Abstract
Long-term callus cultures of Nicotiana tabacum cv. Wisconsin 38 were utilized to examine the auxin and cytokinin requirements, as well as the light requirement for somatic embryogenesis. Initial observations indicated that somatic embryos were formed in callus cultured on Murashige and Skoog (MS) medium supplemented with indoleacetic acid (IAA) (11.4 μM) and ienzylaminopurine (BA) (22.1 μM) in continuous light of ca. 100 μE/m2/sec at 25°C. After 4 weeks of culture, these embryos were transferred to either MS without hormones, or to MS with IAA (11.4 μM) and BA (0.04 μM), the latter being the maintenance medium for the normally dark-grown callus. The somatic embryos were then placed in culture under a 16:8 light:dark cycle at 25°C and regenerated into whole plantlets. Several plantlets grew to maturity, flowered, and set seed. The genetic stability of this seed has not yet been tested. This basic experiment has been successfully repeated 4 times.
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