Summary
Specific stress treatments applied to isolated tobacco (Nicotiana tabacum L.) microspores efficiently induced haploid embryo formation in vitro. A heat shock at 33 or 37°C in the presence of sugar, as well as sucrose-starvation at 25°C, resulted in the formation of embryogenic microspores. A combination of both treatments had an additive effect. Under optimal induction conditions all viable microspores in the culture were embryogenic and developed subsequently into pollen embryos by culture at 25°C in a sugar-containing medium, with induction frequencies of more than 70% with respect to the initial microspore population. A high fraction of the early pollen embryos continued their development in vitro, giving rise to haploid plants. In contrast to other available systems for microspore/pollen embryogenesis, the new protocol allows the production of homogeneous populations of embryogenic microspores and early globular embryos in large-scale cultures, without any purification step, and is therefore well suited for biochemical and molecular work.
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Abbreviations
- EDTA:
-
ethylenediaminetetraacetate
- DAPI:
-
4,6-diamidino-2-phenylindole
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Communicated by I. Potrykus
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Touraev, A., Ilham, A., Vicente, O. et al. Stress-induced microspore embryogenesis in tobacco: an optimized system for molecular studies. Plant Cell Reports 15, 561–565 (1996). https://doi.org/10.1007/BF00232453
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DOI: https://doi.org/10.1007/BF00232453