رویانزائی بدنی از قطعات ساقه Hyoscyamus arachnoideus Pojark. و بررسی کمی و کیفی پروتئینها در مراحل مختلف آن
محورهای موضوعی : ژنتیکمهدیس ابراهیم‌زاده معبود 1 , مه‌لقا قربانلی 2 , حسن ابراهیم‌زاده معبود 3
1 - گروه علوم گیاهی، دانشکده علوم، دانشگاه آزاد اسلامی واحد علوم وتحقیقات
2 - گروه زیست‌شناسی، دانشگاه آزاد اسلامی واحد گرگان
3 - گروه علوم گیاهی، دانشکده زیست‌شناسی، پردیس علوم
کلید واژه: پروتئین, الکتروفورز, بنگ, رویانزانی بدنی,
چکیده مقاله :
گیاهان سردة بنگ (Hyoscyamus) با داشتن خواص داروئی به واسطه وجود تروپان آلکالوئیدهای مختلف موجود در آنها همواره مورد توجه خاصی بوده‌اند. کاربرد روش‌های مهندسی ژنتیک برای تولید گیاهانی از این سرده با خواص داروئی ارزشمندتر و ازدیاد این گیاهان در زمان کوتاه و باصرف هزینه اندک یکی از جنبه های مورد توجه می‌باشد. رویانزائی بدنی و شناخت آن کمک شایانی در این زمینه خواهد بود. هدف از کار حاضر تلاش در جهت انجام رویانزائی بدنی و شناخت سازوکار آن از طریق بررسی پروتئینها در مراحل مختلف تشکیل رویان بود. در این پژوهش کشت قطعات ساقه در محیط کشت MS واجد BAP و IAA منجر به رویانزائی موفقیت آمیزی شد. بررسی کمّی پروتئینهای محلول در مراحل مختلف نمو رویان تغییرات محتوای پروتئین را در مراحل مختلف تشکیل رویان نشان داد. بررسی کیفی پروتئینها بااستفاده از الکتروفورز بر روی دو نوع ژل پلی آکریل آمید در سیستم ناپیوسته (PAGE و SDS PAGE) انجام شد. جرم مولکولی باندهای پروتئینی و پپتیدی تفکیک شده محاسبه گردید و معلوم شد در طی رویانزائی پروتئینها و پپتیدهای مختلف دچار پدیده های فراتنظیمی، فروتنظیمی و تعدیل می‌شوند. به علاوه پروتئینها و پپتیدهای نهادی و اصلی نیز شناسائی گردیدند.
The Hyoscyamus genus has been always regarded because of the therapeutic effects and its contained tropan alkaloids. Utilizing genetic engineering methods for production of plants for production of plant belonging to this genus with higher medical values and propagation of this plant in a fast and cost-effective manner is very important. Somatic embryogenesis is a great help in this field. The aim of the current study is to gain knowledge about mechanism of somatic embryogenesis via protein changes' assessment in different stages of embryo formation. In this study stem culture on MS medium containing BAP and IAA lead to embryogenesis. Quantitative protein evaluation in different stages of embryo development showed changes in protein content. The qualitative evaluation of protein was performed using electrophoresis of two types of polyacryl amide gel in discontinued system (PAGE and SDS-PAGE). Molecular mass of peptide as well as peptide separated bonds were calculated, and it cleared that during embryogenesis different proteins and peptides undergo up-regulation, down-regulation and modulation. In addition, main and fundamental proteins and peptides were known.
Arnold S., Sabala I., Bozhkov P., Dyachok J. and Filonova L. (2002). Developmental pathways of somatic embryogenesis, Plant Cell, Tissue and Organ Culture. 69: 233-249.
Arrigoni O., Gara L., Tommasi F., and Liso R. (1992). Change in ascorbate system during seed development of Vicia faba. Plant Physiol. 99: 235-238.
Baeuerl P.A. (1991) Reactive oxygen intermediates as apparently widely used messengers in the activation of the NF-kB transcription factor and HIV-1 EMBO J. 10: 2247-2258.
Blanckaert A., Belingheri L., Sautiere P-E., Vasseur J. and Hilbert J-L. (2002). 9-KDa acidic and basic nsLTP-like proteins are secreted in the culture-medium conditioned by somatic embryogenesis in Cichorium. Plant Physiology and Biochemistry. Vol: 40. Issue 4. P: 339-345.
