Cowpea is a
major source of dietary protein in most African diets. It suffers
from a wide range of production constraints including viral diseases.
Cowpea aphid-borne mosaic potyvirus (CABMV) is the most important
viral pathogen of cowpea in major cowpea growing regions of the
world. While breeding for resistance is an ongoing activity is
various laboratories, the availability of resistant germplasm is
limited. Our strategy has been to use pathogen derived resistance to
introduce viral genes into cowpea. However, this effort has met with
limited success due to the lack of an efficient, reliable and
reproducible transformation and regeneration system. Our present goal
is to attempt the transformation of cowpea seedlings using
electrophoresis. The coat protein gene of CABMV was cloned into the
binary vector pBI 121 in various forms designed to confer coat
protein-mediated resistance and sense- and antisense- RNA mediated
resistances. The DNA constructs were electrophoresed into the apical
meristem of developing cowpea seedlings under various conditions
including different voltage and current settings as well as
pretreatment of seedlings with acid or plant growth regulators.
Preliminary screening was done using GUS assays, and the plants
allowed to set seed. The seed was harvested and will be replanted
in-vitro on kanamycin. Further analysis will be done using the
polymerase chain reaction (PCR) and Southern blot hybridization to
determine the most effective regimen.