افزایش تولید ترکیبات لیگنانی و فنلی تحت تاثیر دیواره قارچ Fusarium graminearum در کشت تعلیقی کتان سفید Linum album

نوع مقاله: مقاله پژوهشی

نویسندگان

1 دانشیار گروه علوم گیاهی، دانشکده علوم زیستی، دانشگاه تربیت مدرس

2 دانشیار گروه بیماری های گیاهی، دانشکده کشاورزی، دانشگاه تربیت مدرس

3 دانشگاه تربیت مدرس

4 دانشیار گروه ژنتیک، دانشکده علوم زیستی، دانشگاه تربیت مدرس،

چکیده

کتان سفید (Linum album Kotschy ex Boiss.) یکی از گونه های بومی ایران، دارای پودوفیلوتوکسین و دیگر ترکیبات لیگنانی است. پودوفیلوتوکسین، لیگنانی است که در گونه های گیاهی معدودی حضور دارد و به دلیل خواص ضد سرطانی اهمیت دارویی زیادی پیدا کرده است. دست ورزی محیط های کشت سلولی با الیسیتورها یکی از راهکارهای مهم جهت القای تولید متابولیت های ارزشمند می باشد. در این پژوهش، دیواره جدا شده قارچ Fusarium graminearum پس از تعیین غلظت بهینه، در روز 7 بعد از واکشت به کشت سلولی کتان سفید با غلظت 1 درصد (v/v) اضافه گردید. پس از برداشت سلول ها در روزهای 3 و 5 و 7 بعد از تیمار، میزان ترکیبات لیگنانی با HPLC بررسی گردید. همچنین میزان ترکیبات فنلی، فلاونوئیدی، فلاوونول ها و لیگنین بررسی شد. نتایج نشان داد میزان پودوفیلوتوکسین و لاریسی رزینول بعد از تیمار در مقایسه با نمونه های شاهد افزایش قابل ملاحظه ای داشته است. میزان ترکیبات فنلی در روز 5 بعد از تیمار و میزان ترکیبات فلاونوئیدی و فلاونولی در روز 3 و 7 بعد از تیمار افزایش داشته است. لیگنین نیز به عنوان یک ترکیب دفاعی بعد از تیمار با دیواره قارچ افزایش یافت.

کلیدواژه‌ها

عنوان مقاله [English]

Effect of Fusarium graminearum cell wall on lignans and phenolic compounds in cell culture of Linum album

نویسندگان [English]

  • Jaleh Tahsili 1
  • Mozafar Sharifi 2
  • Naser Safaei 3
  • Mehrdad Behmanesh 4

1 MSc. graduated Department of Plant Biology, Faculty of Biological Sciences, Tarbiat Modares University

2 Assistant professor Department of Plant Pathology, Faculty of Agriculture, Tarbiat Modares University

4 Research assistant Department of Genetic, Faculty of Biological Sciences, Tarbiat Modares University

چکیده [English]

Linum album Kotschy ex Boiss., one of endemic species in Iran, produces podophyllotoxin (PTOX) and other lignans. Podophyllotoxin which occurs in a few plant species and has pharmacological significance for it's anticancer activities. Manipulation of cell culture media by elicitors is one of the important strategies for inducing secondary metabolism and production of valuable metabolites. In this study, first we isolated cell wall of Fusarium graminearumas described by Di Pietro et al. (2009). We investigated the different concentrations effect of fungal cell wall on cell growth and PTOX and lariciresinol production in cell suspension cultures of L. album. Then, the optimal concentration of fungal cell wall (1% v/v) was selected for further experiments and thus after 7 days of pre-culture, the cells were treated with 1 % (v/v) of cell wall. Lignan analysis with HPLC demonstrated that production of podophyllotoxin and lariciresinol significantly increased. Also phenolic compounds have reached the most accumulation at 5 day after treatment. But flavnoids and flavones had increased at 3 and 7 day after treatment. As defensive compound, lignin increased after treatment.

کلیدواژه‌ها [English]

  • : Linum album
  • Cell culture
  • fungal elicitors
  • Lignans

اسمعیل زاده ص. (1390). بررسی اثر الیسیتورهای قارچی بر میزان پودوفیلوتوکسین و بیان ژن و فعالیت آنزیم‌های فنیل آلانین آمونیا لیاز و سینامیل الکل دهیدروژناز در Linum album. رساله دکتری، دانشگاه تربیت مدرس، دانشکده علوم زیستی.  

فخاری ص. (1389). بررسی تغییرات میزان پودوفیلوتوکسین در پاسخ به تنظیم کننده های رشدگیاهی و متیل جاسمونات در کشت کالوس و گیاهچهLinum album . پایان نامة کارشناسی ارشد. دانشگاه تربیت مدرس، دانشکدة علوم زیستی.

