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Journal of Experimental Botany, Vol. 53, No. 371, pp. 1119-1129, May 2002
© 2002 Oxford University Press

Original Papers

Antirrhinum majus microspore maturation and transient transformation in vitro

Ioulia Barinova1, Manshuk Zhexembekova1, Ekaterina Barsova2, Sergei Lukyanov2, Erwin Heberle-Bors1 and Alisher Touraev1 3

1Vienna Biocenter, Institute of Microbiology and Genetics, Vienna University, Dr Bohrgasse 9, A-1030 Vienna, Austria
2Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117871 Moscow, Russia

The male gametophyte of higher plants represents an excellent system to study gene regulation, cell fate determination and cellular differentiation in plants because of its relative simplicity compared to the sporophyte and its accessibility for cytological and molecular analysis. Unicellular plant microspores are single haploid cells, which can be isolated in large amounts at a defined developmental stage. Microspores cultured in vitro in a rich medium develop into mature pollen grains, which are fertile upon pollination in vivo. It is reported here that isolated Antirrhinum majus microspores when cultured in an optimal medium develop to form mature, fertile pollen. Their development closely resembled that of pollen formed in vivo. Isolated microspores were bombarded with Aquorea victoria Green Fluorescent Protein (GFP), Discosoma Red Fluorescent Protein (dsRFP) and ß-glucuronidase (GUS) reporter genes under the control of various promoters and transient expression was observed throughout pollen development in vitro. Bombarded and not bombarded in vitro-matured pollen grains were able to germinate both in vitro and on receptive stigmas and to set seed. The protocol of maturation, transient transformation and germination of Antirrhinum majus pollen in vitro described here provides a valuable tool for basic and applied research.

Key words: Antirrhinum majus L., Green Fluorescent Protein (GFP), in vitro maturation, microspores, Red Fluorescent Protein (dsRFP), transformation.


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