PHYTOCHEMICAL AND MOLECULAR ANALYSIS OF SOME MEDICINAL PLANTS OF LABIATAE FAMILY GROWING AT DIF-FERENT ALTITUDES ON SAINT KATHERINE MOUNTAIN, SOUTH SINAI, EGYPT
Abstract
The present investigation was carried out to study the effect of altitudinal gradients on genetic and phytochemicals contents of three medicinal plant species belong to family Labiatae (Nepeta septemcrenata, Ballota undulata and Teucrium polium) in Saint Katherine Mountain under natural conditions. All analyses were carried out through three different altitudes viz., 1800 m a.s.l., 2200 m a.s.l. and 2600 m a.s.l. for the three species. Phytochemicals such as phenols, tannins, alkaloids, flavonoids and saponins were present in the methanolic extracts of aerial parts of three studied plant species but their quantity varied significantly across the different altitudes. The different species under study showed different values of total flavonoids, total phenolic acids, total saponins, total alkaloids and total tannins under the same environmental conditions. Meanwhile the same species exhibited different values of these metabolites under different elevation ranks. In general, total flavonoids and total phenolic acids were strongly increased with the increase of elevation from 1800 m a.s.l. until 2600 m a.s.l., however total saponins, total alkaloids and total tannins were slightly changed. The results are encouraging but scientific validation is necessary before being put into practice. RAPD and ISSR markers were successfully applied to assess the genetic diversity of the three medicinal plant species at the three different altitudes under natural conditions. Each of the five RAPD primers and the five ISSR primers used for analysis amplified different number of fragments. Each of the ten primers yielded from 5 to 12 DNA fragments whose molecular size ranged from approximately 200 to 1890 bp. The total number of amplified was 76 bands with an average of 7.6 fragments / primer and the total number of polymorphic fragments was 64, thus, representing a level of polymorphism of 84%. The highest number of amplified fragments (60) after using all the primers was detected in N. septemcrenata at 2200 m a.s.l with an average of 6 fragments per primer while the lowest number (35) was detected in T. polium at 1800 m a.s.l with an average of 3.5 fragments/primer. The highest similarity value (0.897) was found between B. undulate at 1800 m a.s.l and at 2200 m a.s.l while the lowest value (0.420) was found between B. undulate at 1800 m a.s.l and T. polium at 2200 m a.s.l. The dendrogram separated the three medicinal plant species into two clusters. First cluster included B. undulate and T. polium while second cluster included N. septemcrenata. The dendrogram separated the three different altitudes into two clusters. First cluster included 1800 m a.s.l and 2200 m a.s.l while second cluster included 2600 m a.s.l. Genetic polymorphism, the qualitative and quantitative phytochemical among the species are related to an altitudinal gradient. Assessing of genetic and phytochemical content of plants at varying altitudes can help to select elite genotype and reflect the best suited altitude for commercial cultivation of the species as these phytochemicals are considered as the basis for their medicinal activity.References
Ahmad, B., A. K. Naeem, A. Ghufran and A. Innamudin (2005). Pharmacological Investigation of Cassia sophera, Linn. Var. purpurea, Roxb. Medical Journal of Islamic World Academy of Sciences, 15: 105-109.
Ali, A. A., S. A. Ross, M. K. Mesbah and S. A. El Moghazy (1991). Phytochemical study of Limonium axillare (Forssk.). Bull. Fac. Pharm., 29: 59-62.
Arnold, N., B. Bellomaria, G. Volentiri, C. Yanninou and H. J. Arnald (1993). Plantes Medicinaleset-Phytotherpie, 26: 52-63.
Arzate-Fernández, A. M., M. Miwa, T. Shimada and T. Yonekura (2005). Genetic diversity of miyama-sukashi-yuri (Lilium maculatum Thunb. var. Bukosanense), an endemic endangered species at Mount Buko, Saitama, Japan. Plant Species Biol., 20: 57-65.
Balbaa, S. I., S. H. Hilal and A. Y. Zaki (1981). In "Medicinal Plants Constituents." 3rd Ed. General Organization for Univ. Books, Cairo, Egypt, pp. 644.
Boopathi, C. A. and R. Sivakumar (2011). Phytochemical screening studies on the leaves and stem of Andrographis neesianan Wightan endemic medicinal plants from India. World Appl. Sci. J., 12: 307-311.
Brieskorn, C. H. and P. W. Klinger-Hand (1961). Triterpenes and sterols in leaves of Salvia trioloba and Pyrus malus. Arch. Pharm., 294: 380-391.
