Transgenic peach plants (Prunus persica L.) produced by genetic transformation of embryo sections using the green fluorescent protein (GFP) as an in vivo marker
Purchase on Springer.com
$39.95 / €34.95 / £29.95*
Rent the article at a discountRent now
* Final gross prices may vary according to local VAT.
The main obstacle to genetic engineering of fruit tree species is the regeneration of transformed plantlets. Transformation events in peach (Prunus persica L.) have been reported using particle bombardment or Agrobacteriummediated transformation of immature embryos. However, the regeneration of plants from transgenic tissues is still difficult and the recovery of non-chimeric plants has not been reported to date. In this paper we describe an efficient, reliable transformation and regeneration system to produce transgenic peach plants using embryo sections of mature seeds as starting material. This represents an important advantage due to the availability of such material throughout the year. A. tumefaciens strain C58 (pMP90) containing the binary plasmid pBin19 was used as vector system for transformation. We used the Nospro-nptII-Noster cassette as a selectable marker and the CaMV35Spro-sgfp-CaMV35Ster cassette as a vital reporter gene coding for an improved version of the green fluorescent protein (sGFP). In vitro cultured embryo sections were Agrobacterium-cocultivated and, after selection, transgenic shoots were regenerated. Shoots that survived exhibited high-level of sGFP expression mainly visible in the young leaves of the apex. In vivo monitoring of GFP expression permitted an early, rapid and easy discrimination of both transgenic and escape buds. After elimination of escapes, transgenic shoots were rooted in vitro and the recovered plantlets were screened using PCR amplification. Southern analysis confirmed stable genomic integration of the sgfp transgene. The high levels of GFP expression were also maintained in the second generation of transgenic peach plants.
- Bhansali R. R., Driver JA. and Durzan D. J. 1990. Rapid multiplication of adventitious somatic embryos in peach and nectarine by secondary embryogenesis. Plant Cell Rep. 9: 280-284.
- Callahan A. M., Scorza R., Morgens P. H., Mante S., Cordts J. and Cohen R. 1991. Breeding for cold hardiness: searching for genes to improve fruit quality in cold hardy peach germplam. HortScience 26: 522-526.
- Callahan A. M., Morgens P. H., Wright P. and Nichols K. E. 1992. Comparison of pch313 (pTom13 homolog) accumulation during fruit softening and wounding of two phenotypically different peach cultivars. Plant Physiol. 100: 482-488.
- Callahan A. M., Cohen R. A., Dunn L. J. and Morgens P. H. 1993. Isolation of genes affecting peach fruit ripening. Acta Hort. 336: 47-50.
- Chiu W., Niwa Y., Zeng W. and Hirano T. 1996. Engineered GFP as a vital reporter in plants. Curr. Biol. 6: 325-330.
- Dellaporta S. L., Wood J. and Hicks J. B. 1983. A plant DNA minipreparation: Version II. Plant Mol. Biol. Rep. 4: 19-21.
- Duncan P. B. 1955. Multiple range and multiple F tests. Biometrics 11: 1-42.
- Gentile A., Monticelli S. and Damiano C. 2002. Adventitious shoot regeneration in peach (Prunus persica L. Batsch). Plant Cell Rep. 20: 1011-1016.
- Ghorbel R., Juárez J., Navarro L. and Peña L. 1999. Green fluorescent protein as a screenable marker to increase the efficiency of generating transgenic woody fruit plants. Theor. Appl. Genet. 99: 350-358.
- Hammerschlag F. A., Bauchan G. and Scorza R. 1985. Regeneration of peach plants from callus derived from immature embryos. Theor. Appl. Genet. 70: 248-251.
- Hood E. E., Gelvin S. B., Melchers L. S. and Hoekema A. 1993. New Agrobacterium helper plasmids for gene transfer to lants. Transgenic Res. 2: 208-218.
- Koncz C. and Schell J. 1986. The promoter of TL-DNA gene 5 controls the tissue-specific expression of chimaeric genes carried by a novel type of Agrobacterium binary vector. Mol. Gen. Genet. 204: 383-396.
