Summary
The effects of different factors were investigated in the process of isolated microspore culture of Zea mays L., Using donor plants grown in standard conditions and an efficient isolation technology, homogeneous populations of viable microspores at specific developmental stages were obtained and tested in culture. The cytological evolution of the microspores during the first week of culture was monitored using a DNA-specific fluorochrome. It was found that developmental stages of microspores, number of days of pretreatment at 7°C of the tassel, and culture density greatly influenced the number of microspore-derived embryos. Optimal conditions required for embryo and plant production are described.
Similar content being viewed by others
Abbreviations
- ISO:
-
isolation medium
- MS:
-
Murashige and Skoog
- Na2EDTA:
-
ethylene diaminetetraacetic aciddisodium salt
- AS:
-
androgenic structures
- CFA:
-
correspondence factorial analysis
- FDA:
-
fluorescein diacetate
- IM:
-
induced microspores
References
Alexander MP (1969) Differential staining of aborted and non -aborted pollen. Stain Technol. 44: 117–122
Barloy D, Denis L, Beckert M (1989) Comparison of the aptitude for anther culture in some androgenetic doubled haploid maize lines. Maydica 34: 303–308
Bedinger PA, Edgerton MD (1990) Developmental staging of maize microspores reveals a transition in developing microspore proteins. Plant Physiol. 92: 474–479
Coumans MP, Sohota S, Swanson EB (1989) Plant development from isolated microspores of Zea mays L. Plant Cell Rep. 7: 618–621
Dieu P, Beckert M (1986) Further studies of androgenetic embryo production and plant regeneration from in vitro cultured anthers in maize (Zea mays L.) Maydica 31: 245–259
Genovesi AD, Collins GB (1982) In vitro production of haploid plants of corn via anther culture. Crop Sci. 22: 1137–1144
Heslop-Harrison J, Heslop Harrison Y (1970) Evaluation of pollen viability by enzymatically induced fluorescence; intracellular hydrolysis of fluorescein diacetate. Stain Technol. 45: 115–120
Huang B, Bird S, Kemble R, Simmonds D, Keller W, Miki B (1990) Effects of culture density, conditioned medium and feeder cultures on microspore embryogenesis in Brassica napus L. cv. Topas. Plant Cell Rep. 8: 594–597
Ku MK, Cheng WC, Juo LC, Kuan YL, An HP, Huang CH (1978) In: Proc. Symp. Plant Tissue Culture. Science Press, Peking, pp 35–42
Kyo M, Harada H (1986) Control of the developmental pathway of tobacco pollen in vitro. Planta 168: 427–432
Mandaron P, Niogret MF, Mache R, Moneger F (1990) In vitro protein synthesis in isolated microspores of Zea mays at several stages of development. Theor. Appl. Genet. 80: 134–138
Murashige T, Skoog F (1962) A revised medium for a rapid growth and bioassay with tobacco tissue culture. Physiol. Plant. 15: 473–479
Pechan PM, Keller WA (1988) Identification of potentially embryogenic microspores in Brassica napus. Physiol. Plant. 74: 377–384
Pescitelli SM, Mitchell JC, Jones AM, Pareddy DR, Petolino JF (1989) High frequency androgenesis from isolated microspores of maize. Plant Cell Rep. 7: 673–676
Pescitelli SM, Johnson CD, Petolino JF (1990) Isolated microspore culture of maize: effects of isolation technique, reduced temperature and sucrose level. Plant Cell Rep. 8: 628–631
Petolino JF, Jones AM (1986) Anther culture of elite genotypes of maize. Crop Sci. 26: 1072–1074
Vain P, Flament P, Sondan P (1989) Role of ethylene in embryogenic callus initiation and regeneration in Zea mays L. Plant Physiol. 135: 537–540
Vergne P, Delvallée I, Dumas C (1987) Rapid assessment of microspore and pollen development stage in wheat and maize using DAPI and membrane permeabilization. Stain Technol. 62: 299–304
Vergne P, Gaillard A, Beckert M (1991) Isolation of viable microspores and immature pollen grains from cereal inflorescences. In: Negrutiu I. (ed) BioMethods. Birkhäuser, Basel Boston (in press)
Wei ZM, Kyo M, Harada H (1986) Callus formation and plant regeneration through direct culture of isolated pollen of Hordeum vulgare. Theor. Appl. Genet. 72: 252–255
Author information
Authors and Affiliations
Additional information
Communicated by H. Lörz
Rights and permissions
About this article
Cite this article
Gaillard, A., Vergne, P. & Beckert, M. Optimization of maize microspore isolation and culture conditions for reliable plant regeneration. Plant Cell Reports 10, 55–58 (1991). https://doi.org/10.1007/BF00236456
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00236456