Somatic Embryo Germination and Plant Development from Immature Zygotic Embryos in Cotton
Syed Sarfraz Hussain,
Successful cotton plant transformation depends on regeneration of plants from transformed cells. Recalcitrance of cotton to tissue culture has not only slowed the development of transgenic cotton but also narrowed its genetic base. Keeping this in view, an efficient in vitro plant regeneration system characterized by bulk, rapid and continuous production of somatic embryos using immature zygotic embryos, taken at various stages of development from ovules as explant has been developed in cotton. One of the drawbacks of using this is the requirement of large number of high quality immature zygotic embryos. To address this problem, we have developed a procedure that generates highly homogeneous populations of embryogenic calli by selectively propagating a small number of regeneration proficient calli derived from immature zygotic embryos. Stages showing the early cotyledon development cultured on modified Murashige and Skoog media proliferated intensely. Rapid callogenesis was observed from these immature zygotic embryos. To induce germination and plantlet growth, embryoids were placed on sterile processed cotton, saturated with Stewart and Hsu media. Upon development of roots and leaves, plantlets of 3-4 cms were potted in 1:1:1 mixture of sand, silt and peat moss under high humidity and further hardened under green house conditions. Using this system, we have been able to regenerate approximately 70% of healthy plantlets.
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