Begonia rex Tissue Culture Protocol

JungleBotanics Plant Science | Protocol review 28 September 2026

Begonia rex petiole tissue culture by direct shoot regeneration

This is a literature-based method for Begonia rex cv. DS-EYWA petiole tissue. It is not a validated protocol for JungleBotanics Ginny Galaxy, Snow Capped or B. cucullata. Those plants may respond differently, and variegated offspring need assessment after acclimatisation.

Primary evidence

A 2024 study tested thin petiole sections, BAP, NAA and silver nitrate. The authors reported their best direct-shoot treatment at 1.5 mg/L BAP, 0.5 mg/L NAA and 25 mg/L AgNO₃; lower BAP supported elongation, basal MS supported rooting and peat moss/perlite 1:1 supported acclimatisation. Read the study. A separate B. rex leaf study reported direct buds with 1 mg/L each of BA and NAA, but leaf and petiole protocols must not be combined as if they were one experiment: leaf study.

Materials and media

Use a clean identified mother plant, fresh petioles, sterile tools and vessels, MS salts and vitamins, 30 g/L sucrose and 5 g/L agar, pH 5.8. The study used 1% sodium hypochlorite solution plus 0.5% Tween 20 for disinfection, then sterile water rinses. Check whether your supplier expresses hypochlorite as NaOCl or available chlorine. Silver nitrate is light sensitive and is not in the current JungleBotanics reagent list; reproduce the reported optimum only after obtaining and correctly handling it. Without it, label the batch an adaptation.

Method

1. Choose and document. Select healthy, fully developed petioles; record cultivar, donor and sampling date. Avoid visibly damaged or infected tissue.
2. Surface clean. The researchers washed petioles under running water for 15 minutes, immersed them in 1% NaOCl and 0.5% Tween 20 for 10 minutes, and rinsed three times with sterile distilled water. They allowed 5, 10 and 15 minutes between rinses. Blot dry on sterile paper. Trial a smaller batch first if the tissue is unusually tender.
3. Prepare explants. In the cabinet, cut transverse petiole sections about 1 mm thick with a sterile blade. The published method also examined leaf sections; keep explant type separate in records. Distribute explants individually so contamination can be traced.
4. Establish. Place the sections on basal MS, with or without the selected AgNO₃ treatment. Incubate near 23 ± 2 °C and assess infection and living tissue after four weeks. The paper reports a light intensity around 37.5 µmol m⁻² s⁻¹; confirm its photoperiod wording in the original methods when reproducing its chamber schedule.
5. Induce and elongate. Transfer clean explants to the direct-shoot medium above and subculture onto fresh organogenesis medium every six weeks, as in the paper. Score buds arising directly from tissue, separating them from callus. Move distinct shoots to MS + 0.5 mg/L BAP without auxin for elongation.
6. Root and acclimatise. Transfer elongated shoots with three to four leaves to basal MS without regulators. Once rooted, wash off medium and transfer to sterile 1:1 peat moss/perlite. In the study, plants remained around 25 °C and 80% relative humidity under transparent covers for one month, then moved to a greenhouse around 25 °C and 60% relative humidity. Measure survival and avoid a sudden humidity drop.

How JungleBotanics would validate a new cultivar

Start with at least 10–20 explants per medium and retain a growth-regulator-free control. Record contamination, living explants, direct buds, rooted shoots and plants surviving acclimatisation; include photographs of the donor and each regenerated line. A cultivar's leaf pattern may not be faithfully reproduced by adventitious shoot regeneration. Keep shoot-tip or conventional cutting controls when possible and do not publish multiplication rates from another cultivar as your own.

Source: Davoudipahnekolayi and colleagues, Horticulturae 10, 986 (2024), DOI 10.3390/horticulturae10090986. Adaptation and recordkeeping: JungleBotanics Ltd.