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بهینه‌سازی روش استخراجRNA از بافت پوست شاخه درختان خانواده مخروطیان | ||
| تحقیقات ژنتیک و اصلاح گیاهان مرتعی و جنگلی ایران | ||
| مقاله 9، دوره 24، شماره 1، اردیبهشت 1395، صفحه 102-113 اصل مقاله (253.09 K) | ||
| نوع مقاله: مقاله علمی - پژوهشی | ||
| شناسه دیجیتال (DOI): 10.22092/ijrfpbgr.2016.106171 | ||
| نویسندگان | ||
| فائزه حائری زاده1؛ اسد معصومی اصل* 2؛ پژمان آزادی3؛ حمید رجبی معماری4؛ نفیسه ابوفاضلی5 | ||
| 1دانشجوی کارشناسی ارشد، اصلاح نباتات، دانشگاه یاسوج، یاسوج | ||
| 2نویسنده مسئول مکاتبات، استادیار، دانشکده کشاورزی، دانشگاه یاسوج، یاسوج پست الکترونیک: Masoumiasl@yu.ac.ir | ||
| 3استادیار، پژوهشکده بیوتکنولوژی کشاورزی ایران، کرج | ||
| 4استادیار، دانشکده کشاورزی، دانشگاه شهید چمران، اهواز | ||
| 5عضو شرکت دانش بنیان زیست فناوران نوین گیتی ژن، تهران | ||
| چکیده | ||
| فرآیند استخراج RNA از بافتهای سخت چوبی و بهطور خاص درختان خانواده مخروطیان بهدلیل داشتن غلظتهای بالایی از ترکیبات فنولیک، پلیساکاریدها، متابولیتهای ثانویه و لیگنین، بسیار مشکل است و تا بهحال هیچگونه کیت تجاری برای این منظور ساخته نشده است. در این مطالعه مشکلات فوق برطرف گردیده و روشی سریع، ساده و اقتصادی برای استخراج RNA از بافت ساقه گونه‌های Cedrus deodara و Abies grandis که هر دو از خانواده مخروطیان میباشند، ارائه شده است. نسبت جذب نوری در طول موج‌های 260 به 280 و 260 به 230 نانومتر، بهترتیب 88/1 و 1/1 برای گونه Cedrus deodara و 8/1 و 9/0 برای گونه Abies grandis بهدست آمد. تغییرات صورت گرفته در جهت بهینه‌سازی استخراج RNA نیز شامل استفاده از تیمار پیش‌گرما روی بافر استخراج، کاهش دور سانتریفیوژ حذف آلودگی DNA و حذف DEPC می‌باشد. ساخت cDNA با استفاده از RNA استخراج شده با موفقیت انجام و واکنش RT-PCR باندهای مورد انتظار را تولید نمود. لذا بهنظر میرسد این روش برای استخراج RNA از بافتهای چوبی مناسب است. | ||
| کلیدواژه‌ها | ||
| بافت چوبی؛ ترکیبات فنولیک؛ درخت نراد؛ DEPC | ||
| عنوان مقاله [English] | ||
| Optimization of RNA extraction protocol for trees bark tissue of coniferous family | ||
| نویسندگان [English] | ||
| faeezeh haeerizadeh1؛ asad masoomi asl2؛ pezhman azadi3؛ hamid rajabi-memari4؛ nafiseh aboofazeli5 | ||
| چکیده [English] | ||
| RNA extraction from hard wood tissues, especially in coniferous family trees, is very hard because they have high concentrations of phenolic compounds, polysaccharides, secondary metabolites and lignin. Moreover, up to now, no commercial kit is made for the mentioned purpose. In this study, the problems were resolved and a rapid, simple, and economic protocol is presented for RNA extraction from bark tissue of Cedrus deodara and Abies grandis, which belong to conifer family. Ratios of optical density of 260/280 nm and 260/230 nm were respectively obtained as 1.88 and 1.1 for Cedrus deodara and 1.8 and 0.9 for Abies grandis. Modifications for optimization of RNA extraction were preheating treatment of extraction buffer, and decreasing centrifuge rounds to eliminate DNA contaminations and DEPC. Synthesis of cDNA was performed using extracted RNA. RT-PCR successfully produced expected bands. Therefore, it seems that the presented protocol is suitable for RNA extraction from woody tissues. | ||
| کلیدواژه‌ها [English] | ||
| Abies grandis, bark tissue, DEPC, Phenolic compounds | ||
| مراجع | ||
|
- Asif, M.H., Dhauan, P. and Nath, P., 2000. A simple procedure for the isolation of high quality RNA from ripening banana fruit plant. Molecular Biology Reporter, 18: 109-115.
