A rapid shoot regeneration protocol from the cotyledons of hemp (Cannabis sativa L.)

Autor: Peng Ding-xiang, Guo Xinbo, Zhao Lining, Chen Jianhua, Cheng Chaohua, Tang Qing, Gao Chunsheng, Su Jianguang, Zang Gonggu
Rok vydání: 2016
Předmět:
Zdroj: Industrial Crops and Products. 83:61-65
ISSN: 0926-6690
DOI: 10.1016/j.indcrop.2015.12.035
Popis: Hemp (Cannabis sativa) is an annual multipurpose crop that is distributed worldwide. An efficient regeneration protocol is needed for hemp genetic transformation, micropropagation, and germplasm conservation. We describe here a rapid protocol for in vitro shoot regeneration that uses cotyledons as explants. We concluded that TDZ in MS medium is more efficient in inducing in vitro shoots from cotyledons than BA and ZT. The best result, 51.7% induction frequency and 3.0 shoots per shoot explant, was recorded in MS medium containing 0.4 mg l−1 TDZ and 0.2 mg l−1 NAA (T4N2). The in vitro shoots grew to the height of 1.5–2 cm in 3–4 weeks after culture initiation. At that time, approximately 80% of the shoots were rooted well in half-strength Ms medium combined with 0.5–2 mg l−1 IBA for 4–5 weeks before acclimation. It was observed that younger cotyledons (2–3 days after planting (DAP) were more useful as explants than older ones (5–6 DAP) because they had a significantly higher regeneration frequency. We cultured 3 DAP cotyledons of eight cultivars in T4N2 medium to test the efficiency. The regeneration frequency varied from 35.7% to 54.8%, which indicated that this regeneration protocol was effective, although partly genotype dependent. We regard this protocol as an alternative method for micropropagation and germplasm conservation, and the in vitro plantlets may be suitable to set up a transformation system.
Databáze: OpenAIRE
Externí odkaz: