In Vitro Propagation and Production of Cardiotonic Glycosides in Shoot Cultures of Digitalis Purpurea L

Total Page:16

File Type:pdf, Size:1020Kb

In Vitro Propagation and Production of Cardiotonic Glycosides in Shoot Cultures of Digitalis Purpurea L Appl Microbiol Biotechnol DOI 10.1007/s00253-012-4489-y BIOTECHNOLOGICAL PRODUCTS AND PROCESS ENGINEERING In vitro propagation and production of cardiotonic glycosides in shoot cultures of Digitalis purpurea L. by elicitation and precursor feeding Jitendra Gopichand Patil & Mahendra Laxman Ahire & Kirti Manik Nitnaware & Sayantan Panda & Vijay P. Bhatt & Polavarapu B. Kavi Kishor & Tukaram Dayaram Nikam Received: 18 July 2012 /Revised: 16 September 2012 /Accepted: 1 October 2012 # Springer-Verlag Berlin Heidelberg 2012 Abstract Digitalis purpurea L. (Scrophulariaceae; Fox To further maintain the multiple shoot induction, mother glove) is a source of cardiotonic glycosides such as digitox- tissue was cut into four equal parts and repeatedly sub- in and digoxin which are commercially applied in the treat- cultured on fresh shoot induction liquid medium after each ment to strengthen cardiac diffusion and to regulate heart harvest. On adaptation of this strategy, an average of 18 shoots rhythm. This investigation deals with in vitro propagation per explant could be produced. This strategy was applied for and elicited production of cardiotonic glycosides digitoxin the production of biomass and glycosides digitoxin and di- and digoxin in shoot cultures of D. purpurea L. In vitro goxin in shoot cultures on MS medium supplemented with germinated seedlings were used as a primary source of 7.5 μM BA and several treatments with plant growth regu- explants. Multiple shoot formation was achieved for three lators, incubation period, abiotic (salicylic acid, mannitol, explant types (nodal, internodal, and leaf) cultured on sorbitol, PEG-6000, NaCl, and KCl), biotic (Aspergillus Murashige and Skoog (MS) medium with several treatments niger, Helminthosporium sp., Alternaria sp., chitin, and of cytokinins (6-benzyladenine—BA; kinetin—Kin; and yeast extract) elicitors, and precursors (progesterone, cho- thidiazuron—TDZ) and auxins (indole-3-acetic acid— lesterol, and squalene). The treatment of KCl, mycelial IAA; α-naphthaleneacetic acid—NAA; and 2,4-dichloro- mass of Helminthosporium sp., and progesterone were phenoxy acetic acid—2,4-D). Maximum multiple shoots highly effective for the production of cardenolides. In the (12.7±0.6) were produced from nodal explants on MS+ presence of progesterone (200 to 300 mg/l), digitoxin and 7.5 μM BA. Shoots were rooted in vitro on MS containing digoxin accumulation was enhanced by 9.1- and 11.9-folds 15 μM IAA. Rooted plantlets were successfully acclimatized. respectively. : : : : Keyword Cardiotonic glycosides . Digitalis purpurea . J. G. Patil M. L. Ahire K. M. Nitnaware S. Panda Digitoxin . Digoxin . Shoot cultures T. D. Nikam (*) Department of Botany, University of Pune, Pune 411 007, Maharashtra, India e-mail: tdnikam@unipune.ac.in Introduction V. P. Bhatt Herbal Research and Development Institute, Digitalis purpurea L. belongs to the family Scrophula Mandal, Gopeshwar, riaceae and is a biennial or perennial herb widely distributed 246 401, Chamoli, Uttarakhand, India in the Western Europe, Asia, Northwest Africa, South America, New Zealand, and Canada (Hultén 1968). The P. B. K. Kishor Department of Genetics, Osmania University, entire plant including the roots and seeds is poisonous. It Hyderabad 500 007, India contains several deadly physiological and chemically related Appl Microbiol Biotechnol cardiac and steroidal glycosides (Budavari et al. 1989). 