峰值.对长春花总生物碱含量和总产量亦做了研究,结果发现最佳的收获时期为第27 d.

  7. Tryptophan over-producing cell suspensions of Catharanthus roseus (L) G. Don and their up-scaling in stirred tank bioreactor: detection of a phenolic compound with antioxidant potential.

    Verma, Priyanka; Mathur, Ajay K; Masood, Nusrat; Luqman, Suaib; Shanker, Karuna

    2013-02-01

    Five cell suspension lines of Catharanthus roseus resistant to 5-methyl tryptophan (5-MT; an analogue of tryptophan) were selected and characterized for growth, free tryptophan content and terpenoid indole alkaloid accumulation. These lines showed differential tolerance to analogue-induced growth inhibition by 30 to 70 mg/l 5-MT supplementation (LD(50) = 7-15 mg/l). Lines P40, D40, N30, D50 and P70 recorded growth indices (i.e. percent increment over the initial inoculum weight) of 840.9, 765.0, 643.9, 585.7 and 356.5 in the absence and, 656.7, 573.9, 705.8, 489.0 and 236.0 in the presence of 5-MT after 40 days of culture, respectively. A corresponding increment in the free tryptophan level ranging from 46.7 to 160.0 μg/g dry weight in the absence and 168.0 to 468.0 μg/g dry weight in the presence was noted in the variant lines. Higher tryptophan accumulation of 368.0 and 468.0 g/g dry weight in lines N30 and P40 in 5-MT presence also resulted in higher alkaloid accumulation (0.65 to 0.90 % dry weight) in them. High-performance liquid chromatography (HPLC) analysis of the crude alkaloid extracts of the selected lines did not show the presence of any pharmaceutically important monomeric or dimeric alkaloids except catharanthine in traces in the N30 line that was also unique in terms of a chlorophyllous green phenotype. The N30 line under optimized up-scaling conditions in a 7-l stirred tank bioreactor using Murashige and Skoog medium containing 2 mg/l α-naphthalene acetic acid and 0.2 mg/l kinetin attained 18-folds biomass accumulation within 8 weeks. Interestingly, the cell biomass yield was enhanced to 30-folds if 30 mg/l 5-MT was added in the bioreactor vessel one week prior to harvest. Crude alkaloid extract of the cells grown in shake flask and this bioreactor batch also showed the formation of yellow-coloured crystals which upon (1)HNMR and ESI-MS analysis indicated a phenolic identity. This crude alkaloid extract of bioreactor-harvested cells containing

  8. Phytochrome Is Involved in the Light-Regulation of Vindoline Biosynthesis in Catharanthus1

    Aerts, Rob J.; De Luca, Vincenzo

    1992-01-01

    The enzyme acetylcoenzyme A:deacetylvindoline 4-O-acetyl-transferase (DAT) catalyzes the final step in the biosynthesis of the monoterpenoid indole alkaloid, vindoline. Previous studies have shown that the appearance of DAT activity in etiolated seedlings of Catharanthus roseus is induced by exposure of seedlings to light and that enzyme activity is restricted principally to the cotyledons. Evidence is now presented that phytochrome is involved in the light-mediated induction of DAT activity in Catharanthus cotyledons. PMID:16653011

  9. Effects of Exogenously Supplied Nitrogen on Growth and Nitrogen Metabolism of Periwinkle (Catharanthus roseus (l.) g.Don) under Salt Stress%外源氮对盐胁迫条件下长春花生长和氮代谢途径的影响1)

    朱虹; 郭晓瑞

    2015-01-01

    With the periwinkle (Catharanthus roseus (L.) G.Don), we studied the growth characteristics and nitrogen metabolismof plant under exogenous nitrogen in different forms of exogenous nitrogen under salt stress.The effect of nitrogensource in different forms on the periwinkle plants upon non-salt stress was not significant.But nitrate reductase (NR) andsynthetase (GS) activity and total free amino acid amount were significantly higher when nitrate nitrogen and ammoniumnitrogen were mixed to be supplied.Under salt stress, the leaf biomass accumulation was significantly increased in mixtureof nitrate nitrogen and ammonium nitrogen.NR activity was improved significantly .Overall, when the nitrogen source wasmixed to be supplied, plant damage of salt stress was reduced on periwinkle.The key enzyme activity increased was relatedto ammonium metabolism enzymes in nitrogen metabolism.The right amount of ammonium ion can save the energy of thenitrate nitrogen from plant metabolism.%以药用植物长春花( Catharanthus roseus ( L.) G.Don)为对象,研究不同形态外源氮供应条件下,长春花生长发育与体内氮代谢响应盐胁迫的变化特点。结果表明,无盐胁迫条件下,不同形态氮源供应对长春花生长影响不显著,但硝态氮与氨态氮混合供应的情况下,硝酸还原酶( NR)和谷氨酰胺合成酶( GS)的活性及游离氨基酸总质量分数显著高于其他氮源条件下的;在盐胁迫条件下,硝态氮和氨态氮混合供应显著增加了叶片生物量积累,同时,NR活性也显著提高。总体上,混合氮源供应可以减少盐胁迫对长春花的伤害。这种作用可能与氮代谢过程中关键酶活性的增加有关,特别是与氨代谢有关的酶有关,适量的氨离子可以节省植物代谢硝态氮的能量。

  10. 遮光对长春花叶片长春碱含量和相关合成基因表达的影响%Effects of Shading Light on Alkaloids and Related Genes Expression in Leaves of Catharanthus roseus

    刘英; 赵冬梅; 祖元刚; 张衷华

    2010-01-01

    分别在红色滤光膜遮光处理和正常日光灯照条件下,研究长春花(Catharanthus roseus)无菌幼苗叶片中叶绿素含量和生物碱含量以及生物碱合成相关基因的表达变化.结果表明:与对照相比,叶片中叶绿素含量在前30 d先升高,然后明显降低 (P<0.05),叶绿素a/b比率低于对照;文多灵 (vindoline,VIN)和长春质碱 (catharanthine,CAT)的含量稍高于对照后下降,VIN和CAT耦合的重要抗癌产物长春碱 (vinblastine,VBL)的含量逐渐高于对照,在处理第15天达到最高为(0.028 0±0.005 1) mg/g;遮光处理有利于VBL合成路径相关基因 (Tdc、Str、Dat 和 D4h)的表达,表明植物在适应遮光胁迫过程中发挥着积极的调控作用.

  11. Effects of Nutrition and Environmental Factors on Growth and Alkaloid Production of Catharanthus Roseus Hairy-Roots%营养及环境因子对农杆菌诱导的长春花发根生长和生物碱生成的影响

    刘红蕾; 张玉臻; 陶文沂

    2003-01-01

    将A4发根农杆菌感染长春花(Catharanthus roseus(L.)G.Don)后所长出的发根(hairy-root)进行培养,比较不同营养成分、碳源及初始糖浓度、外源激素以及温度和摇床转速对发根生长及产碱的影响.结果表明,当培养基为1/2B5培养基时发根生长良好;一定范围内初始糖浓度增加,有利于发根生长和生物碱生成;加入500mg/L的L-色氨酸利于发根的生长和产碱;将发根在28℃、75r/min的条件下悬浮培养10d可获得较高产量的生物碱.

  12. 外源NO对NaCl胁迫下长春花幼苗生物量和叶绿素荧光的影响%Effects of exogenous nitric oxide on biomass and chlorophyll fluorescence of Catharanthus roseus seedlings under NaCl stress

    胡凡波; 刘玲; 隆小华; 刘兆普

    2011-01-01

    A pot experiment in greenhouse was carried out to study the effects of different concentration exogenous NO donor ( sodium nitroprusside, SNP) on the biomass, chlorophyll content, and chlorophyll fluorescence of Catharanthus roseus seedlings under the stress of 50 mmol NaCl · L-1. Among the treatments with SNP, the seedlings treated with 50 mmol NaCl · L-1+ 0. 1 mmol SNP · L-1 had the highest fresh weight and dry weight, being significantly higher (18. 8% and 13. 9% , respectively) than the control (50 mmol NaCl · L-1). Similar to biomass, the chlorophyll a, chlorophyll b, and carotenoid contents in treatment 50 mmol NaCl · L-1 + 0. 1 mmol SNP · L-1 were the highest, but had no significant differences with the control. The variation trend of chlorophyll a/b was more complicated. Applying 0. 1 mmol · L-1 of SNP had the best effect in mitigating the salt damage to Ps Ⅱ reaction center, which increased the fluorescence ( Fv ) , maximal fluorescence ( Fm ) , light energy transformation efficiency of PS Ⅱ ( Fv/Fm ) , activity potential of PS Ⅱ ( Fv/Fo) , actual photochemical efficiency of PS Ⅱ ( (¢)PSⅡ ) , and photochemical quenching (qp) markedly, and decreased the minimal fluorescence (Fo) and non-photochemical quenching (qN) obviously. To sum up, 0. 1 mmol · L-1 of SNP had the best effect in protecting chlorophyll and PS Ⅱ reaction center against salt damage, and also, could promote the growth of the seedlings and enhance their tolerance to 50 mmol Ⅱ L-1 NaCl stress.%在温室条件下采用盆栽法,研究了50mmol· L-1 NaCl胁迫下5个不同浓度外源NO供体硝普钠(sodium nitroprusside,SNP)对长春花(Catharanthus roseus)幼苗生物量、叶绿素含量和叶绿素荧光的影响.结果表明:(1)鲜重和干重均在S2(50mmol·L-1NaCl+0.1mmol·L-1SNP)处理下达到最高,分别较对照So(50mmol·L-1 NaCl)显著增加18.8%和13.9%.叶绿素a、叶绿素b和类胡萝卜素与生物量相似,均在S2处理下

  13. 硝态氮缓解长春花幼苗海水胁迫效应的研究%ABATING EFFECT OF NITRATE ON SEAWATER-STRESSED OF CATHARANTHUS ROSEUS SEEDLINGS

    侯杰; 张建忠; 刘玲; 刘兆普

    2009-01-01

    长春花[Catharanthus roseus(L.)G.Don]为夹竹桃科长春花属,多年生草本或亚灌木花卉,原产非洲东部及美洲热带地区,喜温暖、阳光充足的环境,现在我国广为栽培。长春花全草均可入药,目前已从长春花植株的不同部位分离出100多种生物碱,其中长春碱和长春新碱经临床验证具有明显的抗癌作用,是国际上研究和应用最多的抗癌植物药源。

  14. 长春花种子油脂肪酸成分的GC-MS分析%Analysis of the fatty acid composition in Catharanthus roseus seed oil by gas chromatography-mass spectrometry

    牛卉颖; 杨磊; 张琳; 李晓薇; 刘玉; 祖元刚

    2008-01-01

    旨在建立长春花Catharanthus roseus种子脂肪酸成分的检测方法.采用正己烷提取长春花种子油,然后将油样甲酯化,以气相色谱.质谱(GC-MS)联用技术测定其中的组分.从分离出的9个峰中确认了5种组分,采用峰面积归一法计算各组分的相对百分含量,所鉴定的组分占总峰面积的99.91%.主要脂肪酸油酸(C19H36O2)、亚油酸(C19H34O2)、棕榈酸(C17H34O2)和花生酸(C21H42O2)的相对含量分别是73.92%,16.93%,5.58%和3.40%.

  15. Catharanthus roseus:a natural source for the synthesis of silver nanoparticles

    Mukunthan KS; Elumalai EK; Trupti N Patel; V Ramachandra Murty

    2011-01-01

    Objective: To develop a simple rapid procedure for bioreduction of silver nanoparticles (AgNPs) using aqueous leaves extracts of Catharanthus roseus (C. roseus). Methods: Characterization were determined by using UV-Vis spectrophotometry, scanning electron microscopy (SEM), energy dispersive X-ray and X-ray diffraction. Results: SEM showed the formation of silver nanoparticles with an average size of 67 nm to 48 nm. X-ray diffraction analysis showed that the particles were crystalline in nature with face centered cubic geometry. Conclusions: C. roseus demonstrates strong potential for synthesis of silver nanoparticles by rapid reduction of silver ions (Ag+ to Ag0). This study provides evidence for developing large scale commercial production of value-added products for biomedical/nanotechnology-based industries.

  16. 增强UV-B辐射及氮水平对长春花生长和生理代谢的影响%The Effects of Supplementary UV-B Radiation and Nitrogen Levels on the Growth and Physiological Metabolism of Catharanthus roseus

    郭晓瑞; 汪红玲; 王化楠; 段喜华

    2011-01-01

    The increased UV-B radiation and nitrogen deposition have become two important environmental factors affecting plant growth. Here, a medical plant Catharantkus roseus was subjected to altered UV-B and nitrogen level to study their influence in a sole or combined way on growth and vinblastine production. Our results showed that the UV-B exerted an obviously adverse effect on plant growth and biomass accumulation. The UV-B stress also led to physiological damages, for instance, the elevated MDA content. The increased supply with nitrogen nutrition could largely alleviate the inhibitory effects and damage symptom induced by UV-B stress. The UV-B absorbing compounds were enhanced remarkably by the supplementary UV-B radiation, and it was increased further when the nitrogen nutrition were supplied. The vinblastine content was increased markedly in the supplementary UV-B radiation, and which were increased further when the nitrogen supply increased. It was concluded that an increased nitrogen supply could not only alleviate the inhibitory effect of UV stress on plant growth, but enhance vinblastine accumulation in plants. We proposed that enrichment of nitrogen nutrition provide more N source for alkaloid synthesis induced by UV-B radiation, eventually resulting in increased alkaloid content.%地表UV-B辐射增强和氮沉降增加目前已成为影响植物生长的重要生态因子.本文以药用植物长春花(Catharanthus roseus)为材料,研究UV-B辐射和氮供应增加对长春花生长、生理及长春碱含量的协同效应.研究结果表明,紫外辐射增加对长春花生长和生物量积累具有显著的抑制作用.同时外源增加氮供应能明显缓解紫外辐射引起的生长抑制效应.紫外辐射引起的叶片膜脂过氧化胁迫导致了长春花叶片丙二醛含量显著增加,但同时增加氮供应能显著降低丙二醛水平.增强UV-B辐射处理显著促进长春花叶片UV-B吸收化合物合成积累,并随氮供应

  17. The seco-iridoid pathway from Catharanthus roseus

    Miettinen, K.; Dong, L.; Navrot, N.; Burlat, V.; Schneider, T.; Pollier, J.; Woittiez, L.S.; Krol, van der A.R.; Lugan, R.; Llc, T.; Verpoorte, R.; Oksman-Caldentey, K.M.; Martinoia, E.; Bouwmeester, H.J.

    2014-01-01

    The (seco)iridoids and their derivatives, the monoterpenoid indole alkaloids (MIAs), form two large families of plant-derived bioactive compounds with a wide spectrum of high-value pharmacological and insect-repellent activities. Vinblastine and vincristine, MIAs used as anticancer drugs, are produc

  18. Elucidation of the secoiridoid pathway in Catharanthus roseus

    Miettinen, Karel

    2013-01-01

    The (seco)iridoids and their derivatives, the monoterpenoid indole alkaloids (MIAs), form two large families of plant-derived bioactive compounds with a wide spectrum of high-value pharmacological and insect-repellent activities. Vinblastine and vincristine, MIAs used as anti-cancer drugs, are produ

  19. A look inside an alkaloid multisite plant: the Catharanthus logistics.

    Courdavault, Vincent; Papon, Nicolas; Clastre, Marc; Giglioli-Guivarc'h, Nathalie; St-Pierre, Benoit; Burlat, Vincent

    2014-06-01

    Environmental pressures forced plants to diversify specialized metabolisms to accumulate noxious molecules such as alkaloids constituting one of the largest classes of defense metabolites. Catharanthus roseus produces monoterpene indole alkaloids via a highly elaborated biosynthetic pathway whose characterization greatly progressed with the recent expansion of transcriptomic resources. The complex architecture of this pathway, sequentially distributed in at least four cell types and further compartmentalized into several organelles, involves partially identified inter-cellular and intra-cellular translocation events acting as potential key-regulators of metabolic fluxes. The description of this spatial organization and the inherent secretion and sequestration of metabolites not only provide new insight into alkaloid cell biology and its involvement in plant defense processes but also present new biotechnological challenges for synthetic biology. PMID:24727073

  20. ОСОБЕННОСТИ СОДЕРЖАНИЯ ТЕРПЕНОВЫХ ИНДОЛЬНЫХ АЛКАЛОИДОВ В КУЛЬТИВИРУЕМОМ В ИСКУССТВЕННЫХ УСЛОВИЯХ CATHARANTHUS ROSEUS

    РОМАШКО С. Н.; ЯНЦЕВИЧ А. В.; Молчан, О.В.; В.М. Юрин

    2012-01-01

    В данной работе проведен качественный анализ терпеновых индольных алкалоидов, содержащихся в листьях и корнях Catharanthusroseus, культивируемого в искусственных условиях. Выявлены особенности накопления непосредственных предшественников фармакологически ценных бис-индольных алкалоидов (виндолина и катарантина) в различных органах указанного растения. Установлено, что в корневой системе Catharanthus roseus содержится значительное количество бис-индольных алкалоидов.Qualitative content of terp...