Byrne, M., C. P. Elliott, C. J. Yates and D. J. Coates (2008). Maintenance of high pollen dispersal in Eucalyptus wandoo, a dominant tree of the fragmented agricultural region in Western Australia. Conserv. Genet., 9: 97-105.
Chung, M. Y., C. W. Park, E. R. Myers and M. G. Chung (2007). Contrasting levels of genetic diversity between the common, self-compatible Liparis kumokiri and rare, self-incompatible Liparis makinoana (Orchidaceae) in South Korea. Bot. J. Linn. Soc., 153: 41-48.
Citoglu, G. S., B. S. Yilmaz, B. Tarikahya and R. Tipirdamaz (2005). Chemotaxnomy of ballota species. Chem. Nat. Comp., 41: 299-302.
Darabpour, E., H. Motamedi and N. S. M. Seyyed (2010). Antimicrobial properties of Teucrium polium against some clinical pathogens. Asian Paci. J. Tropic. Med., 3: 124-127.
Einhellig, F. (1996). Interactions involving allelopathy in cropping systems. Agron. J., 88: 886-893.
Graglia, E., R. Julkunen-Tiitto, G. R. Shaver, I. K. Schmidt, S. Jonasson and A. Michelsen (2001). Environmental control and intersite variations of phenolics in Betula nana in tundra ecosystems. New Phytologist, 151: 227-236.
Hamrick, J. L and M. J. W. Godt (1989). Allozyme Diversity in Plant Species. In: Plant Population Genetics, Breeding, and Genetic Resources (Brown A. H. D, M. T. Clegg, A. L. Kahler and B. S. Weir, eds.). Sinauer Associates Sunderland, 43-63.
Hasanuzzaman, M., A. M. Ramjan, M. Hossain, S. Kuri and M. S. Islam (2013). Evaluation of total phenolic content, free radical scavenging activity and phytochemical screening of different extracts of Averrhoa bilimbi (fruits) Int. Curr. Pharm. J., 4: 92-96.
Hu, J., M. Nakatani, A. G. Lalusin and T. Kuranouchi (2003). Genetic analysis of sweetpotato and wild relatives using intersimple sequence repeats (ISSRs). Breed. Sci., 53: 297-304.
Ibitoye, D. O. and P. E. Akin-Idowu (2010). Marker-assisted-selection (MAS): A fast track to increase genetic gain in horticultural crop breeding. Afr. J. Biotechnol. 9: 8889-8895.
Jordano, P. and J. A. Godoy (2000). RAPD variation and population genetic structure in Prunus mahaleb (Rosaceae), an animal-dispersed tree. Mol. Ecol. 9: 1293-1305.
Khaleefa, A., A. N. Irshad and A. G. Bashir (2015). Altitudinal variation in some phytochemical constituents and stomatal traits of Primula denticulate. International Journal of Advances in Scientific Research, 1: 93-101.
Khandeal, K. R. (2008). Practical Pharmacognocy. Nirali Prakashan, Pune, edition: 19.
Knoop, W. and B. Walker (1985). Interactions of woody and herbaceous vegetation in a southern African savanna. J. Ecol., 73: 235-253.
Kokate, C. K., A. P. Purohit and S. B. Gokhale (2001). Carbohydrate and derived Products, drugs containing glycosides, drugs containing tannins, lipids and protein alkaloids. Text book of Pharmacognosy, 7: 133-166, 167-254, 255-269, 272-310, 428-523.
Kokate, C. K., A. P. Purohit and S. B. Gokhale (2004). Practical Pharmacognosy; 2nd edition. Vallabh Prakashan, New Delhi, 466-470.
Korner, C. (1999). Alpine Plant Life: Functional Plant Ecology of High Mountain Ecosystems. Springer, Berlin, Heidelberg, New York.
Li, H., Z. Wang and Y. Liu (2003). Review in the studies on tannins activity of cancer prevention and anticancer. Zhong-Yao-Cai, 26: 444-448.
Li, S., J. Li, X. L. Yang and Z. Cheng (2011). Genetic diversity and differentiation of cultivated ginseng (Panax ginseng C. A. Meyer) populations in North-East China revealed by inter-simple sequence repeat (ISSR) markers. Genet. Resour. Crop Evol., 58: 815-824.
Liu, D., X. He, G. Liu and B. Huang (2011). Genetic diversity and phylogenetic relationship of Tadehagi in southwest China evaluated by inter-simple sequence repeat (ISSR). Genet. Resour. Crop Evol., 58: 679-688.