- Laimer M., da Camara Machado A., Hanzer V., Weiss H., Regner F., Steinkellner H., Mattanivich D., Plail R., Knapp E., Kalthoff B. and 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.
- Lee E., Speirs J., Gray J. and Brady C. J. 1990. Homologies to the tomato endopolygalacturonase gene in the peach genome. Plant Cell Environ. 13: 513-521.
- Mante S., Scorza R. and Cordts J. M. 1989. Plant regeneration from cotyledons of Prunus persica, Prunus domestica y Prunus cerasus. Plant Cell Tiss. Org. Cult. 19: 1-11.
- Meng X. and Zhou W. 1981. Induction of embryoid and production of plantlets in vitro from endosperm of peach. Acta Agric. Univ. Peking 7: 95-98.
- Miguel C. M. and Oliveira M. M. 1999. Transgenic almond (Prunus dulcis Mill. ) plants obtained by Agrobacterium-mediated transformation of leaf explants. Plant Cell Rep. 18: 387-393.
- Murashige T. and Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473-479.
- Pooler M. R. and Scorza R. 1995. Regeneration of peach (Prunus persica L. Batsch) rootstock cultivars from cotyledons of mature stored seed. HortScience 30: 355-356.
- Quoirin M. and Lepoivre P. 1977. Étude de milieux adaptes aux cultures in vitro de prunus. Acta. Hort. 78: 437-442.
- Sambrook J., Fritsch E. F. and Maniatis T. 1989. Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA.
- Santarém E. R., Trick H. N., Essig J. H. and Finer J. J. 1998. Sonication-assisted Agrobacterium-mediated transformation of soybean immature cotyledons: optimization of transient expression. Plant Cell Rep. 17: 752-759.
- Scorza R., Morgens P. H., Cordts J. M., Mante S. and Callahan A. M. 1990. Agrobacterium-mediated transformation of peach (Prunus persica L. Batch) leaf segments, immature embryos and longterm embryogenic callus. In Vitro Cell Dev. Biol. 26: 829-834.
- Scorza R., Sherman W. B. 1996. Peaches. In Fruit Breeding Vol. 1: Tree and Tropical Fruits. Wiley J. and Sons, New York, USA.
- Sheen J., Hwang S., Niwa Y., Kobayashi H. and Galbraith D. W. 1995. Green fluorescent protein as a new vital marker in plant cells. Plant J. 8: 777-784.
- Smigocki A. C. and Hammerschlag F. A. 1991. Regeneration of plants from peach embryo cells infected with a shooty mutant strain of Agrobacterium. J. Am. Soc. Hortic. Sci. 116: 1092-1097.
- Smulders M. J. M., Rus-Kortekaas W. and Gillissen L. J. W. 1995. Development of polysomaty during differentiation in diploid and tetraploid tomato (Lycopersicon esculentum) plants. Plant Sci. 97: 53-60.
- Trick H. N. and Finer J. J. 1999. Induction of somatic embryogenesis and genetic transformation of Ohio buckeye (Aesculus glabra Willd. In Vitro Cell Dev. Biol. Plant 35: 57-60.
- Ye X., Brown S. K., Scorza R., Cordts J. and Sanford J. C. 1994. Genetic transformation of peach tissues by particle bombardment. J. Am. Soc. Hort. Sci. 119: 367-373.
- Transgenic peach plants (Prunus persica L.) produced by genetic transformation of embryo sections using the green fluorescent protein (GFP) as an in vivo marker
Volume 14, Issue 4 , pp 419-427
- Cover Date
- Print ISSN
- Online ISSN
- Kluwer Academic Publishers
- Additional Links
- Peach (Prunus persica L.)
- in vitro regeneration
- Embryo sections
- Industry Sectors
- Author Affiliations
- 1. Departamento de Biología del Desarrollo, Instituto de Biología Molecular y Celular de Plantas (C.S.I.C.-U.P.V.), Campus de la Universidad Politécnica de Valencia, Av. de los Naranjos s/n., 46022, Valencia, Spain
- 2. Comercial Técnica y Viveros S.L. Ctra, Nal, 340 Km. 873,5. L´Alcudia, 46250, Valencia, Spain