Nucleic acids that control seed and fruit development in plants

Details Nucleic acids that control seed and fruit development in plants Nucleic acids that control seed and fruit development in plants

2000-04-21

2002-11-05

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, C800S266000, C800S294000, C800S298000, C435S469000, C435S419000, C435S468000, C435S320100, C435S069800, C536S023600

Reexamination Certificate

active

06476296

ABSTRACT:

FIELD OF THE INVENTION
This invention is directed to plant genetic engineering. In particular, it relates to modulating seed (and in particular endosperm, embryo and seed coat) development in plants.
BACKGROUND OF THE INVENTION
A fundamental problem in biology is to understand how seed development. In flowering plants, the ovule generates the female gametophyte which is composed of egg, central, synergid and antipodal cells (Reiser, et al.,
Plant Cell
, 1291-1301 (1993)). All are haploid except the central cell which contains two daughter nuclei that fuse prior to fertilization. One sperm nucleus fertilizes the egg to form the zygote, whereas another sperm nucleus fuses with the diploid central cell nucleus to form the triploid endosperm nucleus (van Went, et al.,
Embryology of Angiosperms
, pp. 273-318 (1984)). The two fertilization products undergo distinct patterns of development. In Arabidopsis, the embryo passes through a series of stages that have been defined morphologically as preglobular, globular, heart, cotyledon and maturation (Goldberg, R. B., et al.,
Science
(1994) 266: 605-614; Mansfield, S. G., et al.,
Arabidopsis: An Atlas of Morphology and Development
, pp. 367-383 (1994)). The primary endosperm nucleus undergoes a series of mitotic divisions to produce nuclei that migrate into the expanding central cell (Mansfield, S. G., et al.,
Arab Inf Serv
27: 53-64 (1990); Webb, M. C., et al.,
Planta
184:187-195 (1991)). Cytokinesis sequesters endosperm cytoplasm and nuclei into discrete cells (Mansfield, S. G., et al.,
Arab Inf Serv
27:65-72 (1990)) that produce storage proteins, starch, and lipids which support embryo growth (Lopes, M. A. et al.,
Plant Cell
5:1383-1399 (1993)). Fertilization also activates development of the integument cell layers of the ovule that become the seed coat, and induces the ovary to grow and form the fruit, or silique, in Arabidopsis.
Of particular interest are recent discoveries of genes that control seed, and in particular endosperm, development. For instance, MEDEA (MEA) (also known as FIE1 (see, e.g., copending U.S. patent application Ser. No. 09/071,838) and F644 (see, e.g., Kiyosue T, et al. (1 999)
Proc Natl Acad Sci USA
96(7):4186-91) encodes an Arabidopsis SET domain polycomb protein that appears to play a role in endosperm development. Inheritance of a maternal loss-of-function mea allele results in embryo abortion and prolonged endosperm production, irrespective of the genotype of the paternal allele. Thus, only the maternal wild-type MEA allele is required for proper embryo, endosperm, and seed coat development (Kinoshita T, et al. (1999)
Plant Cell
10:1945-52). These results reveal functions for plant polycomb proteins in the suppression of central cell proliferation and endosperm development (Kiyosue T, et al. supra).
Another gene product that controls seed development is FIE, also known as FIE3 (see, e.g., copending U.S. patent application Ser. No. 09/071,838). The FIE protein is a homolog of the WD motif-containing Polycomb proteins from Drosophila and mammals (Ohad, N. et al.
Plant Cell
11(3):407-16 (1999)). In Drosophila, these proteins function as repressors of homeotic genes. A female gametophyte with a loss-of-function allele of fie undergoes replication of the central cell nucleus and initiates endosperm development without fertilization. These results suggest that the FIE Polycomb protein functions to suppress a critical aspect of early plant reproduction, namely, endosperm development, until fertilization occurs.
Control of the expression of genes that control egg and central cell differentiation, or those that control reproductive development, i.e. embryo, endosperm and seed coat, is useful in the production of plants with a range of desired traits. These and other advantages are provided by the present application.
