Isolated nucleotide sequences responsible for the tomato high pigment-1 mutant phenotypes (hp-1 and hp-1w) and uses thereof

A nucleotide sequence and nucleotide technology, applied in the fields of applications, chemical instruments and methods, botanical equipment and methods, etc., can solve the problem of 9 base pair deletions, etc.

Inactive Publication Date: 2009-04-22
沃尔坎尼中心-以色列农业部农业研究组织
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Problems solved by technology

For the hp-2 mutant, a point mutation directs alternative sp...

Method used

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  • Isolated nucleotide sequences responsible for the tomato high pigment-1 mutant phenotypes (hp-1 and hp-1w) and uses thereof
  • Isolated nucleotide sequences responsible for the tomato high pigment-1 mutant phenotypes (hp-1 and hp-1w) and uses thereof
  • Isolated nucleotide sequences responsible for the tomato high pigment-1 mutant phenotypes (hp-1 and hp-1w) and uses thereof

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Experimental program
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Embodiment 1

[0087] Identification and cloning of DDB1 tomato homologues

[0088] The DDB1 protein is a heterodimer composed of two subunits, DDB1 and DDB2. Unlike rice, chicken, human, mouse, Drosophila and Schizosaccharomycespombe, the A. thaliana genome has two highly homologous copies of the DDB1 gene (Schroeder et al. 2002; Zolezzi et al. 2002 ; Fu et al.2003; Ishibashi et al.2003): DDB1A and DDB1B, both 1088 amino acids in length (Genbank protein accession numbers NP_192451 and NP_193842, respectively). When each of these two protein accession numbers was used as a query for tblastn analysis in the TIGR database (http: / / www.tigr.org / ) containing the tomato expressed sequence tags (ESTs), both revealed Two highly homologous sequences: TC117371 (394bp) and TC117372 (2206bp). A. thaliana accession number NP_192451 was found to be 87 and 86% identical to tomato TC117371 and TC117372 accession numbers, respectively. On the other hand, accession number NP_193842 has 87 and 83% identit...

Embodiment 2

[0090] Mapping of Tomato DDB1 Gene

[0091] Part of the mapping results, including the approximate location of the tomato DDB1 gene, is shown in Figure 2. These results indicate that DDB1 is located on tomato chromosome 2 in introgressed varieties with the HP-1 gene (Yen et al. 1997).

Embodiment 3

[0093] hp-1 and hp-1 w Sequence characterization of tomato DDB1 in mutants

[0094] Several forward and reverse primers (Table 1) complementary to the 3' region of the tomato DDB1 gene (TIGR Accession No. TC117372) were used for direct cDNA preparation from seedling leaves of hp-1 and normal plants of the Ailsa Craig background. sequencing. No polymorphisms between hp-1 and normal plants were acquired in this region. Therefore, the 5' region of the DDB1 gene in the two genotypes was also cloned and fully sequenced. In silico translation of all sequences revealed that tomato DDB1 is a 1090 amino acid protein. Sequence analysis of the DDB1 coding sequence from hp-1 and its near-isogenic normal genotype revealed the presence of a single A in the coding sequence of the DDB1 gene in mutant hp-1 plants 931 to T 931 base transversion. Asparagine 311 to tyrosine 311 replacement (Figure 4).

[0095] Based on the sequence information obtained in the background of Ailsa Crai...

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Abstract

The present invention provides isolated nucleotide sequences responsible for the tomato high pigment 1 (hp-1) and high pigment 1<w> (hp-1<w>) phenotypes, wherein said sequences comprises an altered tomato DDB1 gene sequence or fragment thereof, wherein said the alteration in said altered sequence or fragment comprises an A-to-T transversion at nucleotide 931 of said DDB1 gene sequence in the case of hp-1 and a G-to-A transition at nucleotide 2392 of said DDB1 gene sequence in the case of hp-1<w>.

Description

technical field [0001] The present invention relates to causing tomato to produce high pigment-1 (high pigment-1) and high pigment-1 w (high pigment-1 w ) phenotype altered nucleotide sequence. More specifically, the present invention discloses point mutations in the tomato homologues of the Arabidopsis thaliana and human DDB1 (UV damaged DNA binding protein 1) genes, and the use of the altered nucleotide sequences. Background technique [0002] Plants respond to light intensity, direction, duration and spectral properties by adjusting their developmental processes in a series of interactions called photomorphogenesis. Photomorphogenic mutants have proven to be excellent tools in the study of the complex interplay between light and plant development, and some of them have also been used in several crop breeding programmes. Photomorphogenic mutants have been reported in several species including Arabidopsis, Sorghum, Brassica, tobacco, tomato and pea. Typically, these mut...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12Q1/68A01H1/04C07K14/415C12NC12N15/82
CPCA01H1/04C12N15/8243C07K14/415C12N15/825
Inventor I·勒文M·烈伯曼O·阿米尔塞格弗N·基尔波阿A·拉拉扎
Owner 沃尔坎尼中心-以色列农业部农业研究组织
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