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Modification of sucrose synthase gene expression in plant tissue and uses therefor

A chimeric gene and plant technology, applied in the field of modification of sucrose synthase gene expression in plant tissue and its application

Inactive Publication Date: 2007-07-18
COMMONWEALTH SCI & IND RES ORG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at the time there was no evidence implicating the role of the SuSy gene in cotton fiber / seed development, although it was speculated (Amor et al., 1995) that part of SuSy localized to the fiber might bind to cellulose synthase to exert carbon synthase. Channel action between the enzymes

Method used

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  • Modification of sucrose synthase gene expression in plant tissue and uses therefor
  • Modification of sucrose synthase gene expression in plant tissue and uses therefor
  • Modification of sucrose synthase gene expression in plant tissue and uses therefor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0099] Example 1: Inhibition of SuSy Expression in Cotton

[0100] To provide evidence for the role of SuSy in fiber development, the inventors transformed cotton with cosuppressive and antisense SuSy constructs that target the 3' end of SuSy's cDNA and consist of constitutively three Leaf dwarf virus promoter (see Figure 5 for a diagram) drives. The presence of the transgene in 11 transgenic lines was confirmed by Southern analysis (see Figure 6). Two of the antisense lines (294-147, 292-43) and one co-suppression line (295-82) showed a significant reduction in fiber and seed development, while the remaining lines showed varying degrees of inhibition of fiber growth.

[0101] Immunolocalization analysis of 0-d ovule sections (Fig. 2) revealed that SuSy protein in line 294-147 was reduced to less than 20% of wild-type levels compared to wild-type fibroblasts and at 295 -82 down to undetectable. Further, the number and size of initial fibers were reduced by at least 50% in b...

Embodiment 2

[0116] Example 2. Progeny analysis of cotton lines 82, 147 and 43 with suppressed SuSy activity.

[0117] To confirm the correlation between the presence of the SuSy suppressor transgene and the reduction in fibrogenesis in cotton, further analyzes were performed on the T1 generation. Approximately 24 T1 seeds from 6 TO generations were sown. All germinated seedlings were screened by PCR for the presence or absence of this transgene (nptII; S7 promoter and SuSy repressor transgene). Six PCR-positive and one PCR-negative isolates from six strains were analyzed at the molecular, biochemical and cellular levels. Line 82 was vegetatively propagated as a reference point. The fiberless phenotype in different lines of the T0 generation (arrows in FIG. 6A ) was retained in 3 and 2 segregants in lines 147 and 43, respectively (arrows in FIG. 6B ). The remaining individuals showed varying degrees of fiber / seed suppression. Vegetatively-kept line 82 continued to produce fiber-free se...

Embodiment 3

[0121] Example 3. Overexpression of sucrose synthase in cotton plants.

[0122] The coding sequence of a potato sucrose synthase cDNA (Genbank Ogatori No. M18745) was operably linked to a ground clover dwarf virus promoter (S7; WO9606932) and a 3' Transcription termination signal and polyadenylation signal. The chimeric gene is operably linked to a selectable marker gene and introduced into a T-DNA vector. Cotton plants were transformed using the Agrobacterium-mediated transformation technique described above. Transgenic cotton lines were characterized and analyzed for sucrose synthase activity, fiber length, fluff fiber length, cellulose content, and lint dry weight. A certain positive correlation was found between SuSy activity and increases in fiber length, cellulose content and lint dry weight.

[0123] Summary of Examples

[0124] Unicellular cotton fibers arise from the ovule epithelium at anthesis, extend to 2.5-3.0 cm in about 16 days and subsequently synthesize la...

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Abstract

Methods and means are provided to modulate fibre quality in fibre-producing plants, such as cotton by modulating sucrose synthase activity and / or expression in such plants. The methods and means may also be used to obtain plants with seedless fruits or male-sterile plants.

Description

technical field [0001] The present invention relates to modifying the expression of a target gene in a plant to obtain a desired effect in the plant. Background technique [0002] Sucrose synthase (SuSy) is a key enzyme in the degradation of sucrose in all plant sink tissues, including cereals and fruits, and has been extensively studied in a variety of plants. However, the protein and gene have only recently been identified from cotton. The full-length (2625bp) cotton sucrose synthase (SuSy) was isolated by Perez-Grau, L and Delmer, D at the University of California, Davis in May 1996 (acquisition number U73588). A 2030 bp fragment of the same cDNA with a 595 bp deletion at the 5' end was isolated by the same group in 1994 and provided to Prem Chourey of USDA / ARS for collaborative research. However, at the time there was no evidence implicating the role of the SuSy gene in cotton fiber / seed development, although it was speculated (Amor et al., 1995) that part of SuSy loca...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/29C12N15/54A01H5/00C12N9/10C12N15/82
CPCC12N9/1062C12N15/8261C12N15/8246Y02A40/146
Inventor D·维林R·福尔班克Y·-L·阮
Owner COMMONWEALTH SCI & IND RES ORG