Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or... – The polynucleotide alters plant part growth
Reexamination Certificate
1998-04-07
2001-01-23
Fox, David T. (Department: 1638)
Multicellular living organisms and unmodified parts thereof and
Method of introducing a polynucleotide molecule into or...
The polynucleotide alters plant part growth
C800S286000, C800S320000, C800S320100, C435S412000, C435S419000, C435S069100, C435S440000, C435S468000, C536S023600, C536S024500
Reexamination Certificate
active
06177614
ABSTRACT:
BACKGROUND OF THE INVENTION
Higher plants have a life cycle that consists of a period of vegetative growth followed by reproductive development. Reproduction in angiosperms is a developmental process that begins with floral induction (evocation). This is the point in time at which the shoot apical meristem, the set of dividing cells that gives rise to most of the plant parts above the roots, stops making leaves and starts making flowers. Bernier, G. (1988) The control of floral evocation and morphogenesis.
Ann. Rev. Plant. Physiol. Plant Molec. Biol
. 39:175-219. Almost nothing is known, however, about the molecular and genetic controls that induce a plant to flower.
There is a great need for more information about the regulatory elements in plants. Increased knowledge of these elements would significantly improve our understanding of the underlying mechanism by which genes induce reproductive development in plants.
SUMMARY OF THE INVENTION
This invention identifies and provides isolated DNA which comprises an Id gene of a maize plant, or a portion thereof, which demonstrates Id gene function. The invention further provides RNA encoded by the DNA of the Id or id* alleles and portions thereof, and antisense (complementary) DNA and/or RNA or portions thereof. Nucleic acids, referred to as Id homologues or equivalents, which 1a) show greater than 50% homology (sequence similarity) or that hybridize under moderate stringency conditions to a portion consisting of 20 or more contiguous nucleotide bases of the Id gene or 1b) show a 70% or greater homology or that hybridize under moderate stringency conditions to the Id gene, and 2) demonstrate Id-type (initiation of reproduction phase) function are also encompassed by this invention. Nucleic acid probes and primers to detect and/or amplify regulatory genes in other plants are included as well. Thus, the DNA of this invention comprises an Id gene, or a portion thereof, the Id gene comprising all or a portion of SEQ ID NO:1, or homologous DNA.
The present invention further encompasses polypeptides which are Id proteins or portions of an Id protein of plant origin, including the polypeptides herein described. Id proteins from all plant species or homologues demonstrating a similar regulatory function (reproductive induction) are encompassed by this invention and the term Id protein as used herein. Amino acid sequences that demonstrate 80% or greater homology to the amino acid sequences described herein are considered homologous polypeptides.
In another aspect, this invention relates to antibodies which bind the polypeptides described herein. Such antibodies can be used to locate sites of regulatory activity in plants. Fusion proteins comprising the Id protein and an additional peptide, such as a protein tag, can also be used to detect sites of Id protein/protein interaction in plants.
In a further aspect, this invention provides methods for producing plants with selected times of transition from the vegetative to the flowering stage. Applicants have created a new allele of the id gene, id*, which, when an active Ac transposable element is present, causes plants to stop vegetative growth and to flower earlier than do other id mutants. As shown herein, the id*/id* plants with an active Ac element exhibit fewer vegetative nodes and flower earlier than id*/id* plants without an Ac element or plants encoding the id allele.
The present invention relates to a new mutant of the id gene which encodes a product that alters flower induction in plants and provides a nucleotide sequence of part of the Id SacI 4.2 kb fragment derived from maize Chromosome 1. Also included is DNA which hybridizes under high stringency conditions to the SacI fragment or a portion thereof and an RNA transcribed from or corresponding to either of said aforementioned DNA. Preferably the DNA is that shown in
FIG. 4
(SEQ ID NO:3).
In another aspect, this invention provides methods for producing new id alleles and methods for detecting other Id alleles or other regulatory genes in plants. Homologues of the Id gene can be identified throughout the plant kingdom, including the multicellular and unicellular algae.
In yet another aspect of this invention are provided plants, seeds, plant tissue culture, and plant parts which contain DNA comprising an altered or exogenously introduced Id allele or portion of an Id allele that alters the timing of flower induction in the subsequent growth of the plant, seeds, plant tissue culture, and/or plant part.
The present invention also relates to transgenic plants in which the time of floral evocation is altered. Transgenic plants are provided in which the time period from germination to flowering is shorter than it is in the corresponding naturally-occurring or wild type (native) plant. Alternatively, plants are provided in which flowering is delayed or absent. As used herein, the term transgenic plants includes plants that contain either DNA or RNA which does not naturally occur in the wild type (native) plant or known variants, or additional or inverted copies of the naturally-occurring DNA and which is introduced as described herein, and any of the above-described alterations which result in plants having altered floral evocation times. Such transgenic plants include, in one embodiment, transgenic plants which are angiosperms, both monocotyledons and dicotyledons. Transgenic plants include those into which DNA has been introduced and their progeny, produced from seed, vegetative propagation, cell, tissue or protoplast culture, or the like.
Transgenic plants of the present invention contain DNA which encodes all or a portion of a protein essential for floral evocation and, when present in plant cells, results in altered floral evocation, either earlier cessation of vegetative growth and initiation of flowering than in untransformed plants of the same variety, or in later flowering or the absence of floral induction. The DNA can be exogenous DNA in a sense or antisense orientation which encodes a protein required for floral induction or exogenous DNA which has been altered in such a manner that it encodes an altered form of a protein required for floral induction. Directed or targeted mutagenesis of a plant's endogenous DNA responsible for initiation of flowering can also result in altered floral induction. Exogenous DNA encoding an altered protein required for floral evocation and endogenous DNA required for floral evocation which has been mutated by directed mutagenesis differ from the corresponding wild type (naturally-occurring) DNA in that these sequences contain a substitution, deletion or addition of at least one nucleotide and encode proteins which differ from the corresponding wild type protein by at least one amino acid residue. (As used herein, the term “nucleotide” is used interchangeably with “nucleic acid”.) Insertion of genetic elements, such as Ds sequences with or without active Ac sequences, are of particular use. Exogenous DNA is introduced into plant cells of the target plant by well-known methods, such as Agrobacterium-mediated transformation, microprojectile bombardment, microinjection or electroporation (see below). Such cells carrying the introduced exogenous DNA or endogenous Id DNA mutated by direct mutagenesis can be used to regenerate transgenic plants which have altered floral induction, therefore becoming sources of additional plants either through seed production or non-seed asexual reproductive means (i.e., cuttings, tissue culture, and the like).
The present invention also relates to methods of producing plants with altered floral induction times, exogenous DNA or RNA whose presence in a plant results in altered floral induction, and vectors or constructs which include DNA or RNA useful for producing recombinant plants with altered floral development. Seeds produced by plants which contain exogenous DNA or RNA encoding a protein which is required for floral induction, such as Id DNA in the sense orientation or exogenous DNA which has been altered in such a manner that it encodes an altered form of
Colasanti Joseph J.
Sundaresan Venkatesan
Cold Spring Harbor Laboratory
Fox David T.
Hamilton Brook Smith & Reynolds P.C.
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