Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or...
Reexamination Certificate
2000-05-24
2004-11-23
Shukla, Ram R. (Department: 1635)
Multicellular living organisms and unmodified parts thereof and
Method of introducing a polynucleotide molecule into or...
C435S091100, C435S091310, C435S468000, C536S023100, C536S024500, C800S295000
Reexamination Certificate
active
06822139
ABSTRACT:
DESCRIPTION
The present invention relates to a process for the production of a transgenic plant the seeds of which form embryos that exhibit a modified development and to seeds, plant derived tissues and plants obtained thereby.
The present invention relates to plant genetic engineering. A goal of plant genetic engineering is to introduce desired genes into a plant in such a manner that these genes will be functional in the desired tissue at the correct time. Plant genetic engineering aims for instance to modify the pathways of primary and secondary metabolites of economic importance including the cellular and organic optimisation of compounds. Furthermore, plant genetic engineering aims to insert and develop mechanisms of resistance against physical, chemical and biological stress factors. Finally, plant genetic engineering aims to alter the development of plants and their seeds. Altering the developmental pattern of a plant enables for instance the production of plants with a modified plant or organ morphology. In particular, it is desired to engineer plants and plant seedlings exhibiting a modified, in particular increased number of storage organs such as cotyledons. These storage organs may contain valuable and commercially interesting substances such as proteins, polysaccharides, globoides, and vegetable oils, such as seed storage lipids of higher plants. It is also desirable to provide genetically engineered plants producing abortive seed for use in breeding programs and for agricultural purposes.
Genes involved in cell division, signal transduction pathways, establishment of cell fate and pattern formation have been extensively studied, since they are of general importance to the understanding of plant development. The Arabidopsis SHAGGY-related protein kinase (ASK) multigene family calls for proteins that share a highly conserved catalytic protein kinase domain being about 70% identical to animal genes known to be involved in signal transduction pathways controlling patterning cell fate determination and cytokinases (Dornelas et al., Gene 212 (1998), 249-257). The ASK proteins are believed to be involved in signal transduction pathways that establish cell fate and/or pattern formation in plants. The ASK gene family comprises various members, such as ASK alpha &agr;, gamma &ggr;, dzeta &zgr;, etha &eegr; and iota &igr;.
Although cDNA and genomic DNA sequences of these genes are available, the function of these genes during the development of the plant is still unknown (Dornelas et al., Plant Molecular Biology 39 (1999), 137-147, Tichtinsky et al., Biochimica et Biophysica Acta 1442 (1998), 261-273). In fact, speculations on their function are only based on sequence similarities of the encoded products to animal counterparts. Up until now, it is not known whether these genes might prove useful in plant genetic engineering and, should this be the case, forwhich purpose.
Thus, the technical problem underlying the present invention is to provide plants which exhibit improved properties that increase their commercial value.
The present invention solves this problem by providing a process for the production of a transgenic plant the seeds of which comprise an embryo exhibiting a modified development, wherein at least one plant cell is transformed with at least one DNA construct comprising a nucleic acid sequence derived from at least one ASK-gene of group II and regenerated to a plant whose embryos exhibit the modified development. The problem is also solved by plants and seeds produced by this process, and plants and seeds comprising an expressible DNA construct containing an ASK-gene of group II.
The present invention relates to the unexpected teaching that a particular ASK-gene which encodes a kinase may be used to specifically alter the development of a plant embryo and/or architecture of a plant. Thus, the present invention foresees the use of a DNA construct comprising a nucleic acid sequence derived from at least one ASK-gene of group II for genetically modifying a plant, whereby a plant with advantageous properties is obtained. Such a DNA construct may be a sense or an antisense construct or a construct comprising a transposable element such as En/Spm or Ac/Ds. According to the present invention, the transposable element transformed into a plant cell is capable of inactivating an endogenous ASK-gene of group II so as to generate plants which produce embryos exhibiting a modified development. In the context of the present invention, a sense construct comprises at least one regulatory element being functionally linked in sense orientation, i.e. wild-type orientation, to a nucleic acid sequence derived from at least one ASK-gene of group II. In a particularly preferred embodiment, the ASK-gene derived sequence is the coding sequence of an ASK-gene of group II. Such a construct may be used to overexpress the coding sequence of an ASK-gene of group II. Such a construct may be particularly useful for a co-suppression technology, wherein at least one transgenic copy of an ASK-gene of group II is inserted into the genome of a target plant cell and wherein due to a high copy number of such a transgenic DNA sequence and/or an increased expression rate, down regulation of an endogenous corresponding ASK-gene can be achieved. In the context of the present invention, the term co-suppression construct refers to such a construct comprising at least one regulatory element being functionally linked in sense orientation to a nucleic acid sequence derived from an ASK-gene of group II, in particular a transcribed region, most preferably a coding region.
The invention also relates to down regulation of expression of an endogenous ASKene of group II by using antisense technology. Thus, in one preferred embodiment of the present invention, a DNA construct is used, wherein at least one regulatory element is operably linked in antisense orientation to a nucleic acid sequence derived from at least one ASK-gene of group II, in particular a transcribed region or a part thereof, in particular the coding sequence or a part thereof. In the context of the present invention, antisense orientation refers to a non-wild-type orientation of a 5′ regulatory element, that is a promoter to its coding sequence, in particular an orientation wherein from a given functional regulatory 5′ element, the antisense strand of the ASK-gene of group II is transcribed.
As explained above, the DNA construct may also comprise a transposable element which is capable of being inserted in an endogenous ASK-gene of group II, thereby inactivating this gene.
The process of the present invention enables the production of transgenic plants producing seeds whose embryos exhibit a modified development. In a particularly preferred embodiment of the present invention, the embryo generated within the seed of the transgenic plant of the present invention is unexpectedly characterised by the development of an increased number of cotyledons in contrast to wild-type plants. Accordingly, in that case where the plant cell transformed is a plant cell obtained from a monocotyledonous plant, the present invention enables the production of seeds whose embryos will develop 2, 3, 4 or even more cotyledons. In the case where the plant cell transformed is a plant cell obtained from a dicotyledonous plant, the present invention enables the production of seeds whose embryos will develop 3, 4, 5 or even more cotyledons. Thus, the present invention enables the production of polycotyledonous plants. These plants and in particular their embryos and seedlings are advantageous in so far as they may contain, due to their increased number of cotyledons, an increased amount of valuable and commercially interesting substances, such as proteins or vegetable oils. Thus, the plant of the present invention may advantageously be used as plants producing in their storage organs, in particular their cotyledons, commercially interesting substances. The process of the present invention enabling the generation of polycotyledonous plants is also useful for t
Dornelas Marcelo
Kreis Martin
van Lammeren Andre A. M.
Advanta Seeds B.V.
Rewoldt Dana S.
Shukla Ram R.
Zara Jane
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