Abstract
Premise of the Study Datura stramonium is a pharmacologically and evolutionarily important plant species in the family Solanaceae. Stable transformation methodology of this species would be advantageous for future genetic studies.MethodsIn vitro plant regeneration and Agrobacterium tumefaciens–mediated transformation techniques were developed for D. stramonium based on methods reported for tomato. A binary vector containing pAtUBQ10::erGFP was used for transformation.ResultsWe recovered primary transformants harboring the green fluorescent protein (GFP) transgene that resulted in expression of fluorescence in all tissues analyzed. Transformants were allowed to self‐pollinate, and two of five progeny contained the GFP transgene and displayed fluorescence identical to the primary transformants.DiscussionWe have demonstrated the first stable transformation in the genus Datura. This is a key first step to study the genetic basis of traits in this evolutionarily interesting species.
Highlights
PREMISE OF THE STUDY: Datura stramonium is a pharmacologically and evolutionarily important plant species in the family Solanaceae
Datura L. is a genus of pharmacologically important plants in the family Solanaceae
Studies of polyploidy were undertaken in Datura, and the first production of a haploid plant was reported in Datura stramonium L. (Blakeslee et al, 1922)
Summary
GFP fluorescence and both individuals showed consistent and uniform fluorescence across the leaf epidermis; fluorescence was greater in the vasculature than in the epidermal tissue (Fig. 3). Stamens and pistils showed very strong fluorescence, as did nectaries and pollen. The GFP transgene was not detected in three T1 progeny (T1-1, T1-2, and T1-3), and these failed to show fluorescence above background levels. The two T1 plants that did show PCR amplification of the GFP transgene showed fluorescence similar to the primary transformants. As observed in the primary transformants, GFP fluorescence was very strong in the stamens, pistil, pollen, and nectaries, and moderate fluorescence was consistently observed in the leaf tissue. Wild-type plants did not show fluorescence in leaf, stem, and most reproductive tissues. Background fluorescence was elevated in anthers and stigmatic tissue, identical to that seen in the anthers and stigmas of non-transgenic T1 plants
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