Abstract
Maize-derived sequences from the transposable elements Activator (Ac) and Dissociation (Ds) have enabled studies of gene function via transposon tagging. The characteristics of modified, transgene-containing Ds elements constructed for some of these studies have demonstrated their ability to resolve complex loci, separate transgenes from marker genes and vector sequences, and to support high and heritable levels of transgene expression. To most efficiently design breeding schemes for developing transgenic populations via Ds-mediated transposition, detailed knowledge of the dynamics and characteristics of transposition in barley is necessary. Examination of a barley transposon tagging population (n = 4,954) derived from crosses of lines containing Ds-bar insertions to lines expressing Ac transposase showed that the frequencies of transposition from eight original Ds-bar loci ranged from 5 to 41 % among F2 individuals. Sequence analysis of Ds-bar terminal sequences and of flanking genomic sequences for 107 F2 and F3 individuals indicated precise integrations. Analysis of 173 flanking sequences derived from these populations and from previously produced populations, primarily using sequence-based methods, enabled the mapping of 159 to a specific chromosome and 136 to specific map locations. Of the 156 DsT loci that could be located to specific contigs, most were located in gene-rich areas and approximately 40 % were either in or near (within 1 kb) expressed sequences or predicted proteins. These data will enable the design of optimal breeding schemes for developing and using Ds-based systems for transposon tagging and for transgene delivery that are specific to barley.
Similar content being viewed by others
References
Ayliffe MA, Pallotta M, Langridge P, Pryor AJ (2007) A barley activation tagging system. Plant Mol Biol 64:329–347
Bregitzer P, Brown RH (2013) Long-term assessment of transgene behavior in barley: Ds-mediated delivery of bar results in robust, stable, and heritable expression. In Vitro Cell Dev Biol Plant 49:231–237
Brown RH, Bregitzer P (2011) A Ds insertional mutant of a barley miR172 gene results in indeterminate spikelet development. Crop Sci 51:1664–1672
Brown RH, Dahleen LS, Bregitzer P (2012) An efficient method for flanking sequence isolation in barley. Crop Sci 52:1229–1234
Cooper LD, Marquez-Cedillo L, Singh J, Sturbaum AK, Zhang S, Edwards V, Johnson K, Kleinhofs A, Rangel S, Carollo V, Bregitzer P, Lemaux PG, Hayes PM (2004) Mapping Ds insertions in barley using a sequence-based approach. Mol Gen Genomics 272:181–193
Cotsaftis O, Sallaud C, Breitler JC, Meynard D, Greco R, Pereira A, Guiderdoni E (2002) Transposon-mediated generation of T-DNA and marker-free rice plants expressing a Bt endotoxin gene. Mol Breed 10:165–180
Cowperthwaite M, Park W, Xu Z, Yan X, Maurais SC, Dooner HK (2002) Use of the transposon Ac as a gene-searching engine in the maize genome. Plant Cell 14:713–726
Dai S, Zheng P, Marmey P, Zhang S, Tian W, Chen S, Beachy RN, Fauquet C (2001) Comparative analysis of transgenic rice plants obtained by Agrobacterium-mediated transformation and particle bombardment. Mol Breed 7:25–33
Deng W, Nickle DC, Learn GH, Maust B, Mullins JI (2007) ViroBLAST: a stand-alone BLAST web server for flexible queries of multiple databases and user’s datasets. Bioinformatics 23:2334–2336
Dooner HK, Keller J, Harper E, Ralston E (1991) Variable patterns of transposition of the maize element Activator in tobacco. Plant Cell 3:473–482
Federoff NV (1989) Maize transposable elements. In: Howe M, Berg D (eds) Mobile DNA. ASM Press, Washington, pp 375–411
Franckowiak JD, Foster AE, Peterson VD, Pyler RE (1985) Registration of Bowman barley. Crop Sci 25:883
Goldsbrough AP, Lastrella CN, Yoder JI (1993) Transposition mediated re-positioning and subsequent elimination of marker genes from transgenic tomato. Biotechnology 11:1286–1292
International Barley Genome Sequencing Consortium (2012) A physical, genetic, and functional sequence assembly of the barley genome. Nature 491:711–716