- Baker, J.T., Sanders, A.L., Maccotta, L. and Buckner, R.L., 2001. Neural correlates of verbal memory encoding during semantic and structural processing tasks. Neuro Report, 12: 1251– 1256.
- Bell, D.A. and DeMarini, D., 1991. Excessive cycling converts PCR products to random-length higher molecular weight fragments. Nucleic Acids Research, 19: 5079-5079.
- Beutler, E., Gelbart, T. and Kuhl, W., 1990. Interference of heparin with the polymerase chain reaction. BioTechniques, 9: 166-166.
- Brown, T.A., 2006. Gene cloning an introduction. Translated by: AhmadianTehrani, P., University of Tehran Press. 415pp.
- Budavari, S., 1996. The Merck index- Encyclopedia of chemicals, Drugs and Biologicals. 12th edition.. Merck and Co., Inc., Rahway, N.J. U.S. 1175pp.
- Cara, A., Gambino, G. and Schubert, A., 2007. A cetylmethylammonium bromide based method to extract low-molecular-weight RNA from polysaccharide-rich plant tissue. Analytical Biochemistry, 360: 318-320.
- Chang, S., Puryear, J. and Carney, J., 1993. A simple and efficient method for isolation RNA from pine trees. Plant Molecular Biology, 11: 113-116.
- Claros, M.G. and Canovas, F.M., 1998. Rapid high quality RNA preparation from Pine seedlings. Plant Molecular Biology Reporter, 16:9-18.
- Dean, W.L. and C. Tanford. 1978. Properties of a delipidated detergent-activated Ca-ATPase. Biochemistry, 17: 1683-1690.
- Fu, X., Deng, S., Su, G., Zeng, Q. and Shi, S., 2004. Isolating high-quality RNA from mangroves without liquid nitrogen. Plant Molecular Biology Reporter, 22: 197-198.
- Gasic, K., Hernandez, A. and Korban, S., 2004. RNA extraction from different apple tissues rich in polyphenols and polysaccharides for cDNA library construction. Plant Molecular Biology, 22: 437-437.
- Gelrig, H.H., Winter, K., Cusman, J., Borland, A. and Taybi, T., 2000. Improved RNA isolation method for succulent plant species rich in polyphenol and polysaccharides. Plant Molecular Biology Reprter, 18: 369-376.
- Harsiah, C., Namasivayam, P. and Mohamed, R., 2012. Comparisons of different RNA extraction methods on woody tissues of the tropical tree, Aquilariamalaccensis. Asia-Pacific Journal of Molecular Biology and Biotechnology, 20 : 107-113.
- Hoppe, B.L., Conti-Tronconi, B.M. and Horton R.M., 1992. Gel-loading dyes compatible with PCR. BioTechniques, 12: 679–680.
- Kainz, P., 2000. The PCR plateau phase-towards an understanding of its limitations. Biochemistry and Biophysics Acta, 1494: 23−27.
-Kamaloddini, H., Solooki, M., Riginegad, N. and Haddadi, F., 2006. Plant Biotechnology. Golbon Publication. Tehran, Iran. 238pp.
- Kolosava, N., Miller, B., Ralph, S., Elis, B.E., Douglas, C., Ritland, K. and Bohlmann, J., 2004. Isolation of high-quality RNA from gymnosperm and Angiosperm trees. Biotechniques, 35: 821-824.
- Ky, H., Yeap, S.K. and Napis, S.B., 2012. The best method for isolated total RNA from durian tissues. International Food Research Journal, 19: 1181-1183.
- Le Provast, G., Herrera, R., Pavia, G., Chaumeil, P., Salin, F. and Plomion, C., 2007. A micromethod for high throughput RNA extraction in forest trees. Biological Research, 40: 291-297.
- Lewinsohn, E., Steele, C.L. and Croteau, R., 1994. Simple isolation of functional RNA from woody stem of gymnosperms. Plant Molecular Biology Report, 12:20-25.
- Loomis, W.D., 1974. Overcoming problems of phenolics and quinones in the isolation of plant enzymes and organelles. Methods in Enzymology, 31:528-544.
- Lorenz, W.W., Yu, Y.S. and Dean, J.F.D., 2010. An improved method of RNA isolation from Loblolly Pine (P. taeda L. ) and other conifer species. Journal of Visualized Experiments, 36: 1751-1756.
-Mousazade Moghadam, M., Babavalian, H., Mirnejad, R. and Shakeri, F., 2012. Rapid DNA extraction of bacterial genome of Staphylococcus aureus using laundry detergents and assessment of the efficiency of DNA in downstream process using. Journal of Medical Laboratory Sciences, 6: 35-42.