  1. Auxins Induce Tryptophan Decarboxylase Activity in Radicles of Catharanthus Seedlings 1

    Aerts, Rob J.; Alarco, Anne-Marie; De Luca, Vincenzo

    1992-01-01

    Germinating seedlings of Catharanthus roseus produce monoterpenoid indole alkaloids as a result of a transient increase of tryptophan decarboxylase (TDC) activity. The influence of auxins on this transient rise of TDC activity was studied. External application of indolebutyric acid or 2,4-dichlorophenoxyacetic acid at a concentration of 20 to 40 μm enhanced and prolonged the rise in TDC activity in developing seedlings. Auxin treatment also influenced the morphology of the seedlings; it induced a shortening and thickening of the hypocotyl and the radicle and promoted the initiation of lateral roots in the radicle. During development, the radicles of auxin-treated seedlings displayed a gradual increase in TDC activity that was absent in the radicles of untreated controls. Examination of immunoblots revealed anti-TDC reactive proteins in extracts from radicles of auxin-treated seedlings, but none in extracts from radicles of control seedlings. In contrast, TDC activity and immunoreactive protein levels in the aerial parts of controls and auxin-treated seedlings were comparable. Our results indicate that externally applied auxins induce both abnormal development and TDC activity in the radicles of Catharanthus seedlings. Although auxins slightly delayed the light-mediated induction of the cotyledon-specific last step in vindoline biosynthesis (i.e. acetylcoenzyme A: deacetylvindolin-O-acetyltransferase activity), seedlings still synthesized vindoline, one of the major alkaloid end products. Images Figure 2 PMID:16653009

  2. Physiological Effect Of Peri winkle (C.roseus On Diabetic Albino Rat

    Eman G.E. Helal*, Samia M. Abd-Elwahab*, Atef M. moussa** and Anwaar A. Mohammad

    2012-10-01

    Full Text Available Back ground: Diabetes mellitus (DM is a major health problem worldwide in recent time. Asia and Africa are the most viable areas where the disease is feared to raise 2–3 folds. Many herbal products have been recommended for the treatment of DM in ancient literature of Ayurveda in India and other worldwide.Material and method: Thirty male adult albino rats were used to investigate the effect of Catharanthus rosea (C.roseus on diabetic rats. Rats were divided into three equal groups, control, diabetic non treated and diabetic C.roseus treated groups. After thirty days of treatment all rats of each group were sacrificed. The body weight of each rat was determined at the beginning and the end of each period. Blood glucose, serum insulin, lipid and protein profiles, liver and kidney functions, blood picture and liver glycogen were determined for each rat at the end of each period. Pancreatic samples were obtained and processed for microscopic and quantitative evaluation after staining the prepared sections with heamatoxylin and eosin as well as special stain for demonstration of the different pancreatic cells in the islets of Langerhans.Results: The obtained results showed that the diabetic rats were diagnosed by laboratory assessment to body weight loss, hyperglycemia, and hypoinsulinemia, significant increase in liver and kidney functions, lipid and protein profiles and decreased liver glycogen content. While, C.roseus treatment led to a significant improvement in these parameters except liver function. Microscopically there was definite vaculation, degeneration, karyolysis and pyknosis of beta pancreatic cells in the diabetic group, while other pancreatic cells were not affected (alpha and delta cells. The use of C.roseus treatment of this study greatly improves such cellular changes.Conclusion: It was recommended that the use of the water extract of C. roseus levies as a hypoglycemic agent may offer a new hope to the diabetics in future. It

  3. Comparison of Biomasses Assignment between Catharanthus roseus and Catharanthus roseus cv. Albus%两种长春花生物量配置的比较

    王非; 李雷鸿; 孙家宝

    2007-01-01

    以不同生境(野生、露地栽培)条件下红长春花和白长春花为研究对象,对其生物总量、根冠比、各部位的生物量配置等方面进行了比较分析.结果表明:与白花长春花相比,红花长春花的根、茎、叶生物量分配比例较为合理,生殖配置高于白花,表现出较强的种间优势.2种长春花对茎和根的生物量的分配与植株的生长高度呈负相关.长春花叶的生物量分配与植株生长高度也出现了不协调.2种生境条件中,栽培条件下的长春花植株个体较大,生物量分配比较合理,生长比较稳定,说明在露地栽培条件下,长春花的营养生长和生殖生长间有较为合理的权衡关系,利于长春花的生长.

  4. Improving the content of terpenoid indole alkaloids in catharanthus rose us by . 0Coγ radiation

    Treated seeds of Catharanthus roseus L. with . 0Coγ ray of different dose, to stat plant germination rate, survival plants' grown rate, observed variation rate of character and took regression analysis. We found that the sub-lethal dose (LD50) was 174 Gy and the optimum irradiation dose for the seed ofadzuki bean is 200 Gy. The early seedling leaves of M1 show asymmetry of leaf shape and and curly edge, and that maybe related with the absorbed dose. Results of radiation breeding we obtained the high stroke deformity and the short stroke deformity. The total content of TIAs and the content of three main kinds of TIAs in mutants and controls were measured and analysised by HPLC. The content of three main kinds of TIAs by the high stroke deformity of 24% higher than CK

  5. Candidatus Phytoplasma malaysianum, a novel taxon associated with virescence and phyllody of Madagascar periwinkle (Catharanthus roseus)

    This study addressed the taxonomic position and group classification of a phytoplasma responsible for virescence and phyllody symptoms in naturally diseased Madagascar periwinkle plants in western Malaysia. Unique regions in the 16S rRNA gene from the Malaysian periwinkle virescence (MaPV) phytopla...

  6. 长春花大田栽培技术%Field Cultural Practice for Catharanthus roseus

    赵剑; 李建国

    2009-01-01

    通过育苗、移栽、田间管理以及采收等一系列田间实验,掌握长春花的生长发育、需水、需肥和病虫害发生规律,提出较为有效的栽培方法,基本达到在长春花栽培过程中"可控、安全、优质"的目标.为长春花的大面积种植提供技术参考.

  7. Retrobiosynthetic study of salicylic acid in Catharanthus roseus cell suspension cultures

    Mustafa, Natali Rianika

    2007-01-01

    Salicylic acid (SA) is an important signal compound in systemic acquired resistance in plants. The level of this C6C1 compound in plants increases after a pathogenic attack. There are two biosynthetic pathways of SA, the phenylalanine pathway, which is thought to occur in plants, and the isochorismate pathway, which is found in microorganisms. The biosynthetic pathway of SA in plant is still a matter of debate and some studies showed that the presence of the microbial pathway in plants is als...

  8. Unlocking the diversity of alkaloids in Catharanthus roseus: nuclear localization suggests metabolic channeling in secondary metabolism.

    Stavrinides, Anna; Tatsis, Evangelos C; Foureau, Emilien; Caputi, Lorenzo; Kellner, Franziska; Courdavault, Vincent; O'Connor, Sarah E

    2015-03-19

    The extraordinary chemical diversity of the plant-derived monoterpene indole alkaloids, which include vinblastine, quinine, and strychnine, originates from a single biosynthetic intermediate, strictosidine aglycone. Here we report for the first time the cloning of a biosynthetic gene and characterization of the corresponding enzyme that acts at this crucial branchpoint. This enzyme, an alcohol dehydrogenase homolog, converts strictosidine aglycone to the heteroyohimbine-type alkaloid tetrahydroalstonine. We also demonstrate how this enzyme, which uses a highly reactive substrate, may interact with the upstream enzyme of the pathway. PMID:25772467

  9. The distribution of inole alkaloids in different organs of Catharanthus roseus G. Don. (Vinca rosea L)

    Hassan Ebrahimzadeh; Azra Ataei -Azimi4 Mohammad-Reza Noori-Dafoi

    1996-01-01

    The study of alkaloid extract of the leaf of Vinca rosea by TLC showed the existence of 13 bands, 7 bands of them were: Serpentine, Vincristine, Vinblastine, Ajmaline, Catharantine, Vindoline and Ajmalicine. The existence of these 7 alkaloids was confirmed in the young leaves, old leaves and the roots of the plant by HPLC. Furthermore, two alkaloids with the short retention times and one alkaloid with the medium retention time in the young leaf, three alkaloids with the short retention time a...

  10. Optimization of transient expression procedures in Catharanthus roseus and Arabidopsis thaliana for subcellular localization studies

    Ribeiro, Diana Margarida da Costa

    2010-01-01

    Dissertação de mestrado em Biotecnologia e Bio-Empreendedorismo em Plantas Aromáticas e Medicinais Nowadays, the availability of much gene sequence information demands the development of tools for their fast characterization at the protein level, where function actually resides. Here, the interest in the characterization of certain of the known Arabidopsis class III peroxidase (Prx) genes, as well as the interest in the characterization of candidate genes implicated in the meta...

  11. Unlocking the Diversity of Alkaloids in Catharanthus roseus: Nuclear Localization Suggests Metabolic Channeling in Secondary Metabolism

    Stavrinides, Anna; Tatsis, Evangelos C.; Foureau, Emilien; Caputi, Lorenzo; Kellner, Franziska; Courdavault, Vincent; O’Connor, Sarah E.

    2015-01-01

    Summary The extraordinary chemical diversity of the plant-derived monoterpene indole alkaloids, which include vinblastine, quinine, and strychnine, originates from a single biosynthetic intermediate, strictosidine aglycone. Here we report for the first time the cloning of a biosynthetic gene and characterization of the corresponding enzyme that acts at this crucial branchpoint. This enzyme, an alcohol dehydrogenase homolog, converts strictosidine aglycone to the heteroyohimbine-type alkaloid ...

  12. The distribution of inole alkaloids in different organs of Catharanthus roseus G. Don. (Vinca rosea L

    Hassan Ebrahimzadeh

    1996-08-01

    Full Text Available The study of alkaloid extract of the leaf of Vinca rosea by TLC showed the existence of 13 bands, 7 bands of them were: Serpentine, Vincristine, Vinblastine, Ajmaline, Catharantine, Vindoline and Ajmalicine. The existence of these 7 alkaloids was confirmed in the young leaves, old leaves and the roots of the plant by HPLC. Furthermore, two alkaloids with the short retention times and one alkaloid with the medium retention time in the young leaf, three alkaloids with the short retention time and one alkaloid with long retention time in the old leaf, and one alkaioid with the short retention time and two other alkaloids with the long retention times in the root, were observed."nThe study of absorption spectrum of alkaloid extract and its comparison with absorption spectrum of ajmalicine showed that the latter can be used for drawing a standard curve and detecting the amount of total alkaloids. The total alkaloids in fresh root, old leaf and young leaf were 2.1 mg , 1.3 mg and 0.9 mg/g respectively, Vindoline and Catharantine were major alkaloids in all of them. However, their amount were more in the root (1.2 mg/ g of fresh material, 0.85 in the old leaf and 0.38 in the young leaf. Vinblastine existed in all three samples but its amount was more in the root {0.22 mg per gramme of the fresh material in the root, 0.26 in the young leaf and 0.003 in the old leaf."nAjmalicine was detected only in the leaf and its amount was more in young leaf than the old leaf (0.141 mg/g of the fresh material in the young leaf and 0.013 in the old leaf. Serpentine was seen in all of the studied organs but its amount was more in the young leaf than other organs (0.192 mg/ g of the fresh material in the young leaf, 0.11 in the root and 0.07 in the old leaf. Ajmaline existed in all of the organs in minute amount but in the old leaf was more (0 .07 mg/g of the fresh material in the old !eaf,0.044 in the young leaf and 0.04 in the root . The percentage of unknown alkaloids were low, except in the root (20.5% of total alkaloid.

  13. Evaluation of the anti-diabetic potential of Catharanthus roseus extracts

    Silva, Dina Maria Morais da

    2009-01-01

    Dissertação de mestrado em Biotecnologia e Bio-Empreendedorismo em Plantas Aromáticas e Medicinais Type 2 DM is a metabolic disorder that results from genetic and environmental factors and affects 100 million people worldwide. It is characterized by hyperglycemia due to: i) decreased secretion of insulin by the pancreas and ii) resistance to the action of insulin in tissues as muscle, liver and adipocytes, leading to impaired glucose uptake. The objectives of the present wor...

  14. A Stereoselective Hydroxylation Step of Alkaloid Biosynthesis by a Unique Cytochrome P450 in Catharanthus roseus*

    Giddings, Lesley-Ann; Liscombe, David K.; Hamilton, John P; Childs, Kevin L.; DellaPenna, Dean; Buell, C. Robin; O'Connor, Sarah E.

    2011-01-01

    Plant cytochrome P450s are involved in the production of over a hundred thousand metabolites such as alkaloids, terpenoids, and phenylpropanoids. Although cytochrome P450 genes constitute one of the largest superfamilies in plants, many of the catalytic functions of the enzymes they encode remain unknown. Here, we report the identification and functional characterization of a cytochrome P450 gene in a new subfamily of CYP71, CYP71BJ1, involved in alkaloid biosynthesis. Co-expression analysis ...

  15. Gene-to-metabolite networks for terpenoid indole alkaloid biosynthesis in Catharanthus roseus cells

    Rischer, Heiko; Orešič, Matej; Seppänen-Laakso, Tuulikki; Katajamaa, Mikko; Lammertyn, Freya; Ardiles-Diaz, Wilson; Van Montagu, Marc C. E.; Inzé, Dirk; Oksman-Caldentey, Kirsi-Marja; Goossens, Alain

    2006-01-01

    Rational engineering of complicated metabolic networks involved in the production of biologically active plant compounds has been greatly impeded by our poor understanding of the regulatory and metabolic pathways underlying the biosynthesis of these compounds. Whereas comprehensive genome-wide functional genomics approaches can be successfully applied to analyze a select number of model plants, these holistic approaches are not yet available for the study of nonmodel plants that include most,...

  16. Production of Annual Vinca (Catharanthus roseus) in WholeTree Substrates

    The objective of this study was to evaluate the potential use of container substrates composed of processed whole pine trees (WholeTree). Three species [loblolly pine (Pinus taeda), slash pine (Pinus elliottii) and longleaf pine (Pinus palustris)] of 8-10 year old pine trees were harvested at groun...

  17. Retrobiosynthetic study of salicylic acid in Catharanthus roseus cell suspension cultures

    Mustafa, Natali Rianika

    2007-01-01

    Salicylic acid (SA) is an important signal compound in systemic acquired resistance in plants. The level of this C6C1 compound in plants increases after a pathogenic attack. There are two biosynthetic pathways of SA, the phenylalanine pathway, which is thought to occur in plants, and the isochorisma

  18. Phytoremediation of TNT: C. roseus hairy roots as a model system

    Lauritzen, J.R.; Hughes, J.B.; Shanks, J.V. [Rice Univ., Houston, TX (United States)

    1996-12-31

    Widespread contamination by 2,4,6-trinitrotoluene (TNT) of Soil exists at former munitions production and handling facilities. Phytoremediation may be an effective alternative to existing methods of TNT remediation: incineration is highly expensive and recalcitrant reduction products are formed in composting. Recently, the intrinsic ability of plants to transform TNT has been demonstrated using hairy root cultures of Catharanthus roseus as a model system. Kinetic studies were performed at concentrations of 30 and 50 mg/L TNT in growth medium. The pseudo-first order rate constants for disappearance ranged from 0.0103 to 0.0161 (L/g-day); TNT disappears completely within seven to ten days of exposure. The fate of the TNT molecule in plants is also currently under study, mass balance studies were performed with 1-{sup 14}C TNT. After a seven day exposure period, 72% of the label was associated with the roots and 30% was associated with the medium. However, HPLC analysis shows that less than 5% (wt%) of the TNT added is recoverable from both the plants and the media in the form of reduction products. 11 refs., 2 figs.

  19. Sonication-Assisted Agrobacterium-Mediated Transformation of Catharanthus roseus%超声波辅助农杆菌介导的长春花(Catharanthus roseus)遗传转化

    王荃; 潘琪芳; 袁芳; 侯晓晔; 赵静雅; 唐克轩

    2009-01-01

    研究以根癌农杆菌介导结合超声波处理和乙酰丁香酮诱导,通过对GUS基因瞬时表达来研究影响长春花转化效率的因素.对转化方法包括超声波处理的功率和时间、乙酰丁香酮(3,5-methoxy-4-hydroxyacetophenone,AS)的浓度、共培养时间进行了系统优化.结果表明:在添加AS 100 p,mol·L-1的1/2 MS培养液中,超声波处理功率80 W,处理时间10 rain,共培养2 d,根癌农杆菌介导的长春花下胚轴遗传转化能获得最佳的GUS基因瞬间表达率.本研究为进一步开展根癌农杆菌介导稳定转化长春花奠定了基础.