Islam AKMR, Shepherd KW, Sparrow DHB (1981) Isolation and characterization of euplasmic wheat-barley chromosome addition lines. Heredity 46:161–174
Ito T, Motohashi R, Kuromori T, Mizukado S, Sakurai T, Kanahara H, Seki M, Shinozaki K (2002) A new resource of locally transposed Dissociation elements for screening gene-knockout lines in silico on the Arabidopsis genome. Plant Physiol 129:1695–1699
Jones JDG, Gilbert DE, Grady KL, Jorgenson RA (1987) T-DNA structure and gene expression in petunia plants transformed by Agrobacterium C58 derivatives. Mol Gen Genet 207:478–485
Jorgensen R, Snyder C, Jones JDG (1987) T-DNA is organized predominantly in inverted repeat structures in plants transformed with Agrobacterium tumefaciens C58 derivatives. Mol Gen Genet 207:471–477
Kim CM, Piao HL, Park SJ, Chon NS, Je BI, Sun B, Park SH, Park JY, Lee EJ, Kim MJ, Chung WS, Lee KH, Lee YS, Lee JJ, Won YJ, Yi G, Nam MH, Cha YS, Yun DW, Eun MY, Han C (2004) Rapid, large-scale generation of Ds transposant lines and analysis of the Ds insertion sites in rice. Plant J 39:252–263
Kohli A, Twyman RM, Abranches R, Wegel E, Stoger E, Christou P (2003) Transgene integration, organization, and interaction in plants. Plant Mol Biol 52:247–258
Kolesnik T, Szeverenyi I, Bachmann D, Kumar CS, Jian S, Ramamoorthy R, Cai M, Ma ZG, Sundaresan V, Ramachandran S (2004) Establishing an efficient Ac/Ds tagging system in rice: large-scale analysis of Ds flanking sequences. Plant J 37:265–274
Koprek T, McElroy D, Louwerse J, Williams-Carrier R, Lemaux PG (2000) An efficient method for dispersing Ds elements in the barley genome as a tool for determining gene function. Plant J 24:253–263
Koprek T, Rangel S, McElroy D, Louwerse JD, Williams-Carrier RE, Lemaux PG (2001) Transposon-mediated single-copy gene delivery leads to increased transgene expression stability in barley. Plant Physiol 125:1354–1362
Krens FA, Mans RMW, van Slogteren TMS, Hoge JHC, Wullems GJ, Shilperoort RA (1985) Structure and expression of DNA transferred to tobacco via transformation of protoplasts with Ti-plasmid DNA: co-transfer of T-DNA and non T-DNA sequences. Plant Mol Biol 5:223–234
Lange M, Vincze E, Møller MG, Holm PB (2006) Molecular analysis of transgene and vector backbone integration into the barley genome following Agrobacterium-mediated transformation. Plant Cell Rep 25:815–820
Lebel EG, Masson H, Bogucki A, Paszkoski J (1995) Transposable elements as plant transformation vectors for long stretches of foreign DNA. Theor Appl Genet 91:899–906
Makarevitch I, Svitashev SK, Somers DA (2003) Complete sequence analysis of transgene loci from plants transformed via microprojectile bombardment. Plant Mol Biol 52:421–432
Mascher M, Muehlbauer GJ, Rokhsar DS, Chapman J, Schmutz J, Barry K, Muñoz-Amatriaín M, Close TJ, Wise RP, Schulman AH, Himmelbach A, Mayer KFX, Scholz U, Poland JA, Stein N, Waugh R (2013) Anchoring and ordering NGS contig assemblies by population sequencing (POPSEQ). Plant J 76:718–727
McElroy D, Louwerse JD, McElroy SM, Lemaux PG (1997) Development of a simple transient assay for Ac/Ds activity in cells of intact barley tissue. Plant J 11:157–165
Meissner R, Chague V, Zhu Q, Emmanuel E, Elkind Y, Levy AA (2000) A high throughput system for transposon tagging and promoter trapping in tomato. Plant J 22:265–274
Meng L, Ziv M, Lemaux PG (2006) Nature of stress and transgene locus influences transgene expression stability in barley. Plant Mol Biol 62:15–28
Morino K, Olsen O-A, Shimamoto K (1999) Silencing of an aleurone-specific gene in transgenic rice is caused by a rearranged transgene. Plant J 17:275–285
Morita R, Kusaba M, Iida S, Yamaguchi H, Nishio T, Nishimura M (2009) Molecular characterization of mutations induced by gamma irradiation in rice. Genes Genet Syst 84:361–370
Nussbaumer T, Martis MM, Roessner SK, Pfeifer M, Bader KC, Sharma S, Gundlach H, Spannagl M (2013) MIPS PlantsDB: a database framework for comparative plant genome research. Nucleic Acids Res 41:D1144–D1151
