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Development
of an electrotransformation protocol for cowpea (Vigna
unguiculata)
I. Sithole-Niang,
R. Mundembe & R. Allison
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.
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