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WUS and STM-based reporter genes for studying meristem development in poplar

  • Genetics and Genomics
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Abstract

We describe the development of a reporter system for monitoring meristem initiation in poplar using promoters of poplar homologs to the meristem-active regulatory genes WUSCHEL (WUS) and SHOOTMERISTEMLESS (STM). When ~3 kb of the 5′ flanking regions of close homologs were used to drive expression of the GUSPlus gene, 50–60% of the transgenic events showed expression in apical and axillary meristems. However, expression was also common in other organs, including in leaf veins (40 and 46% of WUS and STM transgenic events, respectively) and hydathodes (56% of WUS transgenic events). Histochemical GUS staining of explants during callogenesis and shoot regeneration using in vitro stems as explants showed that expression was detectable prior to visible shoot development, starting 3–15 days after explants were placed onto callus inducing medium. A minority of WUS and STM events also showed expression in the cambium, phloem, or xylem of regenerated, greenhouse grown plants undergoing secondary growth. Based on microarray gene expression data, a paralog of poplar WUS was detectably up-regulated during shoot initiation, but the other paralog was not. Both paralogs of poplar STM were down-regulated threefold to sixfold during early callus initiation. We identified 15–35 copies of cytokinin response regulator binding motifs (ARR1AT) and one copy of the auxin response element (AuxRE) in both promoters. Several of the events recovered may be useful for studying the process of primary and secondary meristem development, including treatments intended to stimulate meristem development to promote clonal propagation and genetic transformation.

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Abbreviations

ARR:

Arabidopsis response regulator

AuxRE:

Auxin response element

CIM:

Callus induction medium

CLV :

CLAVATA

CZ:

Central zone

PLACE:

Plant Cis-acting regulatory DNA elements

RAM:

Root apical meristem

SAM:

Shoot apical meristem

SAUR :

Small auxin-up RNA

SIM:

Shoot induction medium

STM :

SHOOTMERISTEMLESS

SURE:

Sulfur-responsive element

TAIR:

The Arabidopsis information resource

VC:

Vascular cambium

WUS :

WUSCHEL

X-Gluc:

5-Bromo-4-chloro-3-indolyl-β-d-glucuronic acid

References

  • Aloni R, Schwalm K, Langhans M, Ullrich CI (2003) Gradual shifts in sites of free-auxin production during leaf-primordium development and their role in vascular differentiation and leaf morphogenesis in Arabidopsis. Planta 216:841–853

    CAS  PubMed  Google Scholar 

  • Arias RS, Filichkin SA, Strauss SH (2006) Divide and conquer: development and cell cycle genes in plant transformation. Trends Biotechnol 24:267–273

    Article  CAS  PubMed  Google Scholar 

  • Bao Y (2008) Genome scale transcriptome analysis and development of reporter systems for studying shoot organogenesis in poplar. M.S. in genetics. Oregon State University, Corvallis

    Google Scholar 

  • Baumann K, De Paolis A, Costantino P, Gualberti G (1999) The DNA binding site of the Dof protein NtBBF1 is essential for tissue-specific and auxin-regulated expression of the rolB oncogene in plants. Plant Cell 11:323–334

    Article  CAS  PubMed  Google Scholar 

  • Baurle I, Laux T (2005) Regulation of WUSCHEL transcription in the stem cell niche of the Arabidopsis shoot meristem. Plant Cell 17:2271–2280

    Article  PubMed  Google Scholar 

  • Bhalla PL, Singh MB (2006) Molecular control of stem cell maintenance in shoot apical meristem. Plant Cell Rep 25:249–256

    Article  CAS  PubMed  Google Scholar 

  • Brunner AM, DiFazio SP, Crasta O, Fei Z, Mane SP, Sobral B, Dharmawardhana P (2007) Microarray expression analysis of poplar development. In: Plant and animal genomes XV conference, San Diego, CA, USA

  • Byrne ME, Kidner CA, Martienssen RA (2003) Plant stem cells: divergent pathways and common themes in shoots and roots. Curr Opin Genet Dev 13:551–557

    Article  CAS  PubMed  Google Scholar 

  • Che P, Lall S, Nettleton D, Howell SH (2006) Gene expression programs during shoot, root, and callus development in Arabidopsis tissue culture. Plant Physiol 141:620–637

