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An improved selection strategy and the use of acetosyringone in shoot induction medium increase almond transformation efficiency by 100-fold

Abstract

An improved almond transformation system was developed with a 100× increase in efficiency (12.3%) as compared to the existing transformation method (0.1%) in spite of the lower regeneration ability of the explants used. Leaf transformation was performed with Agrobacterium EHA105/p35 SGUSINT. Main modifications introduced in the transformation protocol were the use of 150 μM acetosyringone (AS) during the 21-days induction period, and a different selection strategy. Transformed shoots were assayed using PCR, GUS analyses and Southern blotting. The improved methodology is being applied for transformation of in vitro propagated cultivars and opens the possibility of using almond as model for functional studies in Prunoideae.

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Abbreviations

AS:

acetosyringone

GUS:

β-glucuronidase

Kan:

kanamycin

SC:

subculture

References

  1. Druart P, Delporte F, Brazda M, Ugarte-Ballon C, Machado AC, Machado MLC, Jacquemin J, Watillon B, (1998) Genetic transformation of cherry trees Acta Hortic. 468: 71–76

  2. James DJ, Passey AJ, Barbara DJ, Bevan MW, (1989) Genetic transformation of apple (Malus pumila Mill.) using a disarmed Ti-binary vector Plant Cell Rep. 7: 658–666

  3. Machado LCM, Machado AC, Hanzer V, Weiss H, Regner F, Steinkellner H, Mattanovich D, Plail R, Knapp E, Kalthoff B, Katinger H, (1992) Regeneration of transgenic plants of Prunus armeniaca containing the coat protein gene of plum pox virus Plant Cell Rep. 11: 25–29

  4. Mante S, Morgens PH, Scorza R, Cordts JM, Callahan AM, (1991) Agrobacterium-mediated transformation of plum (Prunus domestica L.) hypocotyl slices and regeneration of transgenic plants Bio/Technol. 9: 853–857

  5. Martins M, Sarmento D, Oliveira MM, (2004) Genetic stability of micropropagated almond plantlets, as assessed by RAPD and ISSR markers Plant Cell Rep. 23: 492–496

  6. Miguel CM, Oliveira MM, (1999) Transgenic almond (Prunus dulcis Mill.) plants obtained by Agrobacterium-mediated transformation of leaf explants Plant Cell Rep. 18: 387–393

  7. Miguel CM, Druart P, Oliveira MM, (1996) Shoot regeneration from adventitious buds induced on juvenile and adult almond (Prunus dulcis Mill.) explants In Vitro Cell Dev Biol.-Plant 32: 148–153

  8. Padilla IMG, Webb K, Scorza R, (2003) Early antibiotic selection and efficient rooting and acclimatization improve the production of transgenic plum plants (Prunus domestica L.) Plant Cell Rep. 22: 38–45

  9. Petri C, Albuquerque N, Burgos L, (2005) The effect of aminoglycoside antibiotics on the adventitious regeneration from apricot leaves and selection of nptII-transformed leaf tissues. Plant Cell Tissue Organ Cult. 80: 271–276

  10. Pérez-Clemente RM, Pérez-Sanjuán A, García-Férriz L, Beltrán J-P, Cañas LA, (2004) Transgenic peach plants (Prunus persica L.) produced by genetic transformation of embryo sections using the green fluorescent protein, (GFP) as an in vivo marker Mol. Breed. 14: 419–427

  11. Santos PAG, Figueiredo AC, Oliveira MM, Barroso JG, Pedro LG, Deans AG, Scheffer AJC, (2005) Growth and essential oil composition of hairy root cultures of Levisticum officinale W.D.J. Koch, (lovage) Plant Sci. 168: 1089–1096

  12. Silva C, Garcia-Mas J, Sánchez A M, Arús P, Oliveira MM., (2005) Looking into flowering time in almond (Prunus dulcis, (Mill) D. A. Webb): The candidate gene approach Theor. Appl. Genet. 110: 959 – 968

  13. Smigocki AC, Hammerschlag FA, (1991) Regeneration of plants from peach embryo cells infected with a shooty mutant strain of Agrobacterium J. Am. Soc. Hortic. Sci. 116: 1092–1097

  14. Yancheva SD, Druart P, Watillon B, (2002) Agrobacterium-mediated transformation of plum (Prunus domestica L.) Acta Hortic. 577: 215–217

  15. Yepes LM, Aldwinckle HS, (1994) Factors that affect leaf regeneration efficiency in apple, and effect of antibiotics in morphogenesis Plant Cell Tissue Organ Cult. 37: 257–269

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Acknowledgements

Fundação para a Ciência e a Tecnologia is acknowledged for funding Project POCTI/AGR/38507/2001.

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Correspondence to M. Margarida Oliveira.

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Costa, M.S., Miguel, C. & Oliveira, M.M. An improved selection strategy and the use of acetosyringone in shoot induction medium increase almond transformation efficiency by 100-fold . Plant Cell Tiss Organ Cult 85, 205–209 (2006). https://doi.org/10.1007/s11240-005-9073-z

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Keywords

  • adventitious regeneration
  • Agrobacterium-mediated transformation
  • kanamycin resistance
  • Prunus dulcis
  • Rosaceae