Optimisation of in Vitro Techniques for Cassava Brown Streak Virus Elimination from Infected Cassava Clones

Cassava brown streak disease (CBSD), caused by Cassava brown streak virus (CBSV), is an economically important disease of cassava (Manihot esculenta Crantz) in East Africa. The objective of this study was to optimise in vitro techniques for CBSV elimination from infected Ugandan cassava cultivars. Using semi-solid halfstrength Murashige and Skoog (MS) basal medium, hormone concentration and heat treatment regimes were optimised for micropropagation of farmer’s preferred cassava cultivars and CBSV elimination. Single nodes from young cassava stems were cultured for four weeks on 1⁄2MS medium supplemented with 6-benzyl amino purine (BAP) and 2,4-dichlorophenoxy acetic acid (2,4-D). The BAP and 2,4-D concentration (mg l-1) combinations used were 0.0 and 0.0, 0.5 and 0.1, 1.0 and 0.2, 1.5 and 0.3, and 2.0 and 0.4, respectively. The optimum medium was used for in vitro thermotherapy using four temperature regimes, namely 30-34, 34-38, 36-40 and 38-42 oC for 8 hours darkness and 16 hours light, respectively, for four weeks. The best plantlet growth in terms of height was observed on MS medium supplemented with 0.5 mg l-1 BAP and 0.1 mg l-1 2,4-D. Highest CBSV elimination efficiency of 40%, with 49% plantlet survival was observed at 36 oC for 8 hours darkness and 40 oC for 16 hours light. These results indicate that in vitro techniques can greatly enhance CBSV elimination and, thus, provide a means of CBSD management through dissemination and conservation of popular but CBSD susceptible cultivars.