1992), and shoot cultures (Hagimori et al. 1983; Seitz and However, it is commercially cultivated to obtain cardiotonic Gärtner 1994; Gurel et al. 2011). Further, it is shown that the glycosides namely digitoxin and digoxin (Mastenbroek cardenolide biosynthesis is basically dependent on morpho- 1985). Both glycosides are the best known products to logical differentiation (Eisenbeiβ et al. 1999; Pérez-Alonso strengthen cardiac diffusion and to regulate heart rhythm et al. 2009). To date, to the best of our knowledge, no (Navarro et al. 2000; Pérez-Bermúdez et al. 2010; Sharma information is available on in vitro propagation protocol and Purkait 2012). Actually, they function by inhibiting the and on the influence of elicitors and precursors on the sodium–potassium ATPase activity which results in in- production of Digitalis cardenolides on the in vitro grown creased intracellular concentration of sodium. This helps in shoot cultures. the elevation of intracellular calcium and creation of posi- The aim of the present investigation was to develop in tive inotropic effect (Rahimtoola and Tak 1996; Xie and vitro propagation protocol using different explants of in Askari 2002; Mohammadi et al. 2003; López-Lázaro 2007; vitro germinated seedlings. Attempts were made to test the Kuate et al. 2008; Wu et al. 2012). The Digitalis glycoside is influence of plant growth regulators, different biotic and still the only safe inotropic drug treatment for oral use, abiotic elicitors, and precursors on the accumulation of which improves hemodynamics in patients with a compro- medicinally important cardiotonic glycosides, digitoxin mised cardiac function (Schwinger et al. 2003; Pérez- and digoxin in shoot cultures of D. purpurea L. Alonso et al. 2009). Recently, it is also noted that both the glycosides were effective against several types of cancer (Haux 1999; Stenkvist 2001;Hauxetal.2001; López- Materials and methods Lázaro et al. 2003; López-Lázaro 2007; Sharma and Purkait 2012; Wu et al. 2012). The chemical synthesis of Plant material and culture conditions these glycosides is so far not cost effective due to complex chemical nature (Pérez-Bermúdez et al. 2010). The pharma- Seeds of D. purpurea were obtained from Herbal Research ceutical industries are still relying on natural sources of the and Development Institute, Mandal-Gopeswar, Chamoli, plant material which contains very low amounts of these Uttarakhand. Seeds were washed in running tap water for glycosides. In such a situation, it necessitates the application 5 min followed by five rinses in sterilized distilled water. of biotechnological approaches to enhance the yield of Then, seeds were surface-sterilized with 0.1 % (w/v)mercuric digitoxin and digoxin. chloride (HgCl2) solution for 5 min and washed five to six Plant cell and tissue culture can be used as an alternative times with sterilized distilled water to remove the traces of source for the production of medicinally important metabo- HgCl2. Surface-sterilized seeds were inoculated onto the MS lites (Verpoorte and Memelink 2002; Vanisree and Tsay (Murashige and Skoog 1962) medium for germination. Node, 2004). In vitro grown cell and organ cultures furnish a ready internode, and leaf explants were excised from 21-day-old source of uniform, sterile, and compatible material for bio- aseptic seedlings and used as explants. The explants were chemical characterization and identification of active con- placed on MS medium supplemented with different concen- stituents (Sajc et al. 2000). This process may condense trations (0.0–15.0 μM) of 6-benzyladenine (BA), kinetin biosynthetic cycle and facilitates the higher rate of metabo- (Kin), or thidiazuron (TDZ) individually or in combination lism than field grown plants (Rao and Ravishankar 2002; with auxins indole-3-acetic acid (IAA), α-naphthalene acetic Naik et al. 2011). Production of digitoxin and digoxin has acid (NAA), or 2,4-dichlorophenoxyacetic acid (2,4-D) and been investigated from cultured cells from Digitalis,but 30 gl−1 sucrose. pH of the medium was adjusted to 5.8 and most of the workers reported that undifferentiated cells solidified with 0.8 % (w/v)agar–agar (Hi Media, Mumbai, either did not produce cardenolides (Graves and Smith India) prior to autoclaving at 121 °C for 15 min. Cultures were 1967; Elze et al. 1974; Hirotani and Furuya 1977) or pro- incubated under controlled conditions such as 25±2 °C tem- duce only in trace amounts (Rücker et al. 1976; Lui and perature, 60±10 % relative humidity, and 8-h photoperiod Staba 1979). Attempts were made to use differentiated tis- (PFD 40 μmolm−2s−1) provided by white fluorescent tubes sues or organ cultures and showed the production of medic- (Philips, Kolkata, India). inally important compounds (Wilken et al. 2005;Pérez- Alonso et al. 2009). Redifferentiating organ cultures Maintenance of multiple shoot culture reported to accumulate considerable amount of cardinolides in Digitalis (Hirotani and Furuya 1977; Lui and Staba 1979; In preliminary experiments, shoot cultures showed signifi- Grave et al. 1980; Hagimori et al. 1982). Efforts were made cantgrowthin50mlliquidMSmediumfortifiedwith for in vitro production of cardiotonic glycosides from the 7.5 μM BA in culture glass bottles (400 ml capacity). genus Digitalis using leaf cell (Kreis et al. 1986), embryonic Therefore, multiple shoot cultures were maintained by sub- cell (Lindemann and Luckner 1997), root (Shimomura et al. culturing onto the same fresh medium after 28 days of Appl Microbiol Biotechnol interval. After separation of individual shoots, the basal potato-dextrose medium under controlled conditions as de- clump of mother explants was divided into four pieces and scribed earlier. After 28 days of growth, cultures were were inoculated separately in a bottle containing 50 ml MS autoclaved at 121 °C for 12 min. Cultures were allowed to medium fortified with 7.5 μM BA. The control was main- cool to room temperature, mycelial mats were harvested, tained on medium lacking BA. After 4 weeks of culture, washed five times with sterile distilled water, and dried in shoots were harvested and
Recommended publications
  • Cytotoxic Effect of Some Digitalis Species; a Study of Selectivity
    FABAD J. Pharm. Sci., 43(1), 25-29, 2018 RESEARCH ARTICLE Cytotoxic Effect of Some Digitalis Species; A Study of Selectivity Vahap Murat KUTLUAY *°, İclal SARAÇOĞLU* Cytotoxic Effect of Some Digitalis Species; A Study of Bazı Digitalis Türlerinin Sitotoksik Etkisi; Bir Selektivite Selectivity Çalışması SUMMARY ÖZET Our previous research, on the aerial parts and roots of three Daha önceki çalışmamızda üç farklı Digitalis türünün (D.davisiana different Digitalis species; D.davisiana Heywood, D.viridiflora Heywood, D.viridiflora Lindley ve D.grandiflora Miller) toprak üstü Lindley and D.grandiflora Miller against HEp-2 and HepG2 kısımlarının ve köklerinin HEp-2 ve HepG2 kanser hücre serileri üzerinde, cancer cell lines showed that aerial parts were more cytotoxic than toprak üstü kısımların köklere göre daha sitotoksik olduğu gösterilmiştir. roots. So for further research a comparative cytotoxic activity study Bu nedenle D.davisiana, D.viridiflora ve D.grandiflora’nın toprak üstü was performed on the aerial parts of D.davisiana, D.viridiflora kısımların üzerinde gerçekleştirilen araştırma ilerletilerek HeLa kanser and D.grandiflora against HeLa cancer cell line. 3Y1 non- hücresi üzerindeki sitotoksik etkisi incelenmiştir. Sağlıklı bir hücre cancerous cell line was used to determine the selectivity. All three olan 3Y1, selektivitenin değerlendirilebilmesi için kullanılmıştır. Her üç ekstre de HeLa hücreleri üzerinde kuvvetli sitotoksik etki göstermiş extracts showed strong cytotoxic activity on HeLa cells with IC50 values less than 15 μg/ml. However, observed cytotoxic activity on ve IC50 değerleri 15 μg/ml’nin altında hesaplanmıştır. Ekstrelerin 3Y1 hücreleri üzerinde sitotoksik etkileri daha düşük olmakla birlikte 3Y1 cells was found to be lower than that of HeLa cells with IC50 values in a range of 94.7-772.3 μg/ml.