Bollage D.M. and Edelstein S.J. (1991). Protein Methods. third edition, Wiley-Liss, Inc. New York.
Bradford M.M. (1976). A. rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248-254.
Campalans A., Messeguer R., Goday A. and Pages M. (1999). Plant responses to drought, from ABA signal transduction events to the action of the induced proteins. Plant Physiology and Biochemistry. 37(5): 327-340.
Chapman A., Blervacq A.S., Vasseur J. and Hilbert J-L. (2000). Arabinogalactan proteins in Cichorium somatic embryogenesis: effect of b-glucosyl Yariv reagent and epitope localization during embryo development. Planta 211: 305-314.
Chappell J., Levin A., Tenhaken R., Lusso M. and Lamb C. (1997). Characterization of a diffusible signal capable of inducing defense gene expression in tobacco. Plant Physiol. 113: 621-629.
Cui K.R., Xing G.S., Liu X.M., Xing G.M. and Wang Y.F. (1999). Effect of hydrogen peroxide on somatic embryogenesis of Lycium barbarum L. Plant Sci. 146: 9-16.
Cui K.R., Li J., Xing G.M., Wang L. and Wang Y.F. (2002). Effect of hydrogen peroxide on synthesis of proteins during somatic embryogenesis in Lycium barbarum L. Plant Cell Tissue and Organ Culture. 68: 187-193.
Dahmer M.L., Hilderbrand D.F., Collins G.B. (1992). Comparative protein accumulation patterns in soybean somatic and zygotic embryos. In Vitro Cell. dev. Biol. Plant. 28: 106-114.
De Jong A.J. Cordewener J., Lo Shiuo F., Terzi M., Vande kerckhove J., van Kammen A. and de Vries S.C. (1992). A Daucus carota somatic embryo mutant is rescued by chitinoise Plant Cell. 4: 425-433.
De Jong A.J., Heidstra R., Spaink H.P., Hartog M.V. Meijer E.A., Hendriks T., Lo Shiavo F., Terzi M., Bisseling T., van Kammen A., and de Vries S.C. (1993). Rhizobium lipo-oligosaccharides rescue a Daucus carota somatic embryo variant. Plant Cell. 5: 615-620.
Dodeman V. L., Ducreux G. and Kreis M. (1997). Zygotic embryogenesis versus somatic embryogenesis. J. Exp. Bot. 48: 1493-1509.
Dyachok J.V., Tobin A.E., Price N.P.J. and von Arnold S. (2000). Rhizobial nod factors stimulate somatic embryo development in Picea abies. Plant Cell Rep. 19: 290-297.
Egertsdotter U. and van Arnold S. (1998). Development of somatic embryos in Norway spruce. J. Exp. Bot. 49: 155-162.
Egertsdotter U. and van Arnold S. (1995). Importance of arabinogalactan proteins for the development of somatic embryos of Norway spruce (Picea abies). Physiol. Plant. 93: 334-345.
Gaj M.D. (2004). Factors influencing somatic embryogenesis induction and plant regeneration with particular reference to Arabidopsis thaliana (L.) Heynh. Plant Growth Regulation. 43: 27-47.
Haggman H., Vuosku J., Sarjala T., Jokela A. and Niemi K. (2006). Somatic embryogenesis of pine species : from functional genomics to plantation forestry. Plant Cell Monogrophs. 2: 1861-1370.
Hames B.D. and Richwood D. (1999). Gel Electrophoresis of Proteins: A practical Approach second edition, Oxford university. Press, New York.
Kairong C., Gengsheng X., Xinmin L., Gengmei X., Yafu W., (1999). Effect of hydrogenperoxide on somatic embryogenesis of Lycium barbarum L., Plant science. 149: 9-16.
Kreuger M. and van Holst G.J. (1993). Arabinogalactan proteins are essential in somatic embryogenesis of Pacus carota L. Planta 189: 243-248.
Kwaaitaal M., Vries S. and Russinava E. (2005) Arabidopsis thaliana somatic embryogenesis receptor kinase 1 protein is present in sporophytic and gametophytic cells and undergoes endocytosis. Plant and Fungal Endocytosis. 226(1-2) 55-65.
Laemmeli U.K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685.
Li R., Reed D.W., Liu E., Nowak J., Pelcher L.E., Page J.E. and cevello P. S. (2006). Functional genomic Analysis in Hyoscyamus niger Reveals a Cytochrome P450 Involved in Littorine Rearrangement. Chemistry and Biology. 13(5): 513-520.