  20. Isolation, Purification and Characterization of Vinblastine and Vincristine from Endophytic Fungus Fusarium oxysporum Isolated from Catharanthus roseus

    Ashutosh Kumar; Deepak Patil; Pattuparambil Ramanpillai Rajamohanan; Absar Ahmad

    2013-01-01

    Endophytic fungi reside in a symbiotic fashion inside their host plants, mimic their chemistry and interestingly, produce the same natural products as their hosts and are thus being screened for the production of valuable compounds like taxol, camptothecin, podophyllotoxin, etc. Vinblastine and vincristine are excellent anti-cancer drugs but their current production using plants is non-abundant and expensive. In order to make these drugs readily available to the patients at affordable prices,...

  1. Detrimental effect of quercetin on phytoplasma-infected Catharanthus roseus (L.) G. Don shoots grown in vitro

    Ćurković-Perica, Mirna; Ježić, Marin

    2010-01-01

    Quercetin is known to possess antimicrobial activity against bacteria, fungi, and viruses. The activity of this flavonoid against phytoplasmas, non-cultivable plant pathogenic bacteria that cause numerous plant diseases, has never been examined before. The aim of this research was to examine the effect of different concentrations of quercetin (10 mM, 100 mM and 1 mM) on 'Candidatus Phytoplasma asteris' and on phytoplasma- nfected periwinkle shoots grown in vitro. The addition of quercetin nei...

  2. Expression Patterns of Genes Involved in the Defense and Stress Response of Spiroplasma citri Infected Madagascar Periwinkle Catharanthus roseus

    Naghmeh Nejat

    2012-02-01

    Full Text Available Madagascar periwinkle is an ornamental and a medicinal plant, and is also an indicator plant that is highly susceptible to phytoplasma and spiroplasma infections from different crops. Periwinkle lethal yellows, caused by Spiroplasma citri, is one of the most devastating diseases of periwinkle. The response of plants to S. citri infection is very little known at the transcriptome level. In this study, quantitative real-time PCR (RT-qPCR was used to investigate the expression levels of four selected genes involved in defense and stress responses in naturally and experimentally Spiroplasma citri infected periwinkles. Strictosidine β-glucosidase involved in terpenoid indole alkaloids (TIAs biosynthesis pathway showed significant upregulation in experimentally and naturally infected periwinkles. The transcript level of extensin increased in leaves of periwinkles experimentally infected by S. citri in comparison to healthy ones. A similar level of heat shock protein 90 and metallothionein expression was observed in healthy, naturally and experimentally spiroplasma-diseased periwinkles. Overexpression of Strictosidine β-glucosidase demonstrates the potential utility of this gene as a host biomarker to increase the fidelity of S. citri detection and can also be used in breeding programs to develop stable disease-resistance varieties.

  3. Hydrangea virescence. I.- Description of the disease and its transmission to differential host plant Catharanthus Roseus by Cucusta Subinclusa

    Cousin, M.T.; Sharma, A. K.; Rousseau, J.; Poitevin, J.P.; Savoure, A.

    1986-01-01

    A virescence disease has recently been observed in France on Hydrangea macrophylla. This ornamental plant is cultivated in a rather particular way, comprising two successive phases : "production of mature plants" and "production of flowering plants". The possible period of disease inception and symptom appearance has been studied with respect of these two phases. Symptoms have been compared with those already described from different localities in Europe and America. Transmission of the p...

  4. Evaluation of antimicrobial activity of extracts of in vivo and in vitro grown Vinca rosea L. (Catharanthus roseus) against pathogens.

    Naz, Shagufta; Haq, Rukhama; Aslam, Farah; Ilyas, Saiqa

    2015-05-01

    The antimicrobial activity of Vinca rosea was evaluated against pathogenic bacterial strains (Bacillus subtilis, B. licheniformis and Azotobacter sp.) and fungal strains (Asprgillus niger, Alternaria solani and Rhizopus oryzae) using agar well diffusion method. Methanolic extracts of in vivo leaf, in vitro leaf, in vitro calluses of leaf, nodal and fruit explants were used and exhibited antimicrobial activity as indicated by minimum inhibitory concentration (MIC). In vitro extracts showed better results as compared to the in vivo extracts for both the antibacterial as well as the antifungal activity. Among all the extracts, maximum zone of inhibition (30.3 mm ± 0.58(a)) was formed by in vitro leaf callus extract concentration of 2.0mg/ml against B. licheniformis. Similarly in case of antifungal activity, maximum zone of inhibition (34.6mm ± 0.57(a)) was formed by in vitro leaf callus extract and MIC value is 6.0mg/ml against A. niger. Hence these results clearly depicts that V. rosea possess a great strength to fight against the microbial activity and can be used against various infections. PMID:26004716

  5. Expression Patterns of Genes Involved in the Defense and Stress Response of Spiroplasma citri Infected Madagascar Periwinkle Catharanthus roseus

    Nejat, Naghmeh; Vadamalai, Ganesan; Dickinson, Matthew

    2012-01-01

    Madagascar periwinkle is an ornamental and a medicinal plant, and is also an indicator plant that is highly susceptible to phytoplasma and spiroplasma infections from different crops. Periwinkle lethal yellows, caused by Spiroplasma citri, is one of the most devastating diseases of periwinkle. The response of plants to S. citri infection is very little known at the transcriptome level. In this study, quantitative real-time PCR (RT-qPCR) was used to investigate the expression levels of four selected genes involved in defense and stress responses in naturally and experimentally Spiroplasma citri infected periwinkles. Strictosidine β-glucosidase involved in terpenoid indole alkaloids (TIAs) biosynthesis pathway showed significant upregulation in experimentally and naturally infected periwinkles. The transcript level of extensin increased in leaves of periwinkles experimentally infected by S. citri in comparison to healthy ones. A similar level of heat shock protein 90 and metallothionein expression was observed in healthy, naturally and experimentally spiroplasma-diseased periwinkles. Overexpression of Strictosidine β-glucosidase demonstrates the potential utility of this gene as a host biomarker to increase the fidelity of S. citri detection and can also be used in breeding programs to develop stable disease-resistance varieties. PMID:22408455

  6. Negative-Pressure Cavitation Extraction of Four Main Vinca Alkaloids from Catharanthus roseus Leaves

    Yuangang Zu

    2012-07-01

    Full Text Available In the present study, an improved method termed negative-pressure cavitation extraction (NPCE followed by reverse phase high-performance liquid chromatography (RP-HPLC was developed for the extraction and quantification of vindoline (VDL, catharanthine (CTR, vincristine (VCR and vinblastine (VLB from Catharanthus roseus leaves. The optimized method employed 60-mesh particles, 80% ethanol, a negative pressure of −0.075 MPa, a solid to liquid ratio of 1:20, 30 min of extraction and three extraction cycles. Under these optimized conditions, the extraction yields of VDL, CTR, VCR and VLB are 0.5783, 0.2843, 0.018 and 0.126 mg/g DW, respectively. These extraction yields are equivalent to those from the well-known ultrasonic extraction method and higher than the yields from maceration extraction and heating reflux extraction. Our results suggest that NPCE-RP-HPLC represents an excellent alternative for the extraction and quantification of vinca alkaloids for pilot- and industrial-scale applications.

  7. Catharanthus mosaic virus: A potyvirus from a gymnosperm, Welwitschia mirabilis.

    Koh, Shu Hui; Li, Hua; Admiraal, Ryan; Jones, Michael G K; Wylie, Stephen J

    2015-05-01

    A virus from a symptomatic plant of the gymnosperm Welwitschia mirabilis Hook. growing as an ornamental plant in a domestic garden in Western Australia was inoculated to a plant of Nicotiana benthamiana where it established a systemic infection. The complete genome sequence of 9636 nucleotides was determined using high-throughput and Sanger sequencing technologies. The genome sequence shared greatest identity (83% nucleotides and 91% amino acids) with available partial sequences of catharanthus mosaic virus, indicating that the new isolate belonged to that taxon. Analysis of the phylogeny of the complete virus sequence placed it in a monotypic group in the genus Potyvirus. This is the first record of a virus from W. mirabilis, the first complete genome sequence of catharanthus mosaic virus determined, and the first record from Australia. This finding illustrates the risk to natural and managed systems posed by the international trade in live plants and propagules, which enables viruses to establish in new regions and infect new hosts. PMID:25804761

  8. Biodegradation of a synthetic lubricant by Micrococcus roseus.

    Wright, M. A.; Taylor, F.; Randles, S J; Brown, D E; Higgins, I J

    1993-01-01

    A bacterium that was able to utilize Emkarate 1550 (E1550), a synthetic lubricant ester, as the sole source of carbon was isolated. The isolate was tentatively identified as Micrococcus roseus. The components of the E1550 ester, octanoate, decanoate, and 1,1,1-tris(hydroxymethyl)propane (TMP), were detected in the culture medium of cells growing on the ester. The TMP tertiary alcohol accumulated during growth and was not utilized by this isolate. The detection of the components of the ester i...

  9. Screening molecules for control of citrus huanglongbing using an optimized regeneration system for 'Candidatus Liberibacter asiaticus'-infected periwinkle (Catharanthus roseus) cuttings.

    Zhang, Muqing; Duan, Yongping; Zhou, Lijuan; Turechek, William W; Stover, Ed; Powell, Charles A

    2010-03-01

    Citrus huanglongbing is one of the most destructive diseases of citrus worldwide. The disease is associated with three different species of 'Candidatus Liberibacter', of which 'Ca. L. asiaticus' is the most widely distributed. An optimized system using 'Ca. L. asiaticus'-infected periwinkle cuttings was developed to screen chemical compounds effective for controlling the bacterial population while simultaneously assessing their phytotoxicity. The optimal regeneration conditions were determined to be the use of vermiculite as a growth medium for the cuttings, and a fertilization routine using half-strength Murashige and Tucker medium supplemented with both naphthalene acetic acid (4 microg/ml) and indole-3-butyric acid (4 microg/ml). This system allowed a plant regeneration rate of 60.6% for 'Ca. L. asiaticus'-infected cuttings in contrast to the DBNPA), were found to be effective at eliminating or suppressing the 'Ca. L. asiaticus' bacterium in this periwinkle regeneration system. When treated with penicillin G sodium at 50 microg/ml, all plants regenerated from 'Ca. L. asiaticus'-infected cuttings were 'Ca. L. asiaticus' negative as determined by both nested polymerase chain reaction (PCR) and quantitative real-time PCR. In addition, DBNPA was also able to significantly reduce the percentage of 'Ca. L. asiaticus'-positive plants and the titer of the 'Ca. L. asiaticus' bacterium at 200 microl/liter. PMID:20128697

  10. Analysis of iridoids content and expression studies of genes encoding early enzymes in the indol terpenoid biosynthesis pathway in Catharanthus roseus

    Natalia Palacios-Rojas; Mark Leech

    2007-01-01

    Terpenoid indole alkaloids (TIA) are of pharmaceutical importance, however the industrial use of these compouds is very limited because its accumulation is very low in plant tissues. TIA are derived f rom the shikimate and terpenoid pathways, which supply secologanin and tryptamine, the indole and iridoid moieties, respectively. Secololganin is a terpenoid which is belived to be synthesised the MEP pathway rather than by the acetate/mevalonic acid pathway. Secologanin is thought to be a limit...

  11. Enhancing accumulation of indole alkaloids in Catharanthus roseus leaves%长春花叶片中吲哚生物碱增产的研究

    张秀省; 张荣涛; 聂莉莉; 郭玉海; 翟志席

    2005-01-01

    目的探讨乙烯利、乙酰水杨酸、色氨酸3种试剂对长春花植株中药用成分积累的影响.方法用不同质量浓度的3种试剂溶液分别处理温室盆栽长春花植株,2 d后,测定叶片中的总吲哚生物碱含量,并采用RP-HPLC法测定长春碱和长春质碱的含量.结果3种试剂处理后吲哚总碱、长春碱、长春质碱的积累均有明显的提高,并确定了最适的处理质量浓度.结论3种试剂对长春花植物中吲哚生物碱的产生有明显的促进作用.

  12. Physiological characters of Catharanthus roseus mutant cells%长春花突变细胞生理特征的研究

    张秀省; 聂莉莉; 向蓓蓓; 王淑芳; 朱晔荣; 王勇

    2007-01-01

    目的 研究以秋水仙碱处理的长春花突变细胞在继代培养过程中的生长规律、吲哚生物碱积累的规律及营养成分的适宜浓度等方面的特征,期望得到适于工业化细胞培养的理想材料.方法 将突变细胞培养于MS固体培养基上,定期称取其鲜质量,用有机溶剂萃取吲哚生物碱,应用RP-HPLC法检测药用成分阿玛碱和长春质碱.结果 突变细胞培养至第30代时,生长速度和吲哚总碱积累得最快,培养至第45代时,两者均明显下降.而药用成分的量于第20代达到最高;在培养基中添加适量的色氨酸能提高药用成分的量;培养基中Ca2+和Zn2+的质量浓度分别为1 760和12.6 mg/L时,明显促进了药用成分的积累.结论 长春花突变细胞可能是一种适于工业化细胞培养的理想材料.

  13. Accumulation of Monoterpenoid Indole Alkaloids in Periwinkle Seedlings ("Catharanthus roseus") as a Model for the Study of Plant-Environment Interactions

    Miranda-Ham, Maria de Lourdes; Islas-Flores, Ignacio; Vazquez-Flota, Felipe

    2007-01-01

    Alkaloids are part of the chemical arsenal designed to protect plants against an adverse environment. Therefore, their synthesis and accumulation are frequently induced in response to certain environmental conditions and are mediated by chemical signals, which are formed as the first responses to the external stimulus. A set of experiments using…

  14. Simple and rapid biosynthesis of stable silver nanoparticles using dried leaves of Catharanthus roseus. Linn. G. Donn and its anti microbial activity.

    Kotakadi, Venkata Subbaiah; Rao, Y Subba; Gaddam, Susmila Aparna; Prasad, T N V K V; Reddy, A Varada; Gopal, D V R Sai

    2013-05-01

    Nanoparticles have been used to alter and improve the pharmacokinetic and pharmacodynamic properties of various types of drug molecules. The plant extracts are eco-friendly, economical and cost effective for synthesis of large scale of nanoparticles. In this paper we represent the synthesis of silver nanoparticles (AgNPs) from room dried leaves of Vinca rosea. The AgNPs were characterized by UV-vis spectroscopy. The AgNPs are crystalline in nature, were determined from scanning electron microscopy (SEM) coupled with energy dispersive X-ray (EDX), X-ray diffraction patterns (XRD), and also the size of the NPs was calculated by using Hariba Nanoparticle analyzer and the stability was calculated by using the Zetapotential. The nanoparticles obtained from leaf extracts were of size 27±2 and 30±2 respectively and Zetapotential of AgNPs was found to be -63.1 mV, so it indicates the dispersion and stability. The synthesized AgNPs have very good antimicrobial activity. PMID:23376746

  15. Application of biotechnology in studies of Catharanthus roseus%生物技术在长春花研究中的应用

    陈敏; 廖志华; 孙敏; 罗学

    2001-01-01

    长春花为重要的抗癌药用植物,对细胞组织培养、Ti和Ri质粒遗传转化等生物技术在长春花研究中的应用作一综述,并对长春花的开发利用和长春花生物碱的工业化生产存在问题和前景作了分析。

  16. ISSR Analysis of Genetic Relationships between Forty Varieties of Catharanthus roseus%长春花种质资源遗传多样性的ISSR分析

    姚丹青; 潘彬也; 热娜·多里坤; 关媛; 潘俊松; 何欢乐; 唐克轩; 蔡润

    2009-01-01

    本研究利用ISSR分子标记技术对40个长春花种质资源进行了遗传关系分析,多态性PCR扩增结果表明:在115个位点,其中多态位点78个,多态位点百分率为67.8%,多态性较高;POPGENE32软件计算结果表明,40个长春花品种平均有效等位基因数为1.6384,Nei遗传多样性指数平均为0.3735,平均Shannon信息指数为0.5546;应用NCSYS软件计算得到品种间的遗传相似系数介于0.150~0.900,平均为0.584.通过聚类分析将这40个长春花种质分成7组,该聚类结果与以前根据形态进行的分类结果有极大的相似性.

