Rapid Micro-Propagation of Disease-Free Hemp (Cannabis sativa L.) to Improve Industrial Production

Abstract

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 quantity 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 root 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. GRAPHICAL ABSTRACT