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Generation of new cisgenic apple lines resistant to either apple scab or fire blight.

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Cisgenic apple trees; development, characterization, and performance.

Frans A Krens1, Jan G Schaart1, Aranka M van der Burgh1

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Summary

Two novel methods generated cisgenic apples, one marker-free and one using marker excision. These cisgenic apples exhibit desirable traits like red flesh and scab resistance, paving the way for improved apple varieties.

Keywords:
Malus × domesticaMdMYB10Rvi6anthocyaninsapplecisgenesisfield trialscab resistance

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Area of Science:

  • Plant Biotechnology
  • Agricultural Science
  • Genetics

Background:

  • Developing cisgenic apples is crucial for crop improvement, offering traits like enhanced fruit quality and disease resistance.
  • Traditional genetic modification often involves selectable marker genes, which can raise regulatory concerns and are undesirable in the final product.

Purpose of the Study:

  • To develop and validate two distinct methods for generating cisgenic apples.
  • To introduce desirable traits, such as red flesh color and scab resistance, into apple cultivars using cisgenesis.

Main Methods:

  • Method 1: Gene-of-interest integration between T-DNA borders, avoiding foreign selectable markers. Used MdMYB10 for red flesh.
  • Method 2: Recombinase-based marker excision using the pMF vector system. Introduced scab resistance gene Rvi6.
  • Both methods involved Agrobacterium-mediated transformation, selection, and regeneration of plantlets, followed by grafting and field evaluation.

Main Results:

  • The MdMYB10 cisgene successfully produced red-fleshed apples with red petals and fruit.
  • The Rvi6 cisgene conferred scab resistance in both leaves and fruits, performing comparably to resistant controls.
  • Marker-free cisgenic and intragenic apple lines were successfully generated and verified.

Conclusions:

  • Both developed methods are effective for producing cisgenic apples with commercially relevant traits.
  • The marker-free approach and marker excision strategy offer viable pathways for developing improved apple varieties with enhanced quality and disease resistance.
  • Cisgenic apples demonstrated significant improvements in fruit characteristics and disease management, validating the potential of these biotechnological approaches.