    Article  CAS  PubMed  Google Scholar 

  • Filichkin SA, Meilan R, Busov VB, Ma C, Brunner AM, Strauss SH (2006) Alcohol-inducible gene expression in transgenic Populus. Plant Cell Rep 25:660–667

    Article  CAS  PubMed  Google Scholar 

  • Fusada N, Masuda T, Kuroda H, Shimada H, Ohta H, Takamiya K (2005) Identification of a novel cis-element exhibiting cytokinin-dependent protein binding in vitro in the 5′-region of NADPH-protochlorophyllide oxidoreductase gene in cucumber. Plant Mol Biol 59:631–645

    Article  CAS  PubMed  Google Scholar 

  • Gallie DR (1998) Controlling gene expression in transgenics. Curr Opin Plant Biol 1:166–172

    Article  CAS  PubMed  Google Scholar 

  • Goda H, Sawa S, Asami T, Fujioka S, Shimada Y, Yoshida S (2004) Comprehensive comparison of auxin-regulated and brassinosteroid-regulated genes in Arabidopsis. Plant Physiol 134:1555–1573

    Article  CAS  PubMed  Google Scholar 

  • Groover AT (2005) What genes make a tree a tree? Trends Plant Sci 10:210–214

    Article  CAS  PubMed  Google Scholar 

  • Groover AT, Pattishall A, Jones AM (2003) IAA8 expression during vascular cell differentiation. Plant Mol Biol 51:427–435

    Article  CAS  PubMed  Google Scholar 

  • Groover A, Fontana JR, Dupper G, Ma C, Martienssen R, Strauss S, Meilan R (2004) Gene and enhancer trap tagging of vascular-expressed genes in poplar trees. Plant Physiol 134:1742–1751

    Article  CAS  PubMed  Google Scholar 

  • Groover AT, Mansfield SD, DiFazio SP, Dupper G, Fontana JR, Millar R, Wang Y (2006) The Populus homeobox gene ARBORKNOX1 reveals overlapping mechanisms regulating the shoot apical meristem and the vascular cambium. Plant Mol Biol 61:917–932

    Article  CAS  PubMed  Google Scholar 

  • Haecker A, Gross-Hardt R, Geiges B, Sarkar A, Breuninger H, Herrmann M, Laux T (2004) Expression dynamics of WOX genes mark cell fate decisions during early embryonic patterning in Arabidopsis thaliana. Development 131:657–668

    Article  CAS  PubMed  Google Scholar 

  • Higo K, Ugawa Y, Iwamoto M, Korenaga T (1999) Plant cis-acting regulatory DNA elements (PLACE) database: 1999. Nucleic Acids Res 27:297–300

    Article  CAS  PubMed  Google Scholar 

  • Jansson S, Douglas CJ (2007) Populus: a model system for plant biology. Annu Rev Plant Biol 58:435–458

    Article  CAS  PubMed  Google Scholar 

  • Kamiya N, Nagasaki H, Morikami A, Sato Y, Matsuoka M (2003) Isolation and characterization of a rice WUSCHEL-type homeobox gene that is specifically expressed in the central cells of a quiescent center in the root apical meristem. Plant J 35:429–441

    Article  CAS  PubMed  Google Scholar 

  • Laux T (2003) The stem cell concept in plants: a matter of debate. Cell 113:281–283

    Article  CAS  PubMed  Google Scholar 

  • Laux T, Mayer KF, Berger J, Jurgens G (1996) The WUSCHEL gene is required for shoot and floral meristem integrity in Arabidopsis. Development 122:87–96

    CAS  PubMed  Google Scholar 

  • Lee-Stadelmann OY, Lee SW, Hackett WP (1989) The formation of adventitious buds in vitro on micro-cross sections of hybrid Populus leaf midveins. Plant Sci 61:263–272

    Article  Google Scholar 

  • Leibfried A, To JP, Busch W, Stehling S, Kehle A, Demar M, Kieber JJ, Lohmann JU (2005) WUSCHEL controls meristem function by direct regulation of cytokinin-inducible response regulators. Nature 438:1172–1175

