Cloning and Expression Analysis of a Cupro-Zinc Superoxide Dismutase Gene from Wheat Photo-thermosensitive Genic Male Sterile Line BS366
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    Abstract:

    Superoxide dismutase (SOD) is a major antioxidant enzymes, which is important for plants to cope with adversity stress and anti-aging effect. A cDNA of Cu/Zn-SOD gene defining by the EST sequence assembly from Microarray data, was cloned from the photo-thermosensitive male sterile wheat (PTGMS) line BS366. The length of ORF sequence is 495bp, which encoded a 164-amino-acid polypeptide. Protein sequence contained a typical Cu/Zn-SOD gene family functional domain and 3 dimensional structures. Besides, the protein was located in the cytoplasm by ubcellular localization analysis. Evolution and clustering analysis showed that the Cu/Zn-SOD protein of wheat shared 89% and 94% sequence similarities with the Cu/Zn-SOD protein of Brachypodium distachyon and Hordeum vulgare, respectively. The methods of quantitative real-time PCR (qRT-PCR) were utilized to analyze the expression patterns in different wheat tissues and abiotic stresses. It was showed that the Cu/Zn-SOD gene expressed in root, stem, leaf, pistil, stamen and glume, belonging to constitutive expression. The highest expression level was found in chlorenchyma such as leaf, stem and glume. In addition, the gene expression was induced by a variety of stressesrevealing that the Cu/Zn-SOD gene of wheat might involve in multiple stress regulation pathway. To illuminate the regulation of Cu/Zn-SOD in wheat fertility, expression profile of Cu/Zn-SOD in PTGMS line BS366 pollen during fertility transformation was performed. The results showed that the relative expression of Cu/Zn-SOD at microsporocyte stage and meiosis stage in fertile environment were 8 times and 16 times higher than control, respectively, while there was no significant changes in sterility environment, suggesting that Cu/Zn-SOD might be involved in the regulation of fertility transformation of wheat PTGMS line BS366. This information lays a foundation for further functional study of Cu/Zn-SOD in wheat.

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History
  • Received:January 02,2017
  • Revised:March 02,2017
  • Adopted:April 11,2017
  • Online: September 14,2017
  • Published:
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