Chemistry: molecular biology and microbiology – Treatment of micro-organisms or enzymes with electrical or... – Modification of viruses
Patent
1995-03-15
1997-10-21
Benzion, Gary
Chemistry: molecular biology and microbiology
Treatment of micro-organisms or enzymes with electrical or...
Modification of viruses
4351721, 800205, C12N 1500, C12N 1582
Patent
active
056795580
DESCRIPTION:
BRIEF SUMMARY
This invention relates to a rapid and efficient method for transforming walled cells of monocotyledonous plants, especially gramineous plants, particularly rice, wheat, corn, barley and other major cereals.
This invention also relates to novel transgenic monocotyledonous plants, particularly gramineous plants, obtainable by this method.
BACKGROUND OF THE INVENTION
In recent years, there has been a tremendous expansion of the capabilities for the genetic engineering of plants. Transgenic plants of many dicotyledonous plant species have been produced. However, many species of plants, especially those belonging to the Monocotyledonae and particularly the Gramineae including economically important species such as corn, wheat and rice, have proven to be very recalcitrant to stable genetic transformation.
Difficulties that have been encountered have resided principally in the inability to combine integrative transformation of monocot plant cells (i.e., the stable insertion of foreign DNA in the nuclear genome of the plant cells) with the regeneration of fertile adult plants from those transformed cells. It has been suggested that such difficulties have been predominantly due to the nonavailability of cells that are competent with respect to 1) DNA uptake, 2) integration of DNA in the genome, and 3) regeneration capability (Potrykus I. (1990) Bio/Technology 9:535). Various methods used to transform cereals have been reviewed in the light of the criteria necessary to assess the stable transformation (Potrykus I. (1990) Bio/Technology 9:535; Potrykus (1991) Annu.Rev.Plant Physiol. Plant Mol. Biol. 42:205). In general, direct gene transfer into protoplasts (by polyethyleneglycol (PEG) treatment and/or electroporation) seams to have had the best potential but has nevertheless been hampered by the fact that regeneration from protoplasts has boon difficult to achieve for most genotypes. In practice, protoplasts have most often been obtained from cell suspension cultures (Lazzeri and Lorz (1988) Advances in Cell Culture Vol. 6, Academic press, p. 291Ozias-Akins and Lorz (1994) Trends in Biotechnology 2:119, Hodges et al (1991) In "Rice Biotechnology" ed. Khush end Toenniessen, C.A.B. International, United Kingdom, p. 157; Lynch et al, (1991) In "Rice Biotechnology" ed. Khush and Toenniessen, C.A.B. International, United Kingdom, p. 135).
As plant regeneration from protoplasts has generally been limited to a relatively small number of genotypes for various species, it has been difficult to develop a generally-effective protoplast-based procedure. Therefore, other approaches have recently been explored, particularly in rice.
Lee et al (1991) Proc. Natl. Acad. Sci. USA ("PNAS") 88:6389 have reported the PEG-mediated transformation of small rice cell groups, obtained from rice suspension cultures. Plantlets that had been stably transformed with the gone encoding .beta.-glucuronidase (gus) and a gone encoding neomycin phosphotransferase II (neo) could be regenerated.
Christou et al (1991) Biotechnology 9:957 have reported the transformation of cells of immature zygotic embryos by bombarding the embryos with DNA-coated gold particles. Transgenic rice plants containing the gus gone with either a gene conferring resistance to phosphinothricin (bar) or to hygromycin (hyg) could be regenerated. The introduced genes were reported to segregate in a normal Mendelian ratio in the progeny.
Introduction of DNA into intact plant cells by means of electroporation--a process that is often referred to as electroinjection (see review by Moriwaka et al (1988).
SUMMARY OF THE INVENTION
The present invention provides a method for genetically transforming the genome, particularly the nuclear genome, of cells of a monocotyledonous plant. The genome is stably transformed. The method is preferably applied to an aggregate of the cells and can be applied to cells which can be cultured in suspension. The cells to be transformed are cells which retain at least a portion of their cell wall. The cells of any plant from which regenerable suspension
REFERENCES:
Lindsey et al., Physiologia Plantarum 79:168-172 Copenhagen 1990.
Lindsey et al., Plant Molecular Biology 10;43-52 (1987).
Proc. Ann. Meeting & 28th Symposium of the Jap. Soc. Plant Physiologist (1988) (Translation attached).
Morikawa et al., Gene, 41:121-124 (1986).
Morikawa et al., Biotechnology in Agriculture, 175-202 (1988).
Lee et al., Korean J. Genetics 11-2:65-72 (1989).
Yang et al. Biological abstracts No. 028273 ACTA BOT SIN vol. 33 No. 11 1991.
Li et al. "Introduction of foreign genes into seed embryo cells of rice by electroinjectioon and the regeneration of transgenic rice plants" Science in china Series B vol. 34 No. 8 1991.
Xu et al. "Gene transfer into intact plant cells by electroporation" ACTA BOT SIN vol.32 No. 10, pp. 759-765, abstract 1991.
Tada et al. "Expression of a monocot LHCP promoter in transgenic rice" vol. 10, No. 7 pp. 1803-1809 1991.
Gobel Elke
Nakakido Fumio
Benzion Gary
Plant Genetic Systems N.V.
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