    [Show full text]
  • Molecular Characterization and Genetic Diversity of the Macaw Palm Ex Situ Germplasm Collection Revealed by Microsatellite Markers
    diversity Article Molecular Characterization and Genetic Diversity of the Macaw Palm Ex Situ Germplasm Collection Revealed by Microsatellite Markers Fekadu G. Mengistu 1,*, Sérgio Y. Motoike 2 and Cosme D. Cruz 3 1 Kulumsa Agricultural Research Center (KARC), Ethiopian Institute of Agricultural Research (EIAR), P.O.Box 489, Asella, Ethiopia 2 Departamento de Fitotecnia, Universidade Federal de Viçosa, Av. P.H. Rolfs, Campus, Viçosa, MG 36570-000, Brazil; motoike@ufv.br 3 Departamento de Biologia Geral, Universidade Federal de Viçosa, Av. P.H. Rolfs, Campus, Viçosa, MG 36570-000, Brazil; cdcruz@ufv.br * Correspondence: fgebretensay@yahoo.com; Tel.: +251-0968-23-55-27; Fax: +251-022-331-1508 Academic Editor: Mario A. Pagnotta Received: 29 June 2016; Accepted: 9 October 2016; Published: 13 October 2016 Abstract: Macaw palm (Acrocomia aculeata) is native to tropical forests in South America and highly abundant in Brazil. It is cited as a highly productive oleaginous palm tree presenting high potential for biodiesel production. The aim of this work was to characterize and study the genetic diversity of A. aculeata ex situ collections from different geographical states in Brazil using microsatellite (Simple Sequence Repeats, SSR) markers. A total of 192 accessions from 10 provenances were analyzed with 10 SSR, and variations were detected in allelic diversity, polymorphism, and heterozygosity in the collections. Three major groups of accessions were formed using PCoA—principal coordinate analysis, UPGMA—unweighted pair-group method with arithmetic mean, and Tocher. The Mantel test revealed a weak correlation (r = 0.07) between genetic and geographic distances among the provenances reaffirming the result of the grouping.
    [Show full text]
  • List of 735 Prioritised Plant Taxa of CARE-MEDIFLORA Project
    List of 735 prioritised plant taxa of CARE-MEDIFLORA project In situ and/or ex situ conservation actions were implemented during CARE-MEDIFLORA for 436 of the prioritised plant taxa. Island(s) of occurrence: Balearic Islands (Ba), Corsica (Co), Sardinia (Sa), Sicily (Si), Crete (Cr), Cyprus (Cy) Occurrence: P = present; A = alien (not native to a specific island); D = doubtful presence Distribution type: ENE = Extremely Narrow Endemic (only one population) NE = Narrow Endemic (≤ five populations) RE = Regional Endemic (only one Island) IE = Insular Endemic (more than one island) W = distributed in more islands or in a wider area. Distribution type defines the "regional responsibility" of an Island on a plant species. Criteria: Red Lists (RL): plant species selected is included in the red list (the plant should be EN, CR or VU in order to justify a conservation action); Regional Responsibility (RR): plant species selected plays a key role for the island; the "regional responsibility" criterion is the first order of priority at local level, because it establishes a high priority to plants whose distribution is endemic to the study area (an island in our specific case). Habitats Directive (HD): plant species selected is listed in the Annexes II and V of the Habitat Directive. Wetland plant (WP): plant species selected is a wetland species or grows in wetland habitat. Island(s) where Distribution Island(s) where Taxon (local checklists) Island(s) of occurrence conservation action(s) type taxon prioritised were implemented Ba Co Sa Si Cr Cy RL RR HD WP Ex situ In situ Acer granatense Boiss. P W 1 Ba Ba Acer obtusatum Willd.
    [Show full text]
  • In Vitro Propagation of Digitalis Trojana Ivanina., an Endemic Medicinal Plant of Turkey Nurşen Çördük and Cüneyt Aki
    Chapter In Vitro Propagation of Digitalis trojana Ivanina., an Endemic Medicinal Plant of Turkey Nurşen Çördük and Cüneyt Aki Abstract Digitalis trojana Ivanina is a member of the Plantaginaceae family and known by its common name, Helen of Troy foxglove. It is perennial endemic to Çanakkale and Balıkesir, northwestern Turkey. In order to develop an efficient shoot regen- eration protocol, the leaf explants of D. trojana were cultured on Murashige and Skoog (MS) medium containing 6-benzyl adenine (0.1, 0.5, 1.0, 3.0, 5.0 mg/L) and α-naphthalene acetic acid (0.1, 0.5, 1.0 mg/L), 3% (w/v) sucrose and 0.8% (w/v) agar. The highest number of regenerated shoots was obtained from leaf explants that were cultured on MS medium with 3.0 mg/L BA+0.1 mg/L NAA. Regenerated shoots were rooted on MS medium without plant growth regulators. Rooted plants (2–3 cm) were separately transferred to pots containing a mixture of peat and perlite (2:1 v/v) and acclimatized successfully in a growth chamber. Keywords: endemic, foxglove, in vitro, propagation, regeneration 1. Introduction Turkey has a rich biodiversity as a result of its location, its geological struc- ture and different climatic zones. Turkey hosts three biogeographical regions: Mediterranean, Euro-Siberian and Irano-Turanian. As a result of located on the meeting point of these three different regions, Turkey is one of the most important areas in the world in terms of biological diversity. It is one of the world’s richest countries with regard to diversity of plant species, hosting 167 families, 1320 genera and 9996 species [1].
