Hemp (Cannabis sativa L.) is widely used in industrial production due to its pharmacological properties, particularly cannabidiol (CBD). For commercial production, it is essential to have a large-scale propagation that is consistently disease-free and rapid. Thus, this study aims to establish a protocol for exponentially increasing the amount of high CBD hemp variety CA-UP1 by nodal segment via tissue culture method. Initially, with 10-minute surface sterilization with 0.1 % HgCl2, the microbial contamination rate was reduced to its lowest level (approximately 16.67 %) within 7 days.
The nodal segments exhibited optimal response to MS media supplemented with 0.5 mg/L TDZ, resulting in a 100 % shoot multiplication rate. The average number of shoots per explant was 4.7 ± 1.57, and the average shoot height was 2.30 ± 0.42 cm at 4 weeks. Following the induction of elongation in MS medium supplemented with charcoal, the seedlings were acclimatized and dipped in a 1,000 ppm IBA solution mixed with talcum powder. The cultivation resulted in a 100 % rooting rate, with an average of 5.6 ± 1.94 roots/explant, an average root length of 5.78 ± 1.96 cm, and an average explant height of 4.32 ± 1.52 cm.
The tissue culture protocol can produce over 190 times the number of hemp plants compared with the stem-cutting technique. The researchers anticipate that this micropropagation protocol will be useful as a rapid and economical alternative for the industrial production of effective C. sativa L. HIGHLIGHTS A 10-minute surface cleaning protocol with a minimal microbiological contamination rate of 16.67 % in the nodal segment. Thidiazuron (TDZ) can stimulate the most significant increase in shoot growth in Cannabis sativa L. variety CA-UP1 when grown on a semi-solid medium.
Dispensing talcum powder combined with IBA results in a 70 - 100 % survival percentage by effectively preventing roots during the acclimation stage. The following procedures can be implemented micropropagation of hemp for the production of a large quantity of high-quality, disease-free hemp plants with approximately 190 times more than stem cutting method.
Wongchaya, Panuphon & Bansra, Tatiya & Mahadtanapuk, Supuk. (2024). Rapid Micro-Propagation of Disease-Free Hemp (Cannabis sativa L.) to Improve Industrial Production. Trends in Sciences. 22. 9057. DOI: 10.48048/tis.2025.9057.
Source: Research Gate