نوع مقاله : مقاله کامل پژوهشی

نویسندگان

1 استادیار گروه بیوتکنولوژی کشاورزی، دانشکده علوم کشاورزی، دانشگاه شاهد

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

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

10.22041/ijbme.2012.13108

چکیده

در این تحقیق رشد سلولی، برخی پارامترهای فیزیولوژیک، تولید الکالوئید ضد سرطان تاکسول و بیان ژن در کشت سلولی فندق تحت اثر میدان مغناطیسی بررسی شد. سلولها در کشت تعلیقی با میدان مغناطیسی ایستا با شدت 30 میلی تسلا و در روزهای 8-11 بعد از واکشت، روزی 4 ساعت تیمار شدند. نتایج نشان داد میزان رشد و زنده‌مانی سلولها تحت اثر میدان قرار نگرفت؛ اما تولید H2O2و میزان پراکسیداسیون لیپیدهای غشایی افزایش یافت. فعالیت آنزیمهای فنیل آلانین آمونیا لیاز، پلی فنل اکسیداز و پراکسیداز تحت اعمال میدان مغناطیسی در مقایسه با شاهد افزایش یافت. تولید ترکیبات فنلی و تاکسول نیز در سلولهای تیمار شده در مقایسه با شاهد افزایش یافت. میدان مغناطیسی تاکسول درون سلولی را در مقایسه با تاکسول برون سلولی بیشتر افزایش داد و در کشتهای تیمار شده تولید تاکسول کل در مقایسه با کشتهای شاهد 9/2 برابر بود. بیان ژن 1- دئوکسی-D-زایلولوز -5- فسفات ردوکتوایزومراز نیز- که در تولید پیش‌سازهای تاکسول و بیوسنتز آن دخالت دارد- در سلولهای تیمار شده در مقایسه با شاهد افزایش یافت. به نظر می‌رسد میدان مغناطیسی با تحریک پاسخهای دفاعی سلول و القای بیان ژن دخیل در بیوسنتز تاکسول باعث افزایش تولید آن شده است.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Stimulation of Taxol Production by Magnetic Field in Cell Culture of Hazel (CorylusavellanaL.)

نویسندگان [English]

  • Ayatollah Rezaei 1
  • Faeze Ghanati 2
  • Mehrdad Behmanesh 3

1 Assistant Professor, Department of Agricultural Biotechnology, Faculty of Agricultural Sciences, Shahed University

2 2Associate Professor, Department of Plant Biology, Faculty of Biological Science, TarbiatModares University

3 Associate Professor, Department of Genetics, Faculty of Biological Sciences, TarbiatModares University

چکیده [English]

In this study cell growth, some physiological parameters, production of Taxol and gene expression in cell culture of hazel under effect of the magnetic field were investigated. Cells in suspension culture were treated by a 30 mT static magnetic field on days 8-11 after subculture and 4 hours each day. The results showed that while the growth rate and viability of cells weren’t affected by the magnetic field but membrane lipid peroxidation rate and H2O2 production increased. Activity of phenylalanine ammonia lyase, polyphenol oxidase and peroxidase enzymes was increased by the magnetic field compared with control. Production of phenolic compounds and Taxolin treated cells showed an increase compared to those of control cells. Magnetic field increased intracellular Taxol more than extracellurTaxol, and in treated cultures total taxol production was 2.9-fold compared to control culture. Gene expression of 1- deoxy -D- xylulose -5 - phosphate reductoisomerase involved in producing Taxol precursors and in its biosynthesis was also increased in treated cells compared to control. It appears that magnetic field by stimulating cell defense responses and inducing gene expression involved in Taxol biosynthesis has resulted in improved its production.

کلیدواژه‌ها [English]

  • Cell culture
  • Hazel
  • Taxol
  • Magnetic field
  • Gene expression
[1] Tabata H., Paclitaxel production by plant-cell-culture technology; Adv Biochem Eng Biotechnol, 2004; 87:1–23.
[2] Bestoso F., Ottaggio L., Armirotti A., Balbi A., Damonte G., Degan P., Mazzei M., Cavalli F., Ledda B., Miele M., In vitro cell cultures obtained from different explants of Corylusavellana produce Taxol and taxanes; BMC Biotechnol, 2006; doi:10.1186/1472-6750-6-45.
 [3] Ottaggio L., Bestoso F., Armirotti A., Balbi A., Damonte G., Mazzei M., Sancandi M., Miele M., Taxanes from shells and leaves of Corylusavellana; J Nat Prod, 2008; 7: 58–60.
[4] Rezaei A., Ghanati F., Behmanesh M., Ultrasound potentiated salicylic acid-induced physiological effects and production of taxol in hazelnut (Corylusavellana L.) cell culture; Ultrasound Med Biol, 2011; doi:10.1016/j.ultrasmedbio.2011.06.013.
[5] Dornenburg H., Knorr D., Strategies for the improvement of secondary metabolite production in plant cell cultures; Enzyme Microbial Technol, 1995; 17: 674–684.
[6]  Zhang Q.M., Tokiwa M., Doi T., Nakahara T., Chang P.W., Nakamura N.,  Hori M., Miyakoshi J., Yonei S., Strong static magnetic field and the induction of mutations through elevated production of reactive oxygen species in Escherichia colisoxR; Int J Radiat Biol, 2003; 79:281–286.
[7]  Sahebjamei H., Abdolmaleki P., Ghanati F., Effects of magnetic field on the antioxidant enzyme activities of suspension-cultured tobacco cells; Bioelectromagnetics, 2007; 28: 42–47.
[8]  Green L.M., Miller A.B., Agnew D.A., Greenberg M.L., Li J., Villeneuve J.P., Tibshirani R., Childhood leukemia and personal monitoring of residential exposures to electric and magnetic fields in Ontario; Cancer Cause Control, 1999; 10: 233–243.
[9] Smith M.A.L., Palta J.P., McCown B.H., The measurement of isotonicity and maintenance of osmotic balance in plant protoplast manipulations; Plant SciLett, 1984; 33: 249–258.
[10] De Vos C.H.R., Schat H., De Waal M.A.D., Vooijs R., Ernst W.H.O., Increased resistance to copper-induced damage of the root plasma membrane in copper tolerant Silenecucubalus; Physiol Plant, 1991; 82: 523–528.
[11] Velikova V., Yordanov I., Edreva A., Oxidative stress and some antioxidant systems in acid rain-treated been plants protective role of exogenous polyamines; Plant Sci, 2000; 151: 59–66.
[12] Dornenburg H., Knorr D., Evaluation of elicitor- and high-pressure-induced enzymatic browning utilizing potato (Solanumtuberosum) suspension cultures as a model system for plant tissues; Agri Food Chem, 1997; 45: 4173–4177.
[13] Ochoa-Alejo N., Gomez-Peralta J.E., Activity of enzymes involved in capsaicin biosynthesis in callus tissue and fruits of chili pepper (Capsicum annuum L.); J Plant Physiol, 1993; 141: 147–152.
[14] Yuan Y.J., Wei Z.J., Miao Z.Q., Wu J.C., Acting paths of elicitors on taxol biosynthesis pathway and their synergistic effect; BiochemEng J, 2002; 10: 77–83.
[15] Yano A., Ohashi Y., Hirasaki T., Fujiwara K., Effects of a 60 Hz magnetic field on photosynthetic CO2 uptake and early growth of radish seedlings; Bioelectromagnetics, 2004; 25: 572–581.
[16] Shang G.M., Wu J.C., Yuan Y.J., Improved cell growth and taxol production of suspension-culturedTaxuschinensisvar. maireiin alternating and direct current magnetic fields; Biotechnol Lett, 2004; 26: 875–878.
[17] Yaycili O., Alikamanoglu S., The effect of magnetic field on Paulownia tissue cultures; Plant Cell Tiss Org, 2005; 83: 109–114.
[18] Danilov V., Bas T., Eltez M., Rizakulyeva A., Artificial magnetic field effects on yield and  quality of tomatoes; ActaHortic, 1994; 366: 279–285.
[19] Trebbi G., Borghini F., Lazzarato L., Torrigiani P., Calzoni G.L., Betti L., Extremely low frequency weak magnetic fields enhance resistance of NN tobacco plants to tobacco mosaic virus and elicit stress-related biochemical activities; Bioelectromagnetics, 2007; 28: 214–223.
[20] Dicko M.H., Gruppen H., Barro C., Traore A.S., van Berkel W.J.H., Voragen A.G.J., Impact of phenolic compounds and related enzymes in sorghum varieties for resistance and susceptibility to biotic and abiotic stresses; J ChemEcol, 2005; 31: 2671–2688.
[21] Ghanati F., Abdolmaleki P., Vaezzadeh M., Rajabbeigi E., Yazdani M., Application of magnetic field and iron in order to change medicinal products of Ocimumbasilicum; Environmentalist, 2007; 27: 429–434.
[22] Shoderhall I., Properties of carrot polyphenol oxidase; Phytochemistry, 1995; 39: 33–38.
[23] Ruiz J.M., Garcia P.C., Rivero R.M., Romero L., Response of phenolic metabolism to the application to the carbendazim plus boron in tobacco leaves; Physiol Plant, 1999; 106: 151–157.
[24] Kursevich N.V., Travkin M.P., Effects of magnetic fields with different intensities on some enzymes’ activities in barley seedlings. In: Effects of Natural and Weak Artificial Magnetic Fields on Biological Objects; Belgorod Teacher’s Training College Publishing Co.; Belgorod: Russia; 1973. p. 102–4.
[25] Hiraga S., Sasaki K., Ito H., Ohashi Y., Matsui H., A large family of class III plant peroxidases; Plant Cell Physiol, 2001; 42: 462–468.
[26] Atak C., Emiroglu O., Alikamanoglu S., Rzakoulive A., Stimulation of regeneration by magnetic field in soybean (Glycine max L. Merrill) tissue cultures; J Cell MolBiol, 2003; 2: 113–119.
[27] Abdolmaleki P., Ghanati F., Sahebjamei H., Sarvestani A., Peroxidase activity, lignification and promotion of cell death in tobacco cells exposed to static magnetic field; Environmentalist,  2007; 27: 435–440.
[28] Kato R., Kamada H., Asashama M., Effect of high and very low magnetic field on the growth of hairy roots of  Daucuscarotta and Atropa belladonna; Cell Physiol, 1989; 30: 605–608.
[29] Aukhez S.T., Beharry G.K., Effects of magnetic field on growth and polyphenolic production in callus cultures of Cassia fistula; SciTechnol Res J, 2001; 8: 13–27.
[30] Phillips M.A., Walter M.H., Ralph S.G., Dabrowska P., Luck K., Uros E.M., Boland W., Strack, Rodriguez-Concepcion M., Bohlmann J., Functional identification and differential expression of 1-deoxy-D-xylulose 5-phosphate synthase in induced terpenoid resin formation of Norway spruce (Piceaabies); Plant MolBiol, 2007; 65: 243–257.
[31] Yao H., Gong Y., Zuo K., Ling H., Qiu C., Zhang F., Wang Y., Pi Y., Liu X., Sun X., Tang K., Molecular cloning, expression profiling and functional analysis of a DXR gene encoding 1-deoxy-D-xylulose 5-phosphate reductoisomerase from Camptotheca acuminate; Plant Physiol. 2008; 165: 203–213.