Temporary immersion bioreactors for clonal production of Moringa oleifera tissues

Abstract

Moringa oleifera has excellent value as a medicinal plant due to the various secondary metabolites produced in all its tissues. These secondary metabolites are biologically active and give moringa the properties that make it so useful against many ailments. The presence of such a variety of biologically active compounds gives moringa the potential for discoveries and development of new drugs. One method to produce these metabolites for extraction is producing moringa tissues in tissue culture. Tissue culture provides a controlled environment, and along with clonal reproduction, allows reduced variance between different batches. In this study, clonal proliferation of moringa by using leaf material was investigated. Conventional solidified medium methods were compared to the production in temporary immersion bioreactor systems. Moringa seeds were germinated in vitro, and the seedling leaflets were used as explant material. The first method tested which involved planting leaf material directly onto media or into bioreactors for shoot production was unsuccessful due to high mortality rates of leaf material in the bioreactors. Another method proved more effective and involved first planting all leaf material onto a solidified medium to initiate some callus production before splitting these up between solidified media and bioreactors for shooting. The initiation media consisted of half-strength Murashige and Skoog (MS) basal salts and 0.5 ppm 1-naphthaleneacetic acid (NAA). The shooting media consisted of full-strength MS basal salts and different treatments of kinetin and 6-benzylaminopurine (BA). It was found that there was a difference in the production of differentiated tissue between solidified media and bioreactors, whereas the bioreactors produced more substantial amounts of tissue (wet mass) compared to the solidified media.