Genetic transformation of Robinia pseudoacacia by Agrobacterium tumefaciens.
Transgenic Robinia pseudoacacia plants were obtained by Agrobacterium tumefaciens mediated gene transfer. Agrobacterium strain LBA4404 harbouring a binary vector that contained the chimeric neomycin phosphotransferase II (NPTII) and beta-glucuronidase (GUS) genes was co-cultivated with hypocotyl segments of in vitro raised seedlings of Robinia. Parameters important for high efficiency regeneration and transformation rates included type of explant, pre-conditioning of explants and appropriate length of co-cultivation period with Agrobacterium. A transformation frequency 16.67% was obtained by 48 hr of pre-conditioning followed by 48 hr of co-cultivation. Transformed tissue was selected by the ability to grow on kanamycin containing medium. Successful regeneration was followed after histochemical GUS assay for the detection of transgenic tissue. This transformation procedure has the potential to expand the range of genetic variation in Robinia.
Kanwar
Kamlesh
K
Department of Biotechnology, Dr. Y.S. Parmar University of Horticulture and Forestry, Nauni, Solan 173 230, India. kanwarkamlesh@hotmail.com
Bhardwaj
Anju
A
Agarwal
Sandhya
S
Sharma
D R
DR
D016960:Agrobacterium tumefaciens / Q000502:physiology*; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D019868:Kanamycin Kinase / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D031301:Robinia / Q000201:enzymology / Q000235:genetics* / Q000382:microbiology*; D036226:Seedlings / Q000201:enzymology; D014170:Transformation, Genetic; D019076:Transgenes
D011993:Recombinant Fusion Proteins; D019868:Kanamycin Kinase; D005966:Glucuronidase