Lynch, M. (1990). The similarly index and DNA fingerprinting. Molecular Biology and Evolution, 7: 478-484.
Lynch, M. (1991). Analysis of population genetic structure by DNA fingerprinting. In: DNA Fingerprinting Approaches and Applications (ed. by T. Burke, G. Dolf, A. J. Je،reys & R.Wolf), pp.113-126. Springer Verlag, Basel.
Mahmoud, M., A. H. Salam and A. S. Al-Ameri (2011). Extraction, identification and antimicrobial activity of some phenolic acids as antioxidants in Teucrium polium plant. Ker. J. Pharm. Sci., 2: 197-188.
Maurya, S. and D. Singh (2010). Quantitative analysis of total phenolic content in Adhatoda vasica Nees Extracts. International Journal of PharmTech Research, 4: 2403-2406.
McMahon, J. B., M. J. Currens, R. J. Gulakowski, R. W. J. Buckheit, C. Lackman-Smith, Y. F. Hallock and M. R. Boyd (1995). Michellamine B a novel plant alkaloid, inhibits human immunodeficiency virus-induced all killing by at least two distinct mechanisms. Antimicrobial Agents Chemotherapy, 39: 484-488.
Mossi, A. J., M. Mazutti, N. Paroul, M. L. Corazza, C. Dariva, R. L. Cansian and Oliveira (2009). Chemical variation of tannins and triterpenes in Brazilian populations of Maytenus ilicifolia. Braz. J. Biol., 69:3 39-345.
Naghibi, F., M. Mosaddegh, S. M. Mohamed and A. Ghorbani (2005). Labiateae family in folk medicine in Iran; from ethnobotany to Pharmacology. Iranian J. Pharm. Res. 2: 63-79.
Noble, R. I. (1990). The discovery of Vinca alkaloids chemotherapeutic agents against cancer. Biochem. Cell Biol., 68: 1544-1551.
Nonita, P. P. and M. U. Mylene (2010). Antioxidant and cytotoxic activities and phytochemical screening of four Philippine medicinal plants. Journal of Medicinal Plant Research, 4: 407-414.
Obadoni, B. O. and P. O. Ochuko (2001). Phytochemical studies and comparative efficacy of the crude extracts of some homeostatic plants from Edo and Delta States of Nigeria. Global Journal of Pure and Applied Sciences, 8: 203-208.
Pedrol, N. (2000). Caracterizacin Ecofisiolgica de Gramneas. Competencia y Estrés Hidrico. Thesis dissertation. Vigo, Spain: University of Vigo.
Pennycooke, J. C., S. Cox and C. Stushnoff (2005). Relationship of cold acclimation, total phenolic content and antioxidant capacity with chilling tolerance in petunia. Environ. Exp. Bot., 53: 225-232.
Poulev, A., J. M. O’Neal, S. Logendra, R. B. Pouleva, V. Timeva, A. S. Garvey, D. Gleba, I. S. Jenkins, B. T. Halpern, R. Kneer, G. M. Cragg and I. Raskin (2003). Elicitation, a new window into plant chemodiversity and phytochemical drug discovery. J. Med. Chem., 5: 2542-2547.
Purohit, I. (1998). Eco-physiology and biochemistry of nitrogen fixing tree species of Garhwal Himalaya. PhD. H. N. B. Garhwal Univ. Srinagar (Garhwal).
Rao, R. V. and T. Hodgkin (2002). Genetic diversity and conservation and utilization of plant genetic resources. Plant Cell Tissue Organ Cult., 68: 1-19.
Ricarda, D. S, J. M. Silva, N. Darmon and Y. Fernandez (1991). Oxygen free radical scavenger capacity in aqueous models of different procyanidins from grape seeds. Journal of Agricultural and Food Chemistry, 39: 1549-1552.
Rodríguez-Bernal, A., J. L. Piña-Escutia, L. M. Vázquez-García and A. M. Arzate-Fernández (2013). Genetic diversity of Cosmos species revealed by RAPD and ISSR markers. Genetics and Molecular Research, 12: 6257-6267
Salick, J., Z. Fangb and A. Byg (2009). Eastern Himalayan alpine plant ecology, Tibetan ethnobotany, and climate change. Global. Environ. Korner C. and P. Chochrane. 1983. Influence of leaf physiognomy on leaf temperature on clear midsummer days in the Snowy Mountains, Southeastern Australia. Acta Oecol., 4: 117-124.
