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Analyzing the action of evolutionarily conserved modules on HMW-GS 1Ax1 promoter activity

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The specific and high-level expression of 1Ax1 is determined by different promoter regions. HMW-GS synthesis occurs in aleurone layer cells. Heterologous proteins can be stored in protein bodies.

Abstract

High-molecular-weight glutenin subunit (HMW-GS) is highly expressed in the endosperm of wheat and relative species, where their expression level and allelic variation affect the bread-making quality and nutrient quality of flour. However, the mechanism regulating HMW-GS expression remains elusive. In this study, we analyzed the distribution of cis-acting elements in the 2659-bp promoter region of the HMW-GS gene 1Ax1, which can be divided into five element-enriched regions. Fragments derived from progressive 5′ deletions were used to drive GUS gene expression in transgenic wheat, which was confirmed in aleurone layer cells, inner starchy endosperm cells, starchy endosperm transfer cells, and aleurone transfer cells by histochemical staining. The promoter region ranging from − 297 to − 1 was responsible for tissue-specific expression, while fragments from − 1724 to − 618 and from − 618 to − 297 were responsible for high-level expression. Under the control of the 1Ax1 promoter, heterologous protein could be stored in the form of protein bodies in inner starchy endosperm cells, even without a special location signal. Our findings not only deepen our understanding of glutenin expression regulation, trafficking, and accumulation but also provide a strategy for the utilization of wheat endosperm as a bioreactor for the production of nutrients and metabolic products.

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References

  • Barro F, Rooke L, Bekes F, Gras P, Tatham AS, Fido R, Lazzeri PA, Shewry PR, Barcelo P (1997) Transformation of wheat with high molecular weight subunits genes results in improved functional properties. Nat Biotechnol 15:1295–1299

    Article  CAS  PubMed  Google Scholar 

  • Beasley HL, Uthayakumaran S, Stoddard FL, Partridge SJ, Daqiq L, Chong P, Békés F (2002) Synergistic and additive effects of three high molecular weight glutenin subunit loci. II. Effects on wheat dough functionality and end-use quality. Cereal Chem 79:301–307

    Article  CAS  Google Scholar 

  • Blechl A, Lin J, Nguyen S, Chan R, Anderson OD, Dupont FM (2007) Transgenic wheats with elevated levels of Dx5 and/or Dy10 high-molecular-weight glutenin subunits yield doughs with increased mixing strength and tolerance. J Cereal Sci 45:172–183

    Article  CAS  Google Scholar 

  • Boudet J, Merlino M, Plessis A, Gaudin J, Dardevet M, Perrochon S, Alvarez D, Risacher T, Martre P, Ravel C (2019) The bZIP transcription factor SPA Heterodimerizing Protein represses glutenin synthesis in Triticum aestivum. Plant J 97:858–871

    Article  CAS  PubMed  Google Scholar 

  • Cho K, Jo Y, Lim S, Kim JY, Han O, Lee J (2019) Overexpressing wheat low-molecular-weight glutenin subunits in rice (Oryza sativa L. japonica cv. Koami) seeds. 3 Biotech 9:49

    Article  PubMed  PubMed Central  Google Scholar 

  • Ciolkowski I, Wanke D, Birkenbihl RP, Somssich IE (2008) Studies on DNA-binding selectivity of WRKY transcription factors lend structural clues into WRKY-domain function. Plant Mol Biol 68:81–92

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • De Wilde C, Van Houdt H, De Buck S, Angenon G, De Jaeger G, Depicker A (2000) Plants as bioreactors for protein production: avoiding the problem of transgene silencing. Plant Mol Biol 43:347–359

    Article  PubMed  Google Scholar 

  • Diaz-De-Leon F, Klotz KL, Lagrimini LM (1993) Nucleotide sequence of the tobacco (Nicotiana tabacum) anionic peroxidase gene. Plant Physiol 101:1117–1118

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dickinson CD, Evans RP, Nielsen NC (1988) RY repeats are conserved in the 5’-flanking regions of legume seed-protein genes. Nucleic Acids Res 16:371

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Don C, Mann G, Bekes F, Hamer RJ (2006) HMW-GS affect the properties of glutenin particles in GMP and thus flour quality. J Cereal Sci 44:127–136

