Chemistry: molecular biology and microbiology – Process of mutation – cell fusion – or genetic modification – Introduction of a polynucleotide molecule into or...
Reexamination Certificate
1999-10-22
2002-09-03
Fox, David T. (Department: 1638)
Chemistry: molecular biology and microbiology
Process of mutation, cell fusion, or genetic modification
Introduction of a polynucleotide molecule into or...
C435S412000, C435S419000, C435S430000, C435S430100, C435S424000, C435S431000, C800S278000, C800S298000, C800S320100
Reexamination Certificate
active
06444470
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to the field of genetic engineering. The invention further relates to increasing the embryogenic response of callus, transforming plant cells and regenerating plants from cells and tissues.
BACKGROUND OF THE INVENTION
The development of methods for the introduction of foreign genes into organisms has had a profound impact on fields of medicine and agriculture. While the movement of genes within plant species or between closely related plant species by traditional methods based on sexual reproduction has played an important role in crop improvement for most of this century, the pace of crop improvement by such methods has been slow and limiting due to the reliance on naturally occurring genes. Recent advances in the field of genetic engineering has led to the development of genetic transformation methods that allow the introduction of recombinant DNA, into organisms. The recombinant DNA methods which have been developed have greatly extended the sources from which genetic information can be obtained for crop improvement. Recently, new crop plant varieties, developed through recombinant DNA methods, have reached the marketplace. Genetically engineered soybeans, maize, canola and cotton are now widely utilized by North America farmers.
Rapid progress has been made in developing the tools for manipulating genetic information in plants. Plant genes are being cloned, genetic regulatory signals deciphered, and genes transferred from entirely unrelated organisms to confer new agriculturally useful traits to crop plants. Recombinant DNA methods significantly increase the gene pool available for crop improvement. A variety of methods have been developed for the transformation of plants and plant cells with DNA.
Cereals comprise a commercially valuable group of plant species that could benefit from the introduction and expression of foreign genes controlling improved grain quality and such agronomically important traits as tolerance to disease, insects, herbicides, and stress. The use of microprojectile-bombardment-mediated transformation of embryogenic tissue culture material, with the subsequent regeneration of transgenic plants, has overcome the regeneration problems associated with the production of plants from cereal protoplasts. Using this technology, transgenic plants have been obtained from microprojectile-bombarded tissue cultures of many species.
Many of the recent advances in plant science have resulted from application of the analytical power of recombinant DNA technology coupled with plant transformation. These approaches facilitate studies of the effects of specific gene alterations and additions on plant development and physiology. They also make possible the direct manipulation of genes to bio-engineer improved plant varieties.
While strides have been made in the genetic transformation of plants, it is by no means a routine matter. For many important crop plants, the efficiency of transformation as measured by the number of transformed plants recovered per transformation attempt is very low and highly variable from one genetic line or variety to the next. Often, highly desirable breeding lines exhibit extremely low transformation efficiencies, relative to other genetic lines of the same crop species. Frequently, acceptable levels of transformed plant cells and calli can be obtained from a transformation attempt but such transformed plant cells and calli are recalcitrant to regeneration into transformed embryos and plants. While some success has already been achieved in improving crop plants through the introduction of recombinant DNA, the progress of genetic engineers working to improve many important crop species is impeded by inefficient methods for transforming and regenerating crop plants. Thus, improved methods for transforming plant cells and regenerating transformed plants are desired.
SUMMARY OF THE INVENTION
Methods and compositions are provided for preparing plants for transformation, producing transformed plants and plant embryos and increasing plant transformation efficiency. The methods involve administering to a plant an effective amount of a transformation-enhancing agent prior to attempting to transform a cell from the plant. The transformation-enhancing agent comprises one or more components that when administered to a plant increases transformation efficiency when cells from the plant are subsequently used in plant transformation methods. The compositions comprise a transformation-enhancing agent and a carrier. The methods and compositions of the invention find use in agriculture, particularly in transforming crop plants, more particularly in transforming crop plants that display low transformation efficiencies with existing transformation methods.
Also provided are transgenic plants and seeds thereof.
DETAILED DESCRIPTION OF THE INVENTION
A number of terms used herein are defined and clarified in the following section.
By “increase transformation efficiency” is intended that the number of transformed plants recovered from a transformation attempt is increased preferably at least two fold, preferably at least five fold, more preferably at least ten fold.
By “transformation-enhancing agent” is intended a composition that when applied to a plant prior to and/or during a transformation attempt increases transformation efficiency. Such a “transformation-enhancing agent” may favorably influence one of more of the physiological processes associated with genetic transformation and the subsequent regeneration of transformed plants including, but not limited to, foreign DNA uptake into plant cell, foreign DNA integration in a genome of a plant cell, plant cell proliferation in culture, plant cell culture initiation, proliferation of transformed plant cell, embryogenesis, shoot initiation and root initiation.
By “effective amount of a transformation-enhancing agent” is intended an amount that favorably affects one or more of processes associated with increasing transformation efficiency including, but not limited to, uptake of foreign DNA by a cell, integration of foreign DNA into the genome of a cell, expression of a foreign gene in a transformed cell, proliferation of a transformed cell, callus formation, embryogenesis, root formation and shoot formation. Those of ordinary skill in the art understand that such an “effective amount” depends on a number of factors including, but not limited to, the transformation-enhancing agent employed, the species of plant, the developmental stage of the plant, the method by which the transformation-enhancing agent is administered, environmental conditions and the type of cells to be transformed.
The invention is drawn to compositions and methods for genetically transforming plant cells and regenerating transformed plants. The compositions and methods find use in improving existing methods for producing transformed plants, particular transformed crop plants, more particularly transformed monocot crop plants, most particularly transformed maize plants. The compositions comprise transformation-enhancing agents. When administered to a plant in an effective amount prior to a transformation attempt, such transformation-enhancing agents increase transformation efficiency.
Compositions comprising a transformation-enhancing agent are provided. Additionally, compositions comprising a transformation-enhancing agent and a carrier are provided. The compositions increase transformation efficiency when applied to a plant prior to a transformation attempt. While such transformation-enhancing agents do not depend on any particular biological mechanism for increasing transformation efficiency, it is recognized that the transformation-enhancing agents may alter one or more processes such as, for example, uptake of foreign DNA by a cell, integration of foreign DNA into the genome of a cell, expression of a foreign gene in a transformed cell, proliferation of a transformed cell, cell division, callus formation, embryogenesis, root formation, shoot formation and leaf formation. The transformatio
Church Laura A.
Gordon-Kamm William J.
Ross Margit C.
Fox David T.
Kruse David H.
Pioneer Hi-Bred International , Inc.
Pioneer Hi-Bred International Inc.
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