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Seeds

a seed and seed technology, applied in the field of seeds, can solve the problems of increasing the size of many plant organs, male sterility, and the differences in the size of the ovule, and achieve the effect of more space for growth

Inactive Publication Date: 2008-02-28
UNIVERSITY OF BATH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0034]According to another aspect, the present invention provides a method of modifying cell proliferation in a plant which comprises the step of transforming a plant or plant propagating material with a nucleic acid molecule comprising at least one regulatory sequence, typically a promoter sequence, capable of directing expression within the integuments and / or seed coat of at least one nucleic acid sequence whose expression or transcription product is capable of directly or indirectly modulating cell proliferation, whereby, on expression of that sequence, cell proliferation is modified. Preferably, the overall size of the integuments / seed coat in the plant is modified. This may be useful where a product is produced in the integument / seed coat. In some embodiments, this will be achieved without affecting the growth or development of any part of the plant other than the seed.
[0052]One method of enhancing function is to activate transcription of the gene, resulting in increased levels of mRNA and protein encoded by the gene. This is achieved by linking a plant promoter to the coding region of the gene (either with or without introns) in the sense orientation.

Problems solved by technology

Seed quality is an important factor in the cost of production of commercial seed lots since these must be tested before sale.
Moreover, the authors' inspection of mature unfertilized ovules showed that ovules in Ler were slightly longer than in Cvi, and therefore the authors concluded that ‘ovule size differences could not account for the final Ler / Cvi seed size variation’.
Their overall major conclusion was that ‘the larger size of Cvi seeds compared with Ler is mainly because of the faster and prolonged growth of the integuments and the endosperm’ (i.e. after fertilization); they did not address the question of whether this growth was led by the integuments or the endosperm.
Ectopic ANT expression increases the size of many plant organs including seeds, as well as causing male sterility through failure of anther dehiscence.
However weak overexpressers could generate seeds after hand-pollination with wild-type pollen.
U.S. Pat. No. 6,329,567 describes methods of modulating seed mass using AP2 transgenes, but this patent does not assess any effect of the transgenes on the integuments or seed coat.

Method used

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Examples

Experimental program
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Effect test

example 1

Value of mnt Mutants in Understanding and Modifying Growth of Integuments / Seed Coat

[0135]The mnt mutant seed phenotype demonstrates that there is a correlation between the size of integuments before fertilization and the size of the mature seed in Arabidopsis thaliana (FIGS. 1, 2). Due to the similarities in seed structure among even distantly related groups of flowering plants, this leads to the expectation that modification to integument / seed coat size in other species, and certainly in members of the Brassicaceae such as Brassica napus, will also result in changes to seed size.

[0136]Our knowledge of the mnt mutant phenotype and MNT gene sequence can be exploited in other species through TILLING (‘Targeting induced local lesions in genomes’). In this reverse genetics technique, chemically mutagenized populations are screened for presence of a point mutation in a nucleic acid sequence of interest; this can be done as a high-throughput procedure and is applicable to many species (Ti...

example 2

Modifying Expression of mnt Orthologues in Other Species

[0138]Knowledge of the MNT sequence in Arabidopsis thaliana also allows us to search for orthologues in crop species as a necessary first step in targeted modification of the expression of the gene in these species.

[0139]By way of example, we amplified the putative Brassica napus orthologue (BnARF2) of MNT using primers (SEQ ID NO 7, 8) based on the MNT sequence and on publicly available Brassica oleracea sequence. The BnARF2 cDNA was amplified from total RNA isolated from seedlings of Brassica napus var. Westar. The BnARF2 cDNA from translational start to stop is shown in SEQ ID NO. 9 and is aligned with Arabidopsis thaliana MNT cDNA in FIG. 8. The BnARF2 predicted protein (SEQ ID NO. 10) has 85% identity to Arabidopsis thaliana MNT.

[0140]A family of ARFs has also been characterized in rice and one of these, OsARF2 (accession no. AB071293), is considered to be the orthologue of Arabidopsis thaliana ARF2 (Sato et al., 2001). FI...

example 3

Construction, Transformation, and Analysis of Reporter Vectors to Show where Integument / Seed Coat Promoters are Expressed in Arabidopsis thaliana

[0142]This is to test which promoters are suitable for driving integument / seed coat-specific or -preferred expression of nucleic acids such as MNT antisense or RNAi constructs, or other genes modifying cell proliferation.

[0143]Diagrams of the BJ60, BJ40, pFGC5941, pART7, and BJ36 vectors used in the cloning strategies described in this and following examples are shown in FIG. 10.

[0144]The cloning strategy is shown in FIG. 11.

3a Construction of Reporter Vectors

3a(i) TT8

[0145]A reporter vector based on the promoter of the TT8 gene (Nesi et al., 2000; At4g09820, accession no. AJ277509) is constructed as described below. A 1.7 kb fragment including the TT8 promoter is amplified by the polymerase chain reaction (PCR) from Arabidopsis thaliana genomic DNA 5′ to translational start of the TT8 gene using the primers TT8F and TT8R which introduce a...

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Abstract

This invention relates to a method for modifying cell proliferation in a plant which comprises the step of transforming a plant, or plant propagating material, with a nucleic acid molecule comprising at least one regulatory sequence capable of directing expression within the integuments and / or seed coat and at least one nucleic acid sequence whose expression or transcription product is capable of directly or indirectly modulating cell proliferation, whereby on expression of that nucleic acid sequence cell proliferation is modified. The invention also relates to a plant which includes a nucleic acid molecule comprising at least one regulatory sequence capable of directing expression within integuments and / or seed coat an at least one nucleic acid sequence whose expression or transcription product is capable of directly or indirectly modulated cell proliferation, whereby on expression of that nucleic acid sequence cell proliferation is modified. The invention also relates to reproducible plant material including a nucleic acid molecule comprising at least one regulatory sequence capable of directing expression within integuments and / or seed coat and at least one nucleic acid sequence whose expression or transcription product is capable of directly or indirectly modulated cell proliferation, whereby expression of that nucleic acid sequence cell proliferation is modified. According to another aspect of the invention, there is provided a method for modifying cell proliferation in a plant which comprises the step of modulating the response of the plant to an auxin in which cell proliferation is modified to produce larger or smaller seeds than wild-type.

Description

FIELD OF THE INVENTION[0001]The present invention relates particularly, though not exclusively, to methods for modifying characteristics such as seed size in plants, especially flowering plants, and to plants and reproducible plant material produced by the methods. The invention also relates to nucleic acid constructs for use in such methods, as well as to modified plants and reproducible plant material per se.BACKGROUND OF THE INVENTION[0002]The seeds industry can be split into two high-value, commercial sectors: seeds for field crops such as corn, oil seeds, sugar beet and cereals, and vegetable and flower seed. The scientific improvement of crop plants has gone through a succession of innovations leading to the development of hybrid varieties for many crops and, most recently, to the introduction of genetically enhanced crops. The worldwide commercial seeds market is valued at around $30 billion (International Seed Federation).1. Importance of Seed Size[0003]Yield in crop plants ...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/00A01H5/10C07K14/415
CPCA01H5/10C07K14/415C12N15/8261C12N15/8234C12N15/823Y02A40/146
Inventor SCOTT, RODERICK
Owner UNIVERSITY OF BATH
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