    Article  CAS  Google Scholar 

  • Farré G, Perez-Fons L, Decourcelle M, Breitenbach J, Hem S, Zhu C, Capell T, Christou P, Fraser PD, Sandmann G (2016) Metabolic engineering of astaxanthin biosynthesis in maize endosperm and characterization of a prototype high oil hybrid. Transgenic Res 25:477–489

    Article  PubMed  CAS  Google Scholar 

  • Furtado A, Henry RJ, Takaiwa F (2008) Camparison of promoters in transgenic rice. Plant Biotechnol J 6:679–693

    Article  CAS  PubMed  Google Scholar 

  • Furtado A, Henry RJ, Pellegrineschi A (2009) Analysis of promoters in transgenic barley and wheat. Plant Biotechnol J 7:240–253

    Article  CAS  PubMed  Google Scholar 

  • Gillies SA, Futardo A, Henry RJ (2012) Gene expression in the developing aleurone and starchy endosperm of wheat. Plant Biotechnol J 10:668–679

    Article  CAS  PubMed  Google Scholar 

  • Guo W, Yang H, Liu Y, Gao Y, Ni Z, Peng H, Xin M, Hu Z, Sun Q, Yao Yi (2015) The wheat transcription factor TaGAMyb recruits histone acetyltransferase and activates the expression of a high-molecular-weight glutenin subunit gene. Plant J 84:347–359

    Article  CAS  PubMed  Google Scholar 

  • Halford NG, Field JM, Blair H, Urwin P, Moore K, Robert L, Thompson R, Flavell RB, Tatham AS, Shewry PR (1992) Analysis of HMW glutenin subunits encoded by chromosome 1A of bread wheat (Triticum aestivum L.) indicates quantitative effects on grain quality. Theor Appl Genet 83:373–378

    Article  CAS  PubMed  Google Scholar 

  • Hoffman LM, Donaldson DD (1985) Characterization of two Phaseolus vulgaris phytohemagglutinin genes closely linked on the chromosome. EMBO J 4:883–889

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Jiang PH, Xue JS, Duan LN et al (2019) Effects of high-molecular-weight glutenin subunit combination in common wheat on the quality of crumb structure. J Sci Food Agric 99:1501–1508

    Article  CAS  PubMed  Google Scholar 

  • Kawakatsu T, Takaiwa F (2010) Cereal seed storage protein synthesis: fundamental processes for recombinant protein production in cereal grains. Plant Biotechnol J 8:939–953

    Article  CAS  PubMed  Google Scholar 

  • Lamacchia C, Shewry PR, Fonzo ND, Forsyth JL, Harris N, Lazzeri PA, Napier JA, Halford NG, Barcelo P (2001) Endosperm-specififc activity of a storage protein gene promoter in transgenic wheat seed. J Exp Bot 52:243–250

    Article  CAS  PubMed  Google Scholar 

  • Levanony H, Rubin R, Altschuler Y, Galili G (1992) Evidence for a novel route of wheat storage proteins to vacuoles. J Cell Biol 119(5):1117–1128

    Article  CAS  PubMed  Google Scholar 

  • Li XH, Wang K, Wang SL, Gao LY, Xie XX, Hsam SLK, Zeller FJ, Yan YM (2010) Molecular characterization and comparative transcriptional analysis of LMW-m-type genes from wheat (Triticum aestivum L.) and Aegilops species. Theor Appl Genet 121(5):845–856

    Article  CAS  PubMed  Google Scholar 

  • Li Y, Wang Q, Li X et al (2012) Coexpression of the high molecular weight glutenin subunit 1Ax1 and puroindoline improves dough mixing properties in durum wheat (Triticum turgidum L. ssp. durum). PLoS ONE 7(11):e50057

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li YW, An XL, Yang R et al (2015) Dissecting and enhancing the contributions of high-molecular-weight glutenin subunits to dough functionality and bread quality. Mol Plant 8:332–334

    Article  CAS  PubMed  Google Scholar 

  • Li JH, Wang K, Li GY, Li YL, Zhang Y, Liu ZY, Ye XG, Xia XC, He GH, Cao SH (2018) Dissecting conserved cis-regulatory modules of Glu-1 promoters which confer the highly active endosperm-specific expression via stable wheat transformation. Crop J 321:1–17

