Agrobacterium tumefaciens-mediated transformation of chickpea (Cicer arietinum L.): gene integration, expression and inheritance.
A reproducible method of Agrobacterium-mediated transformation was developed for Cicer arietinum (chickpea). Initial explants consisted of longitudinal slices from embryonic axes of imbibed, mature seed. The plasmid contained a bi-functional fusion gene conferring both beta-glucuronidase and neomycin phosphotransferase activities, under the control of a 35S35SAMV promoter. Using a series of tissue culture media for co-cultivation, shoot initiation and rooting, we recovered transgenic plants from approximately 1.3% of the sliced embryo axes. The addition of a shoot elongation medium to the protocol improved the success rate to 3.1% but increased the time in tissue culture. Inheritance of the gus gene was followed through four generations, both through expression and Southern hybridization assays, and showed the expected Mendelian inheritance pattern.
Polowick
P L
PL
Plant Biotechnology Institute, National Research Council of Canada, 110 Gymnasium Pl., Saskatoon, SK, Canada, S7N 0W9. patricia.polowick@nrc-cnrc.gc.ca
Baliski
D S
DS
Mahon
J D
JD
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D029863:Cicer / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D018744:DNA, Plant / Q000235:genetics; D018506:Gene Expression Regulation, Plant / Q000235:genetics*; D050939:Gene Fusion; D005822:Genetic Vectors / Q000235:genetics*; D018745:Genome, Plant; D005966:Glucuronidase / Q000235:genetics; D040582:Inheritance Patterns / Q000235:genetics; D018520:Plant Shoots / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012639:Seeds / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D014170:Transformation, Genetic / Q000235:genetics*
D018744:DNA, Plant; D011993:Recombinant Fusion Proteins; D005966:Glucuronidase