Abstract
Key message
A strong, stable and root-specific expression system was developed from a rice root-specific GLYCINE - RICH PROTEIN 7 promoter for use as an enabling technology for genetic manipulation of wheat root traits.
Abstract
Root systems play an important role in wheat productivity. Genetic manipulation of wheat root traits often requires a root-specific or root-predominant expression system as an essential enabling technology. In this study, we investigated promoters from rice root-specific or root-predominant expressed genes for development of a root expression system in bread wheat. Transient expression analysis using a GREEN FLUORESCENT PROTEIN (GFP) reporter gene driven by rice promoters identified six promoters that were strongly expressed in wheat roots. Extensive organ specificity analysis of three rice promoters in transgenic wheat revealed that the promoter of rice GLYCINE-RICH PROTEIN 7 (OsGRP7) gene conferred a root-specific expression pattern in wheat. Strong GFP fluorescence in the seminal and branch roots of wheat expressing GFP reporter driven by the OsGRP7 promoter was detected in epidermal, cortical and endodermal cells in mature parts of the root. The GFP reporter driven by the promoter of rice METALLOTHIONEIN-LIKE PROTEIN 1 (OsMTL1) gene was mainly expressed in the roots with essentially no expression in the leaf, stem or seed. However, it was also expressed in floral organs including glume, lemma, palea and awn. In contrast, strong expression of rice RCg2 promoter-driven GFP was found in many tissues. The GFP expression driven by these three rice promoters was stable in transgenic wheat plants through three generations (T1–T3) examined. These data suggest that the OsGRP7 promoter can provide a strong, stable and root-specific expression system for use as an enabling technology for genetic manipulation of wheat root traits.
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Abbreviations
- GFP:
-
Green fluorescent protein
- GRP:
-
Glycine-rich protein
- MTL:
-
Metallothionein-like
References
Barone P, Rosellini D, Lafayette P, Bouton J, Veronesi F, Parrott W (2008) Bacterial citrate synthase expression and soil aluminum tolerance in transgenic alfalfa. Plant Cell Rep 27:893–901
Choi D-W, Song JY, Kwon YM, Kim S-G (1996) Characterization of a cDNA encoding a proline-rich 14 kDa protein in developing cortical cells of the roots of bean (Phaseolus vulgaris) seedlings. Plant Mol Biol 30:973–982
Cogoni C, Macino G (1999) Homology-dependent gene silencing in plants and fungi: a number of variations on the same theme. Curr Opin Microbiol 2:657–662
Dai X, Wang Y, Yang A, Zhang W-H (2012) OsMYB2P-1, an R2R3 MYB transcription factor, is involved in the regulation of phosphate-starvation responses and root architecture in rice. Plant Physiol 159:169–183
Dietz-Pfeilstetter A (2010) Stability of transgene expression as a challenge for genetic engineering. Plant Sci 179:164–167
Fagard M, Vaucheret H (2000) (Trans)gene silencing in plants: how many mechanisms? Annu Rev Plant Physiol Plant Mol Biol 51:167–194
Furtado A, Henry RJ, Takaiwa F (2008) Comparison of promoters in transgenic rice. Plant Biotechnol J 6:679–693
Furtado A, Henry RJ, Pellegrineschi A (2009) Analysis of promoters in transgenic barley and wheat. Plant Biotechnol J 7:240–253
Gao S, Fang J, Xu F, Wang W, Sun X, Chu J, Cai B, Feng Y, Chu C (2014) CYTOKININ OXIDASE/DEHYDROGENASE4 integrates cytokinin and auxin signaling to control rice crown root formation. Plant Physiol 165:1035–1046
Ghanem ME, Hichri I, Smigocki AC, Albacete A, Fauconnier ML, Diatloff E, Martinez-Andujar C, Lutts S, Dodd IC, Pérez-Alfocea F (2011) Root-targeted biotechnology to mediate hormonal signalling and improve crop stress tolerance. Plant Cell Rep 30:807–823
Goddemeier ML, Wulff D, Feix G (1998) Root-specific expression of a Zea mays gene encoding a novel glycine-rich protein, zmGRP3. Plant Mol Biol 36:799–802
Ishida Y, Tsunashima M, Hiei Y, Komari T (2015) Wheat (Triticum aestivum L.) transformation using immature embryos. Methods Mol Biol 1223:189–198
