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Plant chromosome compositions and methods

a technology of chromosome composition and composition, applied in the field of molecular biology, can solve the problems of achieve the effect of severe limiting the use of genetic analysis of plants

Inactive Publication Date: 2005-12-01
UNIVERSITY OF CHICAGO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0016] In one aspect of the invention, a method is provided for the identification of plant centromeres. In one embodiment of the invention, the method may comprise tetrad analysis. Briefly, tetrad analysis measures the recombination frequency between genetic makers and a centromere by analyzing all four products of individual meiosis. A particular advantage arises from the quartet (qrt 1) mutation in Arabidopsis, which causes the four products of pollen mother cell meiosis in Arabidopsis to remain attached. The quartet mutation may also find use in accordance with the invention in species other than Arabidopsis. For example, several naturally occurring plant species are also known to release pollen clusters, including water lilies, cattails, heath (Ericaceae and Epacridceae), evening primrose (Onagraceae), sundews (Droseraceae), orchids (Orchidaceae), and acacias (Mimosaceae) (Preuss 1994; Smyth 1994). None of these species however, has been developed into an experimental systems thus severely limiting their use for genetic analysis. However, it is contemplated by the inventors that a quartet mutation could be introduced into a host plant to enable the use of tetrad analysis in potentially any species. When used to pollinate a flower, one tetrad can result in the formation of four seeds, and the plants from these seeds can be analyzed genetically. With unordered tetrads, however, such as those produced by Arabidopsis, genetic mapping using tetrad analysis requires that two markers be scored simultaneously.

Problems solved by technology

None of these species however, has been developed into an experimental systems thus severely limiting their use for genetic analysis.

Method used

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  • Plant chromosome compositions and methods
  • Plant chromosome compositions and methods
  • Plant chromosome compositions and methods

Examples

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example 1

Generation of an Arabidopsis thaliana Mapping Population

[0350] To generate a pollen donor plant, two parental lines carrying qrtl were crossed to one another. The qrtl-1 allele was in the Landsberg ecotype background and the qrtl-2 allele was in the Columbia ecotype background. The Landsberg ecotype was readily discernible from the Columbia ecotype because it carries a recessive mutation, erecta, which causes the stems to thicken, infloresences to be more compact, and the leaves to be more rounded and small than wildtype. To utilize this as a marker of a donor plant, qrtl-2 pollen was crossed onto a qrtl-1 female stigma. The F1 progeny were heterozygous at all molecular markers yet the progeny retain the quartet phenotype of a tetrad of fused pollen grains. In addition, progeny display the ERECTA phenotype of the Columbia plant. This visible marker serves as an indication that the crossing was successful in generating plants segregating ecotype specific markers. Further testing was...

example 2

Tetrad Pollinations

[0352] Tetrad pollinations were carried out as follows. A mature flower was removed from the donor plant and tapped upon a glass microscope slide to release mature tetrad pollen grains. This slide was then placed under a 20-40× Zeiss dissecting microscope. To isolate individual tetrad pollen grains, a small wooden dowel was used to which an eyebrow hair with rubber cement was mounted. Using the light microscope, a tetrad pollen unit was chosen and touched to the eyebrow hair. The tetrad preferentially adhered to the eyebrow hair and was thus lifted from the microscope slide and transported the recipient plant stigmatic surface. The transfer was carried out without the use of the microscope, and the eyebrow hair with adhering tetrad was then placed against the recipient stigmatic surface and the hair was manually dragged across the stigma surface. The tetrad then preferentially adhered to the stigma of the recipient and the cross pollination was completed.

[0353] ...

example 3

Preparation and Analysis of Centromere-Spanning Contigs

[0354] Previously, DNA fingerprint and hybridization analysis of two bacterial artificial chromosome (BAC) libraries led to the assembly of physical maps covering nearly all single-copy portions of the Arabidopsis genome (Marra et al., 1999). However, the presence of repetitive DNA near the Arabidopsis centromeres, including 180 bp repeats, retroelements, and middle repetitive sequences complicated efforts to anchor centromeric BAC contigs to particular chromosomes (Murata et al., 1997; Heslop-Harrison et al., 1999; Brandes et al., 1997; Franz et al., 1998; Wright et al., 1996; Konieczny et al., 1991; Pelissier et al, 1995; Voytas and Ausubel, 1988; Chye et al., 1997; Tsay et al., 1993; Richards et al., 1991; Simoens et al., 1988; Thompson et al., 1996; Pelissier et al., 1996). The inventors used genetic mapping to unambiguously assign these unanchored contigs to ±20 specific centromeres, scoring polymorphic markers in 48 plant...

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Abstract

The present invention provides for the identification and cloning of functional plant centromeres in Arabidopsis. This will permit construction of stably inherited minichromosomes which can serve as vectors for the construction of transgenic plant and animal cells. In addition, information on the structure and function of these regions will prove valuable in isolating additional centromeric and centromere related genetic elements and polypeptides from other species.

Description

[0001] This application claims the priority of U.S. Provisional Application Ser. No. 60 / 125,219, filed Mar. 18, 1999, U.S. Provisional Application Ser. No. 60 / 127,409, filed Apr. 1, 1999, U.S. Provisional Application Ser. No. 60 / 134,770, filed May 18, 1999, U.S. Provisional Application Ser. No. 60 / 153,584, filed Sep. 13, 1999, U.S. Provisional Application Ser. No. 60 / 154,603, filed Sep. 17, 1999 and U.S. Provisional Application Ser. No ______, filed Dec. 16, 1999, each of which disclosures is specifically incorporated herein by reference in its entirety.[0002] The government owns rights in the invention pursuant to U.S. Department of Agriculture Grant No. 96-35304-3491, National Science Foundation Grant No. 9872641 and Grant No. DOEDE-FG05-920R22072 from the Consortium for Plant Biotechnology.BACKGROUND OF THE INVENTION [0003] I. Field of the Invention [0004] The present invention relates generally to the field of molecular biology. More a particularly, it concerns plant chromosome ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A01H1/00A01H11/00C07H21/02C12N15/79C12N15/81C12N15/82C12N15/87
CPCC12N15/82
Inventor PREUSS, DAPHNECOPENHAVER, GREGORYKEITH, KEVIN
Owner UNIVERSITY OF CHICAGO
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