    Article  CAS  PubMed  Google Scholar 

  • Long JA, Moan EI, Medford JI, Barton MK (1996) A member of the KNOTTED class of homeodomain proteins encoded by the STM gene of Arabidopsis. Nature 379:66–69

    Article  CAS  PubMed  Google Scholar 

  • Maruyama-Nakashita A, Nakamura Y, Watanabe-Takahashi A, Inoue E, Yamaya T, Takahashi H (2005) Identification of a novel cis-acting element conferring sulfur deficiency response in Arabidopsis roots. Plant J 42:305–314

    Article  CAS  PubMed  Google Scholar 

  • Mauseth JD (1988) Plant anatomy. Addison Wesley/Benjamin Cummings, San Francisco

    Google Scholar 

  • Mayer KF, Schoof H, Haecker A, Lenhard M, Jurgens G, Laux T (1998) Role of WUSCHEL in regulating stem cell fate in the Arabidopsis shoot meristem. Cell 95:805–815

    Article  CAS  PubMed  Google Scholar 

  • Nardmann J, Werr W (2006) The shoot stem cell niche in angiosperms: expression patterns of WUS orthologues in rice and maize imply major modifications in the course of mono- and dicot evolution. Mol Biol Evol 23:2492–2504

    Article  CAS  PubMed  Google Scholar 

  • Nehra N, Becwar M, Rottmann W, Pearson L, Chowdhury K, Chang S, Dayton Wilde H, Kodrzycki R, Zhang C, Gause K, Parks D, Hinchee M (2005) Forest biotechnology: innovative methods, emerging opportunities. In Vitro Cell Dev Biol Plant 41:701–717

    Article  CAS  Google Scholar 

  • Nilsson O, Aldén T, Sitbon F, Anthony Little C, Chalupa V, Sandberg G, Olsson O (1992) Spatial pattern of cauliflower mosaic virus 35S promoter-luciferase expression in transgenic hybrid aspen trees monitored by enzymatic assay and non-destructive imaging. Transgenic Res 1:209–220

    Article  CAS  Google Scholar 

  • Ross EJ, Stone JM, Elowsky CG, Arredondo-Peter R, Klucas RV, Sarath G (2004) Activation of the Oryza sativa non-symbiotic haemoglobin-2 promoter by the cytokinin-regulated transcription factor, ARR1. J Exp Bot 55:1721–1731

    Article  CAS  PubMed  Google Scholar 

  • Schrader J, Nilsson J, Mellerowicz E, Berglund A, Nilsson P, Hertzberg M, Sandberg G (2004) A high-resolution transcript profile across the wood-forming meristem of poplar identifies potential regulators of cambial stem cell identity. Plant Cell 16:2278–2292

    Article  CAS  PubMed  Google Scholar 

  • Scofield S, Murray JA (2006) The evolving concept of the meristem. Plant Mol Biol 60:V–VII

    Article  CAS  PubMed  Google Scholar 

  • Sterky F, Bhalerao RR, Unneberg P, Segerman B, Nilsson P, Brunner AM, Charbonnel-Campaa L, Lindvall JJ, Tandre K, Strauss SH, Sundberg B, Gustafsson P, Uhlen M, Bhalerao RP, Nilsson O, Sandberg G, Karlsson J, Lundeberg J, Jansson S (2004) A Populus EST resource for plant functional genomics. Proc Natl Acad Sci USA 101:13951–13956