  17. 两种长春花体内黄酮含量的研究%Comparative study of the flavonoids content in two varieties of Catharanthus roseus

    王非; 孙家宝; 李雷鸿

    2007-01-01

    对野生及露地栽培生境中生长的红长春花和白长春花不同部位中总黄酮含量进行比较研究,研究结果显示:1)长春花各部位中总黄酮含量从高到低为果(含种子)、叶片、花、茎、根;2)野生白长春花体内总黄酮含量低于栽培白长春花;3)野生红长春花体内总黄酮含量与露地栽培红长春花的含量相接近;4)红长春花体内总黄酮含量高于白长春花;5)两种长春花在茎和叶片在不同高度上的总黄酮含量有所差异.长春花体内黄酮物质的这种分布特点将为下一步园林花卉的引种驯化工作提供理论支持.

  18. Optimization of Conditions for Suspension Culture of Catharanthus Roseus%长春花悬浮培养条件的优化

    廖礼佳; 李祥会; 杨军; 伍春莲

    2013-01-01

    以长春花叶片诱导的愈伤组织为材料,悬浮培养在含0.5mg/L的6-BA,40mg/L的Vc 50ml以及0.5mg/的2,4-D的MS培养基中,初始接入量、研究转速、蔗糖浓度和额外添加激素条件对愈伤组织增殖的影响.结果显示,在含有50ml MS液体培养基中接入3g愈伤组织,震荡转速设定为为120rpm,蔗糖浓度为15g/L,以及额外添加0.5mg/L的NAA时,愈伤组织增殖最快.研究结果为建立高效的愈伤组织悬浮培养细胞系奠定了基础.

  19. EMS诱变的长春花细胞系突变研究%Mutation in Catharanthus roseus induced by EMS

    张秀省; 张荣涛; 曹岚; 王勇; 王宁宁; 王淑芳

    2004-01-01

    目的通过化学诱变的手段筛选生长快且吲哚碱含量高的长春花突变细胞系.方法用不同浓度的甲基磺酸乙酯(EMS)处理长春花愈伤组织,挑选成活愈伤组织块继代扩大培养,比较其与对照愈伤组织生长速率、吲哚总碱积累等方面的差异.结果用EMS处理的长春花愈伤组织与对照相比,不仅生长快,并且吲哚总碱含量高.结论EMS处理的愈伤组织发生了变异,变异种是所期望的比较理想的细胞系.

  20. 6-BA对长春花幼苗生长的影响%Effects of 6-BA on the Seedling Growth of Catharanthus roseus

    郭晓燕

    2010-01-01

    本试验用50×10-6,100×10-6,200×10-6的6-BA水溶液对长春花幼苗叶片进行喷施处理,以清水处理为对照,观测不同浓度6-BA水溶液对植株高度和叶片生长量的影响.结果表明,用100×10-6~200×10-6的6-BA水溶液对长春花幼苗进行喷叶处理对幼苗的生长有较好的促进作用.

  1. 长春花内生真菌的分离及鉴定%Isolation and Identification on Endophytic Fungus from Catharanthus roseus

    石岳香; 周敏; 唐忠海; 饶力群

    2008-01-01

    [目的]为研究内生真菌与长春花悬浮细胞互作奠定基础.[方法]从长春花茎杆韧皮部分离出内生真菌,对分离出的菌株进行鉴定,用PDA培养基、PDA培养基+长春花悬浮细胞的水提物对内生真菌进行液体培养,薄板层析检测内生真菌液体培养物的提取物及发酵液.[结果]从常规PDA培养基中分离得到11株菌株,而从其他PDA培养基中没有分离到内生真菌.分离出的11株菌株的大型分生孢子呈纺锤形或镰刀形,其代谢产物通过薄层层析,没有检测到长春花生物碱.表明分离出的内生真菌本身不能直接合成长春花生物碱,而可能对生物碱的合成起促进作用.[结论]从长春花中分离得到的内生真菌属镰刀菌属.

  2. 长春花总生物碱的提取方法研究%On the Extraction Method of Total Alkaloids from Catharanthus Roseus

    王莹; 林长春

    2008-01-01

    采用酸水冷浸法、乙醇冷凝回流法提取长春花中的总生物碱,并通过沉淀反应、显色反应对提取出来的生物碱进行定性检测,通过紫外分光光度法测其相对含量.结果显示酸水冷浸法与乙醇冷凝回流法都能提取长春花中生物总碱,乙醇冷凝回流法比酸水冷浸法提取长春花生物碱含量高,效果更好.

  3. ВОЗДЕЙСТВИЕ ЦИТОКИНИНОВ НА РОСТОВЫЕ ХАРАКТЕРИСТИКИ И НАКОПЛЕНИЕ ФЕНОЛЬНЫХ СОЕДИНЕНИЙ В КАЛЛУСНОЙ КУЛЬТУРЕ CATHARANTHUS ROSEUS: аннотация к дипломной работе/ Кристина Сергеевна Петренко; БГУ, Биологический факультет, Кафедра клеточной биологии и биоинженерии растений; науч. рук. Ромашко С.Н.

    Петренко, Кристина Сергеевна

    2015-01-01

    В данной статье отражены методы экстракции, выделения и очистки алкалоидов индольного ряда из сухих листьев Catharanthus roseus. Целью исследований являлось выявить наиболее эффективный способ экстракции и очистки данных веществ. Показаны существенные отличия в составе выделяемых алкалоидов, полученных различными способами твердофазно – жидкостной и жидкостно – жидкостной экстракциями. Определены оптимальные подходы к выделению алкалоидов индольного ряда из листьев катарантуса розового....

  4. Optimizing the Extraction Process of Four Important Alkaloids from Catharanthus roseu by Ultrasonic Method%四种主要长春花生物碱超声波法提取工艺优化研究

    张静; 隆小华; 刘玲; 刘兆普

    2011-01-01

    In order to explore the efficient and low contaminated extraction process of vindoline, catharanthine, vincristine and vinblastine from Catharanthus roseus, comprehensive optimization of condition in ultrasonic-methanol method were studied. On the basis of single factor experiments, orthogonal test was carried out to study. The optimum parameters for obtaining vindoline and catharanthine were determined as follows:methanol as solvent was used for 180 min,which pH value was 4.0. The ratio of raw material to solvent was 1: 15 (w/v), the extraction temperature was 45 ℃, and extraction frequency was 59 KHz. The yield of vindoline and catharanthine in the conditions was 0.164% and 0.081% ,respectively.%为探索高效、低污染的长春花中文多灵、长春质碱、长春新碱和长春碱的提取工艺,进行了超声波-甲醇法提取工艺条件的优化研究.结果表明:在单因素试验基础上,通过正交试验表明,超声波-甲醇法提取文多灵和长春质碱的最佳工艺条件为:提取溶剂:甲醇(pH4.0);料液比:1∶15;提取温度:45℃,提取时间:180 min,提取频率:59 KHz;文多灵和长春质碱平均得率为0.16%,0.08%.

  5. (phoenicopterus roseus por intoxicación aguda por plomo

    D Romero

    2007-01-01

    Full Text Available La presencia de plomo en el medio ambiente es un hecho indiscutible. Ello es objeto de gran interés sanitario pues repercute tanto en la salud de los animales como en la del ser humano. El plumbismo es una intoxicación que puede afectar a cualquier especie animal. En el ámbito de la fauna silvestre se asocia principalmente a la actividad cinegética en los humedales y a la ingestión de plomo metálico por parte de las aves que frecuentan estos ecosistemas. En España cada año mueren decenas de miles de aves acuáticas como consecuencia de la ingesta de perdigones de plomo. En los individuos capturados enfermos se pueden observar signos clínicos, mientras que a partir de muestras de cadáveres se obtienen datos sobre niveles tisulares de plomo. Sin embargo la evaluación de lesiones anatomopatológicas en animales de vida silvestre es difícil de realizar por la dificultad de encontrar animales recién muertos o afectados de un cuadro agudo mortal. La reproducción de esta patología en animales de experimentación se suele realizar en cisnes, palomas o patos, pero no en flamencos, por lo que no es usual disponer de datos histológicos en esta especie. En este trabajo presentamos la clínica, analítica y estudio microscópico de diferentes tejidos de un flamenco común (Phoenicopterus roseus muerto por plumbismo.

  6. Análisis de iridoides y expresión de genes que codifican enzimas tempranas en la síntesis de alcaloides indol terpenoicos en Catharanthus roseus Analysis of iridoids content and expression studies of genes encoding early enzymes in the indol terpenoid biosynthesis pathway in Catharanthus roseus

    Palacios-Rojas Natalia; Leech Mark

    2004-01-01

    Los alcaloides indol terpenoicos (TIA) son metabolitos secundarios de importancia medicinal por sus propiedades como agentes anticancerígenos, entre otras. Sin embargo, su explotación en la industria farmacéutica se ha visto limitada, ya que la acumulación de estos compuestos en las plantas que los producen es mínima. Dichos alcaloides son biosintetizados por la vía del shikimato y de los terpenoides, los cuales proveen los precursores: secologanina y triptamina, respectivamente. La secologan...

  7. Analysis of iridoids content and expression studies of genes encoding early enzymes in the indol terpenoid biosynthesis pathway in Catharanthus roseus Análisis de iridoides y expresión de genes que codifican enzimas tempranas en la síntesis de alcaloides indol terpenoicos en Catharanthus roseus

    Leech Mark; Palacios-Rojas Natalia

    2004-01-01

    Terpenoid indole alkaloids (TIA) are of pharmaceutical importance, however the industrial use of these compouds is very limited because its accumulation is very low in plant tissues. TIA are derived f rom the shikimate and terpenoid pathways, which supply secologanin and tryptamine, the indole and iridoid moieties, respectively. Secololganin is a terpenoid which is belived to be synthesised the MEP pathway rather than by the acetate/mevalonic acid pathway. Secologanin is thought to be a limit...

  8. Metabolic costs of avian flight in relation to flight velocity : a study in Rose Coloured Starlings (Sturnus roseus, Linnaeus)

    Engel, S; Biebach, H; Visser, GH

    2006-01-01

    The metabolic costs of flight at a natural range of speeds were investigated in Rose Coloured Starlings (Sturnus roseus, Linnaeus) using doubly labelled water. Eight birds flew repeatedly and unrestrained for bouts of 6 h at speeds from 9 to 14 m s(-1) in a low-turbulence wind tunnel, corresponding

  9. The major carotenoid pigment of a psychrotrophic Micrococcus roseus strain: purification, structure, and interaction with synthetic membranes.

    Jagannadham, M. V.; Rao, V J; Shivaji, S.

    1991-01-01

    The major carotenoid pigment of a psychrotrophic Micrococcus roseus strain was purified to homogeneity from methanol extracts of dried cells by reverse-phase liquid chromatography and was designated P-3. On the basis of the UV-visible, infrared, mass, and 1H nuclear magnetic resonance spectra of P-3, it was identified as bisdehydro-beta-carotene-2-carboxylic acid. The pigment interacted with synthetic membranes of phosphatidylcholine and dimyristoyl phosphatidylcholine and stabilized the memb...

  10. Análisis de iridoides y expresión de genes que codifican enzimas tempranas en la síntesis de alcaloides indol terpenoicos en catharanthus roseus

    Palacios-Rojas, Natalia; Leech, Mark

    2007-01-01

    Los alcaloides indol terpenoicos (TIA) son metabolitos secundarios de importancia medicinal por sus propiedades como agentes anticancerígenos, entre otras. Sin embargo, su explotación en la industria farmacéutica se ha visto limitada, ya que la acumulación de estos compuestos en las plantas que los producen es mínima. Dichos alcaloides son biosintetizados por la vía del shikimato y de los terpenoides, los cuales proveen los precursores: secologanina y triptamina, respectivamente. La secologan...

  11. 不同来源长春花药材中文多灵的含量比较%HPLC determination of vindoline extracted from Catharanthus roseus of different sources

    周高丽; 曾令杰

    2009-01-01

    目的 建立长春花中文多灵的含量分析方法,比较不同来源长春花药材中文多灵的含量.方法 采用高效液相色谱法,以依利特C18柱为分析柱,甲醇:水(体积比64:36,以二乙胺和磷酸调pH11)为流动相,流速为1 mL/min;220 nm为检测波长.结果 红色长春花、白色长春花以及恩平、阳江、广州紫色长春花样品中文多灵含量分别是1.561、0.390、0.982、0.622、0.587 mg/g.结论 不同来源的长春花中文多灵的含量差异较大.

  12. Dynamic Variation of Vinblastine Content in Catharanthus roseus by Hand Harvest%人为采收后长春花中长春碱含量的动态变化

    王非; 李雷鸿; 孙家宝; 祖元刚

    2005-01-01

    采用超声提取和高效液相色谱法,对人为采收后的野生长春花中长春碱含量的动态变化进行了比较分析.分析结果表明:人为采收在干扰长春花生长发育的同时也影响到长春碱的合成与积累.采收后20 d与采收前相比,长春花各部位的生物量降低,长春花叶片和茎中的长春碱含量也有所降低.采收后20~40 d,长春花不同部位的生物量均呈上升趋势;采收后40~60 d,长春花各部位的生物量增加相对稳定.采收后40 d时,长春花根、叶片和茎中长春碱含量均显著高于采收前;采收后60 d时,长春花不同部位长春碱含量略低于采收前.人为采收后长春花单株长春碱总量也有所降低.长春碱的这种动态变化可能与其对外界干扰的拮抗作用有关.

  13. Induction and Manipulation of Alkaloid Biosynthesis in Catharanthus roseus by Elicitors%诱导子对长春花生物碱生物合成的诱导与调控

    贺丽虹; 赵淑娟; 胡之璧

    2008-01-01

    真菌诱导子、信号分子、植物生长调节物质等诱导子对长春花生物碱的生物合成具有调控作用.文章介绍了诱导子种类、诱导效果与调控机理等方面的研究进展.

  14. Advances in Research of Regulation and Controlling for Alkaloids Biosynthesis in Catharanthus Roseus%长春花生物碱合成途径调控机制研究进展

    伍春莲; 杨军; 陈卫英; 孙敏

    2010-01-01

    长春花是一种重要的药用植物,其中含多种抗癌生物碱,然而在植物体内的含量甚微.文章对长春花生物碱合成途径中上游、中游、下游3个方面进行分阶段阐述,在每一个阶段对影响生物碱合成的物质,特别是关键酶进行了详细的综述.最后,对长春花生物碱合成途径的复杂调控性、影响因素和人工改造提出了一些设想.

  15. Effects of NaCl on the growth and alkaloid content of Catharanthus roseus seedlings%盐胁迫对长春花幼苗生长和生物碱含量的影响

    王景艳; 刘兆普; 刘玲; 刘冲

    2008-01-01

    以NaCl浓度分别为0、50、100、150、200和250 mmol·L-1的1/2 Hoagland营养液处理长春花幼苗,7 d后测定其鲜质量、干质量、丙二醛(MDA)和叶绿素含量、色氨酸脱羧酶(TDC)和过氧化物酶(POD)活性等生理指标及文多灵、长春质碱、长春新碱和长春碱等生物碱含量.结果表明:NaCl显著地降低长春花幼苗的鲜质量和干质量,提高MDA含量;叶绿素含量在低盐浓度(50 mmol·L-1)下与对照相比差异不显著,在高于50 mmol·L-1时随NaCl浓度的增加而逐渐降低;在NaCl处理下,POD活性与对照相比显著上升;TDC活性在50 mmol·L-1 NaCl处理下活性最高,而后随盐浓度的增加逐渐降低;文多灵、长春质碱、长春新碱和长春碱含量都是在50 mmol·L-1NaCl处理下最高,分别为4.61、3.56、1.19和2.95 mg·g-1,并显著高于对照及其他处理.盐胁迫虽然在一定程度上抑制了长春花幼苗生长,但促进了其生物碱的代谢,提高了生物碱含量;50 mmol·L-1NaCl处理对长春花吲哚生物碱代谢的促进作用最大.

  16. 长春花化学成分和药理作用研究进展%Advances in studies on chemical constituents in Catharanthus roseus and their pharmacological activities

    高贤; 单淇; 辛宁; 侯文彬

    2011-01-01

    长春花属植物是原产于非洲东海岸的野生花卉,现已广泛分布于世界各地.长春花含有萜类生物碱、二聚吲哚生物碱、单吲哚生物碱等多种生物碱类活性成分和黄酮类成分.近年来长春花中单体成分或结构修饰物作为药物被广泛用于治疗多种癌症、糖尿病、高血压及何杰金氏病.综述了近年来对长春花生物碱类和药理活性的研究进展,为更好地开发、利用长春花提供参考.