    [Show full text]
  • Prof. Dr. Faik Yaltirik'in Kizinin Arşivinden Dr. Davis
    PROF. DR. FAİK YALTIRIK’IN KIZININ ARŞİVİNDEN DR. DAVIS “Peter Hadland Davis: outstanding botanist” Rengigül Ural, 23 Nisan 2021 “Sevgili Rengigül Hanım, Dr. Davis'i platform (Kültürel ve Doğal Mirası İzleme) için yazabilirseniz seviniriz. Davis’in külliyatı tam anlamıyla anıtsal bir araştırma. Yaptığı proje inanılmaz. Ruhu şad olsun. Rahmetli Davis ile babanızın ülkemizin botanik kültürüne yaptıkları unutulmaz. Her türlü takdirin üzerinde. Ne yapsak az.” Nezih Başgelen, 22 Nisan 2021 “TERCİHLERE SAYGIYI ALTI YAŞIMDA EDINBURGH'TAKİ İLKOKULUMDA ÖĞRENDİM” konu başlığımla yazım şöyle idi: "Babam, Royal Botanic Garden’daki bilimsel çalışmalarına devam ediyordu. Akademik arkadaşları bizlere yardımcı oluyordu. Hepsi birbirinden iyi insanlardı. Araştırma grubunun başı Dr. Davis’ti. Ian Hedge, Mark Coode ve ailesi. Jennifer ve annesi. Jennifer, güleç yüzlü, uzun boylu, kumral dalgalı saçlı, cana yakın ve zarif bir hanımdı. Hele annesi ne kadar da tatlı, görmüş geçirmiş, gönül alan bir Büyük Hanım’dı. Babam bu şehirdeki ilk Türk bilim adamıydı. Ben de acaba ilk Türk kızı mıydım bu okulda? Annem ve babam ise her gün ne yaptığımızı soruyor, ben de anlatıyordum. Bir gün yine târif ettim sınıfta ne yaptığımızı. Hemen anladılar. Sabah dua ediliyordu. Annem “Sen, Müslümansın, put çıkartma. Duaya iştirak et. Dua her dilde duadır, kabul edilir. Ya da içinden bizim duamızı yap.” dedi. Ben bunu, öğretmenime nasıl anlatacaktım? Tabiî ki babam gelip, izâh etti ve anlayış ile karşılandı. Tercihlere saygıyı, altı yaşımda öğrenmiş oldum. Onlar “cross” işâreti yaptılar, ben avuçlarımı açtım. Çocuk duamız ve Tanrı’mız “bir” idi aslında... Okulun en alt katındaki, büyük salonu Cuma günleri; çarşı, pazar, yaya kaldırımı ile şehirde yaşamı öğreten bir düzene sokuyorlardı biz çocuklar için.
    [Show full text]
  • The Foxgloves (Digitalis) Revisited*
    Reviews The Foxgloves (Digitalis) Revisited* Author Wolfgang Kreis Affiliation Supporting information available online at Lehrstuhl Pharmazeutische Biologie, Department Biology, http://www.thieme-connect.de/products FAU Erlangen-Nürnberg, Erlangen, Germany ABSTRACT Key words Digitalis, Plantaginaceae, cardiac glycosides, plant biotech- This review provides a renewed look at the genus Digitalis. nology, biosynthesis, plant tissue culture, phylogeny Emphasis will be put on those issues that attracted the most attention or even went through paradigmatic changes since received March 17, 2017 the turn of the millennium. PubMed and Google Scholar were “ ” “ ” revised April 27, 2017 used ( Digitalis and Foxglove were the key words) to iden- accepted May 8, 2017 tify research from 2000 till 2017 containing data relevant enough to be presented here. Intriguing new results emerged Bibliography from studies related to the phylogeny and taxonomy of the DOI https://doi.org/10.1055/s-0043-111240 genus as well as to the biosynthesis and potential medicinal Published online May 23, 2017 | Planta Med 2017; 83: 962– uses of the key active compounds, the cardiac glycosides. 976 © Georg Thieme Verlag KG Stuttgart · New York | Several Eastern and Western Foxgloves were studied with re- ISSN 0032‑0943 spect to their propagation in vitro. In this context, molecular biology tools were applied and phytochemical analyses were Correspondence conducted. Structure elucidation and analytical methods, Prof. Dr. Wolfgang Kreis which have experienced less exciting progress, will not be Department Biology, FAU Erlangen-Nürnberg considered here in great detail. Staudtstr. 5, 91058 Erlangen, Germany Phone:+4991318528241,Fax:+4991318528243 wolfgang.kreis@fau.de Taxus species is a prime example [4].
