Journal of Advances in Biology & Biotechnology

Background: Intergeneric hybridization is an important approach for broadening genetic variation and recovering desirable horticultural traits in orchids. However, reproductive barriers and the limited natural germination of orchid seeds can restrict successful progeny recovery.

Aims: To assess the cross-compatibility of the intergeneric hybrid Papilionanda John Clubb with Vanda tessellata through hand-pollination and to document the developmental timeline of the resulting progeny from pollination to plantlet stage.

Study Design: Controlled hand-pollination followed by asymbiotic in vitro seed culture and sequential plantlet regeneration.

Place and Duration of Study: Jawaharlal Nehru Tropical Botanic Garden and Research Institute (JNTBGRI), Thiruvananthapuram, Kerala, India, between March 2025 and May 2026.

Methodology: Papilionanda John Clubb (Papilionanthe Cooperi × V. tessellata; pod parent) was hand-pollinated with V. tessellata (pollen parent) on 4 March 2025. Immature capsules were harvested 105 days after pollination and their dimensions were recorded; seeds were measured microscopically (ImageJ) and inoculated into half-strength Murashige and Skoog (MS) liquid medium with 0.5 mg L⁻¹ α-naphthaleneacetic acid (NAA) and 0.5 mg L⁻¹ 6-benzylaminopurine (BAP). Protocorms were then subcultured on full-strength MS solid medium with NAA and BAP at 0.5 + 0.5, 1.0 + 0.5, and 2.0 + 1.0 mg L⁻¹.

Results: All five pollinated flowers of Papilionanda John Clubb set capsules, whereas the reciprocal cross (V. tessellata as pod parent) did not. Capsules measured 7.0 ± 0.71 cm × 1.0 ± 0.14 cm (n = 5), and seeds measured 406.56 ± 93.20 μm × 23.17 ± 5.87 μm. Embryo swelling and greening occurred at 40 days after inoculation (DAI), chlorophyllous protocorms with leaf primordia by 86 DAI, and vigorous multi-shoot plantlets (shoots 2.0–3.0 cm; 7–9 leaves; 2–3 roots) by 329 DAI on MS with 2.0 mg L⁻¹ NAA + 1.0 mg L⁻¹ BAP.

Conclusion: Papilionanda John Clubb was cross-compatible with V. tessellata as the pod parent, and progeny were recovered from immature capsules through asymbiotic culture. This is the first documentation of the complete pollination-to-plantlet timeline for intergeneric hybridization between Papilionanda John Clubb and Vanda tessellata.