Summary: | As a valuable ornamental plant, <i>Sinningia hybrida</i> ‘Isa’s Murmur’ (<i>S. hybrida</i>) has genetic flower diversity, which has great potential to develop different flower characters in the horticultural market. The present study focuses on establishing a practical approach for the sustainable propagation of <i>S. hybrida</i>. Compared with aseptic seeding leaves explants, field-grown leaves explants are more suitable for adventitious shoot regeneration. Adding 0.1 mg L<sup>−1</sup> NAA and 2.0 mg L<sup>−1</sup> TDZ could obtain the highest adventitious shoot proliferation coefficient (24.5), and the induction rate was 91.7%. The shoot proliferation coefficient (20.7) and the greatest shoot length and induction rate (95.3%) were achieved in 0.1 mg L<sup>−1</sup> NAA and 2.0 mg L<sup>−1</sup> BA medium, accompanied by rooting formation. Adding 0.5 mg L<sup>−1</sup> GA<sub>3</sub>, 1.0 mg L<sup>−1</sup> BA, and 0.2 mg L<sup>−1</sup> IBA to MS medium can effectively prolong the regenerated buds for rooting. The best for rooting was 1/2 MS medium containing 0.3 mg L<sup>−1</sup> IBA, with the maximum number of roots (13.4 per shoot) and survival rate for transplanting (100%). This work aims to build an efficient, definitive, and scalable protocol for <i>S. hybrida</i> regeneration useful for large-scale cultivation and even more protoplast fusion and genetic transformation to develop more colorful or fragrant flowers.
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