  17. A study on the hairy root culture and antitumor alkaloids production of Catharanthus roseus%长春花毛状根培养及抗癌生物碱产生的研究

    孙敏; 曾建军

    2005-01-01

    目的:建立长春花毛状根转化体系,从毛状根中获得生物碱.方法:用发根农杆菌A4和R1000菌株分别感染长春花的不同外植体获得毛状根,并对毛状根的培养条件进行优化.结果:采用A4感染叶片,分别预、共培养2 d及添加100 mg·L-1的乙酰丁香酮(As)可得到最佳转化效果,毛状根的诱导率达86.25%.以蔗糖为碳源及以水解乳蛋白为氮源的1/2MS培养基为毛状根的最佳生长条件.检测结果表明,毛状根中的总生物碱含量高于长春花的原植株和愈伤组织.结论:可通过毛状根的培养来获得抗癌生物碱长春碱和长春新碱等.

  18. Variation in Intergenerational Contents of Alkaloids of Catharanthus roseus by Space Mutation%空间诱变长春花代际生物碱含量变异研究

    于景华; 贾雪莹; 张学科; 唐中华; 原树生; 祖元刚

    2010-01-01

    以经"实践八号"返回式卫星搭载进行空间诱变的长春花种子为材料,研究诱变后代(SP1)及筛选后代(sP2、SP3)长春花叶片文多灵、长春质碱、长春碱等3种吲哚类生物碱的含量变异.结果显示,与留地对照组相比,空间诱变的长春花SP1代形态、生殖等变异增强,叶片生物碱含量变幅加大,变异系数达到留地对照组的2.06倍;经过3轮以长春碱含量为目标的筛选,获得的SP3代4个株系叶片长春碱含量达到300μg·g-1以上,并为留地对照组的3倍以上,株系内变异系数低于15%.上述结果表明,空间环境导致了长春花种子的遗传变异,经过多代筛选,可培育出目的活性物质含量高的优质品种.

  19. Study on variety trends of flavonoids in Catharanthus roseus in greenhouse%温室内长春花总黄酮含量的动态变化研究

    马书荣; 曲笑岩; 祖元刚

    2006-01-01

    以长春花为试材,应用超声提取和分光光度法测定和分析了不同发育阶段长春花不同部位总黄酮含量的动态变化.结果表明:长春花不同时期各部位总黄酮含量由高到低的排列顺序是:果、叶、花、茎、根;不同叶位总黄酮含量是花叶大于果叶,果叶大于营养叶,不同枝位总黄酮含量是果枝大于叶枝,叶枝大于茎,形成由上到下的递减趋势;侧枝各部位总黄酮含量高于主枝,须根总黄酮含量高于主根,形成由外到内的递减趋势.

  20. Improving the Content of Terpenoid Indole Alkaloids in Catharanthus roseus by 60Coγ Radiation%60Coγ射线辐射诱变提高长春花生物碱含量

    赵剑; 李建国

    2009-01-01

    本实验采用长春花种子为材料,用60Coγ射线不同剂量处理,统计处理萌发率和成株率,并观察当代表型变异率和进行回归模型分析.结果分析60Coγ射线辐照长春花种子的半致死剂量平均为174 Gy,照射长春花种子的较适宜剂量为200 Gy.我们观察到长春花M,代生长初期有明显的叶形不对称、叶缘卷曲等致变现象,各种畸形叶出现频率与辐照剂量大小有关.通过辐照,我们得到长春花高杆变异、矮杆变异等变异植株,并利用HPLC检测发现高杆变异株型长春花生物碱含量比未照射的CK多出24%.

  1. 广东地区不同长春花品种的遗传多样性分析%ISSR analysis on genetic diversity of Catharanthus roseus cultivars cultivated in Guangdong

    石林; 何丽贞; 李苏全

    2013-01-01

    利用ISSR分子标记技术对广东地区栽培的25个长春花品种进行遗传多样性分析.结果表明,供试的15条引物共扩增出110条清晰的条带,平均每条引物扩增出7.33条;多态性条带为69条,多态性比率(PPB)为62.7%,说明25个长春花品种间存在较丰富的遗传变异,ISSR标记可用于长春花的遗传多样性分析.品种间的遗传相似系数介于0.69~0.94之间.UPGMA聚类分析将25个长春花品种分成2组,该聚类结果表明同一系列不同颜色的品种具有较高的遗传相似性,各个商业品系的品种间存在广泛的杂交或具有相同的原始育种材料;花色遗传在分类学上的地位比花眼花色遗传重要;偏蓝色系、白色系均与其他色系相比遗传距离较远.

  2. 一株产长春新碱内生真菌的初步研究%Preliminary study of a vincristine-producing endophytic fungus isolated from leaves of Catharanthus roseus

    杨显志; 张玲琪; 郭波; 郭仕平

    2004-01-01

    目的通过长春花叶内生真菌的分离,筛选产生长春新碱的菌株.方法从长春花叶中分离内生真菌,对其发酵提取物进行TLC和HPLC分析.结果从长春花叶中分离筛选到一株无孢菌群菌株:97CY3.它能产生长春新碱,HPLC测定其长春新碱含量约为0.205 μg/L.结论长春花内生真菌中,有的菌株可产生与宿主所产相同的抗癌物质长春新碱.

  3. Advances on Effects of Cultural Factors on Alkaloid Concentration of Catharanthus roseus%栽培因子影响长春花生物碱含量的研究进展

    陈泉; 李建国

    2009-01-01

    长春花为重要的抗癌药用植物,综述长春花栽培过程中,土壤水份、温度、光照等环境因子和施肥、喷施外源激素、喷施生物碱合成前体物质、刈割、采收等人为因子对长春花药用生物碱含量的影响.

  4. Effects of domestic sewage effluents on the growth and physiological characteristics of Catharanthus roseus%生活污水对长春花生长及生理影响的研究

    胡宏友; 卢昌义; 叶勇; 陈登雄

    2005-01-01

    利用不同稀释倍数的生活污水(污水∶清水,V/V,分别为1∶0;1∶1;1∶2;0∶1)浇灌长春花结果表明,与清水灌溉相比,污灌处理土壤NH +4 -N含量由59.01mg / kg提高到64.22~65.89mg / kg,NO-3 -N含量由2.69mg / kg提高到3.03~4.54mg / kg,速效磷含量由32.29mg / kg提高到37.40~42.41mg / kg;增加了土壤酶活性,其中土壤过氧化氢酶、脲酶及蛋白酶活性分别比对照提高10.76%~23.32%、33.41%~103.69%和31.82%~131.82%;植物根系活性由170.25μg / g*h提高到177.88~471.69μg / g*h,叶绿素含量由1.78mg / dm2提高到2.68~2.98mg / dm2,植物干物质量由11.660g / 盆提高到12.130~16.577g / 盆.且以稀释1倍污水灌溉处理效果最佳.用原生生活污水直接灌溉虽降低污水肥效,造成土壤Cl-的积累,但对长春花生长及干物质积累无显著影响.

  5. 长春花转化毛状根诱导及培养条件的优化%Induction and Culture of Transformed Hairy Root in Catharanthus Roseus

    孙敏; 汪洪; 王颖; 伍春莲

    2002-01-01

    用A4和R10002种发根农杆菌菌株分别感染长春花的愈伤组织、真叶叶片、叶柄及根,诱导出毛状根.研究了不同外植体、不同菌株以及外植体预培养时间、与发根农杆菌共培养时间、乙酰丁香酮(As)等条件对转化频率的影响,其结果为:A4转化频率高于R1000,真叶叶片转化频率最高,最佳预培养时间为2~3 d;最佳共培养时间2~3 d;以100 mg/mL的As同时加入共培养基中转化频率高达86.25%.不同种类的培养基以及碳源、氮源对毛状根生长也有较大的影响.

  6. Standard Operational Procedure (SOP) for Breeding Original Seeds of Medicinal Catharanthus roseus%药用作物长春花原种生产标准操作规程(SOP)

    袁媛; 庞玉新; 张影波; 王祝年

    2009-01-01

    根据(GAP)的指导原则,对药用作物长春花原种生产的生产区划、生态环境、生产技术、采收加工、贮藏、质量分级、包装运输、文件和档案管理进行了研究和规定,制定了药用作物长春花原种生产标准操作规程(SOP),这对于保证药用长春花优良品种的纯度和优良种性,提供足够数量的原种或大田用种,促进中药材规范化生产具有重要意义.

  7. Dicty_cDB: Contig-U14930-1 [Dicty_cDB

    Full Text Available ful extensions: 7805 Number of sequences better than 10.0: 1165 length of query: 1570 length of database: 8,...ns: 13632520 Number of sequences better than 10.0: 266 Length of query: 1570 Length of databa...pid:none) Medicago truncatula clone mth2-17k... 74 2e-11 FJ647194_1( FJ647194 |pid:none) Catharanthus roseus tabersoni...72 7e-11 EU541505_1( EU541505 |pid:none) Catharanthus roseus cytochrome P45... 72...92573_1( AY192573 |pid:none) Catharanthus roseus cytochrome P45... 72 7e-11 BT059557_1( BT059557 |pid:none)

  8. Pedobacter roseus sp. nov., isolated from a hypertrophic pond, and emended description of the genus Pedobacter.

    Hwang, Chung Yeon; Choi, Dong Han; Cho, Byung Cheol

    2006-08-01

    A Gram-negative, pink-coloured, rod-shaped, non-flagellated bacterium, designated CL-GP80(T), was isolated from a hypertrophic pond located within the campus of Seoul National University, Korea. Analysis of its 16S rRNA gene sequence revealed that strain CL-GP80(T) belongs to the family Sphingobacteriaceae and is closely related to Pedobacter heparinus ATCC 13125(T) (95.8 % sequence similarity) and to other members of the genus Pedobacter (90.8-95.3 % similarity). Temperature and pH ranges for growth were 5-33 degrees C and pH 6-8, respectively. The DNA G+C content was 41.3 mol%. The major fatty acids were iso-C(15 : 0) (37.0 %), iso-C(15 : 0) 2-OH and/or C(16 : 1)omega7c (24.5 %), and iso-C(17 : 0) 3-OH (11.3 %). Phenotypic, chemotaxonomic and phylogenetic analyses indicated that strain CL-GP80(T) could be assigned to the genus Pedobacter, but distinguished from recognized species of the genus. Strain CL-GP80(T) (=KCCM 42272(T)=JCM 13399(T)) is therefore proposed as the type strain of a novel species, for which the name Pedobacter roseus sp. nov. is proposed. PMID:16902016

  9. Stable and High Ajmalicine or Serpentine Production of Gamma Radiation Induction Mutant Catharantus Roseus

    Catharantus roseus Mutant have been selected by gamma irradiation with 20 krad doses of radiation and characterized as biochemical mutant with anti-feed back inhibition mechanism of tritophan decarboxylase (TDR) enzyme in biosynthetic path way of indole alkaloid. Production of indole alkaloid mainly ajmalicine with high economical values as a pharmaceutical drug for heart attack have been studied by using cell suspension cultures with several variation of medium, elicitors and stress osmosis. This treatment produced variation of indole alkaloid ajmalicine and serpentine. Several induction methods using Murashige and Skoog (MS) medium and polyethylene glycol PEG (6000) 1 to 7%, with hormones concentration of 2,4-D and kinetin as (10 : 1), showed optimal results of ajmalicine range between 20 and 50 nmol/gFW, and serpentine 10 to 60 nmol/gFW. This production increases ten time in mutant (20 Krad) by stress osmotic condition and performed long term stability in culture without subculture. In this paper explanation in detail about the selection methods, stability of mutant and the production of indole alkaloid ajmalicine and serpentine during growth phase, such as adaptation, log, and stationar in suspention culture of mutan cells. (author)

  10. Gamma irradiation effect on the formation of Clonal variation from catharantus roseus plant

    Clonal variation have been found in Catharantus roseus plant after gamma irradiation. Several doses have been used to produce clonal variation. The most effective doses used to perform better clonal variation was 20 krad. About 103 seeds irradiated for every radiation treatment, but only several clones were grown better than wild type. We have success to get (M) seeds the expected mutant. The seeds from selected mutant are bigger when compare to the wild type and growth better on medium containing 5-methyl Tryptophan (5-MT). The chlorophyll content is higher (almost twice) as compared to the wild type. Fulther experiment continue to do in vitro culture in order to develop embryonic callus from leaf tip and leaf base. Several manipulation of auxin and cytokini have been used to differentiate the callus formation. Modified MS medium with kinetin and cytokinin (10:1) can induce globular embryo like structure. Dragendrof alkaloid reagent were used to determine high alkaloid clones from the expected mutant. TLC analysis from callus mutant shows 3 clear bands with subsequence Rf about 0.22, 0.58 while control shows two smearing bands at 0.21 and 0.52

  11. Inactivation of cephapirin sodium by the radiation-resistant strain micrococcus roseus

    The susceptibility of the radioresistant mutants B. firmus, B.megaterium, B, laterosporus, M. roseus and M. luteus to the betalactam antibiotic cephapirin sodium was estimated using the microbiological assay technique. All the studied species were found to be sensitive to the concerned antibiotic except the radioresistant mutant M. rosues. Accordingly, the inactivation of betalactam, antibiotic cephapirin sodium, by this mutant strain was interesting to be investigated. A microbiological assay was used to determine the potency of the studied antibiotic and its degraded compound produced after its incubation with the above mentioned mutant strain for different periods of time in basal salt mineral medium.Results obtained for antibiotic samples extracted after 7-day incubation with the mutant strain indicated that the antibiotic was metabolized by this mutant strain to inactive products. These results were confirmed by chromatograms of the antibiotic samples, extracted from cultures with the mutant incubated for zero, 7 and 14 days. Degraded products were eluted at retention time values different from those observed for the noninucubated antibiotic samples. The inactivation of the antibiotic by the studied mutant starin seems to be due to extracellular enzymes in the surrounding medium.1 tab

  12. Geminicoccus roseus gen. nov., sp. nov., an aerobic phototrophic Alphaproteobacterium isolated from a marine aquaculture biofilter.

    Foesel, Bärbel U; Gössner, Anita S; Drake, Harold L; Schramm, Andreas

    2007-12-01

    A Gram-negative, strictly aerobic, diplococcoid bacterium (strain D2-3(T)) was isolated from the biofilter of a recirculating marine aquaculture system. Phylogenetic analysis of the 16S rRNA gene sequence of D2-3(T) indicated that the new organism occupied a novel lineage within the alpha-1 subclass of Proteobacteria and was related to the genera Rhodothalassium, Azospirillum, Craurococcus, Acidiphilium, and Tistrella. The highest sequence similarity (90.8%) of the 16S rRNA gene sequence of D2-3(T) was to that of Candidatus "Alysiosphaera europaea". D2-3(T) was mesophilic, heterotrophic, required sea salt, and had a pH optimum of 8.0. Growth in the presence of light resulted in the formation of pink colonies, a 25% increased cell yield, and a slightly increased growth rate. D2-3(T) contained carotenoids and low amounts of bacteriochlorophyll a. Membranes of D2-3(T) contained b-type cytochromes. The G+C content of the DNA was 60.3+/-0.1mol%. Phylogenetic, morphological, physiological, and biochemical analyses demonstrated that D2-3(T) represented a new aerobic phototrophic genus, for which the name Geminicoccus roseus gen. nov., sp. nov. is proposed for the type species (D2-3(T)=DSM 18922(T)=ATCC BAA-1445(T)). PMID:17643894

  13. Meiothermus roseus sp. nov., a thermophilic bacterium isolated from a geothermal area.

    Ming, Hong; Duan, Yan-Yan; Guo, Qian-Qian; Yin, Yi-Rui; Zhou, En-Min; Liu, Lan; Li, Shuai; Nie, Guo-Xing; Li, Wen-Jun

    2015-10-01

    Two closely related thermophilic bacterial strains, designated YIM 71031(T) and YIM 71039, were isolated from a hot spring in Tengchong county, Yunnan province, south-western China. The novel isolates were observed to be Gram-negative, aerobic, rod-shaped and yellow-pigmented bacteria. The strains were found to be able to grow at 37-65 °C, pH 6.0-9.0 and with a NaCl tolerance up to 1.0 % (w/v). Phylogenetic analysis based on 16S rRNA gene sequences placed these two isolates in the genus Meiothermus. They were found to be closely related to Meiothermus timidus DSM 17022(T) (98.6 % similarity), and formed a cluster with this species. The predominant menaquinone was identified as MK-8 and the major fatty acids (>10 %) as anteiso-C15:0, iso-C15:0, anteiso-C17:0, iso-C16:0 and C16:0. The genomic DNA G+C contents of strains YIM 71031(T) and YIM 71039 were determined to be 64.0 and 65.4 mol%, respectively. DNA-DNA hybridizations showed low values between strains YIM 71031(T) and YIM 71039 and their closely related neighbour M. timidus DSM 17022(T). Morphological phylogenetic and chemotaxonomic results suggest that strains YIM 71031(T) and YIM 71039 are representatives of a new species within the genus Meiothermus, for which the name Meiothermus roseus sp. nov. is proposed. The type strain is YIM 71031(T) (=KCTC 42495(T) =NBRC 110900(T)). PMID:26219565

  14. Kõige tõsisem taim vähktõve vastu : [roosa lõunahali

    2011-01-01

    Roosa lõunahali (Catharanthus roseus (L.) G. Don. Sünonüümid: Ammocallis rosea (L.) Small, Lochnera rosea (L.) Rchb., Vinca rosea L. Kasutamisest rahvameditsiinis ja farmakoteraapias (ravimpreparaadid vinblastiin, vinkristiin)

  15. Structural determinants of reductive terpene cyclization in iridoid biosynthesis

    Kries, Hajo; Caputi, Lorenzo; Stevenson, Clare E M;

    2016-01-01

    The carbon skeleton of ecologically and pharmacologically important iridoid monoterpenes is formed in a reductive cyclization reaction unrelated to canonical terpene cyclization. Here we report the crystal structure of the recently discovered iridoid cyclase (from Catharanthus roseus) bound to a...

