Abstract
Approximately 2,000 embryogenic uninuclear microspores of rapeseed (Brassica napus) cv. Topas were intranuclearly injected with a chimaeric β-glucuronidase (Escherichia coli Uid A) gene. Stable integration had not occurred among 55 plants that were regenerated. Coinjection of the dye Lucifer Yellow and detection of injected DNA by the polymerase chain reaction revealed high frequencies of transfer. However, the amount of DNA injected was less than 20 copies, which may have been insufficient for stable transformation of microspores.
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Abbreviations
- PCR:
-
polymerase chain reaction
- GUS:
-
β-glucuronidase
References
Bolik M & Koop HU (1991) Identification of embryogenic microspores of barley (Hordeum vulgare L.) by individual selection and culture and their potential for transformation by microin-jection. Protoplasma 162: 61–68
Crossway A, Oakes JV, Irvine JM, Ward B, Knauf VC & Shewmaker CK (1986) Integration of foreign DNA following microinjection of tobacco mesophyll protoplasts. Mol. Gen. Genet. 202: 179–185
Gaillard A, Matthys-Rochon E & Dumas C (1992) Selection of microspore derived embryogenic structures in maize related to transformation potential by microinjection. Bot. Acta 105: 313–318
Gilliland G, Perrin S & Franklin Bunn H (1990) Competitive PCR for quantitation of mRNA. In: Innis MA, Gelfand DH, Sninsky JJ & White TJ (Eds) PCR Protocols: A Guide to Methods and Applications (pp 60–69) Academic Press, Toronto
Huang B & Keller WA (1989) Microspore culture technology. J. Tissue Cult. Meth. 12: 171–178
Jefferson RA, Kavanaugh TA & Bevan MW (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in high plants. EMBO J. 6: 3901–3907
Miki BL, Huang B, Bird S, Kemble R, Simmonds D & Keller W (1989) A procedure for the microinjection of plant cells and protoplasts. J. Tissue Cult. Meth. 12: 139–144
Neuhaus G, Spangenberg G, Mittelsten Scheid O & Schwieger H-G (1987) Transgenic rapeseed plants obtained by the microinjection of DNA into microspore-derived embryos. Theor. Appl. Genet. 75: 30–36
Nomura K & Komamine A (1986) Embryogenesis from microin-jected single cells in a carrot cell suspension culture. Plant Sci. 44: 53–58
Reich TJ, Iyer VN & Miki BL (1986a) Efficient transformation of alfalfa protoplasts by the intranuclear microinjection of Ti plasmids. Bio/Technol 4: 1001–1004
Reich T, Iyer VN, Scobie B & Miki BL (1986b) A detailed procedure for the intranuclear microinjection of plant protoplasts. Can J. Bot. 64: 1255–1258
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Jones-Villeneuve, E., Huang, B., Prudhomme, I. et al. Assessment of microinjection for introducing DNA into uninuclear microspores of rapeseed. Plant Cell Tiss Organ Cult 40, 97–100 (1995). https://doi.org/10.1007/BF00041124
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DOI: https://doi.org/10.1007/BF00041124