    Article  PubMed  Google Scholar 

  • Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599

    Article  CAS  PubMed  Google Scholar 

  • Tuskan GA, Difazio S, Jansson S, Bohlmann J, Grigoriev I, Hellsten U, Putnam N, Ralph S, Rombauts S, Salamov A, Schein J, Sterck L, Aerts A, Bhalerao RR, Bhalerao RP, Blaudez D, Boerjan W, Brun A, Brunner A, Busov V, Campbell M, Carlson J, Chalot M, Chapman J, Chen GL, Cooper D, Coutinho PM, Couturier J, Covert S, Cronk Q, Cunningham R, Davis J, Degroeve S, Dejardin A, Depamphilis C, Detter J, Dirks B, Dubchak I, Duplessis S, Ehlting J, Ellis B, Gendler K, Goodstein D, Gribskov M, Grimwood J, Groover A, Gunter L, Hamberger B, Heinze B, Helariutta Y, Henrissat B, Holligan D, Holt R, Huang W, Islam-Faridi N, Jones S, Jones-Rhoades M, Jorgensen R, Joshi C, Kangasjarvi J, Karlsson J, Kelleher C, Kirkpatrick R, Kirst M, Kohler A, Kalluri U, Larimer F, Leebens-Mack J, Leple JC, Locascio P, Lou Y, Lucas S, Martin F, Montanini B, Napoli C, Nelson DR, Nelson C, Nieminen K, Nilsson O, Pereda V, Peter G, Philippe R, Pilate G, Poliakov A, Razumovskaya J, Richardson P, Rinaldi C, Ritland K, Rouze P, Ryaboy D, Schmutz J, Schrader J, Segerman B, Shin H, Siddiqui A, Sterky F, Terry A, Tsai CJ, Uberbacher E, Unneberg P, Vahala J, Wall K, Wessler S, Yang G, Yin T, Douglas C, Marra M, Sandberg G, Van de Peer Y, Rokhsar D (2006) The genome of black cottonwood, Populus trichocarpa (Torr. & Gray). Science 313:1596–1604

    Article  CAS  PubMed  Google Scholar 

  • Uchida N, Townsley B, Chung KH, Sinha N (2007) Regulation of SHOOT MERISTEMLESS genes via an upstream-conserved noncoding sequence coordinates leaf development. Proc Natl Acad Sci USA 104:15953–15958

    Article  CAS  PubMed  Google Scholar 

  • van der Hoeven C, Dietz A, Landsmann J (1994) Variability of organ-specific gene expression in transgenic tobacco plants. Transgenic Res 3:159–166

    Article  Google Scholar 

  • Vernoux T, Benfey PN (2005) Signals that regulate stem cell activity during plant development. Curr Opin Genet Dev 15:388–394

    Article  CAS  PubMed  Google Scholar 

  • Wang X, He H, Li L, Chen R, Deng XW, Li S (2006) NMPP: a user-customized NimbleGen microarray data processing pipeline. Bioinformatics 22:2955–2957

    Article  CAS  PubMed  Google Scholar 

  • Wei H, Meilan R, Brunner A, Skinner J, Ma C, Gandhi H, Strauss S (2007) Field trial detects incomplete barstar attenuation of vegetative cytotoxicity in Populus trees containing a poplar LEAFY promoter::barnase sterility transgene. Mol Breed 19:69–85

    Article  CAS  Google Scholar 

  • Weigel D, Glazebrook J (2002) Arabidopsis: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor

    Google Scholar 

  • Williams L, Fletcher JC (2005) Stem cell regulation in the Arabidopsis shoot apical meristem. Curr Opin Plant Biol 8:582–586

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

We would like to thank Rhonda Pham, and other high school and undergraduate student assistants working under Cathleen Ma, who performed transformation and in vitro propagation. We thank the USDA ARS Nursery and Floriculture Initiative, and the industrial members of the TBGRC Research Cooperative at Oregon State University for their support.

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Author notes

  1. P. Dharmawardhana

    Present address: Department of Botany and Plant Pathology, Oregon State University, Corvallis, OR, 97331-2902, USA

  2. R. Arias

    Present address: Genomics Laboratory, USDA-ARS, Stoneville, MS, 38776-0038, USA

  3. M. B. Allen

    Present address: College of Pharmacy, Oregon State University, Corvallis, OR, 97331-3507, USA

Authors and Affiliations

  1. Department of Forest Ecosystems and Society, Oregon State University, Corvallis, OR, 97331-5752, USA

    Y. Bao, P. Dharmawardhana, C. Ma & Steven H. Strauss

  2. Genetics Graduate Program, Oregon State University, Corvallis, OR, 97331-7303, USA

    Y. Bao & Steven H. Strauss

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  1. Y. Bao
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Correspondence to Steven H. Strauss.

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Communicated by S. Merkle.

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Bao, Y., Dharmawardhana, P., Arias, R. et al. WUS and STM-based reporter genes for studying meristem development in poplar. Plant Cell Rep 28, 947–962 (2009). https://doi.org/10.1007/s00299-009-0685-3

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