Abstract
Alfalfa is the most important forage legume worldwide. However, similar to other minor forage crops, it is usually harvested along with weeds, which decrease its nutrient quality and thus reduce its high value in the market. In addition, weeds reduce alfalfa yield by about 50 %. Although weeds are the limiting factor for alfalfa production, little progress has been made in the incorporation of herbicide-tolerant traits into commercial alfalfa. This is partially due to the high times and costs needed for the production of vast numbers of transgenic alfalfa events as an empirical approach to bypass the random transgenic silencing and for the identification of an event with optimal transgene expression. In this focus article, we report the complete sequence of pPZP200BAR and the extremely high efficiency of this binary vector in alfalfa transformation, opening the way for rapid and inexpensive production of transgenic events for alfalfa improvement public programs.
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References
D’Halluin K, Botterman J, De Greef W (1990) Engineering of herbicide-resistant alfalfa and evaluation under field conditions. Crop Sci 30:866–871
Garcia AN, Ayub ND, Fox AR, Gomez MC, Dieguez MJ, Pagano EM, Berini CA, Muschietti JP, Soto G (2014) Alfalfa snakin-1 prevents fungal colonization and probably coevolved with rhizobia. BMC Plant Biol 14:248
Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 41:95–98
Heap I (2014) Global perspective of herbicide-resistant weeds. Pest Manag Sci 70:1306–1310
Laguia-Becher M, Martin V, Kraemer M, Corigliano M, Yacono ML, Goldman A, Clemente M (2010) Effect of codon optimization and subcellular targeting on Toxoplasma gondii antigen SAG1 expression in tobacco leaves to use in subcutaneous and oral immunization in mice. BMC Biotechnol 10:52
Meek S (2011) A review of the environmental safety of the PAT protein. Environ Biosafety Res 10:73–101
Montague A, Ziauddin A, Lee R, Ainley WM, Strommer J (2007) High-efficiency phosphinothricin-based selection for alfalfa transformation. Plant Cell Tissue Organ Cult 91:29–36
Rivero M, Furman N, Mencacci N, Picca P, Toum L, Lentz E, Bravo-Almonacid F, Mentaberry A (2012) Stacking of antimicrobial genes in potato transgenic plants confers increased resistance to bacterial and fungal pathogens. J Biotechnol 157:334–343
Rogan G, Fitzpatrick S (2004) Petition for determination of nonregulated status: roundup ready alfalfa (Medicago sativa L.) events J101 and J163. USDA Petition Number 04-110-101
Rubiales D (2014) Alfalfa: back to the future. J Int Legume Soc. ISSN: 2340-1559
Schubert D, Lechtenberg B, Forsbach A, Gils M, Bahadur S, Schmidt R (2004) Silencing in Arabidopsis T-DNA transformants: the predominant role of a gene-specific RNA sensing mechanism versus position effects. Plant Cell 16:2561–2572
Shen WJ, Forde BG (1989) Efficient transformation of Agrobacterium spp. by high voltage electroporation. Nucleic Acids Res 17:8385
Soto G, Stritzler M, Lisi C, Alleva K, Pagano ME, Ardila F, Mozzicafreddo M, Cuccioloni M, Angeletti M, Ayub ND (2011) Acetoacetyl-CoA thiolase regulates the mevalonate pathway during abiotic stress adaptation. J Exp Bot 62:5699–5711
Thompson CJ, Movva NR, Tizard R, Crameri R, Davies JE, Lauwereys M, Botterman J (1987) Characterization of the herbicide-resistance gene bar from Streptomyces hygroscopicus. EMBO J 6:2519–2523
Tuteja N, Verma S, Sahoo RK, Raveendar S, Reddy IN (2012) Recent advances in development of marker-free transgenic plants: regulation and biosafety concern. J Biosci 37:167–197
White J, Chang SY, Bibb MJ (1990) A cassette containing the bar gene of Streptomyces hygroscopicus: a selectable marker for plant transformation. Nucleic Acids Res 18:1062
Acknowledgments
We are grateful to Dr. Esteban Hopp (IB-INTA) for his help in this work, especially during the initial phases. This work was supported by grants INTACD648/06, INTACD1016/12 and PICT Start-Up 2014-3759 to G.S.
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Communicated by N. Stewart.
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Jozefkowicz, C., Bottero, E., Pascuan, C. et al. Minimizing the time and cost of production of transgenic alfalfa libraries using the highly efficient completely sequenced vector pPZP200BAR. Plant Cell Rep 35, 1987–1990 (2016). https://doi.org/10.1007/s00299-016-2026-7
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DOI: https://doi.org/10.1007/s00299-016-2026-7