بیان گذرای ژن در گیاهان و کاربردهای آن در کشاورزی مولکولی و شناسایی عملکرد ژن‌ها

نوع مقاله: مروری

نویسنده

استادیار پژوهشی، بخش مهندسی ژنتیک و ایمنی زیستی، پژوهشکده بیوتکنولوژی کشاورزی ایران، سازمان تحقیقات، آموزش و ترویج کشاورزی، کرج، ایران

چکیده

تراریختی گذرای گیاهان روشی مفید و در برخی موارد جایگزینی مناسب برای تراریختی پایدار به ویژه در غلات، گیاهان درختی و گیاهانی است که باززایی و یا تراریختی آنها با مشکل مواجه است. در این روش در زمانی کوتاه تعداد زیادی از نسخه‌های تراژن وارد سلول گیاهی شده، رونویسی و ترجمه می‌شوند و چون توالی DNA در ژنوم میزبان وارد نمی‌شود، بیان ژن تحت تاثیر محل درج و تغییرات اپی‌ژنتیکی قرار نمیگیرد. تراریختی گذرا روشی مفید و کارآمد در مطالعات بررسی عملکرد ژن‌ها مانند بیش بیان ژن، خاموش کردن ژن، شبکه بیانی ژن‌ها، تعیین محل استقرار پروتئین، بررسی پروموتر، شناسایی مسیرهای بیوسنتزی و ... است. همچنین استفاده از این روش در صنایع بیوتکنولوژی همچون تولید پروتئین‌های نوترکیب (پلنتی‌بادی‌ها، واکسن‏ها و ترکیبات دارویی) رشد فزاینده‌ای داشته است. انتقال ژن در گیاهان با روش‌های مختلفی انجام می‌شود که در بسیاری از موارد این روش‌ها با هم همپوشانی داشته و از تلفیق آنها برای بیان‌گذرا استفاده می‌شود. در مقاله حاضر انواع روش‌های انتقال و بیان‌گذرای ژن، مزایا و معایب آنها و آخرین پیشرفت‌ها در ارتباط با بیان‌گذرا در گیاهان مدل و زراعی مطرح شده است. همچنین قابلیت‌ها و محدودیت‌های این روش‌ها در تحقیقات و صنعت مورد بررسی قرار گرفته است.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Transient gene expression in plants and its application in molecular farming and functional genomics

نویسنده [English]

  • Katayoun Zamani
Assistant Professor, Department of Genetic Engineering and Biosafety, Agricultural Biotechnology Research Institute of Iran, Agricultural Research, Education, and Extension Organization, Karaj, Iran
چکیده [English]

Transient expression is widely used as an alternative method for stable transformation of plants, especially cereals, trees and recalcitrants. In this method, multiple copies of transgene are introduced in to the plant cell and then are transcribed and translated. As introduced gene is not integrated in the genome, its expression is not affected by epigenetic factors and the location of insertion. Transient expression is a convenient method for functional analysis of genes including overexpression, silencing, gene expression networks, protein localization, promoter analysis, identification of biosynthesis pathways and so on. Additionally, this method is applied in biotechnological industries like production of recombinant proteins. Transient transformation is performed by different methods, however, many of them have overlap. So, a combination of these methods can be used for transient expression. In this review, various transient transformation techniques, their advantages and disadvantages, latest progress regarding transient expression in model and crop plants and potential and limitations regarding the application of transient expression technique in science and industry will be discussed.

کلیدواژه‌ها [English]

  • transient expression
  • gene function
  • Transformation
  • agroinfiltration
  • recombinant protein
Andersson M, Turesson H, Nicolia A, Fält AS, Samuelsson M, Hofvander P (2017) Efficient targeted multiallelic mutagenesis in tetraploid potato (Solanum tuberosum) by transient CRISPR-Cas9 expression in protoplasts. Plant. Cell. Rep. 36(1):117-28.

Baltes NJ, Gil-Humanes J, Cermak T, Atkins PA, Voytas DF (2014) DNA replicons for plant genome engineering. Plant. Cell. 26(1): 151-63.

Bond DM, Albert NW, Lee RH, Gillard GB, Brown CM, Hellens RP, Macknight RC (2016) Infiltration-RNAseq: transcriptome profiling of Agrobacterium-mediated infiltration of transcription factors to discover gene function and expression networks in plants. Plant methods. 12(1):41.