    [Show full text]
  • Full Text (PDF)
    Environmental and Experimental Biology (2014) 12: 167–178 Original Paper Direct organogenesis, phytochemical screening and assessment of genetic stability in clonally raised Chlorophytum borivilianum Sautrik Basu1, Timir Baran Jha2* 1Department of Botany, Maulana Azad College, 8 Rafi Ahmed Kidwai Road, Kolkata 13, India 2P.G Department of Botany, Barasat Govt. College, Barasat, Kolkata 700124, India *Corresponding author, E-mail: tbjha2000@yahoo.co.in Abstract Keeping in mind the immense commercial and pharmacological importance of the rare aphrodisiac herb safed musli (Chlorophytum borivilianum Sant et Fern), the present investigation describes the development of a rapid and efficient protocol for direct plant regeneration from leaf explants of two populations of C. borivilianum collected from different agroclimatic zones of India. The induction of shoot initials was achieved on both Murashige and Skoog’s (MS) and Gamborg’s basal media supplemented with varying levels of 6-benzyladenine (1.11 to 8.90 μM) and thidiazuron (1.14 to 9.08 μM). MS basal medium supplemented with 4.54 μM thidiazuron was found ideal for shoot induction and gave rise to numerous shoot primordia within three weeks without any callus phase. Seperation of shoot clumps and their subsequent transfer to MS basal medium supplemented with 4.54 μM thidiazuron resulted in differentiation of more than 90% of the shoot initials into well developed shoots. Healthy shoots were rooted on half strength MS supplemented with varying concentrations of indole-3-acetic acid (2.28 to 11.42 μM), indole-3-butyric acid (1.97 to 9.80 μM) and naphthalene acetic acid (2.7 to 10.8 μM).
    [Show full text]
  • Munibe Monographs. Nature Series, 4 Le Programme Partenarial Espagne-France- Uzten Dituzte Agerian
    Liburu honetan, Botanika Piriniotar- Sarrera - Introducción - Introduction. Kantabriarraren XI. Nazioarteko Biltzarrean Iñaki Aizpuru. Batzorde zientifikoa/Comité científico/ Comité scientifique aurkeztutako lanak biltzen dira, zeina Bertizko Jaurerria Natur Parkean (Nafarroa) Treinta años de estudios botánicos pirenaico- cantábricos. Fundamento, situación y egin baitzen. Lan guztiek azken urteetan perspectivas. Luis Villar mendikate honetan egindako ikerketak Munibe Monographs. Nature Series, 4 Le programme partenarial Espagne-France- uzten dituzte agerian. Biltzarrak, gainera, Andorre FLORAPYR: Maintenir et développer les testuinguru egoki bat eskaini zuen klima- bases de connaissance sur la Flore des Pyrénées et les indicateurs de suivi en lien avec le change- aldaketaren aurrean floraren eta habitaten ment climatique. Gérard Largier (coordination) kontserbazioaren erronkari buru egingo Conservación ex situ de Lilium pyrenaicum Gouan: dioten egitasmo berriak eta elkarlanerako un endemismo pirenaico-cantábrico. Asier Jáñez, zubiak sortzeko. FLORA ETA HABITAT PIRINIAR-KANTABRIARRAK Agustí Agut, José Ignacio García-Plazaola Flora amenazada y vegetación del monte Jaizkibel. ALDAKETA KLIMATIKOAREN ERRONKAREN AURREAN Anaïs Mitxelena, Leire Oreja, Yoana García, Mari Azpiroz Munibe Monographs. Nature Series, 4 Les Atlas de la biodiversité Communale (ABC) dans Este libro recoge los trabajos presentados le Parc national des Pyrénées: premiers résultats sur en el XI. Coloquio Internacional de La flora y los hábitats pirenaico-cantábricos l’amélioration
    [Show full text]
  • Mesophyll Conductance to CO2: Current Knowledge and Future Prospects