- Paula, M.F.B., Sagio, S.A., Lazzari, F., Barreto, H.G., Pavia, L.V. and Chalfun-Junior, A., 2012. Efficiency of extraction protocols in different types of coffee plant tissues. Coffee Science, 7:284-293.
- Powell, L.M., Wallis, S.C., Pease, R.G., Edwards, Y.H., Knott, T.J. and Scott, J., 1987. A novel form of tissue-specific RNA processing produces apolipoprotein-B48 in intestine. Cell, 50: 831–840.
- Reid, K.E., Olsson, N., Schlosser, J., Peng, F. and Lund, S.T., 2006. An optimized grapevine RNA isolation procedure and statistical determination of reference genes for real-time RT-PCR during berry development. Plant Biology, 6: 27-37.
- Rodrigues, S.M., Soares, V.L., De Oliveira, T.M., Gesteira, A.S., Otoni, W.C. and Costa, M.G., 2007. Isolation and purification of RNA from tissues rich in polyphenols, polysaccharides, and pigments of annatto (Bixa orellana L.). Molecular Biotechnology, 37: 220-224.
- Roux, K.H., 2009. Optimization and Trouble shooting in PCR. Cold Spring Harbor Laboratory Press. Vol. 4, Issue 4. 7pp.
- Rubio-Pina, J. and Zapata-Perez, O., 2011. Isolation of total RNA from tissues rich in polyphenols and polysaccharides of mangrove plants. Plant Biotechnology, 14:-11-11.
- Saiki, R.K., Scharf, S., Faloona, F., Mullis, K., Glenn, B., Horn, T., Erlich, H.A. and Arnheim, N.,1985. Enzymatic Amplification of ß-Globin genomic sequence and restriction site analysis for diagnosis of Sickle Cell Anemia. Science, 230: 1350-1354.
- Sambrook, J. and Russel, D.W., 2001. Molecular Cloning: A laboratory manual. Third edition. New York: Cold Spring Harbor Laoratory Press. 2100p.
- Sarkar, G., Kapelner, S. and Sommer, S.S., 1990. Formamid can dramatically improve the specificity of PCR . Nucleic Acid Research, 18:7465-7465.
- Schneiderbauer, A., Sanderman H.Jr. and Ernst, D., 1991. Isolation of functional RNA from plant tissues rich in phenolic compounds. Annual Biochemistry, 197: 91-95.
- Steele, C.L., Katoh, S., Bolmann, J. and Croteau, R.,1998. Regulation of oleorsinosis in Grand fir (Abies grandis) differential transcriptional control of monoterpene, sesquiterpene, and diterpene synthase gene in response to wounding. Plant Physiology. 116: 1497-504.
- Sullivan, D., Fahey, B. and Titus, D., 2006. Fast PCR: General considerations for minimizing run times and maximizing throughput. Bio-Rad Laboratories, Inc. Hercules, CA, USA. 36pp.
- Sun, Y., Hegamyer, G. and Colburn, N.H.,1993. PCR-directed sequencing of a GC-rich region by inclusion of 10% DMSO: application to c-jun. Biotechniques, 15: 372–374.
- Tattersall, E.A.R., Ergul, A., Alkayal, F., Deluc, L., Cushman, J.C. and Cramer, G.R., 2005. Comparison of methods for isolating high-quality RNA from leaves of grapevine. American Journal of Enology and Viticulture, 56: 400-406.
- Vasanthaiah, H.K.N., Katam, R. and Sheikh, M.B., 2008. Efficient protocol for isolation of functional RNA from different grape tissue rich in polyphenols and polysaccharides for gene expression studies. Electronic Journal of Biotechnology,11:1-8.
- Wang, T., Zhang, N. and Du, L., 2005. Isolation of RNA of high quality and yield from Ginkgo biloba leaves. Biotechnology Letters, 27: 629-633.
- Warner, M.A. and Harper, J.V., 1985. Cardiac dysrhythmias associated with chemical peeling with phenol. Anesthesiology, 62: 366–367.
- Weyant, R.S., Edmonds, P. and Swaminathan, B., 1990. Effect of ionic and nonionic detergents on the Taq polymerase. BioTechniques, 9: 308–309.
- Yang, G., Zhou, R., Tang, T. and Shi, S., 2008. Simple and efficient isolation of high-quality total RNA from Hibiscus tiliaceus, a mangrove associate and its relatives. Preparative Biochemistry and Biotechnology, 38: 257-264. | ||
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