Burris KP, Dlugosz EM, Collins AG, Stewart CN, Lenaghan SC (2016) Development of a rapid, low-cost protoplast transfection system for switchgrass (Panicum virgatum L.). Plant. Cell. Rep. 35(3):693-704.

Canto T (2016) Transient Expression Systems in Plants: Potentialities and Constraints." Advanced Technologies for Protein Complex Production and Characterization. Springer International Publishing, 287-301.

Chen Q, Lai H, Hurtado J, Stahnke J, Leuzinger K, Dent M (2013) Agroinfiltration as an effective and scalable strategy of gene delivery for production of pharmaceutical proteins. Adv. Tech. Biol. Med.1(1).

Chujo T, Yoshikawa M, Ariga H, Endo M, Toki S, Ishibashi K (2017) A removable virus vector suitable for plant genome editing. Plant. J. 91(3):558-561.

Dawson WO, Folimonova SY (2013) Virus-based transient expression vectors for woody crops: a new frontier for vector design and use. Annu. Rev. Phytopathol. 51:321-37.

Dhandapani S, Jin J, Sridhar V, Sarojam R, Chua NH, Jang IC (2017) Integrated metabolome and transcriptome analysis of Magnolia champaca identifies biosynthetic pathways for floral volatile organic compounds. BMC genomics. 18(1):463.

Dobnik D, Lazar A, Stare T, Gruden K, Vleeshouwers VG, Žel J (2016) Solanum venturii, a suitable model system for virus-induced gene silencing studies in potato reveals St MKK6 as an important player in plant immunity. Plant methods. 12(1):29.

Duarte P, Ribeiro D, Carqueijeiro I, Bettencourt S, Sottomayor M (2016) Protoplast transformation as a plant-transferable transient expression system. Biotechnology of Plant Secondary Metabolism: Meth. Protoc. 137-48.

Fischer R, Vaquero‐Martin C, Sack M, Drossard J, Emans N, Commandeur U (1999) Towards molecular farming in the future: transient protein expression in plants. Biotechnol. Appl. Biochem. 30(2):113-6.

Fister AS, Shi Z, Zhang Y, Helliwell EE, Maximova SN, Guiltinan MJ (2016) Protocol: transient expression system for functional genomics in the tropical tree Theobroma cacao L. Plant methods.12(1):19.

Fujiuchi N, Matoba N, Matsuda R (2016) Environment control to improve recombinant protein yields in plants based on Agrobacterium-mediated transient gene expression. Front. Bioeng. Biotechnol. 4.  

Gallón SM, Elejalde-Palmett C, Daudu D, Liesecke F, Jullien F, Papon N, de Bernonville TD, Courdavault V, Lanoue A, Oudin A, Glévarec G (2017) Virus-induced gene silencing of the two squalene synthase isoforms of apple tree (Malus× domestica L.) negatively impacts phytosterol biosynthesis, plastid pigmentation and leaf growth. Planta. 27:1-6.

Gao C, Nielsen KK (2013) Comparison between Agrobacterium-mediated and direct gene transfer using the gene gun. Biolistic DNA Delivery: Meth. Protoc. 3-16.

Gil-Humanes J, Wang Y, Liang Z, Shan Q, Ozuna CV, Sánchez-León S, Baltes NJ, Starker C, Barro F, Gao C, Voytas DF (2017) High-efficiency gene targeting in hexaploid wheat using DNA replicons and CRISPR/Cas9. Plant. J. 89(6):1251-62.

Gleba Y, Klimyuk V, Marillonnet S (2007) Viral vectors for the expression of proteins in plants. Curr. Opin. Biotechnol.18(2):134-41.

Hayashi S, Wakasa Y, Ozawa K, Takaiwa F (2016) Characterization of IRE1 ribonuclease‐mediated mRNA decay in plants using transient expression analyses in rice protoplasts. New. Phytol. 210(4):1259-68.

Huo A, Chen Z, Wang P, Yang L, Wang G, Wang D, Liao S, Cheng T, Chen J, Shi J (2017) Establishment of transient gene expression systems in protoplasts from Liriodendron hybrid mesophyll cells. PloS One. 12(3): e0172475.

