Genetic transformation of cowpea (Vigna unguiculata L.) and stable transmission of the transgenes to progeny.
Cowpeas are nutritious grains that provide the main source of protein, highly digestible energy and vitamins to some of the world's poorest people. The demand for cowpeas is high but yields remain critically low, largely because of insect pests. Cowpea germplasm contains little or no resistance to major insect pests and a gene technology approach to adding insect protection traits is now a high priority. We have adapted features of several legume and other transformation systems and reproducibly obtained transgenic cowpeas that obey Mendelian rules in transmitting the transgene to their progeny. Critical parameters in this transformation system include the choice of cotyledonary nodes from developing or mature seeds as explants and a tissue culture medium devoid of auxins in the early stages, but including the cytokinin BAP at low levels during shoot initiation and elongation. Addition of thiol-compounds during infection and co-culture with Agrobacterium and the choice of the bar gene for selection with phosphinothricin were also important. Transgenic cowpeas that transmit the transgenes to their progeny can be recovered at a rate of one fertile plant per thousand explants. These results pave the way for the introduction of new traits into cowpea and the first genes to be trialled will include those with potential to protect against insect pests.
Popelka
J Carlos
JC
CSIRO Plant Industry, GPO Box 1600, Canberra, ACT 2601, Australia.
Gollasch
Stephanie
S
Moore
Andy
A
Molvig
Lisa
L
Higgins
Thomas J V
TJ
D003470:Culture Media; D007887:Fabaceae / Q000235:genetics* / Q000502:physiology*; D018506:Gene Expression Regulation, Plant; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D012098:Reproduction / Q000502:physiology; D012231:Rhizobium; D012641:Selection, Genetic; D046509:Tissue Culture Techniques; D014170:Transformation, Genetic
D003470:Culture Media; D010940:Plant Proteins