Abstract
An ovule culture technique has been established for barley that allows the regeneration of plants from zygotes. An average of 1.3 plantlets per ovule could be regenerated from more than 60% of the cultured ovules and about 75% of the regenerated plantlets developed into normal, fertile plants. The same regeneration frequencies were obtained in intact ovules and in ovules where the two integuments had been removed from the micropylar region. Unfertilized ovules and ovules where the fertilized eggs had been destroyed by a microinjection needle did not give rise to embryo-like structures. Plants could be regenerated from the zygote at the same frequency at developmental stages from immediately after fertilization until the formation of bicellular embryos. This tissue culture system appeared to be largely independent of genotype since similar regeneration frequencies were obtained in two different barley cultivars, Igri and Alexis, that in anther and microspore culture behave differently. The same technique has also been applied successfully in the wheat cultivar Walter.
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References
Cameron-Mills V, Duffus CM (1977) The in vitro culture of immature barley embryos on different culture media. Ann Bot 41:1117–1127
Campenot MK, Zhang G, Cutler AJ, Cass DD (1992) Zea mays embryo sacs in culture. I. Plant regeneration from 1 day after pollination embryos. Am J Bot 79:1369–1373
Castillo AM, Cistué L (1993) Production of gynogenic haploids of Hordeum vulgare L. Plant Cell Rep 12:139–143
Comeau A, Nadeau P, Plourde A, Simard R, Maes O, Kelly S, Harper L, Lettre J, Landry B, St-Pierre CA (1992) Media for the in ovulo culture of proembryos of wheat and wheat-derived interspecific hybrids or haploids. Plant Scie 81:117–125
Engell K (1989) Embryology of barley: time course and analysis of controlled fertilization and early embryo formation based on serial sections. Nord J Bot 9:265–280
Holm PB, Knudsen S, Mouritzen P, Negri D, Olsen FL, Roué C (1994) Regeneration of fertile barley plants from mechanically isolated protoplasts of the fertilized egg cell. Plant Cell 6:531–543
Huang B-Q, Russell SD (1992) Female germ unit: organization, isolation and function. Int Rev Cytol 140:233–293
Kao KN, Michayluk MR (1975) Nutritional requirements for growth of Vicia hajastana cells and protoplasts at a very low population density in liquid media. Planta 126:105–110
Keijzer CJ, Reinders MC, Leferik-ten Kloster HB (1988) A micromanipulation method for artificial fertilization in Torenia. In: Cresti M, Gori P, Pacini E (eds) Sexual reproduction in higher plants. Springer, Berlin Heidelberg New York, pp 119–124
Kranz E, Lörz H (1993) In vitro fertilization with isolated single gametes results in zygotic embryogenesis and fertile maize plants. Plant Cell 5:739–746
Kranz E, Bautor J, Lörz H (1991) In vitro fertilization of single isolated gametes of maize mediated by electrofusion. Sex Plant Reprod 4:12–16
Maheshwari P (1950) An introduction to the embryology of angiosperms. McGraw-Hill, New York
Mathias RJ (1987) Plant microinjection techniques. In: Setlow JK (ed) Genetic engineering principles and methods. Vol 9, Plenum Press, New York, pp 199–277
Mòl R, Matthys-Rochon E, Dumas C (1993) In vitro culture of fertilized embryo sacs of maize: zygotes and two-celled embryos can develop into plants. Planta 189:213–217
Murashige T, Skoog F (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:472–497
Norstog K (1961) The growth and differentiation of cultured barley embryos. Am J Bot 48:876–884
Norstog K (1970) Induction of embryolike structures by kinetin in cultured barley embryos. Dev Biol 23:665–670
Norstog K (1973) New synthetic medium for the culture of premature barley embryos. In Vitro 8:307–308
Norstog K, Klein RM (1972) Development of cultured barley embryos. II. Precocious germination and dormancy. Can J Bot 50:1887–1894
Olsen FL (1987) Induction of microspore embryogenesis in cultured anthers of Hordeum vulgare. The effects of ammonium nitrate, glutamine and asparagine as nitrogen sources. Carlsberg Res Commun 52:393–404
Rangan TS (1982) Ovary ovule and nucellus culture In: Johri BM (ed) Experimental embryology of vascular plants. Springer, New York, pp 105–129
San Noeum L-H (1979) In vitro induction of gynogenesis in higher plants. Proceedings Conference on Broadening of the Genetic Base of Crops, Wageningen, 1978. Pudoc, Wageningen, pp 327–329
Spurr AR (1969) A low-viscosity epoxy resin embedding medium for electron microscopy. J Ultrastruct Res 26:31–43
Steinbiss H-H, Stabel P, Töpfer R, Hirtz RD, Schell J (1985) Transformation of plant cells by microinjection of DNA. In: Chapmann GP, Mantell SH, Daniels RW (eds) The experimental manipulation of ovule tissue. Longman, New York, pp 64–75
Töpfer R, Steinbiss H-H (1985) Plant regeneration from cultured fertilized barley ovules. Plant Sci 41:49–54
Umbeck PF, Norstog K (1979) Effects of abscisic acid and ammonium ion on morphogenesis of cultured barley embryos. Bull Torrey Bot Club 106:110–116
Wagner VT, Dumas C, Mogensen HL (1990) Quantitative threedimensional study on the position of the female gametophyte and its constitutive cells as a prerequisite for corn (Zea mays) transformation. Theor Appl Genet 79:72–76
Yang HY, Zhou C (1982) In vitro induction of haploid plants from unpollinated ovaries and ovules. Theor Appl Genet 63:97–104
Ziebur NC, Brink RA, Graff LH, Stahmann MA (1950) The effect of casein hydrolysate on the growth in vitro of immature Hordeum embryos. Am J Bot 37:144–148
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Holm, P.B., Knudsen, S., Mouritzen, P. et al. Regeneration of the barley zygote in ovule culture. Sexual Plant Reprod 8, 49–59 (1995). https://doi.org/10.1007/BF00228763
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DOI: https://doi.org/10.1007/BF00228763