    Google Scholar 

  • Liu HY, Wang K, Xiao LL, Wang SL, Du LP, Cao XY, Zhang XX, Zhou Y, Yan YM, Ye XG (2016) Comprehensive identification and bread-making quality evaluation of common wheat somatic variation line AS208 on glutenin composition. PLoS ONE 11:e0146933

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Logemann E, Parniske M, Hahlbrock K (1995) Modes of expression and common structural features of the complete phenylalanine ammonia-lyase gene family in parsley. Proc Natl Acad Sci USA 92:5905–5909

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Loussert C, Popineau Y, Mangavel C (2008) Protein bodies ontogeny and localization of prolmain components in the developing endosperom of wheat caryopses. J Cereal Sci 47:445–456

    Article  CAS  Google Scholar 

  • Makai S, Éva C, Tamás L, Juhász A (2015) Multiple elements controlling the expression of wheat high molecular weight glutenin paralogs. Funct Integr Genom 15:661

    Article  CAS  Google Scholar 

  • Mangel N, Fudge JB, Li K, Wu TY, Tohge T, Fernie AR, Szurek B, Fitzpatrick TB, Gruissem W, Vanderschuren H (2019) Enhancement of vitamin B6 levels in rice expressing Arabidopsis vitamin B6 biosynthesis de novo genes. Plant J 99:1047–1065

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mu J, Chen L, Gu Y, Duan L, Han S, Li Y, Yan Y, Li X (2019) Genome-wide identification of internal reference genes for normalization of gene expression values during endosperm development in wheat. J Appl Genet 60:233–241

    Article  CAS  PubMed  Google Scholar 

  • Norre F, Peyrot C, Garcia C, Rance I, Drevet J, Theisen M, Gruber V (2002) Powerful efect of an atypical bifactorial endosperm box from wheat HMWG-Dx5 promoter in maize endosperm. Plant Mol Biol 50:699–712

    Article  CAS  PubMed  Google Scholar 

  • Onodera Y, Suzuki A, Wu CY, Washida H, Takaiwa F (2001) A rice functional transcriptional activator, RISBZ1, responsible for endosperm-specific expression of storage protein genes through GCN4 motif. J Biol Chem 276:14139–14152

    Article  CAS  PubMed  Google Scholar 

  • Paine JA, Shipton CA, Chaggar S et al (2005) Improving the nutritional value of Golden Rice through increased pro-vitamin A content. Nat Biotechnol 23:482–487

    Article  CAS  PubMed  Google Scholar 

  • Parker ML (1982) Protein accumulation in the developing endosperm of a high protein line of Triticum dicoccoides. Plant Cell Environ 5:37–43

    CAS  Google Scholar 

  • Payne PI, Nightingale MA, Krattiger AF, Holt LM (1987) The relationship between HMW glutenin subunit composition and the bread-making quality of British-grown wheat varieties. J Sci Food Agric 40:51–65

    Article  CAS  Google Scholar 

  • Payne PI, Holt LM, Krattiger AF, Carrillo JM (1988) Relationships between seed quality characteristics and HMW glutenin subunit composition determined using wheat grown in Spain. J Cereal Sci 7:229–235

    Article  CAS  Google Scholar 

  • Pearce S, Huttly AK, Prosser IM et al (2015) Heterologous expression and transcript analysis of gibberellin biosynthetic genes of grasses reveals novel functionality in the GA3ox family. BMC Plant Biol 15:130

    Article  PubMed  PubMed Central  Google Scholar 

  • Perret SJ, Valentine J, Leggett M, Morris P (2003) Integration, expression and inheritance of transgenes in hexaploid oat (Avena sativa L.). J Plant Physiol 160:931–943

    Article  CAS  PubMed  Google Scholar 

  • Pfeifer M, Kugler KG, Sandve SR, Zhan B, Rudi H, Hvidsten TR, Mayer KFX, Olsen O-A (2014) Genome interplay in the grain transcriptome of hexaploid bread wheat. Science 345(6194):1250091

