Abstract
Grapevine embryogenic cultures are ideal target tissues for inserting desired traits of interest and improving existing cultivars via precision breeding (PB). PB is a new approach that, like conventional breeding, utilizes only DNA fragments obtained from sexually compatible grapevine plants. Embryogenic culture induction occurs by placing leaves or stamens and pistils on induction medium with a dark/light photoperiod cycle for 12–16 weeks. Resulting cultures produce sectors of embryogenic and non-embryogenic callus, which can be identified on the basis of callus morphology and color. Somatic embryo development occurs following transfer of embryogenic callus to development medium and cultures can be maintained for extended periods of time by transfer of the proliferating proembryonic masses to fresh medium at 4–6-week intervals. To demonstrate plant recovery via PB, somatic embryos at the mid-cotyledonary stage are cocultivated with Agrobacterium containing the desired gene of interest along with a, non-PB, enhanced green fluorescent protein/neomycin phosphotransferase II (egfp/nptII) fusion gene. Modified cultures are grown on proliferation and development medium to produce uniformly modified somatic embryos via secondary embryogenesis. Modified embryos identified on the basis of green fluorescence and kanamycin resistance are transferred to germination medium for plant development. The resulting plants are considered to prototype examples of the PB approach, since they contain egfp/nptII, a non-grapevine-derived fusion gene. Uniform green fluorescent protein (GFP) fluorescence can be observed in all tissues of regenerated plants.
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References
Bertelli A, Das D (2009) Grapes, wines, resveratrol and heart health. J Cardiovasc Pharmacol 54:468–476
Anderson K, Aryal N (2014) Database of regional, national and global winegrape bearing areas by variety, 2000 and 2010. Wine Econ. Res. Ctr., Univ. Adelaide, Dec 2013. http://www.adelaide.edu.au/wine-econ/databases/winegrapes/
Gray DJ, Jayasankar S, Li Z (2005) Vitis spp. Grape (Chapter 22). In: Litz RE (ed) Biotechnology of fruit and nut crops. CAB International, Wallingford, Oxfordshire, pp 672–706
Einset J, Pratt C (1975) Grapes. In: Moore JN, Janick J (eds) Advances in fruit breeding. Purdue University Press, Lafayette, IN, pp 130–153
Winkler AJ, Cook JA, Kliewer WM, Lider LA (1974) General viticulture. University of California Press, Berkeley, pp 16–28
Vivier MA, Pretorius IS (2000) Genetic improvement of grapevine: tailoring grape varieties for the third millennium – a review. S Afr J Enol Vitic 21:5–26
Gray DJ, Li ZT, Dhekney SA (2014) Precision breeding of grapevine for improved traits. Plant Sci. doi:10.1016/j.plantsci.2014.03.023, Online First
Li ZT, Dhekney SA, Gray DJ (2011) Use of the VvMybA1 gene for non-destructive quantification of promoter activity via color histogram analysis in grapevine (Vitis vinifera) and tobacco. Transgenic Res 20:1087–1097
Li ZT, Kim KH, Jasinski JR, Creech MR, Gray DJ (2012) Large-scale characterization of promoters from grapevine (Vitis spp.) using quantitative anthocyanin and GUS assay. Plant Sci 196:132–142
Kikkert JR, Vidal MJ, Reisch BI (2004) Stable transformation of plant cells by particle bombardment/biolistics. In: Pena L (ed) Transgenic plants: methods and protocols, 286th edn. Humana, Totowa, NJ, pp 61–78
Gray DJ, Dhekney SA, Li ZT, Cordts JM (2012) Genetic engineering of grapevine and progress towards commercial deployment. In: Scorza R, Mou B (eds) Transgenic horticultural crops: challenges and opportunities – essays by experts. CRC, Taylor and Francis, Boca Raton, FL, pp 317–331
Gray DJ (1995) Somatic embryogenesis in grape. In: Jain SM, Gupta PK, Newton RJ (eds) Somatic embryogenesis in woody plants, vol 2. Kluwer Academic, Dordrecht, pp 191–217
Dhekney SA, Li ZT, Compton ME, Gray DJ (2009) Optimizing initiation and maintenance of Vitis embryogenic cultures. HortSci 44:1400–1406
Dhekney SA, Li ZT, Gray DJ (2011) Factors influencing induction and maintenance of Vitis rotundifolia Michx. embryogenic cultures. Plant Cell Tissue Organ Cult 105:175–180
Dhekney SA, Li ZT, Dutt M, Gray DJ (2008) Agrobacterium-mediated transformation of embryogenic cultures and regeneration of transgenic plants in Vitis routundifolia Michx. (muscadine grape). Plant Cell Rep 27:865–872
Dhekney SA, Li ZT, Zimmerman TW, Gray DJ (2009) Factors influencing genetic transformation and plant regeneration of Vitis. Am J Enol Vitic 60:285–292
Dhekney SA, Li ZT, Gray DJ (2011) Grapevines engineered to express cisgenic Vitis vinifera thaumatin-like protein exhibit fungal disease resistance. In Vitro Cell Dev Biol Plant 47:458–466
Li ZT, Dhekney S, Dutt M, Van Aman M, Tattersall J, Kelley KT, Gray DJ (2006) Optimizing Agrobacterium-mediated transformation of grapevine. In Vitro Cell Dev Biol Plant 42:220–227
Li ZT, Dhekney SA, Dutt M, Gray DJ (2008) An improved protocol for Agrobacterium-mediated transformation of grapevine. Plant Cell Tissue Organ Cult 93:311–321
Jayasankar S, Gray DJ, Litz RE (1999) High frequency somatic embryogenesis and plant regeneration from suspension cultures of grapevine. Plant Cell Rep 18:533–537
Nitsch JP, Nitsch C (1969) Haploid plants from pollen grains. Science 163:85–87
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Franks T, He DG, Thomas M (1998) Regeneration of transgenic Vitis vinifera L. Sultana plants: genotypic and phenotypic analysis. Mol Breed 4:321–333
Driver JA, Kuniyuki AH (1984) In vitro propagation of paradox walnut rootstock. HortSci 19:507–509
Gray DJ, Benton CM (1991) In vitro micropropagation and plant establishment of muscadine grape cultivars (Vitis rotundifolia). Plant Cell Tissue Organ Cult 27:7–14
Li ZT, Kim KH, Dhekney SA, Jasinski JR, Creech MR, Gray DJ (2014) An optimized procedure for plant recovery from somatic embryos significantly facilitates the genetic improvement of Vitis. Hort Res 1:1–7
Acknowledgements
S.A. Dhekney holds the E.A. Whitney Endowed Professorship in the UW Department of Plant Sciences. The research that enabled the Precision Breeding approach for grapevine was fostered by long-term support from the Florida Department of Agriculture and Consumer Services Viticulture Trust Fund, the USDA Specialty Crops Research Initiative Grant Program, and the Florida Agricultural Experiment Station, UF/IFAS.
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Dhekney, S.A., Li, Z.T., Grant, T.N.L., Gray, D.J. (2016). Somatic Embryogenesis and Genetic Modification of Vitis . In: Germana, M., Lambardi, M. (eds) In Vitro Embryogenesis in Higher Plants. Methods in Molecular Biology, vol 1359. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3061-6_11
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DOI: https://doi.org/10.1007/978-1-4939-3061-6_11
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