Expression analysis of the fatty acid desaturase 2-4 and 2-3 genes from Gossypium hirsutum in transformed yeast cells and transgenic Arabidopsis plants. Metadata

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Title

  • Main Title Expression analysis of the fatty acid desaturase 2-4 and 2-3 genes from Gossypium hirsutum in transformed yeast cells and transgenic Arabidopsis plants.

Creator

  • Author: Zhang, Daiyuan
    Creator Type: Personal

Contributor

  • Chair: Pirtle, Robert M.
    Contributor Type: Personal
    Contributor Info: Major Professor
  • Committee Member: Ayre, Brian G.
    Contributor Type: Personal
  • Committee Member: Chapman, Kent D.
    Contributor Type: Personal
  • Committee Member: Conrad-Webb, Heather
    Contributor Type: Personal
  • Committee Member: Root, Douglas D.
    Contributor Type: Personal

Publisher

  • Name: University of North Texas
    Place of Publication: Denton, Texas

Date

  • Creation: 2008-08
  • Digitized: 2009-04-16

Language

  • English

Description

  • Content Description: Fatty acid desaturase 2 (FAD2) enzymes are phosphatidylcholine desaturases occurring as integral membrane proteins in the endoplasmic reticulum membrane and convert monounsaturated oleic acid into polyunsaturated linoleic acid. The major objective of this research was to study the expression and function of two cotton FAD2 genes (the FAD2-3 and FAD2-4 genes) and their possible role in plant sensitivity to environmental stress, since plants may increase the polyunsaturated phospholipids in membranes under environmental stress events, such as low temperature and osmotic stress. Two FAD2 cDNA clones corresponding to the two FAD2 genes have been isolated from a cotton cDNA library, indicating both genes are truly expressed in cotton. Model yeast cells transformed with two cotton FAD2 genes were used to study the chilling sensitivity, ethanol tolerance, and growth rate of yeast cells. The expression patterns of the two FAD2 genes were analyzed by reverse transcription polymerase chain reactions (RT-PCR) and Western blot analyses in cotton plants under different treatment conditions. The coding regions of both FAD2 genes were inserted downstream from the CaMV 35S promoter in the pMDC gateway binary vector system. Five different FAD2/pMDC constructs were transformed into the Arabidopsis fad2 knockout mutant background, and multiple potential transgenic Arabidopsis plant lines harboring the cotton FAD2 genes were generated. The cotton FAD2 genes were amplified by the polymerase chain reaction (PCR) from the genomic DNAs isolated from the transgenic Arabidopsis T1 plant lines. Complementation of the putative transgenic Arabidopsis plants with the two cotton FAD2 genes was demonstrated by gas chromatography analyses of the fatty acid profiles of leaf tissues. The cellular localization of cotton FAD2-4 polypeptides with N-terminal green fluorescence protein (GFP) was visualized by confocal fluorescence microscopy. The phenotype of transgenic Arabidopsis plants transformed with the cotton FAD2-4 gene was compared to Arabidopsis knockout fad2 mutant plants and wild type Arabidopsis plants regarding their sensitivity to low temperature, and the size and height of the plants.

Subject

  • Keyword: Fatty acid desaturase
  • Keyword: cotton
  • Keyword: Arabidopsis
  • Keyword: yeast
  • Library of Congress Subject Headings: Cotton -- Genetics.
  • Library of Congress Subject Headings: Unsaturated fatty acids.
  • Library of Congress Subject Headings: Yeast fungi.
  • Library of Congress Subject Headings: Arabidopsis.

Collection

  • Name: UNT Theses and Dissertations
    Code: UNTETD

Institution

  • Name: UNT Libraries
    Code: UNT

Rights

  • Rights Access: public
  • Rights License: copyright
  • Rights Holder: Zhang, Daiyuan
  • Rights Statement: Copyright is held by the author, unless otherwise noted. All rights reserved.

Resource Type

  • Thesis or Dissertation

Format

  • Text

Identifier

  • OCLC: 369166765
  • Archival Resource Key: ark:/67531/metadc9002

Degree

  • Degree Name: Doctor of Philosophy
  • Degree Level: Doctoral
  • Degree Discipline: Molecular Biology
  • Academic Department: Department of Biological Sciences
  • Degree Grantor: University of North Texas

Note

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