    Article  PubMed  CAS  Google Scholar 

  • Qu LQ, Xing YP, Liu WX, Xu XP, Song YR (2008) Expression pattern and activity of six glutelin gene promoters in transgenic rice. J Exp Bot 59:2417–2424

    Article  CAS  PubMed Central  Google Scholar 

  • Ravel C, Fiquet S, Boudet J, Dardevet M, Vincent J, Merlino M, Michard R, Martre P (2014) Conserved cis-regulatory modules in promoters of genes encoding wheat high-molecular-weight glutenin subunits. Front Plant Sci 5:621

    Article  PubMed  PubMed Central  Google Scholar 

  • Rushton PJ, Torres JT, Parniske M, Wernert P, Hahlbrock K, Somssich IE (1996) Interaction of elicitor-induced DNA-binding proteins with elicitor response elements in the promoters of parsley PR1 genes. EMBO J 15:5690–5700

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Seilmeier W, Belitz H, Wieser H (1991) Separation and quantitative determination of high-molecular-weight subunits of glutenin from different wheat varieties and genetic variants of the variety Sicco. Z Lebensm Unters Forsch A 192:124–129

    Article  CAS  Google Scholar 

  • Sun F, Liu X, Wei Q, Liu J, Yang T, Jia L, Wang Y, Yang G, He G (2017) Functional characterization of TaFUSCA3, a B3-superfamily transcription factor gene in the wheat. Front Plant Sci 8:1133

    Article  PubMed  PubMed Central  Google Scholar 

  • Takaiwa F (2007) A rice-based edible vaccine expressing multiple T-cell epitopes to induce oral tolerance and inhibit allergy. Immunol Allergy Clin N Am 27:129–139

    Article  Google Scholar 

  • Takaiwa F, Yamanouchi U, Yoshihara T, Washida H, Tanabe F, Kato A, Yamada K (1996) Characterization of common cis-regulatory elements responsible for the endosperm-specific expression of members of the rice glutelin multigene family. Plant Mol Biol 30:1207–1221

    Article  CAS  PubMed  Google Scholar 

  • Thilmony R, Guttman ME, Lin JW, Blechl AE (2014) The wheat HMW-glutenin 1Dy10 gene promoter controls endosperm expression in Brachypodium distachyon GM Crops Food 5:36–43

    Article  PubMed  Google Scholar 

  • Thomas MS, Flavell RB (1990) Identification of an enhancer element for the endosperm-specific expression of high molecular weight glutenin. Plant Cell 2:1171–1180

    CAS  PubMed  PubMed Central  Google Scholar 

  • Tosi P, Parker M, Gritsch CS, Carzaniga R, Martin B, Shewry PR (2009) Trafficking of storage proteins in developing grain of wheat. J Exp Bot 60:979–911

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Uthayakumaran S, Beasley HL, Stoddard FL, Keentok M, Phan-Thien N, Tanner RI, Békés F (2002) Synergistic and additive effects of three high molecular weight glutenin subunit loci. I. Effects on wheat dough rheology. Cereal Chem 79:294–300

    Article  CAS  Google Scholar 

  • Wang HL, Patrick JW, Offler CE (1995) The cellular pathway of photosynthates transfer in the developing wheat grain. III. A structural analysis and physiological studies of the pathway from the endosperm cavity to the starchy endosperm. Plant Cell Environ 18:373–388

    Article  Google Scholar 

  • Wang HH, Wang F, Liu DT, Gu YJ, Wang G (2011) Structure and development of endosperm transfer cells in wheat. J Titiceae Crops 31:944–952

    Google Scholar 

  • Wang K, Zhang X, Zhao Y, Chen FG, Xia GM (2013) Structure, variation and expression analysis of glutenin gene promoters from Triticum aestivum cultivar Chinese Spring shows the distal region of promoter 1Bx7 is key regulatory sequence. Gene 527:484–490

    Article  CAS  PubMed  Google Scholar 

  • Wang ZJ, Li YW, Yang YS, Liu X, Qin HJ, Dong ZY, Zheng SH, Zhang KP, Wang DW (2017) New insight into the function of wheat glutenin proteins as investigated with two series of genetic mutants. Sci Rep 7:3428

