"Transformation of F. oxysporum was done with pCAMBIA 1302 vector harboring sGFP gene under the control of CaMV35S promoter and hygromycin B phosphotransferase (HPT) as the selection markerene (generously provided by Dr. Richard A. Jefferson, CAMBIA, Canberra, Australia). Two different protocols were followed- a) F. oxysporum spores were germinated in carboxy methyl cellulose (CMC) broth (15 g carboxymethyl cellulose, 1 g yeast extract, 0.5 gMgSO4, 1 g NH4NO3 and 1 g KH2PO4, for 1 litre broth) at 100 rpm, at28ÂºC for 1 week. The culture was filtered through two layers of cheesecloth and spores were harvested at 4,500 rpm, 4ÂºC for 5 min. Usinghaemocytometer, spores were counted and diluted to 106 spores/ml ofspore suspension. Agrobacterium-mediated transformation (AMT) method and, b)glass-bead method.
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Last date updated on July, 2014