Page DR, Köhler C, da Costa-Nunes J, Baroux C, Moore JM, Grossniklaus U (2004) Intrachromosomal excision of a hybrid Ds element induces large genomic deletions in Arabidopsis. Proc Natl Acad Sci USA 101:2969–2974
Rasmusson DC, Wilcoxson RW (1979) Registration of Morex barley. Crop Sci 19:293
Rinehart TA, Dean C, Weil CF (1997) Comparative analysis of non-random DNA repair following Ac transposon excision in maize and Arabidopsis. Plant J 12:1419–1427
Rubin E, Lithwick G, Levy AA (2001) The structure and evolution of the hAT transposon superfamily. Genetics 158:949–957
Scholz S, Lörz H, Lütticke S (2001) Transposition of the maize transposable element Ac in barley (Hordeum vulgare L.). Mol Gen Genet 264:653–661
Scott L, LaFoe D, Weil CF (1996) Adjacent sequences influence DNA repair accompanying transposon excision in maize. Genetics 142:237–246
Sigurbjorsson B, Micke A (1969) Progress in mutation breeding. In: Induced mutation in plants. Proceedings of an international symposium on the nature, induction, and utilization of mutation in plants. International Atomic Energy Agency, Vienna, pp 673–697
Singh J, Zhang S, Chen C, Cooper L, Bregitzer P, Sturbaum A, Hayes PM, Lemaux PG (2006) High-frequency Ds remobilization over multiple generations in barley facilitates gene tagging in large genome cereals. Plant Mol Biol 62:93–950
Singh S, Tan HQ, Singh J (2012) Mutagenesis of barley malting quality QTLs with Ds transposons. Funct Integr Genomics 12:131–141
Somers DA, Makarevitch I (2004) Transgene integration in plants: poking or patching holes in promiscuous genomes. Curr Opin Biotechnol 15:126–131
Springer PS (2000) Gene traps: tools for plant development and genomics. Plant Cell 12:1007–1020
Takano M, Egawa H, Ikeda J-E, Wakasa K (1997) The structures of integration sites in transgenic rice. Plant J 11:353–361
Travella S, Ross SM, Harden J, Everett C, Snape JW, Harwood WA (2005) A comparison of transgenic barley lines produced by particle bombardment and Agrobacterium-mediated techniques. Plant Cell Rep 23:780–789
Upadhyaya NM, Zhu Q-H, Zhou X-R, Eamens AL, Hoque MS, Ramm K, Shivakkumar R, Smith KF, Pan S-T, Li S, Peng K, Kim SJ, Dennis ES (2006) Dissociation (Ds) constructs, mapped Ds launch pads and a transiently expressed transposase system suitable for localized insertional mutagenesis in rice. Theor Appl Genet 112:1326–1341
Vollbrecht E, Duvick J, Schares JP, Ahern KR, Deewatthanawong P, Xu L, Conrad LJ, Kikuchi K, Kubinec TA, Hall BD, Weeks R, Unger-Wallace E, Muszynski M, Brendel VP, Brutnell TP (2010) Genome-wide distribution of transposed Dissociation elements in maize. Plant Cell 22:1667–1685
Walbot V (1992) Strategies for mutagenesis and gene cloning using transposon tagging and T-DNA insertional mutagenesis. Annu Rev Plant Physiol Mol Biol 43:49–82
Wang Y, Yau Y-Y, Perkins-Balding D, Thomson JG (2011) Recombinase technology: applications and possibilities. Plant Cell Rep 30:267–285
Wicker T, Mayer KFX, Gundlach H, Martis M, Steuemagel B, Scholz U, Šmiková H, Kubaláková M, Choulet F, Taudien S, Platzer M, Feuillet C, Fahima T, Budak H, Doležel J, Keller B, Stein N (2011) Frequent gene movement and pseudogene evolution is common to the large and complex genomes of wheat, barley, and their relatives. Plant Cell 23:1706–1718
Xiao Y-L, Peterson T (2002) Ac transposition is impaired by a small terminal deletion. Mol Genet Genomics 266:720–731
Zhao T, Palotta M, Langridge P, Prasad M, Graner A, Schulze-Lefert P, Koprek T (2006) Mapped Ds/T-DNA launch pads for functional genomics in barley. Plant J 47:811–826
Acknowledgments
Funding for this research was provided by the Agricultural Research Service, USDA, projects 5366-21000-031-00D and 5442-21000-036-00D. The USDA-ARS is an equal opportunity employer.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Brown, R.H., Singh, J., Singh, S. et al. Behavior of a modified Dissociation element in barley: a tool for genetic studies and for breeding transgenic barley. Mol Breeding 35, 85 (2015). https://doi.org/10.1007/s11032-015-0193-9
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s11032-015-0193-9