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Washida H, Wu CY, Suzuki A, Yamanouchi U, Akihama T, Harada K, Takaiwa F (1999) Identification of cis-regulatory elements required for endosperm expression of the rice storage protein glutelin gene GluB-1. Plant Mol Biol 40:1–12

    Article  CAS  PubMed  Google Scholar 

  • Wu CY, Suzuki A, Washida H, Takaiwa F (1998) The GCN4 motif in a rice glutelin gene is essential for endosperm-specific gene expression and is activated by Opaque-2 in transgenic rice plants. Plant J 14:673–683

    Article  CAS  PubMed  Google Scholar 

  • Wu CY, Washida H, Onodera Y, Harada K, Takaiwa F (2000) Quantitative nature of the Prolamin-box, ACGT and AACA motifs in a rice glutelin gene promoter: minimal cis-element requirements for endosperm-specific gene expression. Plant J 23:415–421

    Article  CAS  PubMed  Google Scholar 

  • Wu J, Yu L, Li L, Hu J, Zhou J, Zhou X (2007) Oral immunization with transgenic rice seeds expressing VP2 protein of infectious bursal disease virus induces protective immune responses in chickens. Plant Biotech J 5:570–578

    Article  CAS  Google Scholar 

  • Xiong F, Yu XR, Zhou L, Wang Z, Wang F, Xiong AS (2013) Structural development of aleurone and its function in common wheat. Mol Biol Rep 40:6785

    Article  CAS  PubMed  Google Scholar 

  • Yang YH, Li S, Zhang KP et al (2014) Efficient isolation of ion beam-induced mutants for homoeologous loci in common wheat and comparison of the contributions of Glu-1 loci to gluten functionality. Theor Appl Genet 127:359–372

    Article  CAS  PubMed  Google Scholar 

  • Zhang PP, Jondiko O, Tilleyc M, Awika JM (2014) Effect of high molecular weight glutenin subunit composition in common wheat on dough properties and steamed bread quality. J Sci Food Agric 94:2801–2806

    Article  CAS  PubMed  Google Scholar 

  • Zhu Q, Yu S, Zeng D et al (2017) Development of ‘‘Purple Endosperm Rice’’ by engineering anthocyanin biosynthesis in the endosperm with a high-efficiency transgene stacking system. Mol Plant 10:918–929

    Article  CAS  PubMed  Google Scholar 

  • Zhu J, Fang L, Yu J, Zhao Y, Chen F, Xia G (2018a) 5-azacytidine treatment and tapbf-d, over-expression increases glutenin accumulation within the wheat grain by hypomethylating the glu-1, promoters. Theor Appl Genet 131(3):735–746

    Article  CAS  PubMed  Google Scholar 

  • Zhu Q, Zeng D, Yu S et al (2018b) From golden rice to aSTARice: bioengineering Astaxanthin biosynthesis in rice endosperm. Mol Plant 11:1440–1448

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This research was supported by grants from National Key R&D Program of China (2016ZX08009003-004), National Natural Science Foundation of China (31571652), the Natural Science Foundation of Beijing (6162002), and the Youth Innovative Research Team of Capital Normal University.

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  1. Luning Duan and Shichen Han have contributed equally to this work.

Authors and Affiliations

  1. Key Laboratory of Genetics and Biotechnology, College of Life Science, Capital Normal University, Beijing, 100048, China

    Luning Duan, Shichen Han, Peihong Jiang, Yunsong Gu, Lin Chen, Junyi Mu, Yaxuan Li, Yueming Yan & Xiaohui Li

  2. National Wheat Improvement Center, Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100081, China

    Ke Wang & Xingguo Ye

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  1. Luning Duan
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Contributions

XHL conceived and designed the experiment. LND, SCH, KW, PHJ, YSG, LC, and JYM performed the experiment. XGY, YXL, and YMY provided technical assistance and scientific discussion. LND and XHL wrote the paper. All authors read and approved the final manuscript.

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Correspondence to Xiaohui Li.

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Duan, L., Han, S., Wang, K. et al. Analyzing the action of evolutionarily conserved modules on HMW-GS 1Ax1 promoter activity. Plant Mol Biol 102, 225–237 (2020). https://doi.org/10.1007/s11103-019-00943-6

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