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Krens FA, Schaart JG, van der Burgh AM, et al. Cisgenic apple trees; development, characterization, and performance. Front Plant Sci. 2015;6:286doi: 10.3389/fpls.2015.00286.
Krens, F. A., Schaart, J. G., van der Burgh, A. M., Tinnenbroek-Capel, I. E., Groenwold, R., Kodde, L. P., Broggini, G. A., Gessler, C., & Schouten, H. J. (2015). Cisgenic apple trees; development, characterization, and performance. Frontiers in plant science, 6286. https://doi.org/10.3389/fpls.2015.00286
Krens, Frans A, et al. "Cisgenic apple trees; development, characterization, and performance." Frontiers in plant science vol. 6 (2015): 286. doi: https://doi.org/10.3389/fpls.2015.00286
Krens FA, Schaart JG, van der Burgh AM, Tinnenbroek-Capel IE, Groenwold R, Kodde LP, Broggini GA, Gessler C, Schouten HJ. Cisgenic apple trees; development, characterization, and performance. Front Plant Sci. 2015 Apr 27;6:286. doi: 10.3389/fpls.2015.00286. eCollection 2015. PMID: 25964793; PMCID: PMC4410516.
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Front Plant Sci. 2015 Apr 27;6:286. doi: 10.3389/fpls.2015.00286. eCollection 2015.

Cisgenic apple trees; development, characterization, and performance.

Frontiers in plant science

Frans A Krens, Jan G Schaart, Aranka M van der Burgh, Iris E M Tinnenbroek-Capel, Remmelt Groenwold, Linda P Kodde, Giovanni A L Broggini, Cesare Gessler, Henk J Schouten

Affiliations

  1. Wageningen UR Plant Breeding, Wageningen University and Research Centre Wageningen, Netherlands.
  2. Plant Pathology, Institute of Integrative Biology, ETH Zürich Zürich, Switzerland.

PMID: 25964793 PMCID: PMC4410516 DOI: 10.3389/fpls.2015.00286

Abstract

Two methods were developed for the generation of cisgenic apples. Both have been successfully applied producing trees. The first method avoids the use of any foreign selectable marker genes; only the gene-of-interest is integrated between the T-DNA border sequences. The second method makes use of recombinase-based marker excision. For the first method we used the MdMYB10 gene from a red-fleshed apple coding for a transcription factor involved in regulating anthocyanin biosynthesis. Red plantlets were obtained and presence of the cisgene was confirmed. Plantlets were grafted and grown in a greenhouse. After 3 years, the first flowers appeared, showing red petals. Pollination led to production of red-fleshed cisgenic apples. The second method used the pM(arker)F(ree) vector system, introducing the scab resistance gene Rvi6, derived from apple. Agrobacterium-mediated transformation, followed by selection on kanamycin, produced genetically modified apple lines. Next, leaves from in vitro material were treated to activate the recombinase leading to excision of selection genes. Subsequently, the leaf explants were subjected to negative selection for marker-free plantlets by inducing regeneration on medium containing 5-fluorocytosine. After verification of the marker-free nature, the obtained plants were grafted onto rootstocks. Young trees from four cisgenic lines and one intragenic line, all containing Rvi6, were planted in an orchard. Appropriate controls were incorporated in this trial. We scored scab incidence for three consecutive years on leaves after inoculations with Rvi6-avirulent strains. One cisgenic line and the intragenic line performed as well as the resistant control. In 2014 trees started to overcome their juvenile character and formed flowers and fruits. The first results of scoring scab symptoms on apple fruits were obtained. Apple fruits from susceptible controls showed scab symptoms, while fruits from cisgenic and intragenic lines were free of scab.

Keywords: Malus × domestica; MdMYB10; Rvi6; anthocyanins; apple; cisgenesis; field trial; scab resistance

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