  16. Effect of composites based nickel foam anode in microbial fuel cell using Acetobacter aceti and Gluconobacter roseus as a biocatalysts.

    Karthikeyan, Rengasamy; Krishnaraj, Navanietha; Selvam, Ammaiyappan; Wong, Jonathan Woon-Chung; Lee, Patrick K H; Leung, Michael K H; Berchmans, Sheela

    2016-10-01

    This study explores the use of materials such as chitosan (chit), polyaniline (PANI) and titanium carbide (TC) as anode materials for microbial fuel cells. Nickel foam (NF) was used as the base anode substrate. Four different types of anodes (NF, NF/PANI, NF/PANI/TC, NF/PANI/TC/Chit) are thus prepared and used in batch type microbial fuel cells operated with a mixed consortium of Acetobacter aceti and Gluconobacter roseus as the biocatalysts and bad wine as a feedstock. A maximum power density of 18.8Wm(-3) (≈2.3 times higher than NF) was obtained in the case of the anode modified with a composite of PANI/TC/Chit. The MFCs running under a constant external resistance of (50Ω) yielded 14.7% coulombic efficiency with a maximum chemical oxygen demand (COD) removal of 87-93%. The overall results suggest that the catalytic materials embedded in the chitosan matrix show the best performance and have potentials for further development. PMID:26970695

  17. 产新型类胡萝卜素微球菌的鉴定%Identification of New Carotenoids-Producing Micrococcus roseus

    袁婀娜; 石贵阳; 蔡宇杰

    2005-01-01

    从野外土壤中分离得到1株产类胡萝卜素菌株.根据其形态特征、生理生化特征的测定结果分析,该菌株应归为微球菌属(Micrococcus)玫瑰色微球菌(Micrococcus roseus).对其所产类胡萝卜素进行了吸收光谱扫描及高效液相分析,研究表明为新型类胡萝卜素.

  18. High quality draft genome sequence and analysis of Pontibacter roseus type strain SRC-1T (DSM 17521T) isolated from muddy waters of a drainage system in Chandigarh, India

    Mukherjee, Supratim; Lapidus, Alla; Shapiro, Nicole; Cheng, Jan-Fang; Han, James; Reddy, T. B.K.; Huntemann, Marcel; Ivanova, Natalia; Mikhailova, Natalia; Chen, Amy; Palaniappan, Krishna; Spring, Stefan; G& #246; ker, Markus; Woyke, Tanja; Tindall, Brian J.; Klenk, Hans-Peter; Pati, Amrita

    2014-11-26

    Pontibacter roseus Suresh et al 2006 is a member of genus Pontibacter family Cytophagaceae, class Cytophagia. While the type species of the genus Pontibacter actiniarum was isolated in 2005 from a marine environment, subsequent species of the same genus have been found in different types of habitats ranging from seawater, sediment, desert soil, rhizosphere, contaminated sites, solar saltern and muddy water. Here we describe the features of Pontibacter roseus strain SRC-1T along with its complete genome sequence and annotation from a culture of DSM 17521T. The 4,581,480 bp long draft genome consists of 12 scaffolds with 4,003 protein-coding and 50 RNA genes and is a part of Genomic encyclopedia of Type Strains, Phase I: the one thousand microbial genomes (KMG-I) project.

  19. Metabolic reprogramming of periwinkle plant culture

    Runguphan, Weerawat; O’Connor, Sarah E.

    2009-01-01

    We transformed an alkaloid biosynthetic gene with reengineered substrate specificity into Catharanthus roseus. The resulting transgenic plant cell culture produced a variety of unnatural alkaloid compounds when cocultured with simple, achiral, commercially available precursors that the reengineered enzyme was designed to accept. This work demonstrates the power of genetic engineering to retailor the structures of complex alkaloid natural products in plant culture.

  20. Seed transmission of Candidatus Liberibacter asiaticus in periwinkle and dodder resulted in low bacterial titer and very mild disease in periwinkle

    Canadidatus Liberibacter asiaticus (Las) is the most widely-distributed of three species of Liberibacter that are associated with citrus Huanglongbing (HLB), a lethal disease of citrus worldwide. In addition to citrus, periwinkle (Catharanthus roseus) and dodder (Cuscuta pentagona) are two experime...

  1. UniProt search blastx result: AK287483 [KOME

    Full Text Available AK287483 J043025M01 Q8W013|COMT 1_CATRO Caffeic acid 3-O-methyltransferase (EC 2.1.1.68) (S-adeno ... -L-methionine:caffeic acid 3-O-methyltransferase) (COMT ) (CAOMT) - Catharanthus roseus (Rosy periwinkle) ( ...

  2. An assay for secologanin in plant tissues based on enzymatic conversion into strictosidine

    Hallard, Didier; van der Heijden, Robert; Contin, Adriana;

    1998-01-01

    strictosidine, a reaction catalysed by the enzyme strictosidine synthase (STR; E.C. 4.3.3.2). Subsequently, the formation of strictosidine is quantified by HPLC. STR was isolated from transgenic Nicotiana tabacum cells expressing a cDNA-derived gene coding for STR from Catharanthus roseus. The high specificity...

  3. Molecular docking and pharmacogenomics of alkaloids and their monomeric precursors; vindoline and catharanthine

    Sertel, Serkan; Fu, Yujie; Zu, Yuangang; Rebacz, Blanka; Konkimalla, Badireenath; Plinkert, Peter K.; Krämer, Alwin; Gertsch, Jürg; Efferth, Thomas

    2011-01-01

    Abstract Vinblastine and vincristine are dimeric indole alkaloids derived from Catharanthus roseus (formerly: Vinca rosea). Their monomeric precursor molecules are vindoline and catharanthine. While vinblastine and vincristine are well-known mitotic spindle poisons, not much is known about vindoline and catharanthine. Vindoline and catharanthine showed weak cytotoxicity, while vinblastine, vincristine, and the semisynthetic vindesine and vinorelbine revealed high cytotoxicity towar...

  4. Understanding CrRLK1L Function

    Nissen, Karen Stoltenberg; Willats, William George Tycho; Malinovsky, Frederikke Gro

    2016-01-01

    To develop successfully in an ever-changing environment, it is essential for plants to monitor and control their growth. Therefore, cell expansion is carefully regulated to establish correct cell shape and size. In this review, we explore the role of the Catharanthus roseus receptor-like kinase (Cr...

  5. Receptor-like kinase complexes in plant innate immunity

    Greeff, Michael Christiaan; Roux, Milena Edna; Mundy, John;

    2012-01-01

    domain defines ligand specificity, and RLK families are sub-classed according to this domain. The most studied of these subfamilies include those with (1) leucine-rich repeat (LRR) domains, (2) LysM domains (LYM), and (3) the Catharanthus roseus RLK1-like (CrRLK1L) domain. These proteins recognize...

  6. Exogenous Abscisic Acid and Gibberellic Acid Elicit Opposing Effects on Fusarium graminearum Infection in Wheat.

    Buhrow, Leann M; Cram, Dustin; Tulpan, Dan; Foroud, Nora A; Loewen, Michele C

    2016-09-01

    Although the roles of salicylate (SA) and jasmonic acid (JA) have been well-characterized in Fusarium head blight (FHB)-infected cereals, the roles of other phytohormones remain more ambiguous. Here, the association between an array of phytohormones and FHB pathogenesis in wheat is investigated. Comprehensive profiling of endogenous hormones demonstrated altered cytokinin, gibberellic acid (GA), and JA metabolism in a FHB-resistant cultivar, whereas challenge by Fusarium graminearum increased abscisic acid (ABA), JA, and SA in both FHB-susceptible and -resistant cultivars. Subsequent investigation of ABA or GA coapplication with fungal challenge increased and decreased FHB spread, respectively. These phytohormones-induced effects may be attributed to alteration of the F. graminearum transcriptome because ABA promoted expression of early-infection genes, including hydrolases and cytoskeletal reorganization genes, while GA suppressed nitrogen metabolic gene expression. Neither ABA nor GA elicited significant effects on F. graminearum fungal growth or sporulation in axenic conditions, nor do these phytohormones affect trichothecene gene expression, deoxynivalenol mycotoxin accumulation, or SA/JA biosynthesis in F. graminearum-challenged wheat spikes. Finally, the combined application of GA and paclobutrazol, a Fusarium fungicide, provided additive effects on reducing FHB severity, highlighting the potential for combining fungicidal agents with select phytohormone-related treatments for management of FHB infection in wheat. PMID:27135677

  7. Current Population Status and Activity Pattern of Lesser Flamingos (Phoeniconaias minor and Greater Flamingo (Phoenicopterus roseus in Abijata-Shalla Lakes National Park (ASLNP, Ethiopia

    Tewodros Kumssa

    2014-01-01

    Full Text Available A study of the population status, habitat preference, and activity pattern of nonbreeding flamingos was carried out in Lakes Abijata, Shalla, and Chitu, part of the Great Rift Valley, Ethiopia, from 2011 to 2013. The current population status and habitat preference of flamingos in the area are still poorly known. Likewise, data on diurnal and seasonal activity pattern of the species are scarce and this leads to the misunderstanding of how Flamingos use local wetlands throughout the different seasons. Data regarding population size and activity pattern were gathered during the wet and dry seasons. Point-count method was used to estimate the population size. Behaviors were recorded using scan sampling techniques. A total of 53671 individuals representing two species of flamingo were counted during both wet and dry seasons from the three lakes. There were more flamingos during the dry season than the wet season in Lake Abijata contrary to Lakes Shalla and Chitu during the wet season. Lesser flamingos (Phoeniconaias minor were the most abundant species comprising 95.39%, while Greater Flamingos (Phoenicopterus roseus accounted for 4.61% of the total population. Lake Abijata is the major stronghold of Lesser Flamingos in the area. There was significant variation in the mean number of both species during the wet and dry season in the different study sites of the lake, respectively. The species were known to use varied habitats within the lakes. The Lesser Flamingo mainly preferred the shoreline and mudflat areas of the lakes. However, Greater Flamingo on several occasions showed preference to offshore area of the lakes. Seasonal average flock sizes were not similar between the species. There was a strong relationship between time allocated to each activity and time of day. Feeding activity varied among daylight hours and was higher in the evening (76.5% and late morning (74.56% and least during midday (54%. Some variations in activity breakdown were

  8. Evaluación del uso de señales visuales y de localización por el colibrí cola-ancha (Selasphorus platycercus al visitar flores de Penstemon roseus Evaluation of the use of visual and location cues by the Broad-tailed hummingbird (Selasphorus platycercusforaging in flowers of Penstemon roseus

    Guillermo Pérez

    2012-03-01

    Full Text Available En los colibríes la memoria espacial desempeña un papel importante durante el forrajeo. Éste se basa en el uso de señales específicas (visuales o en señales espaciales (localización de flores y plantas con néctar. Sin embargo, el uso de estas señales por los colibríes puede variar de acuerdo con la escala espacial que enfrentan cuando visitan flores de una o más plantas durante el forrajeo; ésto se puso a prueba con individuos del colibrí cola-ancha Selasphorus platycercus. Por otro lado, para evaluar la posible variación en el uso de las señales, se llevaron a cabo experimentos en condiciones semi-naturales utilizando flores de la planta Penstemon roseus, nativa del sitio de estudio. A través de la manipulación de la presencia/ausencia de una recompensa (néctar y señales visuales, evaluamos el uso de la memoria espacial durante el forrajeo entre 2 plantas (experimento 1 y dentro de una sola planta (experimento 2. Los resultados demostraron que los colibríes utilizaron la memoria de localización de la planta de cuyas flores obtuvieron recompensa, independientemente de la presencia de señales visuales. Por el contrario, en flores individuales de una sola planta, después de un corto periodo de aprendizaje los colibríes pueden utilizar las señales visuales para guiar su forrajeo y discriminar las flores sin recompensa. Asimismo, en ausencia de señales visuales los individuos basaron su forrajeo en la memoria de localización de la flor con recompensa visitada previamente. Estos resultados sugieren plasticidad en el comportamiento de forrajeo de los colibríes influenciada por la escala espacial y por la información adquirida en visitas previas.In hummingbirds spatial memory plays an important role during foraging. It is based in use of specific cues (visual or spatial cues (location of flowers and plants with nectar. However, use of these cues by hummingbirds may change according to the spatial scale they face when visit

  9. Synthesis of Xylooligosaccharides of Daidzein and Their Anti-Oxidant and Anti-Allergic Activities

    Hatsuyuki Hamada; Kei Shimoda; Hiroki Hamada

    2011-01-01

    The biocatalytic synthesis of xylooligosaccharides of daidzein was investigated using cultured cells of Catharanthus roseus and Aspergillus sp. β-xylosidase. The cultured cells of C. roseus converted daidzein into its 4′-O-β-glucoside, 7-O-β-glucoside, and 7-O-β-primeveroside, which was a new compound. The 7-O-β-primeveroside of daidzein was further xylosylated by Aspergillus sp. β-xylosidase to daidzein trisaccharide, i.e., 7-O-[6-O-(4-O-(β-d-xylopyranosyl))-β-d-xylopyranosyl]-β-d-glucopyran...

  10. Phosphatidylinositol species of suspension cultured plant cells

    Heim, S.; Wagner, K.G.

    Suspension cultured Nicotiana tabacum and Catharanthus roseus cells were labeled with (/sup 3/H)inositol, the phospholipid fraction extracted and separated by thin layer chromatography. Three different solvent systems and reference compounds were used to assign the different /sup 3/H-labeled species by autoradiography. The ratio of (/sup 3/H)inositol incorporation into PI, PIP and PIP/sub 2/ was found to be 95:4:1; with some preparations a lyso-PI band was obtained which incorporated about a tenth of the label of the PIP band. With Catharanthus roseus cells a very faint band between PI and lyso-PI was detected which could not be assigned to a reference compound.

  11. Dicty_cDB: Contig-U00527-1 [Dicty_cDB

    Full Text Available ions: 2109 Number of sequences better than 10.0: 256 length of query: 820 length of database: 8,075,542 effective HSP length...49264 ) RecName: Full=Cytochrome P450 71B1; EC=1.14.-.... 75 2e-17 AJ251269_1( AJ251269 |pid:none) Catharanth...ytochro... 80 3e-17 AY192573_1( AY192573 |pid:none) Catharanthus roseus cytochrome P45... 73 3e-17 FM208178_...8 5e-17 EU541505_1( EU541505 |pid:none) Catharanthus roseus cytochrome P45... 72 5e-17 BT065841_1( BT065841 ...1( AY064051 |pid:none) Arabidopsis thaliana putative cyto... 70 2e-16 AP008209_913( AP008209 |pid:none) Oryza sativa (japoni

  12. AcEST: DK958562 [AcEST

    Full Text Available ... 292 4e-92 sp|Q9ZRS4|CHSY_CATRO Chalcone synthase OS=Catharanthus roseus GN... 294 8e-92 sp|Q9FUB7|CHSY_HYPAN Chalcone syntha...se 1 OS=Cicer arietinum GN=C... 283 1e-88 sp|P23569|CHSY_PUELO Chalcone synthase OS=Pueraria lobat...lcone synthase OS=Catharanthus roseus GN=CHS PE=2 SV=1 Length = 389 Score = 294 bits (7...se OS=Senna ... 293 6e-91 tr|Q2WFX2|Q2WFX2_IRIGE Chalcone synthase OS=Iris germani...se OS=Hypericum... 289 3e-89 tr|B7SQE0|B7SQE0_ABEMA Chalcone synthase OS=Abelmoschus manihot ... 289 3e-

  13. Triple subcellular targeting of isopentenyl diphosphate isomerases encoded by a single gene

    Guirimand, Grégory; Guihur, Anthony; Phillips, Michael A.; Oudin, Audrey; Glévarec, Gaëlle; Mahroug, Samira; Melin, Céline; Papon, Nicolas; Clastre, Marc; Giglioli-Guivarc'h, Nathalie; St-Pierre, Benoit; Rodríguez-Concepción, Manuel; Burlat, Vincent; Courdavault, Vincent

    2012-01-01

    Isopentenyl diphosphate isomerase (IDI) is a key enzyme of the isoprenoid pathway, catalyzing the interconversion of isopentenyl diphosphate and dimethylallyl diphosphate, the universal precursors of all isoprenoids. In plants, several subcellular compartments, including cytosol/ER, peroxisomes, mitochondria and plastids, are involved in isoprenoid biosynthesis. Here, we report on the unique triple targeting of two Catharanthus roseus IDI isoforms encoded by a single gene (CrIDI1). The triple...