SUMMARY OF THE INVENTION
This invention provides isolated nucleic acids comprising a polynucleotide sequence, or its complement, encoding an ATR polypeptide exhibiting at least 60% sequence identity to SEQ ID NO:2. For instance, the nucleic acid can encode the ATR polypeptide displayed in SEQ ID NO:2. In one aspect, the polynucleotide sequence comprises SEQ ID NO:5 or SEQ ID NO:1. In some aspects of the invention, the nucleic acid further comprises a promoter operably linked to the polynucleotide. In some embodiments, the promoter is constitutive. In other embodiments, the promoter is from an ATR gene. For example, the promoter can comprise a polynucleotide at least 70% identical to SEQ ID NO:3. In some aspects, the promoter comprises SEQ ID NO:3. In some aspects of this invention, the promoter further comprises a polynucleotide at least 70% identical to SEQ ID NO:4. For example, in some aspects the promoter comprises SEQ ID NO:4. In some aspects, the polynucleotide sequence is linked to the promoter in an antisense orientation.
The invention also provides an isolated nucleic acid molecule comprising a polynucleotide sequence exhibiting at least 60% sequence identity to SEQ ID NO:1.
The invention also provides an expression cassette comprising a promoter operably linked to a heterologous polynucleotide sequence, or complement thereof, encoding an ATR polypeptide exhibiting at least 60% sequence identity to SEQ ID NO:2. For instance, the nucleic acid can encode the ATR polypeptide displayed in SEQ ID NO:2. In some aspects, the polynucleotide sequence comprises SEQ ID NO:5 or SEQ ID NO:1. In some aspects of the invention, the nucleic acid further comprises a promoter operably linked to the polynucleotide. In some embodiments, the promoter is constitutive. In other embodiments, the promoter is from an ATR gene. For example, the promoter can comprise a polynucleotide at least 70% identical to SEQ ID NO:3. In some aspects, the promoter comprises SEQ ID NO:3. In some aspects of this invention, the promoter further comprises a polynucleotide at least 70% identical to SEQ ID NO:4. For example, in some aspects the promoter comprises SEQ ID NO:4. In some aspects, the polynucleotide sequence is linked to the promoter in an antisense orientation.
The invention also provides an expression cassette for the expression of a heterologous polynucleotide in a plant cell. In some aspects, the expression cassette comprises a promoter polynucleotide at least 70% identical to SEQ ID NO:3 that is operably linked to a heterologous polynucleotide. In some aspects, the promoter comprises SEQ ID NO:3. In some aspects, the promoter further comprises a polynucleotide at least 70% identical to SEQ ID NO:4. For instance, in some embodiments, the promoter comprises SEQ ID NO:4.
The present invention also provides a host cell comprising an exogenous polynucleotide sequence comprising a polynucleotide sequence, or complement thereof, encoding an ATR polypeptide exhibiting at least 60% sequence identity to SEQ ID NO:2. In some aspects of the invention, the nucleic acid further comprises a promoter operably linked to the polynucleotide sequence. In some aspects, the promoter is constitutive. In some aspects, the promoter comprises a polynucleotide at least 70% identical to SEQ ID NO:3. The promoter, for instance, can comprise SEQ ID NO:3. In some aspects, the promoter further comprises a polynucleotide at least 70% identical to SEQ ID NO:4. For instance, in some embodiments, the promoter comprises SEQ ID NO:4. In some aspects, the promoter is operably linked to the exogenous polynucleotide sequence in an antisense orientation.
The present invention also provides an isolated polypeptide comprising an amino acid sequence at least 60% identical to SEQ ID NO:2 and capable of exhibiting at least one biological activity of the polypeptide displayed in SEQ ID NO:2, or fragment thereof. The present invention also provides for an antibody capable of binding such polypeptides.
The present invention also provides a method of introducing an isolated nucleic acid into a host cell comprising, (a) providing an isolated nucleic acid or its complement, encoding an ATR polypeptide exhibiting at least 60% sequence identity to SEQ ID NO:2.

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