    Plant, Cell and Environment (2008) 31, 602–621 doi: 10.1111/j.1365-3040.2007.01757.x Mesophyll conductance to CO2: current knowledge and future prospects JAUME FLEXAS1, MIQUEL RIBAS-CARBÓ1, ANTONIO DIAZ-ESPEJO2, JERONI GALMÉS1 & HIPÓLITO MEDRANO1 1Grup de Recerca en Biologia de les Plantes en Condicions Mediterrànies, Departament de Biologia, Universitat de les Illes Balears, Carretera de Valldemossa Km 7.5, 07122 Palma de Mallorca, Balears, Spain and 2Instituto de Recursos Naturales y Agrobiología, CSIC, Apartado 1052, 41080 Sevilla, Spain ABSTRACT INTRODUCTION: THE CONCEPT OF MESOPHYLL CONDUCTANCE TO CO2 AND During photosynthesis, CO2 moves from the atmosphere THE EVOLUTION OF ITS PERCEPTION IN (Ca) surrounding the leaf to the sub-stomatal internal cavi- PLANT SCIENCE ties (Ci) through stomata, and from there to the site of carboxylation inside the chloroplast stroma (Cc) through During photosynthesis, CO2 has to move from the atmo- the leaf mesophyll. The latter CO2 diffusion component is sphere surrounding the leaf across a boundary layer in the called mesophyll conductance (gm), and can be divided in air above the foliage surface to the sub-stomatal internal at least three components, that is, conductance through cavities through the stomata (Fig. 1a), and from there to the intercellular air spaces (gias), through cell wall (gw) and site of carboxylation inside the stroma through the leaf through the liquid phase inside cells (gliq). A large body of mesophyll (Fig. 1b). From Fick’s first law of diffusion, evidence has accumulated in the past two decades indicat- the net photosynthetic flux at steady state (AN) can be ing that gm is sufficiently small as to significantly decrease expressed as: AN = gs (Ca - Ci) = gm (Ci - Cc), where gs and Cc relative to Ci, therefore limiting photosynthesis.
    [Show full text]
  • Uncorrected Proof
    Industrial Crops & Products xxx (2018) xxx-xxx Contents lists available at ScienceDirect Industrial Crops & Products journal homepage: www.elsevier.com Efficient direct shoot organogenesis, genetic stability and secondary metabolite production of micropropagated Digitalis purpurea L. Naivy Pérez-Alonsoa⁠ ,⁠ g⁠ ,⁠ 1⁠ , Randel Martínb⁠ , Alina Capotea⁠ , Anabel Péreza⁠ , Elizabeth Kairúz Hernández-Díazb⁠ , Luis Rojasa⁠ , Elio Jiménezc⁠ , Elisa Quialad⁠ , Geert Angenone⁠ , Rolando Garcia-Gonzalesf⁠ , Borys Chong-Péreza⁠ ,⁠ f⁠ ,⁠ ⁎⁠ ,⁠ 1⁠ a Instituto de Biotecnología de las Plantas, Universidad Central “Marta Abreu” de Las Villas, Carretera a Camajuaní km 5,5, Santa Clara, Villa Clara, CP 54830, Cuba b Universidad Central “Marta Abreu” de Las Villas, Facultad de Ciencias Agropecuarias, Carretera a Camajuaní km 5,5, Santa Clara, Villa Clara, CP 54830, Cuba c Florida Crystals Corp, 25550 State Road 880 Atlantic Sugar Mill Rd, Belle Glade, FL 33430, USA d Instituto Nacional de Investigaciones Agropecuarias, INIAP Av. Eloy Alfaro N30-350 y Amazonas, Quito, Ecuador PROOF e Laboratory of Plant Genetics, Vrije Universiteit Brussel, Pleinlaan 2, B-1050 Brussels, Belgium f Sociedad de Investigación y Servicios BioTECNOS Ltda, Camino a Pangal km 2,5, San Javier, Chile g Botanical Solutions SpA, Ave. Quilin 3550, Santiago de Chile, Chile ARTICLE INFO ABSTRACT Keywords: Cardiovascular and cancer diseases are the first causes of death in the world. Digitalis purpurea L. is a medicinal Cardenolides plant that produces secondary metabolites, like digoxin and digitoxin, which are employed in therapies against Direct organogenesis heart failure. Moreover, anticancer and antiviral properties of these metabolites have recently been described. Foxglove The present work details a method to obtain in vitro plants of D.