Janssen BJ, Gardner RC (1990) Localized transient expression of GUS in leaf discs following cocultivation with Agrobacterium. Plant Mol. Boil. 14(1):61-72.

Jelly NS, Valat L, Walter B, Maillot P (2014) Transient expression assays in grapevine: a step towards genetic improvement. Plant biotechnol. J. 12(9):1231-45.

Kanagarajan S, Muthusamy S, Gliszczyńska A, Lundgren A, Brodelius PE (2012) Functional expression and characterization of sesquiterpene synthases from Artemisia annua L. using transient expression system in Nicotiana benthamiana. Plant. Cell. Rep. 31(7):1309-19.

Koroleva OA, Tomlinson ML, Leader D, Shaw P, Doonan JH (2005) High‐throughput protein localization in Arabidopsis using Agrobacterium‐mediated transient expression of GFP‐ORF fusions. Plant. J. 41(1):162-74.

Kudo M, Kidokoro S, Yoshida T, Mizoi J, Todaka D, Fernie AR, Shinozaki K, Yamaguchi‐Shinozaki K (2017) Double overexpression of DREB and PIF transcription factors improves drought stress tolerance and cell elongation in transgenic plants. Plant. Biotechnol. J. 15(4):458-71.

Lacroix B, Citovsky V (2013) The roles of bacterial and host plant factors in Agrobacterium-mediated genetic transformation. Int. J. Dev. Biol. 57(6-7-8):467-81.

Leonelli L, Erickson E, Lyska D, Niyogi KK (2016) Transient expression in Nicotiana benthamiana for rapid functional analysis of genes involved in non‐photochemical quenching and carotenoid biosynthesis. Plant. J. 88(3):375-86.

Li JF, Park E, von Arnim AG, Nebenführ A (2009) The FAST technique: a simplified Agrobacterium-based transformation method for transient gene expression analysis in seedlings of Arabidopsis and other plant species. Plant Methods. 5(1):6.

Liang Z, Chen K, Li T, Zhang Y, Wang Y, Zhao Q, Liu J, Zhang H, Liu C, Ran Y, Gao C (2017) Efficient DNA-free genome editing of bread wheat using CRISPR/Cas9 ribonucleoprotein complexes. Nat. Commun. 8.

Loh HS, Green BJ, Yusibov V (2017) Using transgenic plants and modified plant viruses for the development of treatments for human diseases. Cur. Opin. Virol. 31; 26:81-9.

Marion J, Bach L, Bellec Y, Meyer C, Gissot L, Faure JD (2008) Systematic analysis of protein subcellular localization and interaction using high‐throughput transient transformation of Arabidopsis seedlings. Plant J. 56(1):169-79.

Martin K, Kopperud K, Chakrabarty R, Banerjee R, Brooks R, Goodin MM (2009) Transient expression in Nicotiana benthamiana fluorescent marker lines provides enhanced definition of protein localization, movement and interactions in planta. Plant J. 59(1):150-62.

Mortimer CL, Dugdale B, Dale JL (2015) Updates in inducible transgene expression using viral vectors: from transient to stable expression. Cur. Opin. Biotechnol. 32:85-92.

Mugford ST, Louveau T, Melton R, Qi X, Bakht S, Hill L, Tsurushima T, Honkanen S, Rosser SJ, Lomonossoff GP, Osbourn A (2013) Modularity of plant metabolic gene clusters: a trio of linked genes that are collectively required for acylation of triterpenes in oat. Plant. Cell. 25(3):1078-92.

Mugford ST, Qi X, Bakht S, Hill L, Wegel E, Hughes RK, Papadopoulou K, Melton R, Philo M, Sainsbury F, Lomonossoff GP (2009) A serine carboxypeptidase-like acyltransferase is required for synthesis of antimicrobial compounds and disease resistance in oats. Plant. Cell. 21(8):2473-84.

Nanjareddy K, Arthikala MK, Blanco L, Arellano ES, Lara M (2016) Protoplast isolation, transient transformation of leaf mesophyll protoplasts and improved Agrobacterium-mediated leaf disc infiltration of Phaseolus vulgaris: tools for rapid gene expression analysis. BMC biotechnol.16(1):53.

