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Functional and Metabolic Characterization of a Stilbene Synthase Gene (Sbsts1) from Sorghum: (English)

Functional and Metabolic Characterization of a Stilbene Synthase Gene (Sbsts1) from Sorghum: (English)

          
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About the Book

This dissertation, "Functional and Metabolic Characterization of a Stilbene Synthase Gene (SbSTS1) From Sorghum" by Ka-yin, Yu, 余家燕, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any way. We have altered the formatting in order to facilitate the ease of printing and reading of the dissertation. All rights not granted by the above license are retained by the author. Abstract: Abstract of thesis entitled FUNCTIONAL AND METABOLIC CHARACTERIZATION OF A STILBENE SYNTHASE GENE (SbSTS1) FROM SORGHUM submitted by YU Ka Yin for the Degree of Doctor of Philosophy at The University of Hong Kong in May 2007 A chalcone synthase-like gene, SbCHS8, with high expressed sequence tag abundance in a pathogen-induced cDNA library was identified previously in sorghum. Genomic Southern blot analysis revealed that SbCHS8 represents a single copy gene. The expression of SbCHS8 was induced in sorghum mesocotyls following inoculation with Cochliobolus heterotrophus and Colletotrichum sublineolum, which are non-pathogenic and pathogenic to sorghum, respectively. However, the induction 2 was delayed by approximately 24 h when compared to the expression of other SbCHS genes. In addition, SbCHS8 expression was not induced by light and did not occur in a tissue-specific manner. To elucidate their biochemical functions, recombinant SbCHS2 and SbCHS8 proteins were expressed in Escherichia coli and assayed with different phenylpropanoid-CoA esters. Chalcones and stilbenes were detected as the reaction products of SbCHS2 and SbCHS8, respectively. In addition, SbCHS2 and SbCHS8 were over-expressed in transgenic Arabidopsis tt4 mutants defective in CHS activities. SbCHS2 rescued the ability of these mutants to accumulate flavonoids in seed coats and seedlings. In contrast, SbCHS8 failed to complement the tt mutation. Results from HPLC-UV and LC-ESI-MS/MS analyses demonstrated that cis-piceid (a resveratrol glucoside) accumulated as the predominant stilbene metabolite in transgenic Arabidopsis lines overexpressing SbCHS8. Up to 580 μg/g FW of cis-piceid were detected in 2-week-old seedlings, potentially serving as a convenient source of the cis-stilbene isomer (which is less common) for pharmacological investigations. TM A novel, specific and sensitive MS/MS method using the QTRAP instrument has also been developed to identify stilbene-related metabolites in both transgenic Arabidopsis plants and infected sorghum seedlings. The results demonstrated the 3 presence of three additional resveratrol derivatives including resveratrol diglucoside, trans- and cis-resveratrol acetylhexoside in the transgenic Arabidopsis lines. On the other hand, trans-piceid accumulated as the major stilbene in infected sorghum, which may serve as an additional phytoalexin distinct from the well-characterized 3-deoxyanthocyainidins. In addition, trace amounts of trans-resveratrol, resveratrol acetylhexoside and cis-piceid have been identified in infected sorghum. In conclusion, SbCHS8, now re-termed SbSTS1, encodes an enzyme with stilbene synthase activity. Accumulations of resveratrol-related metabolites were detected in both transgenic Arabidopsis plants overexpressing SbSTS1 and sorghum seedlings infected with fungal pathogens. DOI: 10.5353/th_b3868940 Subjects: Sorghum - Genetics Plant genetics - Technique


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Product Details
  • ISBN-13: 9781361422403
  • Publisher: Open Dissertation Press
  • Publisher Imprint: Open Dissertation Press
  • Height: 279 mm
  • No of Pages: 178
  • Spine Width: 10 mm
  • Width: 216 mm
  • ISBN-10: 1361422408
  • Publisher Date: 27 Jan 2017
  • Binding: Paperback
  • Language: English
  • Series Title: English
  • Weight: 426 gr


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