Liang C., Xi Y., Shu J., Li J., Yang J., Che K., Jin D., Liu X., Weng M., He Y. and Wang B. (2004). Construction of a BAC library of physcomitrella patens and isolation of a LEA gene. Plant Science. 167(3): 491-498.
Lubaretz O. and Nieden U. (2002) Accumulation of plant small heat-stress proteins in storage organs. Planta. 215(2): 220-228.
Luo J. P., Jiang S. T. and Pan L. J. (2001). Enhanced somatic embryogenesis by salicylic acid of Astragalus adsurgens Pall. : relationship with H2O2 production and H2O2 –metabolizing enzyme activities. Plant Science. 161: 125-132.
Majewska-Sawka A. and Nothnagel E.A. (2000). The multiple roles of arabinogalactan proteins in plant development. Plant Physiol. 122: 3-9.
Mordhorst A.P., Toonen M. A.J. and de Vries S.C. (1997). Plant embryogenesis. Crit. Rev. Plant Sci. Vol: 16. P: 535-576.
Mordhorst A.P., Voerman K.J., Hartog M. V., Meijer E.A., van Went J., Koorneef M. and de Vries S.C. (1998). Somatic embryogenesis in Arabidopsis thaliana is facilitated by mutation in genes repressing meristematic cell divisions. Genetics. 149: 549-563.
Novozhilova O.A., Grinash M.N., Arefeva L.P. and Semikhov V.F. (2004). Protein biosynthesis in the embryo and endosperm during embryogenesis in Pinus sylvestris L. 35(2) 69-75.
Reinbothe C., Tewes A., Lehmann J. Parthier B. and Reinbothe S. (1994). Induction by methyl jasmonat of embryogenesis-related proteins and mRNA in Nicotiana; plumbaginifolia. Plant Science. 104(1): 59-70.
Roberts D.R., Flinn B.S., Webb D.T., Webster F.B., Sutton B.C.S. (1989). Characterization of immature embryos of interior spruce by SDS-PAGE and microscopy in relation to their competence for somatic embryogenesis. Plant.Cell. Rep. 8: 285-288.
Roja Rani A., Reddy D., Prakash Babu P. and Padmaja G. (2005). Changes in protein profiles associated with somatic embryogenesis in peanut. Biologia Plantarum. 49(3): 347-354.
Schmidt M.A., Tucker D.M. and Cahoon E.B. (2005). Towards normalization of soybean somatic embryo maturation. Plant Cell Rep. 24: 383-391.
Silveira V., Floh E.S., Handro W. and Guerra M.P. (2004). Effect of plant growth regulators on the cellular growth and levels of intracellular protein, starch and polyamines in embryogenic suspension cultures of Pinus taeda. Plant Cell, Tissue and Organ Culture. 76(1): 53-60.
Tan S.K., Kamada H. (2000). Initial identification of a phosphoprotein that appears to be involved in the induction of somatic embryogenesis in carrot. Plant Cell Reports. 19: 739-747.
Thompson H.J.M. and Knox J.P. (1998). stage-specific responses of embryogenic carrot cell suspension cultures to arabinogalactan protein-binding B-glucosyl Yarin reagent. Planta. 205: 32-38.
Van Hengel A.J., Tadesse Z., Immerzeel P., Schols H., Van Kammen A., and de Vries S.C. (2001). N-acetylglucosamine and glucosamine-containing arabinogalactan proteins control somatic embryogenesis. Plant Physiol. 125: 1880-18X.
Vroemen C., de Vries S. and Quatrano R. (1999) Signalling in plants embyos during the establishment of the polar axis. Seminars in Cell and Developmental Biology. 10(2): 157-164.
Wilekens H., Langebartels C., Tire C., Montagu M.V. Inze D., Camp W.V. (1994). Differential expression of catalase genes in Nicotiana plumbaginifolia (L.). Proc Natl. Acad. Sci. USA. 91: 10450-10454.
_||_Arnold S., Sabala I., Bozhkov P., Dyachok J. and Filonova L. (2002). Developmental pathways of somatic embryogenesis, Plant Cell, Tissue and Organ Culture. 69: 233-249.
Arrigoni O., Gara L., Tommasi F., and Liso R. (1992). Change in ascorbate system during seed development of Vicia faba. Plant Physiol. 99: 235-238.
Baeuerl P.A. (1991) Reactive oxygen intermediates as apparently widely used messengers in the activation of the NF-kB transcription factor and HIV-1 EMBO J. 10: 2247-2258.
