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<p>Blueberry is an important fruit crop with a unique flavor and high nutritional value. The integration of modern biotechnological tools with traditional methods is essential for accelerating the generation of new cultivars; however, the implementation of such approaches in Southern Highbush Blueberry (SHB) has been hindered due to the genotype-specific nature of shoot regeneration. Therefore, the current study aims to develop shoot regeneration protocols for six important SHB cultivars (‘Albus’, ‘Arcadia’, ‘Colossus’, ‘FL 14–242’, ‘Keecrisp’, and ‘Optimus’) representing the genetic diversity of the University of Florida (UF) blueberry germplasm collection. Varying concentrations of cytokinins (trans-Zeatin [tZ] and Thidiazuron [TDZ]) and an auxin (Naphthalene Acetic Acid [NAA]) were tested using leaf explants. Different responses to shoot regeneration with minimal auxin effects were observed, with efficiencies ranging from 38 to 92%. For ‘Colossus’, the optimal hormone concentration was 2.2&#xa0;mg/l tZ, while 8.9&#xa0;mg/l tZ was most effective for ‘Albus’, ‘Keecrisp’, and ‘Optimus’. In ‘FL-14-242’, a combination of 0.22&#xa0;mg/l TDZ and 2.2&#xa0;mg/l tZ yielded similar shoot regeneration rates. Although ‘Arcadia’ showed low regeneration efficiency overall, 4&#xa0;mg/l tZ produced the highest response for this cultivar. To demonstrate the applicability of the established protocols, the cultivar ‘Albus’ was subjected to <i>Agrobacterium</i>-mediated transformation using a vector carrying <i>AmROSEA1</i>, <i>eGFP</i>, and <i>nptII</i> genes. Putative transgenic lines were visually confirmed and further validated by PCR, resulting in a 2% transformation efficiency. This study provides a foundation for implementing novel breeding strategies in important SHB blueberry cultivars.</p>

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Establishment of in vitro shoot regeneration and genetic transformation system for Southern Highbush Blueberry (SHB)

  • 期刊论文

  • 发表日期：

    2025年11月12日

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  • Anandi Karn,
  • Felix Enciso-Rodriguez,
  • Yaiphabi Kumam,
  • Paul Adunola,
  • Tao Jiang,
  • Camila Azevedo,
  • Heqiang Huo,
  • Patricio Munoz

摘要

Blueberry is an important fruit crop with a unique flavor and high nutritional value. The integration of modern biotechnological tools with traditional methods is essential for accelerating the generation of new cultivars; however, the implementation of such approaches in Southern Highbush Blueberry (SHB) has been hindered due to the genotype-specific nature of shoot regeneration. Therefore, the current study aims to develop shoot regeneration protocols for six important SHB cultivars (‘Albus’, ‘Arcadia’, ‘Colossus’, ‘FL 14–242’, ‘Keecrisp’, and ‘Optimus’) representing the genetic diversity of the University of Florida (UF) blueberry germplasm collection. Varying concentrations of cytokinins (trans-Zeatin [tZ] and Thidiazuron [TDZ]) and an auxin (Naphthalene Acetic Acid [NAA]) were tested using leaf explants. Different responses to shoot regeneration with minimal auxin effects were observed, with efficiencies ranging from 38 to 92%. For ‘Colossus’, the optimal hormone concentration was 2.2 mg/l tZ, while 8.9 mg/l tZ was most effective for ‘Albus’, ‘Keecrisp’, and ‘Optimus’. In ‘FL-14-242’, a combination of 0.22 mg/l TDZ and 2.2 mg/l tZ yielded similar shoot regeneration rates. Although ‘Arcadia’ showed low regeneration efficiency overall, 4 mg/l tZ produced the highest response for this cultivar. To demonstrate the applicability of the established protocols, the cultivar ‘Albus’ was subjected to Agrobacterium-mediated transformation using a vector carrying AmROSEA1, eGFP, and nptII genes. Putative transgenic lines were visually confirmed and further validated by PCR, resulting in a 2% transformation efficiency. This study provides a foundation for implementing novel breeding strategies in important SHB blueberry cultivars.