  14. Structural determinants of reductive terpene cyclization in iridoid biosynthesis.

    Kries, Hajo; Caputi, Lorenzo; Stevenson, Clare E M; Kamileen, Mohammed O; Sherden, Nathaniel H; Geu-Flores, Fernando; Lawson, David M; O'Connor, Sarah E

    2016-01-01

    The carbon skeleton of ecologically and pharmacologically important iridoid monoterpenes is formed in a reductive cyclization reaction unrelated to canonical terpene cyclization. Here we report the crystal structure of the recently discovered iridoid cyclase (from Catharanthus roseus) bound to a mechanism-inspired inhibitor that illuminates substrate binding and catalytic function of the enzyme. Key features that distinguish iridoid synthase from its close homolog progesterone 5β-reductase are highlighted. PMID:26551396

  15. 盆栽长春花

    载宇光

    2003-01-01

    长春花(Catharanthus roseus)又名日日春,雁来红,四时花等,为夹竹桃科长春花属植物。原产于南亚,非洲东南部及美洲热带,我国长江以南各地均有栽培,长春花在原产地呈亚灌木状多年生草

  16. Evaluation of the Larvicidal Efficacy of Five Indigenous Weeds against an Indian Strain of Dengue Vector, Aedes aegypti L. (Diptera: Culicidae)

    Aarti Sharma; Sarita Kumar; Pushplata Tripathi

    2016-01-01

    Background and Objectives. Aedes aegypti, dengue fever mosquito, is primarily associated with the transmission of dengue and chikungunya in tropical and subtropical regions of the world. The present investigations were carried out to assess the larvicidal efficiency of five indigenous weeds against Ae. aegypti. Methods. The 1,000 ppm hexane and ethanol extracts prepared from the leaves and stem of five plants (Achyranthes aspera, Cassia occidentalis, Catharanthus roseus, Lantana camara, and X...

  17. L-Amino Acids Elicit Diverse Response Patterns in Taste Sensory Cells: A Role for Multiple Receptors.

    Shreoshi Pal Choudhuri

    Full Text Available Umami, the fifth basic taste, is elicited by the L-amino acid, glutamate. A unique characteristic of umami taste is the response potentiation by 5' ribonucleotide monophosphates, which are also capable of eliciting an umami taste. Initial reports using human embryonic kidney (HEK cells suggested that there is one broadly tuned receptor heterodimer, T1r1+T1r3, which detects L-glutamate and all other L-amino acids. However, there is growing evidence that multiple receptors detect glutamate in the oral cavity. While much is understood about glutamate transduction, the mechanisms for detecting the tastes of other L-amino acids are less well understood. We used calcium imaging of isolated taste sensory cells and taste cell clusters from the circumvallate and foliate papillae of C57BL/6J and T1r3 knockout mice to determine if other receptors might also be involved in detection of L-amino acids. Ratiometric imaging with Fura-2 was used to study calcium responses to monopotassium L-glutamate, L-serine, L-arginine, and L-glutamine, with and without inosine 5' monophosphate (IMP. The results of these experiments showed that the response patterns elicited by L-amino acids varied significantly across taste sensory cells. L-amino acids other than glutamate also elicited synergistic responses in a subset of taste sensory cells. Along with its role in synergism, IMP alone elicited a response in a large number of taste sensory cells. Our data indicate that synergistic and non-synergistic responses to L-amino acids and IMP are mediated by multiple receptors or possibly a receptor complex.

  18. Isolation of Cells Specialized in Anticancer Alkaloid Metabolism by Fluorescence-Activated Cell Sorting.

    Carqueijeiro, Inês; Guimarães, Ana Luísa; Bettencourt, Sara; Martínez-Cortés, Teresa; Guedes, Joana G; Gardner, Rui; Lopes, Telma; Andrade, Cláudia; Bispo, Cláudia; Martins, Nuno Pimpão; Andrade, Paula; Valentão, Patrícia; Valente, Inês M; Rodrigues, José A; Duarte, Patrícia; Sottomayor, Mariana

    2016-08-01

    Plant specialized metabolism often presents a complex cell-specific compartmentation essential to accomplish the biosynthesis of valuable plant natural products. Hence, the disclosure and potential manipulation of such pathways may depend on the capacity to isolate and characterize specific cell types. Catharanthus roseus is the source of several medicinal terpenoid indole alkaloids, including the low-level anticancer vinblastine and vincristine, for which the late biosynthetic steps occur in specialized mesophyll cells called idioblasts. Here, the optical, fluorescence, and alkaloid-accumulating properties of C. roseus leaf idioblasts are characterized, and a methodology for the isolation of idioblast protoplasts by fluorescence-activated cell sorting is established, taking advantage of the distinctive autofluorescence of these cells. This achievement represents a crucial step for the development of differential omic strategies leading to the identification of candidate genes putatively involved in the biosynthesis, pathway regulation, and transmembrane transport leading to the anticancer alkaloids from C. roseus. PMID:27356972

  19. Preliminary Studies on Dynamic Change of Three Alkaloids and Expression of The Associated Biosynthetic Genes In Catharathus Roseus Leaves%长春花叶片三种生物碱含量及合成基因表达的初步研究

    田跃胜; 陆平; 王名雪; 唐克轩; 赵静雅

    2011-01-01

    In order to clarify the dynamic change of three alkaloids (TIAs including vindoline, catharanthine and vinblastine) and the expression of the associated biosynthetic genes in the Catharathus Roseus leaves (young,medium and old) at different growing stages, high performance liquid chromatography was applied to determine the contents of TIAs at the leaf period, pre-floweringperiod, bud stage, flowering period, fully-mature period 1 and fully-mature period 2 in five pairs of leaves from Catharathus Roseus.In addition, the expression of the associated biosynthetic genes was analyzed by molecular biological technique (real time PCR, FQ-PCT).The results showed that highest contents of vindoline and catharanthine were presented in young leaves while the content of vinblastine was highest in old leaves.The best time for collecting the leaves of Catharathus Roseus was between the stage of alabastrum and flowering since the maximum contents of TIAs could be extracted in this period.The data from the FQ-PCR determination indicated that the expression of TIAs contents in C.Roseus leaves was more affected by the six genes inclucding GGPP,G10H ,SLS,STR,D4H and DAT while two genes (ASA, TDC) showed less influence on the production TIAs.%为探明长春花叶片中生物碱的动态变化和生物碱合成途径中的关键酶基因的表达量,利用高效液相色谱技术(HPLC)测定了5对真叶期、开花前期、出现花蕾期、出现开花期、成熟期1和成熟期2等6个生长阶段的长春花幼嫩、中等成熟、成熟三种叶片的生物碱(文多灵、长春质碱、长春碱)的含量.幼嫩叶片中文多灵、长春质碱和成熟叶片中长春碱的含量最高.长春花叶片的最佳采集期是花蕾期到开花期.利用荧光定量PCR技术检测了长春花叶片中生物碱合成途径中10个相关基因的相对表达量,GGPP,G10H,SLS,STR,D4H及DAT这6个基因对生物碱含量贡献较大,ASA及TDC基因对生物碱的增加的作用不明显.

  20. ZCT1 and ZCT2 transcription factors repress the activity of a gene promoter from the methyl erythritol phosphate pathway in Madagascar periwinkle cells.

    Chebbi, Mouadh; Ginis, Olivia; Courdavault, Vincent; Glévarec, Gaëlle; Lanoue, Arnaud; Clastre, Marc; Papon, Nicolas; Gaillard, Cécile; Atanassova, Rossitza; St-Pierre, Benoit; Giglioli-Guivarc'h, Nathalie; Courtois, Martine; Oudin, Audrey

    2014-10-15

    In Catharanthus roseus, accumulating data highlighted the existence of a coordinated transcriptional regulation of structural genes that takes place within the secoiridoid biosynthetic branch, including the methyl erythritol phosphate (MEP) pathway and the following steps leading to secologanin. To identify transcription factors acting in these pathways, we performed a yeast one-hybrid screening using as bait a promoter region of the hydroxymethylbutenyl 4-diphosphate synthase (HDS) gene involved in the responsiveness of C. roseus cells to hormonal signals inducing monoterpene indole alkaloid (MIA) production. We identified that ZCT2, one of the three members of the zinc finger Catharanthus protein (ZCT) family, can bind to a HDS promoter region involved in hormonal responsiveness. By trans-activation assays, we demonstrated that ZCT1 and ZCT2 but not ZCT3 repress the HDS promoter activity. Gene expression analyses in C. roseus cells exposed to methyljasmonate revealed a persistence of induction of ZCT2 gene expression suggesting the existence of feed-back regulatory events acting on HDS gene expression in correlation with the MIA production. PMID:25108262

  1. Review of the taxonomy of the genus Arthrobacter, emendation of the genus Arthrobacter sensu lato, proposal to reclassify selected species of the genus Arthrobacter in the novel genera Glutamicibacter gen. nov., Paeniglutamicibacter gen. nov., Pseudoglutamicibacter gen. nov., Paenarthrobacter gen. nov. and Pseudarthrobacter gen. nov., and emended description of Arthrobacter roseus.

    Busse, Hans-Jürgen

    2016-01-01

    In this paper, the taxonomy of the genus Arthrobacter is discussed, from its first description in 1947 to the present state. Emphasis is given to intrageneric phylogeny and chemotaxonomic characteristics, concentrating on quinone systems, peptidoglycan compositions and polar lipid profiles. Internal groups within the genus Arthrobacter indicated from homogeneous chemotaxonomic traits and corresponding to phylogenetic grouping and/or high 16S rRNA gene sequence similarities are highlighted. Furthermore, polar lipid profiles and quinone systems of selected species are shown, filling some gaps concerning these chemotaxonomic traits. Based on phylogenetic groupings, 16S rRNA gene sequence similarities and homogeneity in peptidoglycan types, quinone systems and polar lipid profiles, a description of the genus Arthrobacter sensu lato and an emended description of Arthrobacter roseus are provided. Furthermore, reclassifications of selected species of the genus Arthrobacter into novel genera are proposed, namely Glutamicibacter gen. nov. (nine species), Paeniglutamicibacter gen. nov. (six species), Pseudoglutamicibacter gen. nov. (two species), Paenarthrobacter gen. nov. (six species) and Pseudarthrobacter gen. nov. (ten species). PMID:26486726

  2. Transfer experiments, light microscopy and electron microscopy studies on the contribution of procaryotes to the novel forest decline patterns. Final report. Uebertragungsversuche, licht- und elektronenmikroskopische Untersuchungen zur Pruefung der ursaechlichen Beteiligung von Prokaryonten (MLO, RLO) an den neuartigen Waldschaeden. Abschlussbericht

    Petzold, H.

    1989-01-01

    The now terminated research project was to clarify if procaryotes are a causal part of the novel damage to forests. For this extensive microscopic analyses on differently damaged spruce trees and several silver firs from the forest damage areas in the Fichtelgebirge and Bayerischer Wald mountains were made. Beyond this it was attempted to transfer possible phytopathogenic agents from damaged forest trees to healthy young trees of the same kind by pruning. In addition it was attempted to transfer with the aid of parasitic plants of the species Cuscuta possibly occurring mycoplasms of trees suffering from acute yellowing to the test plant Catharanthus roseus, the most suitable for this. Additional tests served to transfer defined MLO from Catharanthus onto healthy seedlings of diverse deciduous and coniferous trees to determine if forest trees are susceptible to the mycoplasmoses common in Germany. (orig./MG).

  3. Synthesis of Xylooligosaccharides of Daidzein and Their Anti-Oxidant and Anti-Allergic Activities

    Hatsuyuki Hamada

    2011-08-01

    Full Text Available The biocatalytic synthesis of xylooligosaccharides of daidzein was investigated using cultured cells of Catharanthus roseus and Aspergillus sp. β-xylosidase. The cultured cells of C. roseus converted daidzein into its 4'-O-β-glucoside, 7-O-β-glucoside, and 7-O-β-primeveroside, which was a new compound. The 7-O-β-primeveroside of daidzein was further xylosylated by Aspergillus sp. β-xylosidase to daidzein trisaccharide, i.e., 7-O-[6-O-(4-O-(β-D-xylopyranosyl-β-D-xylopyranosyl]-β-D-glucopyranoside, which was a new compound. The 4'-O-β-glucoside, 7-O-β-glucoside, and 7-O-β-primeveroside of daidzein exerted DPPH free-radical scavenging and superoxide radical scavenging activity. On the other hand, 7-O-β-glucoside and 7-O-β-primeveroside of daidzein showed inhibitory effects on IgE antibody production.

  4. Main: CACGTGMOTIF [PLACE

    Full Text Available ; shoot; Pti4; ERF; PR; tomato (Lycopersicon esculentum); Arabidopsis thaliana; s...TAGTT) and G-box (CACGTG)); A prominent hit by in silico analysis in both induced and repressed phyA-responsive promoters...napdragon (Antirrhinum majus); wheat (Triticum aestivum); parsley: maize (Zea mays); periwinkle (Catharanthus roseus); Brassica napus; bean (Phaseolus vulgaris); CACGTG ... ... of the Str promoter via direct interaction with the G-box; See S000345; Essential for expressi... (Hudson and Quail 2003); Review by Terzaghi WB, Cashmore AR. Light-

  5. In Vivo and Real-time Monitoring of Secondary Metabolites of Living Organisms by Mass Spectrometry

    Hu, Bin; Wang, Lei; Ye, Wen-Cai; Yao, Zhong-Ping

    2013-07-01

    Secondary metabolites are compounds that are important for the survival and propagation of animals and plants. Our current understanding on the roles and secretion mechanism of secondary metabolites is limited by the existing techniques that typically cannot provide transient and dynamic information about the metabolic processes. In this manuscript, by detecting venoms secreted by living scorpion and toad upon attack and variation of alkaloids in living Catharanthus roseus upon stimulation, which represent three different sampling methods for living organisms, we demonstrated that in vivo and real-time monitoring of secondary metabolites released from living animals and plants could be readily achieved by using field-induced direct ionization mass spectrometry.

  6. Effect of jasmonic acid elicitation on the yield, chemical composition, and antioxidant and anti-inflammatory properties of essential oil of lettuce leaf basil (Ocimum basilicum L.).

    Złotek, Urszula; Michalak-Majewska, Monika; Szymanowska, Urszula

    2016-12-15

    The effect of elicitation with jasmonic acid (JA) on the plant yield, the production and composition of essential oils of lettuce leaf basil was evaluated. JA-elicitation slightly affected the yield of plants and significantly increased the amount of essential oils produced by basil - the highest oil yield (0.78±0.005mL/100gdw) was achieved in plants elicited with 100μM JA. The application of the tested elicitor also influenced the chemical composition of basil essential oils - 100μM JA increased the linalool, eugenol, and limonene levels, while 1μM JA caused the highest increase in the methyl eugenol content. Essential oils from JA-elicited basil (especially 1μM and 100μM) exhibited more effective antioxidant and anti-inflammatory potential; therefore, this inducer may be a very useful biochemical tool for improving production and composition of herbal essential oils. PMID:27451148

  7. SuperSAGE analysis of the Nicotiana attenuata transcriptome after fatty acid-amino acid elicitation (FAC: identification of early mediators of insect responses

    Baldwin Ian T

    2010-04-01

    Full Text Available Abstract Background Plants trigger and tailor defense responses after perception of the oral secretions (OS of attacking specialist lepidopteran larvae. Fatty acid-amino acid conjugates (FACs in the OS of the Manduca sexta larvae are necessary and sufficient to elicit the herbivory-specific responses in Nicotiana attenuata, an annual wild tobacco species. How FACs are perceived and activate signal transduction mechanisms is unknown. Results We used SuperSAGE combined with 454 sequencing to quantify the early transcriptional changes elicited by the FAC N-linolenoyl-glutamic acid (18:3-Glu and virus induced gene silencing (VIGS to examine the function of candidate genes in the M. sexta-N. attenuata interaction. The analysis targeted mRNAs encoding regulatory components: rare transcripts with very rapid FAC-elicited kinetics (increases within 60 and declines within 120 min. From 12,744 unique Tag sequences identified (UniTags, 430 and 117 were significantly up- and down-regulated ≥ 2.5-fold, respectively, after 18:3-Glu elicitation compared to wounding. Based on gene ontology classification, more than 25% of the annotated UniTags corresponded to putative regulatory components, including 30 transcriptional regulators and 22 protein kinases. Quantitative PCR analysis was used to analyze the FAC-dependent regulation of a subset of 27 of these UniTags and for most of them a rapid and transient induction was confirmed. Six FAC-regulated genes were functionally characterized by VIGS and two, a putative lipid phosphate phosphatase (LPP and a protein of unknown function, were identified as important mediators of the M. sexta-N. attenuata interaction. Conclusions The analysis of the early changes in the transcriptome of N. attenuata after FAC elicitation using SuperSAGE/454 has identified regulatory genes involved in insect-specific mediated responses in plants. Moreover, it has provided a foundation for the identification of additional novel regulators associated with this process.

