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<p><i>Eucalyptus camaldulensis</i> is an important raw material for the paper and pulp industries. Genetic modifications that improve pulpwood yield could augment the raw material availability for industries. However, the genetic transformation of <i>Eucalyptus</i> is considered challenging. This study has evaluated different explants, co-cultivation media components, and sonication-cum-vacuum infiltration-assisted co-cultivation for improving transformation efficiency. Cotyledonary leaf explants were found to be most readily transformable, showing the highest transient GUS expression (81.33%) than proximal hypocotyl and distal hypocotyl regions (13.33% and 16%, respectively). Further, the optimal series of post-co-cultivation hygromycin selection concentrations to be used during subcultures of co-cultivated clonal explants for regeneration of transformed plants was determined. Sonication of explants in combination with vacuum infiltration-assisted co-cultivation resulted in an increased transformation efficiency as assayed through transient and stable GFP expression. A higher efficiency of regeneration of transformed plants at 0.64 to 3.92% was observed in sonicated cotyledonary leaf explants compared to non-sonicated cotyledonary leaf explants (0 to 0.76%). The lack of CaCl<sub>2</sub> and NaH<sub>2</sub>PO<sub>4</sub> or inclusion of MES or AB salts in co-cultivation medium did not affect transformation efficiency. However, biochar at 2&#xa0;g L<sup>−1</sup> and 4&#xa0;g L<sup>−1</sup> in co-cultivation broth resulted in a higher mean number of GUS spots at 2.0 ± 0.116 and 2.58 ± 0.15, respectively, as compared to 1.91 ± 0.09 observed for 0&#xa0;g L<sup>−1</sup> of biochar. This study is the first report on enhancing the transformation efficiency of <i>E. camaldulensis</i> using biochar and sonication-cum-vacuum infiltration-assisted co-cultivation.</p>

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Effect of explant sources, sonication combined with vacuum infiltration, and biochar on improving Agrobacterium-mediated transformation efficiency in Eucalyptus camaldulensis

  • 期刊论文

  • 发表日期：

    2025年11月5日

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  • M. C. Sandhya,
  • Sreeja Sahadevan,
  • Krishnaraj Shamili,
  • Manoj Kumar Rajendran,
  • Harshita Singh,
  • Aiyar Balasubramanian,
  • Muthukrishnan Arun,
  • Sivakumar Veerasamy,
  • Mathish Nambiar-Veetil

摘要

Eucalyptus camaldulensis is an important raw material for the paper and pulp industries. Genetic modifications that improve pulpwood yield could augment the raw material availability for industries. However, the genetic transformation of Eucalyptus is considered challenging. This study has evaluated different explants, co-cultivation media components, and sonication-cum-vacuum infiltration-assisted co-cultivation for improving transformation efficiency. Cotyledonary leaf explants were found to be most readily transformable, showing the highest transient GUS expression (81.33%) than proximal hypocotyl and distal hypocotyl regions (13.33% and 16%, respectively). Further, the optimal series of post-co-cultivation hygromycin selection concentrations to be used during subcultures of co-cultivated clonal explants for regeneration of transformed plants was determined. Sonication of explants in combination with vacuum infiltration-assisted co-cultivation resulted in an increased transformation efficiency as assayed through transient and stable GFP expression. A higher efficiency of regeneration of transformed plants at 0.64 to 3.92% was observed in sonicated cotyledonary leaf explants compared to non-sonicated cotyledonary leaf explants (0 to 0.76%). The lack of CaCl2 and NaH2PO4 or inclusion of MES or AB salts in co-cultivation medium did not affect transformation efficiency. However, biochar at 2 g L−1 and 4 g L−1 in co-cultivation broth resulted in a higher mean number of GUS spots at 2.0 ± 0.116 and 2.58 ± 0.15, respectively, as compared to 1.91 ± 0.09 observed for 0 g L−1 of biochar. This study is the first report on enhancing the transformation efficiency of E. camaldulensis using biochar and sonication-cum-vacuum infiltration-assisted co-cultivation.