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Cotton fiber transcriptional factors

a transcription factor and cotton fiber technology, applied in the direction of lysine, peptide source, peptide, etc., can solve the problems of plant melanin production, fiber-tissue transcription initiation regions of this invention are typically not readily detectable in other plant tissues,

Inactive Publication Date: 2003-06-05
MONSANTO TECH LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Further, fiber-tissue transcription initiation regions of this invention are typically not readily detectable in other plant tissues.
Presumably, the plants were not able to produce melanin due to deficiency of the required substrates in the plant cell cytosol.

Method used

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  • Cotton fiber transcriptional factors
  • Cotton fiber transcriptional factors

Examples

Experimental program
Comparison scheme
Effect test

example 2

Isolation of cDNA Clones from Cotton

[0068] cDNA to the 4-4 clone was isolated from the cotton fiber library described above, and shown to express in fiber but not other tissues. This sequence was not related to any known protein. Only 400 kb of encoding sequence was present in this clone, so the library was rescreened using the cDNA to obtain full-length clones. The full-length encoding sequence is provided in FIG. 1.

[0069] Another clone was sequenced which showed high homology to animal Rac proteins. This clone, designated Rac13, was not quite full-length, and the library was re-screened using this initial Rac13 DNA segment as probe. Of approximately 130,000 primary plaques screened, 56 screened positive; of these, 14 clones were isolated and sequenced. Of these 14 clones, 12 showed identical sequence homology to the original Rac13 clone and one of these cDNA clones encoded a full length Rac13. One other partial-length cDNA clone, designated Rac9, was clearly related, but distinct ...

example 3

Expression of Cotton Fiber Genes in Developing Fibers

[0071] Expression of the Rac13 and 4-4 genes was assessed using mRNA prepared from various cotton tissues and from fibers at different stages of development. Blots were hybridized with probes derived from 3'-untranslated regions of either the Rac13 or 4-4 genes. The gene for Rac13 exhibits highly-enhanced expression in fibers; virtually no detectable mRNA is present in leaves, roots, or flower parts, even under conditions of extended development time. Rac13 expression is detected in seeds at an age that corresponds to the highest expression levels observed in fiber tissue derived from seeds of this same age. The pattern of Rac13 expression in fibers is very dependent upon the developmental stage. Expression is very low during the stage of primary wall synthesis (0-14 dpa, see Meinert and Delmer, 1977), reaches a maximum during the transition to secondary wall synthesis (about 15-18 dpa), and declining during the stage of maximal s...

example 4

Genomic DNA

[0072] cDNA for both the 4-4 and Rac13 was used to probe for genomic clones. For both, full length genomic DNA was obtained from a library made using the lambda dash 2 vector from Stratagene.TM., which was used to construct a genomic DNA library from cotton variety Coker 130 (Gossypium hirsutum cv. coker 130), using DNA obtained from germinating seedlings.

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Abstract

Novel DNA constructs are provided which may be used as molecular probes or inserted into a plant host to provide for modification of transcription of a DNA sequence of interest in cotton fiber, particularly in very early fiber development. The DNA constructs comprise a cotton fiber transcriptional initiation regulatory region associated with a gene which is expressed in cotton fiber.

Description

[0001] This invention relates to methods of using in vitro constructed DNA transcription or expression cassettes capable of directing fiber-tissue transcription of a DNA sequence of interest in plants to produce fiber cells having an altered phenotype, and to methods of providing for or modifying various characteristics of cotton fiber. The invention is exemplified by methods of using cotton fiber promoters for altering the a phenotype of cotton fiber, and cotton fibers produced by the method.[0002] In general, genetic engineering techniques have been directed to modifying the phenotype of individual prokaryotic and eukaryotic cells, especially in culture. Plant cells have proven more intransigent than other eukaryotic cells, due not only to a lack of suitable vector systems but also as a result of the different goals involved. For many applications, it is desirable to be able to control gene expression at a particular stage in the growth of a plant or in a particular plant part. Fo...

Claims

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Application Information

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IPC IPC(8): C07K14/36C07K14/415C12N9/02C12N9/88C12N15/82
CPCC07K14/36C07K14/415C12N9/0069C12N9/0071C12N15/8261C12N15/8222C12N15/8233C12N15/825C12N9/88Y02A40/146
Inventor MCBRIDE, KEVINSTALKER, DAVID M.PEAR, JULIE R.PEREZ-GRAU, LUIS
Owner MONSANTO TECH LLC
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