  8. Enhancement of anti-inflammatory activity of Aloe vera adventitious root extracts through the alteration of primary and secondary metabolites via salicylic acid elicitation.

    Yun Sun Lee

    Full Text Available Aloe vera (Asphodeloideae is a medicinal plant in which useful secondary metabolites are plentiful. Among the representative secondary metabolites of Aloe vera are the anthraquinones including aloe emodin and chrysophanol, which are tricyclic aromatic quinones synthesized via a plant-specific type III polyketide biosynthesis pathway. However, it is not yet clear which cellular responses can induce the pathway, leading to production of tricyclic aromatic quinones. In this study, we examined the effect of endogenous elicitors on the type III polyketide biosynthesis pathway and identified the metabolic changes induced in elicitor-treated Aloe vera adventitious roots. Salicylic acid, methyl jasmonate, and ethephon were used to treat Aloe vera adventitious roots cultured on MS liquid media with 0.3 mg/L IBA for 35 days. Aloe emodin and chrysophanol were remarkably increased by the SA treatment, more than 10-11 and 5-13 fold as compared with untreated control, respectively. Ultra-performance liquid chromatography-electrospray ionization mass spectrometry analysis identified a total of 37 SA-induced compounds, including aloe emodin and chrysophanol, and 3 of the compounds were tentatively identified as tricyclic aromatic quinones. Transcript accumulation analysis of polyketide synthase genes and gas chromatography mass spectrometry showed that these secondary metabolic changes resulted from increased expression of octaketide synthase genes and decreases in malonyl-CoA, which is the precursor for the tricyclic aromatic quinone biosynthesis pathway. In addition, anti-inflammatory activity was enhanced in extracts of SA-treated adventitious roots. Our results suggest that SA has an important role in activation of the plant specific-type III polyketide biosynthetic pathway, and therefore that the efficacy of Aloe vera as medicinal agent can be improved through SA treatment.

  9. Insecticidal and biological effects of three plant extracts tested against the dengue vector, Stegomyia agyptii (Diptera: Culicidae

    R. Kokila

    2016-04-01

    Full Text Available Development of resistant in vectors especially vector mosquitoes are becoming a challenge for the scientific community for management and control mosquito population. Vector mosquitoes are likely to withstand toxicity and develop resistant mechanism to single active compound hence, combining medicinal plants with rich active compounds stops resistant development and proliferation of mosquitoes. In this study we put effort to evaluate the effect of methanol extract of Tagetes patula, Clerodentron phillomedis, and Catharanthus roseus in individual and in combination against the dengue vector, Stegomyia agyptii. Lethal concentrations (LC50 and LC90 were calculated to find out the effect of the test plants in individual and in combination. T. patula extract showed vaguely higher mortality rate when compared to C. phillomedis, and C. roseus but there was no significant variation among the three test plants. The median LC of combined treatment showed a significant difference between the combined (2.25 µg/mL/3rd instar and individual treatment (6.41 µg/mL/3rd instar for T. patula, 6.85 µg/mL/3rd instar for C. phillomedis and 6.59 µg/mL/3rd instar for C. roseus. The combined efficacy of three test plants was also effective in controlling vector mosquitoes at fields with different agro-climatic conditions. The study proves that the combination of T. patula, C. phillomedis, and C. roseus is effective in different field conditions at lower concentrations.

  10. Phytochemical genomics of the Madagascar periwinkle: Unravelling the last twists of the alkaloid engine.

    Dugé de Bernonville, Thomas; Clastre, Marc; Besseau, Sébastien; Oudin, Audrey; Burlat, Vincent; Glévarec, Gaëlle; Lanoue, Arnaud; Papon, Nicolas; Giglioli-Guivarc'h, Nathalie; St-Pierre, Benoit; Courdavault, Vincent

    2015-05-01

    The Madagascar periwinkle produces a large palette of Monoterpenoid Indole Alkaloids (MIAs), a class of complex alkaloids including some of the most valuable plant natural products with precious therapeutical values. Evolutionary pressure on one of the hotspots of biodiversity has obviously turned this endemic Malagasy plant into an innovative alkaloid engine. Catharanthus is a unique taxon producing vinblastine and vincristine, heterodimeric MIAs with complex stereochemistry, and also manufactures more than 100 different MIAs, some shared with the Apocynaceae, Loganiaceae and Rubiaceae members. For over 60 years, the quest for these powerful anticancer drugs has inspired biologists, chemists, and pharmacists to unravel the chemistry, biochemistry, therapeutic activity, cell and molecular biology of Catharanthus roseus. Recently, the "omics" technologies have fuelled rapid progress in deciphering the last secret of strictosidine biosynthesis, the central precursor opening biosynthetic routes to several thousand MIA compounds. Dedicated C. roseus transcriptome, proteome and metabolome databases, comprising organ-, tissue- and cell-specific libraries, and other phytogenomic resources, were developed for instance by PhytoMetaSyn, Medicinal Plant Genomic Resources and SmartCell consortium. Tissue specific library screening, orthology comparison in species with or without MIA-biochemical engines, clustering of gene expression profiles together with various functional validation strategies, largely contributed to enrich the toolbox for plant synthetic biology and metabolic engineering of MIA biosynthesis. PMID:25146650

  11. A novel method for rapid and non-invasive detection of plants senescence using delayed fluorescence technique

    Zhang, Lingrui; Xing, Da; Wang, Junsheng; Zeng, Lizhang; Li, Qiang

    2007-05-01

    Plants senescence is a phase of plants ontogeny marked by declining photosynthetic activity that is paralleled by a decline in chloroplast function. The photosystem II ( PSII ) in a plant is considered the primary site where light-induced delayed fluorescence (DF) is produced. With the leaves of Catharanthus roseus (Catharanthus roseus (L.) G.Don) as testing models, we have studied the effects of plants senescence induced by dark and/or exogenous hormones treatments on characteristics of DF by using a home-made portable DF detection system, which can enable various DF parameters, such as DF decay kinetic curve and DF intensity, to be rapidly produced for the plants in a short time. The results show that the changes in DF intensity of green plants can truly reflect the changes in photosynthetic capacity and chlorophyll content. Therefore, DF may be used an important means of evaluating in vivo plants senescence physiology. The changes in DF intensity may provide a new approach for the rapid and early detection of plants senescence caused by age or other senescence-related factors. DF technique could be potential useful for high throughput screening and less time-consuming and automated identifying the interesting mutants with genetic modifications that change plants senescence progress.

  12. Apocynaceae species with antiproliferative and/or antiplasmodial properties: a review of ten genera.

    Chan, Eric Wei Chiang; Wong, Siu Kuin; Chan, Hung Tuck

    2016-07-01

    Apocynaceae is a large family of tropical trees, shrubs and vines with most species producing white latex. Major metabolites of species are triterpenoids, iridoids, alkaloids and cardenolides, which are known for a wide range of biological and pharmacological activities such as cardioprotective, hepatoprotective, neuroprotective, anti-inflammatory, anticancer and antimalarial properties. Prompted by their anticancer and antimalarial properties, the current knowledge on ten genera (Allamanda, Alstonia, Calotropis, Catharanthus, Cerbera, Dyera, Kopsia, Nerium, Plumeria and Vallaris) is updated. Major classes of metabolites are described using some species as examples. Species with antiproliferative (APF) and/or antiplasmodial (APM) properties have been identified. With the exception of the genus Dyera, nine genera of 22 species possess APF activity. Seven genera (Alstonia, Calotropis, Catharanthus, Dyera, Kopsia, Plumeria and Vallaris) of 13 species have APM properties. Among these species, Alstonia angustiloba, Alstonia macrophylla, Calotropis gigantea, Calotropis procera, Catharanthus roseus, Plumeria alba and Vallaris glabra displayed both APF and APM properties. The chemical constituents of these seven species are compiled for assessment and further research. PMID:27417173

  13. "Self" and "non-self" in the control of phytoalexin biosynthesis: plant phospholipases A2 with alkaloid-specific molecular fingerprints.

    Heinze, Michael; Brandt, Wolfgang; Marillonnet, Sylvestre; Roos, Werner

    2015-02-01

    The overproduction of specialized metabolites requires plants to manage the inherent burdens, including the risk of self-intoxication. We present a control mechanism that stops the expression of phytoalexin biosynthetic enzymes by blocking the antecedent signal transduction cascade. Cultured cells of Eschscholzia californica (Papaveraceae) and Catharanthus roseus (Apocynaceae) overproduce benzophenanthridine alkaloids and monoterpenoid indole alkaloids, respectively, in response to microbial elicitors. In both plants, an elicitor-responsive phospholipase A2 (PLA2) at the plasma membrane generates signal molecules that initiate the induction of biosynthetic enzymes. The final alkaloids produced in the respective plant inhibit the respective PLA, a negative feedback that prevents continuous overexpression. The selective inhibition by alkaloids from the class produced in the "self" plant could be transferred to leaves of Nicotiana benthamiana via recombinant expression of PLA2. The 3D homology model of each PLA2 displays a binding pocket that specifically accommodates alkaloids of the class produced by the same plant, but not of the other class; for example, C. roseus PLA2 only accommodates C. roseus alkaloids. The interaction energies of docked alkaloids correlate with their selective inhibition of PLA2 activity. The existence in two evolutionary distant plants of phospholipases A2 that discriminate "self-made" from "foreign" alkaloids reveals molecular fingerprints left in signal enzymes during the evolution of species-specific, cytotoxic phytoalexins. PMID:25670767

  14. Genome-guided investigation of plant natural product biosynthesis.

    Kellner, Franziska; Kim, Jeongwoon; Clavijo, Bernardo J; Hamilton, John P; Childs, Kevin L; Vaillancourt, Brieanne; Cepela, Jason; Habermann, Marc; Steuernagel, Burkhard; Clissold, Leah; McLay, Kirsten; Buell, Carol Robin; O'Connor, Sarah E

    2015-05-01

    The medicinal plant Madagascar periwinkle, Catharanthus roseus (L.) G. Don, produces hundreds of biologically active monoterpene-derived indole alkaloid (MIA) metabolites and is the sole source of the potent, expensive anti-cancer compounds vinblastine and vincristine. Access to a genome sequence would enable insights into the biochemistry, control, and evolution of genes responsible for MIA biosynthesis. However, generation of a near-complete, scaffolded genome is prohibitive to small research communities due to the expense, time, and expertise required. In this study, we generated a genome assembly for C. roseus that provides a near-comprehensive representation of the genic space that revealed the genomic context of key points within the MIA biosynthetic pathway including physically clustered genes, tandem gene duplication, expression sub-functionalization, and putative neo-functionalization. The genome sequence also facilitated high resolution co-expression analyses that revealed three distinct clusters of co-expression within the components of the MIA pathway. Coordinated biosynthesis of precursors and intermediates throughout the pathway appear to be a feature of vinblastine/vincristine biosynthesis. The C. roseus genome also revealed localization of enzyme-rich genic regions and transporters near known biosynthetic enzymes, highlighting how even a draft genome sequence can empower the study of high-value specialized metabolites. PMID:25759247

  15. Greater flamingos Phoenicopterus roseus are partial capital breeders

    Rendón-Martos, Manuel; Rendón, Miguel A.; Garrido, Araceli; Amat, Juan A.

    2011-01-01

    Capital breeding refers to a strategy in which birds use body stores for egg formation, whereas income breeders obtain all resources for egg formation at breeding sites. Capital breeding should occur more in large-bodied species because the relative cost of carrying stores for egg formation becomes smaller with increasing body size. Based on a comparison between stable isotopes of carbon and nitrogen in potential prey at wintering sites and eggs, we examined whether greater flamingos use nutr...

  16. AcEST: DK957247 [AcEST

    Full Text Available Definition sp|P48522|TCMO_CATRO Trans-cinnamate 4-monooxygenase OS=Catharanthus roseus Align length 115 Scor...t alignments: (bits) Value sp|P48522|TCMO_CATRO Trans-cinnamate 4-monooxygenase OS=Catharan... 222 1e-57 sp|Q42797|TCMO_SOYBN Tra...ns-cinnamate 4-monooxygenase OS=Glycine ... 220 3e-57 sp|Q43240|TCMO_ZINEL Tra...ns-cinnamate 4-monooxygenase OS=Zinnia e... 219 9e-57 sp|Q04468|TCMO_HELTU Trans-cinnamate... 4-monooxygenase OS=Helianth... 219 9e-57 sp|Q96423|TCMO_GLYEC Trans-cinnamate 4-monooxygenase OS=Glycyrrh..

  17. AcEST: DK962436 [AcEST

    Full Text Available sp_hit_id P48522 Definition sp|P48522|TCMO_CATRO Trans-cinnamate 4-monooxygenase OS=Catharanthus roseus Ali... Miller, and David J. Lipman (1997), Gapped BLAST and PSI-BLAST: a new generation of protein database search program...oducing significant alignments: (bits) Value sp|P48522|TCMO_CATRO Trans-cinnamate 4-monooxygenase OS=Cathara...n... 225 1e-58 sp|Q43240|TCMO_ZINEL Trans-cinnamate 4-monooxygenase OS=Zinnia e...... 224 2e-58 sp|Q42797|TCMO_SOYBN Trans-cinnamate 4-monooxygenase OS=Glycine ... 223 4e-58 sp|Q04468|TCMO_HELTU Trans-cinnama

  18. Mass attenuation coefficients of X-rays in different medicinal plants

    Morabad, R.B. [Department of Post-Graduate Studies and Research in Physics, Gulbarga University, Gulbarga 585106, Karnataka (India); Kerur, B.R. [Department of Post-Graduate Studies and Research in Physics, Gulbarga University, Gulbarga 585106, Karnataka (India)], E-mail: kerurbrk@yahoo.com

    2010-02-15

    The mass attenuation coefficients of specific parts of several plants, (fruits, leaves, stem and seeds) often used as medicines in the Indian herbal system, have been measured employing NaI (TI)) detector. The electronic setup used is a NaI (TI) detector, which is coupled to MCA for analysis of the spectrum. A source of {sup 241}Am is used to get X-rays in the energy range 8-32 keV from Cu, Rb, Mo, Ag and Ba targets. In the present study, the measured mass attenuation coefficient of Ocimum sanctum, Catharanthus roseus, Trigonella foenum-graecum, Azadirachta indica, Aegle marmelos, Zingiber officinalis, Emblica officinalis, Anacardium occidentale, Momordica charantia and Syzygium cumini show a linear relation with the energy.

  19. 海南药用长春花标准化生产技术

    徐三林; 林秋梅; 汪煌

    2010-01-01

    @@ 长春花[Catharanthus roseus(L.)G.Don]别名雁来红,日日新.为夹竹桃科植物长春花,主要以茎、叶作为植物原料药.其茎叶含有50多种生物碱,如长春碱长春新碱和文多林等,对治疗绒癌等恶性肿瘤、淋巴肉瘤及儿童急性白血病都有一定疗效,是目前国际上应用较多的抗癌植物药.市场售价看好,可获得丰厚的经济效益.

  20. Effect of Leaf Structure on the Formaldehyde-resistance Capacity in Five Ornamental Plants%几种花卉叶片的组织结构对其抗甲醛能力的影响

    邸葆; 陈段芬; 果秀敏; 鲁媛; 张成合

    2007-01-01

    以绿萝(Scindapsus aureus)、亮丝草(Aglaonema moclestum)、花烛(Anthurium andreanum)、长寿花(Catharanthus roseus)和非洲菊(Gerbera jamesonii)为试材,浓度为5.0 mg/mL的甲醛处理24 h后,观察其受害状况,并对其叶片进行解剖观察.结果表明:叶片表皮的气孔密度、栅栏组织和海绵组织的细胞排列方式等对植物抗甲醛能力有明显影响,未发现处理前后气孔开度变化与植物抗甲醛能力之间存在明显的正相关.