Culture of Isolated Pollen of Wheat (Triticum aestivum L.)
Pollen culture provides a system with haploid single cells and thus is a promising haplopahase tool for basic and applied biotechnological breeding programs. Anther culture (Nitzsche and Wenzel 1977; de Buyser and Henry 1980, 1986; Maheshwari et al. 1980;
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1 Introduction Pollen culture provides a system with haploid single cells and thus is a promising haplopahase tool for basic and applied biotechnological breeding programs. Anther culture (Nitzsche and Wenzel 1977; de Buyser and Henry 1980, 1986; Maheshwari et al. 1980; Bajaj 1983; Schaeffer et al. 1984; Wenzel and ForoughiWehr 1984; Bajaj and Gosal 1986; Dunwelll986; Han 1986; Keller et al. 1987) is one of the methods of haploid induction most commonly used now for crop improvement. Culture of whole spikes (Wilson 1977; Brettel et al. 1980, Ozias-Akins and Vasil 1982; Datta 1987) may simplify this technique. The third way is the culture of isolated microspores. However, in most reports the microspores are cultured within the anthers. Only a rather limited number of reports have been published on the successful use of isolated microspore culture in cereals (Kohler and Wenzel 1985; Wei et al. 1986; Datta and Wenzel 1987, 1988; Datta and Potrykus 1988). While regeneration of plants from single cells of cereals is still a problem, plant regeneration from in vitro cultures originating from multicellular explants has been achieved from all the major cereal crops (Vasil 1983; Lorz et al. 1988). Plant regeneration from microspores of wheat follows the two general pathways of in vitro culture, viz organogenesis and embryogenesis. The latter one is preferred for its unicellular origin and production of non-chimeric plants.
2 Advantages of Isolated Pollen over Anther Culture The microspore culture technique is the most promising and interesting method as it has the following advantages over anther culture. 1. 2. 3. 4.
Undesired selection due to limited space and nutrition within the anther are overcome. All possible contamination with somatic, diploid anther tissue can be eliminated. Such cultures offer a chance to observe more precisely the development of the micros pores. Microspores can be cultivated as single-cell organisms and therefore microbiological techniques can be used.
1 Swiss Federal Institute of Technology, ETH, Institute of Plant Sciences, 8092 Ziirich, Switzerland 'BBA, Institute for Resistance Genetics, W-8059 Griinbach-Bockhorn, FRG
Biotechnology in Agriculture and Forestry, Vol. 13 Wheat (ed. by Y.P.S. Bajaj) © Springer-Verlag Berlin Heidelberg 1990
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5. 6. 7.
S.K. Datta et al.
As microspore culture is a single cell system, it makes selection amongst single cells possible and furthermore offers new prospects for genetic manipulation (e.g., mutagenesis, transformation). As microspores may develop directly into an embryo, they provide an excellent tool for understanding the physiology and biochemistry of androgenesis. Direct gene transfer by microinjection into isolated multicellular pollen embryoids (N euhaus et al. 1987) offers the possibility oftransgenic plant formation of all cereals including wheat by using culture of isolated pollen having high regeneration efficiency (Potrykus et al. 1985; Potrykus 1988).
3 Brief Review of the Work on Culture of Isolated Pollen Since Nitsch (1974) made he
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