Blanckaert A., Belingheri L., Sautiere P-E., Vasseur J. and Hilbert J-L. (2002). 9-KDa acidic and basic nsLTP-like proteins are secreted in the culture-medium conditioned by somatic embryogenesis in Cichorium. Plant Physiology and Biochemistry. Vol: 40. Issue 4. P: 339-345.
Bollage D.M. and Edelstein S.J. (1991). Protein Methods. third edition, Wiley-Liss, Inc. New York.
Bradford M.M. (1976). A. rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72: 248-254.
Campalans A., Messeguer R., Goday A. and Pages M. (1999). Plant responses to drought, from ABA signal transduction events to the action of the induced proteins. Plant Physiology and Biochemistry. 37(5): 327-340.
Chapman A., Blervacq A.S., Vasseur J. and Hilbert J-L. (2000). Arabinogalactan proteins in Cichorium somatic embryogenesis: effect of b-glucosyl Yariv reagent and epitope localization during embryo development. Planta 211: 305-314.
Chappell J., Levin A., Tenhaken R., Lusso M. and Lamb C. (1997). Characterization of a diffusible signal capable of inducing defense gene expression in tobacco. Plant Physiol. 113: 621-629.
Cui K.R., Xing G.S., Liu X.M., Xing G.M. and Wang Y.F. (1999). Effect of hydrogen peroxide on somatic embryogenesis of Lycium barbarum L. Plant Sci. 146: 9-16.
Cui K.R., Li J., Xing G.M., Wang L. and Wang Y.F. (2002). Effect of hydrogen peroxide on synthesis of proteins during somatic embryogenesis in Lycium barbarum L. Plant Cell Tissue and Organ Culture. 68: 187-193.
Dahmer M.L., Hilderbrand D.F., Collins G.B. (1992). Comparative protein accumulation patterns in soybean somatic and zygotic embryos. In Vitro Cell. dev. Biol. Plant. 28: 106-114.
De Jong A.J. Cordewener J., Lo Shiuo F., Terzi M., Vande kerckhove J., van Kammen A. and de Vries S.C. (1992). A Daucus carota somatic embryo mutant is rescued by chitinoise Plant Cell. 4: 425-433.
De Jong A.J., Heidstra R., Spaink H.P., Hartog M.V. Meijer E.A., Hendriks T., Lo Shiavo F., Terzi M., Bisseling T., van Kammen A., and de Vries S.C. (1993). Rhizobium lipo-oligosaccharides rescue a Daucus carota somatic embryo variant. Plant Cell. 5: 615-620.
Dodeman V. L., Ducreux G. and Kreis M. (1997). Zygotic embryogenesis versus somatic embryogenesis. J. Exp. Bot. 48: 1493-1509.
Dyachok J.V., Tobin A.E., Price N.P.J. and von Arnold S. (2000). Rhizobial nod factors stimulate somatic embryo development in Picea abies. Plant Cell Rep. 19: 290-297.
Egertsdotter U. and van Arnold S. (1998). Development of somatic embryos in Norway spruce. J. Exp. Bot. 49: 155-162.
Egertsdotter U. and van Arnold S. (1995). Importance of arabinogalactan proteins for the development of somatic embryos of Norway spruce (Picea abies). Physiol. Plant. 93: 334-345.
Gaj M.D. (2004). Factors influencing somatic embryogenesis induction and plant regeneration with particular reference to Arabidopsis thaliana (L.) Heynh. Plant Growth Regulation. 43: 27-47.
Haggman H., Vuosku J., Sarjala T., Jokela A. and Niemi K. (2006). Somatic embryogenesis of pine species : from functional genomics to plantation forestry. Plant Cell Monogrophs. 2: 1861-1370.
Hames B.D. and Richwood D. (1999). Gel Electrophoresis of Proteins: A practical Approach second edition, Oxford university. Press, New York.
Kairong C., Gengsheng X., Xinmin L., Gengmei X., Yafu W., (1999). Effect of hydrogenperoxide on somatic embryogenesis of Lycium barbarum L., Plant science. 149: 9-16.
Kreuger M. and van Holst G.J. (1993). Arabinogalactan proteins are essential in somatic embryogenesis of Pacus carota L. Planta 189: 243-248.
