Production of embryogenic tissues and regeneration of transgenic plants in cassava (Manihot esculenta Crantz)

Abstract

Disorganised embryogenic tissues have been utilised as target tissues for transgene insertion and transgenic plant regeneration in cassava (Manihot esculenta). The production of friable embryogenic callus in fourteen geographically diverse cassava cultivars, from which eleven were established as embryogenic suspension cultures, is reported. Embryogenic tissues were similar in nature in all cultivars tested although there was variation in the time required to generate friable callus and the growth rates of suspension cultures. Regeneration of plants has been achieved from eight cultivars but varied significantly in efficiency, with cv. TMS 60444and Line 2 from Zimbabwe being the most responsive. Tissues from the remaining eight cultivars became arrested at globular and torpedo stages of regeneration indicating that they most likely process an inherent ability to produce plants but require further research to allow this to be realised. Significant numbers of transgenic plants containing transgenes for putative resistance to important viral diseases of cassava in addition to visual marker genes have been regenerated. Transgenic plants from three the cultivars TMS 60444, Bonoua Rouge and M.Col 1505 were recovered after particle bombardment of embryogenic suspension cultures. Correlation's have been made between abnormal leaf morphology and plant vigour with the use of embryogenic suspension cultures for transgene insertion. As an result friable embryogenic callus is now being successfully utilsed as the target tissue for genetic transformation and plant regeneration at ILTAB.