Pandey SK, Nookaraju A, Fujino T, Pattathil S, Joshi CP (2016) Virus-induced gene silencing (VIGS)-mediated functional characterization of two genes involved in lignocellulosic secondary cell wall formation. Plant. Cell. Rep. 35(11):2353-67.

Reis PA, Carpinetti PA, Freitas PP, Santos EG, Camargos LF, Oliveira IH, Silva JC, Carvalho HH, Dal-Bianco M, Soares-Ramos JR, Fontes EP (2016) Functional and regulatory conservation of the soybean ER stress-induced DCD/NRP-mediated cell death signaling in plants. BMC Plant. Boil. 16(1):156.

Ruslan SA, Dolhaji NH, Awal A, Osman M, Aziz M, Noor MR (2017) Protoplast Isolation from Hibiscus sabdariffa L. Adv. Sci. Lett. 23(2):1333-6.

Shen J, Fu J, Ma J, Wang X, Gao C, Zhuang C, Wan J, Jiang L (2014) Isolation, culture, and transient transformation of plant protoplasts. Curr. Protoc. Cell. Biol. 3:2-8.

Shen Y, Meng D, McGrouther K, Zhang J, Cheng L (2017) Efficient isolation of Magnolia protoplasts and the application to subcellular localization of MdeHSF1. Plant. Methods. 13(1):44.

Stracke R, Thiedig K, Kuhlmann M, Weisshaar B (2016) Analyzing synthetic promoters using arabidopsis protoplasts. Plant Synthetic Promoters: Meth. Protoc. 67-81.

Tsuda K, Qi Y, Nguyen LV, Bethke G, Tsuda Y, Glazebrook J, Katagiri F (2012) An efficient Agrobacterium‐mediated transient transformation of Arabidopsis. Plant J. 69(4):713-9.

Ueki S, Lacroix B, Krichevsky A, Lazarowitz SG, Citovsky V (2009) Functional transient genetic transformation of Arabidopsis leaves by biolistic bombardment. Nat. Protoc. 4(1):71.

Wu JZ, Liu Q, Geng XS, Li KM, Luo LJ, Liu JP (2017) Highly efficient mesophyll protoplast isolation and PEG-mediated transient gene expression for rapid and large-scale gene characterization in cassava (Manihot esculenta Crantz). BMC Biotechnol. 17(1):29.

Xu X, Xu X, Zhou Y, Zeng S, Kong W (2017) Identification of protoplast-isolation responsive microRNAs in Citrus reticulata Blanco by high-throughput sequencing. PloS one.12(8): e0183524.

Yao J, Weng Y, Dickey A, Wang KY (2015) Plants as factories for human pharmaceuticals: applications and challenges. Int. J. Mol. Sci. 16(12):28549-65.

Yao L, Liao X, Gan Z, Peng X, Wang P, Li S, Li T (2016) Protoplast isolation and development of a transient expression system for sweet cherry (Prunus avium L.). Sci. Hort. 209:14-21.

Yu G, Cheng Q, Xie Z, Xu B, Huang B, Zhao B (2017) An efficient protocol for perennial ryegrass mesophyll protoplast isolation and transformation, and its application on interaction study between LpNOL and LpNYC1. Plant methods. 5;13(1):46.

Zaidi SS, Mansoor S (2017) Viral Vectors for Plant Genome Engineering. Front. Plant. Sci. 8.

Zamani K, Lohrasebi T, Sabet MS, Malboobi MA, Mousavi A (2014) Expression pattern and subcellular localization of Arabidopsis purple acid phosphatase AtPAP9. Gene. Expr. Patterns. 14(1):9-18.

Zamani K, Sabet M, Lohrasebi T, Mousavi A, Malboobi M (2012) Improved phosphate metabolism and biomass production by overexpression of AtPAP18 in tobacco. Biologia. 67(4):713-20.

Zhang Y, Liang Z, Zong Y, Wang Y, Liu J, Chen K, Qiu JL, Gao C (2016) Efficient and transgene-free genome editing in wheat through transient expression of CRISPR/Cas9 DNA or RNA. Nat. Commun. 7:12617.

Zhong L, Zhang Y, Liu H, Sun G, Chen R, Song S (2016) Agrobacterium-mediated transient expression via root absorption in flowering Chinese cabbage. SpringerPlus. 5(1):1825.