    [Show full text]
  • Gene Der Cardenolid-Biosynthese Aus Digitalis - Und Isoplexis -Spezies
    Gene der Cardenolid-Biosynthese aus Digitalis - und Isoplexis -Spezies Den Naturwissenschaftlichen Fakultäten der Friedrich-Alexander-Universität Erlangen-Nürnberg zur Erlangung des Doktorgrades vorgelegt von Vanessa Jeannine Herl aus Nürnberg Gene der Cardenolid-Biosynthese aus Digitalis - und Isoplexis -Spezies Den Naturwissenschaftlichen Fakultäten der Friedrich-Alexander-Universität Erlangen-Nürnberg zur Erlangung des Doktorgrades vorgelegt von Vanessa Jeannine Herl aus Nürnberg Als Dissertation genehmigt von den Naturwissenschaftlichen Fakultäten der Universität Erlangen-Nürnberg Tag der mündlichen Prüfung: 03.11.2006 Vorsitzender der Promotionskommission: Prof. Dr. D.-P. Häder Erstberichterstatter: Prof. Dr. W. Kreis Zweitberichterstatter: Prof. Dr. W. Hillen Drittberichterstatter: Prof. Dr. B. Dräger Für meine Familie Der Mensch hat dreierlei Wege klug zu handeln: erstens durch Nachdenken, das ist der edelste, zweitens durch Nachahmen, das ist der leichteste, und drittens durch Erfahrung, das ist der bitterste. Konfuzius Danksagung Mein Dank gilt im besonderen Maße meinem Doktorvater, Herrn Prof. Dr. Wolfgang Kreis, der mir die Gelegenheit gab, dieses interessante und spannende Thema bearbeiten zu dürfen. Er verstand es durch seine stete Diskussionsbereitschaft meinen Blick auf das Wesentliche zu fokussieren, Ergebnisse auch kritisch zu hinterfragen und hat auf diese Weise entscheidend zum Gelingen dieser Arbeit beigetragen. Ganz besonders möchte ich mich bei Herrn Dr. Frieder Müller-Uri bedanken, der durch seinen Erfahrungsschatz, seine große Gesprächs- und Diskussionsbereitschaft vor und nach Feierabend, sowie durch seine Geduld und Unterstützung meine Begeisterung für die Forschung geweckt hat. Ein riesiges Dankeschön gilt Frau Gab(r)i(ele) Fischer, die sich für ihre exzellente technische Assistenz, unermüdliche Hilfs- und Diskussionsbereitschaft sowie für ihren unvergleichlichen Sinn für Humor mehr als nur einen „Du-bist-sooo-tapfer-Orden“ verdient hätte.
    [Show full text]
  • Morfogénesis in Vitro Y Efecto De Agentes Selectivos Sobre Explantes
    UNIVERSIDAD CENTRAL ʺMARTA ABREUʺ DE LAS VILLAS FACULTAD DE CIENCIAS AGROPECUARIAS DEPARTAMENTO DE BIOLOGÍA Tesis de Diploma Morfogénesis in vitro y efecto de agentes selectivos sobre segmentos foliares de Digitalis purpurea L. Randel Martín Hernández 2015 UNIVERSIDAD CENTRAL ʺMARTA ABREUʺ DE LAS VILLAS FACULTAD DE CIENCIAS AGROPECUARIAS DEPARTAMENTO DE BIOLOGIA Tesis de Diploma Morfogénesis in vitro y efecto de agentes selectivos sobre segmentos foliares de Digitalis purpurea L. Autor: Randel Martín Hernández a Tutores: Dra. C. Naivy Pérez Alonso b Dr. C. Borys Chong Pérez a naivy@ibp.co.cu b boris@ibp.co.cu Instituto de Biotecnología de las Plantas, Universidad Central "Marta Abreu" de las Villas Carretera a Camajuaní km 5.5, Santa Clara, Villa Clara, Cuba 2015 “… y conoceréis la verdad, y la verdad os hará libres” Jn 8:32 A Ramona y René a quienes les debo todo lo que soy, por ser además de mis abuelos, mis padres, mis amigos, mis héroes… Agradecimientos Agradecimientos Agradecer es lo mínimo y a la vez lo más difícil que puedo intentar hacer después de haber recibido tanto, prometo hacer mi mayor esfuerzo. Aunque temo que las siguientes palabras nunca estarán a la altura de a quienes va dirigido…Agradezco a: Luis Bravo mi mejor amigo, quien fue durante todos mis años de universidad mi guía, mi apoyo, como un padre me enseñó a andar… Todos los profesores del Dpto. de Biología de la UCLV y al Instituto de Biotecnología de las Plantas, de quienes obtuve mi formación académica y aprendí casi todo lo que sé… Raúl Barbón, por brindarme
    [Show full text]