In-ovule Embryo Culture: A Novel Method of Cotton Transformation
Syed Sarfraz Hussain,
The study was undertaken to develop a culture technique permitting to grow cotton embryos from fertilization to germination with minimum of manipulation and maximum yield of viable plants. During the study, conditions were optimized for transfer of zygotic ovules after post anthesis and selection of media for the growth of ovules. Forty-eight hours post anthesis was selected as the most appropriate time since >50% of ovules were found alive, floating and developing. Also, ST medium was found suitable as it supported the growth very well and ovules continued to grow till the end of 10 weeks. This in-ovule technique has several advantages over culture of isolated embryos. The aseptic removal of 48 h old ovules from the ovaries was rapid and efficient and minimum damage was observed in this technique. No change of medium was needed during the entire culture period. This technique was used for the first time as a means of cotton transformation using Sonication assisted Agrobacterium mediated transformation (SAAT) method while a marker gene was used to transform cotton ovules.
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