Abstract
Generating transgenic cotton is a time-consuming process which mainly relies on somatic embryogenesis. Moreover recovery of transgenic cotton plants via Agrobacterium mediated transformation is not an easy task. Therefore, the present study reports a simple and effective in planta transformation system for cotton cv.SVPR-2 by utilizing wounding, sonication and vacuum infiltration techniques altogether. In this method, Agrobacterium tumefaciens strain GV3101 carrying pCAMBIA1304–hyg binary vector was utilized for the experiment. The putatively transformed cotton plants were identified by using 15 mg/L hygromycin. The transgenic plants were assessed through β-glucuronidase (GUS) assay, Green fluorescent protein (GFP) assay and polymerase chain reaction (PCR) analysis. After optimizing several factors, maximum of 28.66% transformation frequency was achieved with 18 h pre-culture, sonication for 60s and vacuum infiltration for 90s and 3 days of co-culture on Murashige and Skoog (MS) medium supplemented with 150 μM acetosyringone. The in planta transformation system assisted by sonication and vacuum infiltration techniques can be used for transient expression, genome editing and developing stable cotton transformants with desirable genetic traits.
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