Kwaaitaal M., Vries S. and Russinava E. (2005) Arabidopsis thaliana somatic embryogenesis receptor kinase 1 protein is present in sporophytic and gametophytic cells and undergoes endocytosis. Plant and Fungal Endocytosis. 226(1-2) 55-65.
Laemmeli U.K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685.
Li R., Reed D.W., Liu E., Nowak J., Pelcher L.E., Page J.E. and cevello P. S. (2006). Functional genomic Analysis in Hyoscyamus niger Reveals a Cytochrome P450 Involved in Littorine Rearrangement. Chemistry and Biology. 13(5): 513-520.
Liang C., Xi Y., Shu J., Li J., Yang J., Che K., Jin D., Liu X., Weng M., He Y. and Wang B. (2004). Construction of a BAC library of physcomitrella patens and isolation of a LEA gene. Plant Science. 167(3): 491-498.
Lubaretz O. and Nieden U. (2002) Accumulation of plant small heat-stress proteins in storage organs. Planta. 215(2): 220-228.
Luo J. P., Jiang S. T. and Pan L. J. (2001). Enhanced somatic embryogenesis by salicylic acid of Astragalus adsurgens Pall. : relationship with H2O2 production and H2O2 –metabolizing enzyme activities. Plant Science. 161: 125-132.
Majewska-Sawka A. and Nothnagel E.A. (2000). The multiple roles of arabinogalactan proteins in plant development. Plant Physiol. 122: 3-9.
Mordhorst A.P., Toonen M. A.J. and de Vries S.C. (1997). Plant embryogenesis. Crit. Rev. Plant Sci. Vol: 16. P: 535-576.
Mordhorst A.P., Voerman K.J., Hartog M. V., Meijer E.A., van Went J., Koorneef M. and de Vries S.C. (1998). Somatic embryogenesis in Arabidopsis thaliana is facilitated by mutation in genes repressing meristematic cell divisions. Genetics. 149: 549-563.
Novozhilova O.A., Grinash M.N., Arefeva L.P. and Semikhov V.F. (2004). Protein biosynthesis in the embryo and endosperm during embryogenesis in Pinus sylvestris L. 35(2) 69-75.
Reinbothe C., Tewes A., Lehmann J. Parthier B. and Reinbothe S. (1994). Induction by methyl jasmonat of embryogenesis-related proteins and mRNA in Nicotiana; plumbaginifolia. Plant Science. 104(1): 59-70.
Roberts D.R., Flinn B.S., Webb D.T., Webster F.B., Sutton B.C.S. (1989). Characterization of immature embryos of interior spruce by SDS-PAGE and microscopy in relation to their competence for somatic embryogenesis. Plant.Cell. Rep. 8: 285-288.
Roja Rani A., Reddy D., Prakash Babu P. and Padmaja G. (2005). Changes in protein profiles associated with somatic embryogenesis in peanut. Biologia Plantarum. 49(3): 347-354.
Schmidt M.A., Tucker D.M. and Cahoon E.B. (2005). Towards normalization of soybean somatic embryo maturation. Plant Cell Rep. 24: 383-391.
Silveira V., Floh E.S., Handro W. and Guerra M.P. (2004). Effect of plant growth regulators on the cellular growth and levels of intracellular protein, starch and polyamines in embryogenic suspension cultures of Pinus taeda. Plant Cell, Tissue and Organ Culture. 76(1): 53-60.
Tan S.K., Kamada H. (2000). Initial identification of a phosphoprotein that appears to be involved in the induction of somatic embryogenesis in carrot. Plant Cell Reports. 19: 739-747.
Thompson H.J.M. and Knox J.P. (1998). stage-specific responses of embryogenic carrot cell suspension cultures to arabinogalactan protein-binding B-glucosyl Yarin reagent. Planta. 205: 32-38.
Van Hengel A.J., Tadesse Z., Immerzeel P., Schols H., Van Kammen A., and de Vries S.C. (2001). N-acetylglucosamine and glucosamine-containing arabinogalactan proteins control somatic embryogenesis. Plant Physiol. 125: 1880-18X.
Vroemen C., de Vries S. and Quatrano R. (1999) Signalling in plants embyos during the establishment of the polar axis. Seminars in Cell and Developmental Biology. 10(2): 157-164.
Wilekens H., Langebartels C., Tire C., Montagu M.V. Inze D., Camp W.V. (1994). Differential expression of catalase genes in Nicotiana plumbaginifolia (L.). Proc Natl. Acad. Sci. USA. 91: 10450-10454.