Samatha, R., R. Shyamsundarachary, P. Srinivas, N. R. Swamy and L. Kurz (2012). Phytochemical screening and spectroscopic determination of total phenolic and flavonoid contents of Eclipta alba. Linn, 5: 177-179.
Sathiyamoorthy, P., H. Lugasi-Evgi, P. Schlesinger, I. Kedar, J. Gopas, Y. Pollack and A. Golan-Goldhirsh (1999). Screening for cytotoxic and antimalarial activities in desert plants of the Negev and Bedouin market plant products. Pharm. Biol., 37: 188-195.
Sathiyamoorthy, P., H. Lugasi-Evgi, P. Van-Damme, A. Abu-Rabia, J. Gopas and A. Golan-Goldhirsh (1997). Larvicidal activity in desert plants of the Negev and Bedouin market plant products. Int. J. Pharmacogn, 35: 265-273.
Seladji, M., C. Bekhechi, F. Beddou, H. Dib and N. Bendimerad (2014). Antioxidant activity and phytochemical screening of Nepeta nepetella aqueous and methanolic extracts from Algeria. Journal of Applied Pharmaceutical Science, 4: 012-016.
Sharaf, A. A., A. K. Om-Mohammed, E. H. El-Bialy and M. M. Mohamed (2013). Effect of altitudinal gradients on the content of carbohydrate, protein, proline and total Phenols of some desert plants in Saint Katherine Mountain, South Sinai, Egypt. Middle-East Journal of Scientific Research, 14: 122-129.
Sharma, S., S. A. Ali, A. Khare and T. K. Nailwal (2015). Genetic diversity analysis of Urtica parviflora in Uttarakhand Himalayas by Rapid marker. Biotechnol Biomater, 5: 183-186
Sneath, P. H. A. and R. R. Sokal (1973). Numerical Taxonomy. W. H. Freeman, San Francisco, USA.
Sodipo, O. A., M. A. Akanji, F. B. Kolawole and A. A. Odutuga (1991). Saponin is the active antifungal principle in Garcinia kola, heckle seed. Biosci. Res. Commun., 3: 171-171.
Soladoye, M. O. and E. C. Chukwuma (2012). Quantitative phytochemical profile of the leaves of Cissus populnea Guill. & Perr. (Vitaceae) an important medicinal plant in central Nigeria, Archives of Applied Science Research, 4: 200-206.
Solouki, M., N. R. Nazhad, R. Vignani, and B. A. Siahsar (2007). Polymorphism of some native Sistan grapes assessed by long and short primers for RAPD markers. Pak. J. Biol. Sci., 10: 1996-2001.
Spitaler, R., A. Winkler and I. Lins (2008). Altitudinal variation of phenolic contents in flowering heads of Arnica montana cv. ARBO: a 3-year comparison. J. Chem. Ecol., 34: 369-375.
Takhaolm, V. (1974). Student's Flora of Egypt. 2ed Ed., Published by Cairo Univ., Printed by Cooperative Printing Co., Beirut, pp. 888.
Tanaka-Oda, A., T. Kenzo, S. Kashimura, I. Ninomiya, L. Wang, K. Yoshikawa and K. Fukuda (2010). Physiological and morphological differences in the heterophylly of Sabina vulgaris Ant. In the semiarid environment of Mu Us Desert, Inner Mongolia, China. J. Arid. Environ., 74: 43-48.
Tattini, M., L. Guidi, L. Morassi-Bonzi, P. Pinelli, D. Remorini, E. Innocenti, C. Giordano, R. Massai and G. Agati (2005). On the role of flavonoids in the integrated mechanisms of response of Ligustrum vulgare and Phillyrea latifolia to high solar radiation. N. Phytol., 167: 457-470.
Treare, G. E. and W. C. Evans (1985). Pharmacognosy 17th edition, Bahiv Tinal, London, pp. 149.
Vinoth, S., P. Rajeshkanna, P. Gurusaravanan and N. Jayabalan (2011). Evaluation of phytochemical, antimicrobial and GC-MS analysis of extracts of Indigofera trita L.f. Spp. Subulata (Vahl ex poir), Int. J. Agri. Res., 6: 358-367.
Woo, W. S., H. J. Chi and S. Hye (1977). Alkaloid screening of some Saudi Arabian plants. Saengyak Hakhoe Chi (Hanguk Saengyak Hakhoe), 8: 109-113.