یوسف زادی م. (1389). بررسی اثر تیروزین، تریپتوفان، نور و سالیسیلیک اسید بر میزان پودوفیلوتوکسین، فعالیت آنزیمی و بیان ژن برخی از آنزیم‌های مسیر بیوسنتزی پودوفیلوتوکسین در گیاه Linum album. رساله دکتری، دانشگاه تربیت مدرس. دانشکدة علوم زیستی.

Akkol, E. K., Goger, F., Kosar, M., Baser, K. H. (2008). Phenolic composition and biological activities of Salvia halophila and Salvia virgata from Turkey. Food Chemistry. 108: 942–949.

Arroo, R.R., Alfermann, A.W., Medarde, M., Petersen, M., Woolley, J.G. (2003). Plant cell factories as a source for anti-cancer lignans.Phytochemistry Reviews.1: 27-35.

Baldi, A., Jain A., Bisaria V.C. (2008). Co-culture of arbuscular mycorrhiza-like fungi (Piriformospora indica and Sebacina vermifera) with plant cells of Linum album for enhanced production of podophyllotoxin: a first report. Biotechnology Letter. 30: 1671–1677.

Chen, S.Y., Dickson, D.W., Mitchel, D.J. (2000). Viability of Heterodera glycines exposed to fungal filtrates. Journal of Nematology. 32: 190-197.

Chong, T., Abdulah, M., Lajis, N. (2005). Effective elicitation factors in Morinda elliptica cell suspension culture. Process Biochemistry. 40: 3397- 3405.

Cordell, G.A. (1997). The Alkaloids: Chemistry and Biology. Academic Press, New York, USA. 380-384.

De Alwis, R., Fujita, K., Ashitani, T. (2009). Induced monoterpene and lignin production in mechanically stressed and fungal elicited cultured Cupressus lusitanica cells. Plant Biotechnology Reports. 3:57–65.

De Pietro, A., Rosales, R.P., Luque-Gracia, J. L., Martines lopez, R., Gil, C. (2009). The Fusarium oxysporum cell wall proteome under adhesion-inducing conditions. Proteomocs. 9 : 4755-4769.

Esmaeilzadeh Bahabadi, S., Sharifi, M., Safaie, N., Murata, J., Yamagaki, T., and Satake, H., 2011. Increased lignan biosynthesis in the suspension cultures of Linum album by fungal extracts. Plant Biotechnology Report, 5:367-373.

Funk,C., Brodelius, P.E. (1992). Phenylpropanoid metabolism in suspension cultures of Vanilia planifolia. Plant Physiology. 94: 95–101.

Furden, B.V., Humburg, A., Fuss, E. (2005). Influence of methyl jasmonate on podophyllotoxin and 6-methoxypodophyllotoxin accumulation in Linum album cell suspension cultures. Plant Cell Report. 24: 312–317.

Federolf, K., Alfermann, A.W., Fuss, E. (2004). Aryltetralin-lignan formation in two different cell lines of Linum album: deoxypodophyllotoxin 6-hydroxylase, a key enzyme for the formation of 6-methoxypodophyllotoxin. Phytochemistry. 68:1397-1406.

Fuss, E. (2003). Lignans in plant cell and organ cultures: An overview. Phytochemistry Review.2: 307–320.

Hano, C., Addi, M., Bensaddek, L., Baltora-Rosset, S., Doussot, J., Maury, S., Mesnard, F., Chabbert, B., Hawkins, S., Laine, E., Lamblin, F. (2006). Differential accumulation of monolignol-derived compounds in elicited flax (Linum usitatissimum) cell suspension cultures. Planta: 223: 975–989.

Iiyama, K., Wallis, A. (1988). An improved acetyl bromide procedure for determining lignin in woods and wood pulps. Wood Science Technology: 22: 271-280.

Imbert, T. F. (1998). Discovery of podophyllotoxin. Biochimie. 80: 207-222.

Imre, E., Somssich, I.E., Hahlbrok, K. (1998). Pathogen defence in plant-a paradigm of biological complexity. Trends in Plant Science. 3:86–90.

Ionkova, I. (2007). Biotechnological approaches for the production of lignans. Pharmacognosy Reviews. 1: 427-443.

Kang, S.M., Jung, H.Y., Kang, Y.M., Yun, D.J., Bahk, J.D., Yang, J.K., Choi, M.S. (2004). Effects of methyl jasmonate and salicylic acid on the production of tropane alkaloids and the expression of PMT and H6H in adventitious root cultures of Scopolia parviflora. Plant Science. 166: 745-751.

