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- Title
Histological analysis of direct somatic embryogenesis in Arabidopsis thaliana (L.) Heynh.
- Authors
Kurczyńska, Ewa U.; Gaj, Małgorzata D.; Ujczak, Agnieszka; Mazur, Ewa
- Abstract
In Arabidopsis the in vitro culture of immature zygotic embryos (IZEs) at a late stage of development, on the solid medium containing synthetic auxin, leads to formation of somatic embryos via direct somatic embryogenesis (DSE). The presented results provide evidence that in IZE cells competent for DSE are located in the protodermis and subprotodermis of the adaxial side of cotyledons and somatic embryos displayed a single- or multicellular origin. Transgenic Arabidopsis lines expressing the GUS reporter gene, driven by the DR5 and LEC2 promoters, were used to analyse the distribution of auxin to mark embryogenic cells in cultured explants and develop somatic embryos. The analysis showed that at the start of the culture auxin was accumulated in all explant tissues, but from the fourth day onwards its location shifted to the protodermis and subprotodermis of the explant cotyledons. In globular somatic embryos auxin was detected in all cells, with a higher concentration in the protodermis, and in the heart stage its activity was mainly displayed in the shoot, root pole and cotyledon primordia. The embryogenic nature of dividing protodermal and subprotodermal cells accumulating auxin was confirmed by high expression of promoter activity of LEC2 in these cells. Analysis of symplasmic tracer (CFDA) distribution indicated symplasmic isolation between tissues engaged in DSE and other parts of an explant. Symplasmic isolation of somatic embryos from the explant was also detected.
- Subjects
SOMATIC embryogenesis; PLANT cell culture; PLANT cells &; tissues; ARABIDOPSIS thaliana; AUXIN; PLANT hormones; SYNTHETIC seeds; REPORTER genes; PLANT genetics
- Publication
Planta: An International Journal of Plant Biology, 2007, Vol 226, Issue 3, p619
- ISSN
0032-0935
- Publication type
Article
- DOI
10.1007/s00425-007-0510-6