Direct and indirect in vitro regeneration of Vanilla odorata C. Presl. and V. pompona Schiede, two aromatic species with potential relevance for future vanillin production

• Published in: In vitro cellular & developmental biology. Plant 2023-10, Vol.59 (5), p.621-636
• Main Authors: Warner, Jorge, Camacho-Solís, Yesly, Jiménez, Víctor M.
• Format: Article
• Language: English
• Subjects: Acclimatization; Acetic acid; Acids; Biomedical and Life Sciences; Biotechnology; Butyric acid; Calcium oxalate; Callus; Cell Biology; Developmental Biology; Explants; Flowers & plants; Growth regulators; Indole-3-butyric acid; Life Sciences; Morphology; Naphthalene; Plant Breeding/Biotechnology; Plant Genetics and Genomics; Plant growth; Plant Sciences; Plant Tissue Culture; Plantlets; Propagation; Regeneration; Segments; Shoots; Subculture; Survival; Thidiazuron; Vanilla odorata; Vanilla pompona; Vanillin
• ISSN: 1054-5476; 1475-2689
• DOI: 10.1007/s11627-023-10386-w

Direct and indirect shoot regeneration in Vanilla odorata and indirect for V. pompona are described for the first time. Both species produce fragrant, vanillin-containing seed pods and are closely related to the commercially cultivated V. planifolia . Stem nodal segments cultured on Murashige and Skoog medium containing 2.0 mg l −1 6-benzyl aminopurine (BAP) and 1.0 mg l −1 indole butyric acid (IBA) showed > 75% bud sprouting. Media containing 1.0 mg l −1 BAP or thidiazuron (TDZ) in combination with 0.5–1.0 mg l −1 IBA produced approximately eight shoots/explant. In callus induction experiments, we found that shoot tips from in vitro plantlets were more responsive than leaf and nodal explants with the highest callus induction (47% in V. odorata and 40% in V. pompona ) when cultured in the dark on medium containing 2.0 mg l −1 naphthalene acetic acid (NAA) and 1.0 mg l −1 BAP. Calluses transferred to basal medium containing 3.0 mg l −1 BAP plus 1.0 mg l −1 NAA and grown under a 16 h photoperiod produced an average of 4.1 and 3.5 shoots per callus segment for V. odorata and V. pompona , respectively, eight weeks after culture initiation. Histological analysis revealed the presence of calcium oxalate raphides, as confirmed by Raman microscopy, in callus cultured under both dark and light conditions. Over 90% of the plantlets derived from axillary buds exhibited spontaneous root formation on multiplication media; similar to that observed when shoots originating from callus were cultured on plant growth regulator-free medium. During the subsequent greenhouse acclimatization phase, which took place 12 weeks after the final subculture, these plantlets achieved a survival rate of 95%. The results of the present study will be helpful for further large-scale mass propagation efforts of V. odorata and V. pompona and other biotechnological applications such as protoplast isolation.