Khosroushahi, A., Valizadeh, M., Ghasempour, M. Naghdibadi , H., Dadpour, M.R., Omidi, Y. (2005). Improved Taxol production by combination of inducing factors in suspension cell culture of Taxus baccata. Cell Biology International. 30: 262-269.

Kovacik, J., Backor, M., Strnad, M., Repcak, M. (2009). Salicylic acid-induced changes to growth and phenolic metabolism in Matricaria chamomilla plants. Plant Cell Report. 28:135–143.

Kumar, V., Rajauria, G., Sahai, V., Bisaria, V.S. (2012). Culture filtrate of root endophytic fungus Piriformospora indica promotes the growth and lignan production of Linumalbum hairy root cultures. Process Biochemistry. 47:901–907.

Missawa, M. (1994). Plant tissue culture: an alternative for production of useful metabolites. Daya Publishing House. Toronto, Canada. Chapter 7.

Muranaka, T., Miyata, M, Tachibana S. (1998). Production of podophyllotoxin in Juniperus chinensis callus cultures treated with oligosaccharides and a biogenetic precursor. Phytochemistry. 49: 491–496.

Murashige, T., Skoog, F. (1962). A revised medium for rapid growth and bioassay of tobacco tissue culture. Physiologia Plantarum. 15: 473-497.

Namdeo, A. (2007). Plant cell elicitation for production of secondary metabolites: A review. Pharmacognosy Review. 1: 320-345.

Pu, G.B., Dong-Ming, M., Chen, J.L., Ma, L.Q., Wang, H., Li, G.F. (2009). Salicylic acid activates artemisinin biosynthesis in Artemisia annua L. Plant Cell Report. 28: 1127–1135.

Savitha, B.C., Thimmaraju, R., Bhagyalakshmi, N. (2006). Different biotic and abiotic elicitors influence betalain production inhairy root cultures of Beta vulgaris in shake-flask and bioreactor. Process Biochemistry. 41: 50–60.

Scheel, D., Parker, J.E. (1990). Elicitor recognition and signal transduction in plant defense gene activation. Zeitschrift für Naturforschung C. 45: 569-75.

Schützendübel, A., Schwanz, P., Teichmann, T., Gross, K., Langenfeld-Heyser, R., Godbold, D.L. (2001). Cadmium-Induced changes in antioxidative systems, hydrogen peroxide content, and differentiation in scots pine roots. Plant Physiology. 127: 887-898.

Sok, D.E., Cui, H.S., Kim, M. (2009). Isolation and boactivities of furfuran type lignan compounds from edible plants. Recent Patents on Food, Nutrition & Agriculture. 1: 87-95.

Suzuki, S., Umezawa, T. (2007). Biosynthesis of lignans and norilignans. Journal of Wood Science. 53: 273-284.

Tokunaga N., Sakakibara N., Umezawa T., Ito Y., Fukuda H. and Sato Y. (2005). Involvement of extracellular dilignols in lignification during tracheary element differentiation of isolated Zinnia mesophyll cells. Plant and Cell Physiology, 46:224-232. Umezawa T. (2003) Diversity in lignan biosynthesis. Phytochemistry Review, 2:371390.

Van Uden, W., Pras, N., Visser, J.F., Malingre, T.M. (1989). Detection and identification of podophyllotoxin produced by cell cultures derived from Podophyllum hexandrum Royle. Plant Cell Report. 8:165–168.

Vasconsuelo, A., Boland, R. (2007). Molecular aspects of the early stages of elicitation of secondary metabolites in plants. Plant Science. 172: 861-875.

Vogt, T. (2010). Phenylpropanoid biosynthesis. Molecular Plant. 3: 2-20.

Wang, C.G., Wu. J.Y. (2001) Enhancement of taxol production and excretion in Taxus chinensis cell culture by fungal elicitation and medium renewal. Applied Microbiology and Biotechnology. 55: 404–410.

Westcott, N.D., Muir, A.D. (2003).  Flax seed lignan in disease prevention and health promotion. Phytochemistry  Review. 2: 401-417.

Wojdyo, A., Oszmian, J., Czemerys, R. (2007) Antioxidant activity and phenolic compounds in 32 selected herbs. Food Chemistry. 105: 940–949.

Yousefzadi, M., Sharifi, M., Behmanesh, M., Moyano, E., Palazon, J. (2010). Salicylic acid improves podophyllotoxin production in cell cultures of Linum album by increasing the expression of genes related with its biosynthesis. Biotechnology Letter. 32: 1739–1743.

Zhao, J., Davis, L.C., Verpoorte, R. (2005). Elicitor signal transduction leading to production of plant secondary metabolites. Biotechnolgy Advance. 23: 283–333.