Jeong JS, Kim YS, Baek KH, Jung H, Ha SH, Do Choi Y, Kim M, Reuzeau C, Kim JK (2010) Root-specific expression of OsNAC10 improves drought tolerance and grain yield in rice under field drought conditions. Plant Physiol 153:185–197
Jeong JS, Kim YS, Redillas MCFR, Jang G, Jung H, Bang SW, Choi YD, Ha S-H, Reuzeau C, Kim J-K (2013) OsNAC5 overexpression enlarges root diameter in rice plants leading to enhanced drought tolerance and increased grain yield in the field. Plant Biotechnol J 11:101–114
Kong K, Ntui VO, Makabe S, Khan RS, Mii M, Nakamura I (2014) Transgenic tobacco and tomato plants expressing Wasabi defensin genes driven by root-specific LjNRT2 and AtNRT2.1 promoters confer resistance against Fusarium oxysporum. Plant Biotechnol 31:89–96
Kooiker M, Drenth J, Glassop D, McIntyre CL, Xue GP (2013) TaMYB13-1, a R2R3 MYB transcription factor, regulates the fructan synthetic pathway and contributes to enhanced fructan accumulation in bread wheat. J Exp Bot 64:3681–3696
Li G, Yang S, Li M, Qiao Y, Wang J (2009) Functional analysis of an Aspergillus ficuum phytase gene in Saccharomyces cerevisiae and its root-specific, secretory expression in transgenic soybean plants. Biotechnol Lett 31:1297–1303
Li F, Xing SC, Guo QF, Zhao MR, Zhang J, Gao Q, Wang G, Wang W (2011) Drought tolerance through over-expression of the expansin gene TaEXPB23 in transgenic tobacco. J Plant Physiol 168:960–966
Li Y, Liu S, Yu Z, Liu Y, Wu P (2013) Isolation and characterization of two novel root-specific promoters in rice (Oryza sativa L.). Plant Sci 207:37–44
Li AX, Han YY, Wang X, Chen YH, Zhao MR, Zhou SM, Wang W (2015) Root-specific expression of wheat expansin gene TaEXPB23 enhances root growth and water stress tolerance in tobacco. Environ Exp Bot 110:73–84
Ma XF, Tudor S, Butler T, Ge Y, Xi Y, Bouton J, Harrison M, Wang ZY (2012) Transgenic expression of phytase and acid phosphatase genes in alfalfa (Medicago sativa) leads to improved phosphate uptake in natural soils. Mol Breed 30:377–391
Mangeon A, Junqueira RM, Sachetto-Martins G (2010) Functional diversity of the plant glycine-rich proteins superfamily. Plant Signal Behav 5:99–104
Matsuyama T, Satoh H, Yamada Y, Hashimoto T (1999) A maize glycine-rich protein is synthesized in the lateral root cap and accumulates in the mucilage. Plant Physiol 120:665–674
Meister R, Rajani MS, Ruzicka D, Schachtman DP (2014) Challenges of modifying root traits in crops for agriculture. Trends Plant Sci 19:779–788
Mrízová K, Jiskrová E, Vyroubalová Š, Novák O, Ohnoutková L, Pospíšilová H, Frébort I, Harwood WA, Galuszka P (2013) Overexpression of cytokinin dehydrogenase genes in barley (Hordeum vulgare cv. Golden Promise) fundamentally affects morphology and fertility. PLoS One 8:e79029
Mudge SR, Smith FW, Richardson AE (2003) Root-specific and phosphate-regulated expression of phytase under the control of a phosphate transporter promoter enables Arabidopsis to grow on phytate as a sole P source. Plant Sci 165:871–878
Radi A, Dina P, Guy A (2006) Expression of sarcotoxin IA gene via a root-specific tob promoter enhanced host resistance against parasitic weeds in tomato plants. Plant Cell Rep 25:297–303
Redillas MCFR, Jeong JS, Kim YS, Jung H, Bang SW, Choi YD, Ha S-H, Reuzeau C, Kim J-K (2012) The overexpression of OsNAC9 alters the root architecture of rice plants enhancing drought resistance and grain yield under field conditions. Plant Biotechnol J 10:792–805
Ryan PR, Tyerman SD, Sasaki T, Furuichi T, Yamamoto Y, Zhang WH, Delhaize E (2011) The identification of aluminium-resistance genes provides opportunities for enhancing crop production on acid soils. J Exp Bot 62:9–20
Shaw LM, McIntyre CL, Gresshoff PM, Xue GP (2009) Members of the Dof transcription factor family in Triticum aestivum are associated with light-mediated gene regulation. Funct Integr Genomics 9:485–498
Shrawat AK, Carroll RT, DePauw M, Taylor GJ, Good AG (2008) Genetic engineering of improved nitrogen use efficiency in rice by the tissue-specific expression of alanine aminotransferase. Plant Biotechnol J 6:722–732
Stam M, Mol JNM, Kooter JM (1997) The silence of genes in transgenic plants. Ann Bot 79:3–12
Urwin PE, Lilley CJ, McPherson MJ, Atkinson HJ (1997) Resistance to both cyst and root-knot nematodes conferred by transgenic Arabidopsis expressing a modified plant cystatin. Plant J 12:455–461
Vercruyssen L, Gonzalez N, Werner T, Schmulling T, Inze D (2011) Combining enhanced root and shoot growth reveals cross talk between pathways that control plant organ size in Arabidopsis. Plant Physiol 155:1339–1352
Wang MB, Li ZY, Matthews PR, Upadhyaya NM, Waterhouse PM (1998) Improved vectors for Agrobacterium tumefaciens-mediated transformation of monocot plants. Acta Hortic 461:401–405
Wasson AP, Richards RA, Chatrath R, Misra SC, Prasad SV, Rebetzke GJ, Kirkegaard JA, Christopher J, Watt M (2012) Traits and selection strategies to improve root systems and water uptake in water-limited wheat crops. J Exp Bot 63:3485–3498
Werner T, Nehnevajova E, Köllmer I, Novák O, Strnad M, Krämer U, Schmülling T (2010) Root-specific reduction of cytokinin causes enhanced root growth, drought tolerance, and leaf mineral enrichment in Arabidopsis and tobacco. Plant Cell 22:3905–3920
Xiao FH, Xue GP (2001) Analysis of the promoter activity of late embryogenesis abundant protein genes in barley seedlings under conditions of water deficit. Plant Cell Rep 20:667–673
Xing SC, Li F, Guo QF, Liu DR, Zhao XX, Wang W (2009) The involvement of an expansin gene TaEXPB23 from wheat in regulating plant cell growth. Biol Plantarum 53:429–434
Xu Y, Buchholz WG, DeRose RT, Hall TC (1995) Characterization of a rice gene family encoding root-specific proteins. Plant Mol Biol 27:237–248
Xue GP (2003) The DNA-binding activity of an AP2 transcriptional activator HvCBF2 involved in regulation of low-temperature responsive genes in barley is modulated by temperature. Plant J 33:373–383
Xue GP, Loveridge CW (2004) HvDRF1 is involved in abscisic acid-mediated gene regulation in barley and produces two forms of AP2 transcriptional activators, interacting preferably to a CT-rich element. Plant J 37:326–339
Xue GP, McIntyre CL, Jenkins CLD, Glassop D, van Herwaarden AF, Shorter R (2008) Molecular dissection of variation in carbohydrate metabolism related to water soluble carbohydrate accumulation in stems of wheat (Triticum aestivam L.). Plant Physiol 146:441–454
Xue GP, Kooiker M, Drenth J, McIntyre CL (2011) TaMYB13 is a transcriptional activator of fructosyltransferase genes involved in β-2,6-linked fructan synthesis in wheat. Plant J 68:857–870
Xue GP, Sadat S, Drenth J, McIntyre CL (2014) The heat shock factor family from Triticum aestivum in response to heat and other major abiotic stresses and their role in regulation of heat shock protein genes. J Exp Bot 65:539–557
Xue GP, Drenth J, McIntyre CL (2015) TaHsfA6f is a transcriptional activator that regulates a suite of heat stress protection genes in wheat (Triticum aestivum L.) including previously unknown Hsf targets. J Exp Bot 66:1025–1039
Yang WT, Baek D, Yun D-J, Hwang WH, Park DS, Nam MH, Chung ES, Chung YS, Yi YB, Kim DH (2014) Overexpression of OsMYB4P, an R2R3-type MYB transcriptional activator, increases phosphate acquisition in rice. Plant Physiol Biochem 80:259–267
Acknowledgments
We are grateful to Smitha Louis and Dhara Bhatt for production of transgenic wheat plants and Terry Grant for maintenance of the controlled environment facility for growing wheat plants. We thank Dr. Emmanuel Delhaize for his critical reading of this manuscript.
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Communicated by P. Lakshmanan.
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Xue, GP., Rae, A.L., White, R.G. et al. A strong root-specific expression system for stable transgene expression in bread wheat. Plant Cell Rep 35, 469–481 (2016). https://doi.org/10.1007/s00299-015-1897-3
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DOI: https://doi.org/10.1007/s00299-015-1897-3