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RNAi mediated inhibition of viroid infection in transgenic plants expressing viroid-specific small RNAs derived from various functional domains.

Abstract
Previous attempts to develop RNAi-mediated viroid-resistant transgenic plants using nearly full-length Potato spindle tuber viroid (PSTVd) hairpin RNA (hpRNA) were successful; however unusual phenotypes resembling viroid infection occurred. Therefore, in the present work, transgenic Nicotiana benthamiana lines expressing both partial and truncated versions of PSTVd hpRNA were developed. Specifically, seven partial or truncated versions of PSTVd sequences were selected according to the hotspots of both PSTVd-sRNAs and functional domains of the PSTVd. A total of 21 transgenic lines Nicotiana benthamiana were developed under the control of either the CaMV-35S or the CoYMV promoters. All of the transgenic lines established here were monitored for the induction of phenotypic changes, for PSTVd-sRNA expression and for the resistance against PSTVd infection. Additionally, this study demonstrates the use of inverted repeat construct sequences as short as 26- to -49 nucleotides for both the efficient expression of the PSTVd-sRNA and the inhibition of PSTVd infection.
AuthorsCharith Raj Adkar-Purushothama, Atsushi Kasai, Kohei Sugawara, Hideki Yamamoto, Yuto Yamazaki, Ying-Hong He, Nobuyuki Takada, Hideki Goto, Sahori Shindo, Takeo Harada, Teruo Sano
JournalScientific reports (Sci Rep) Vol. 5 Pg. 17949 (Dec 14 2015) ISSN: 2045-2322 [Electronic] England
PMID26656294 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • RNA, Small Interfering
  • RNA, Viral
Topics
  • Base Sequence
  • Binding Sites
  • Disease Resistance (genetics)
  • Gene Expression
  • Inverted Repeat Sequences
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • Plant Diseases (genetics, virology)
  • Plant Viruses (physiology)
  • Plants, Genetically Modified (genetics)
  • Promoter Regions, Genetic
  • RNA Interference
  • RNA, Small Interfering (chemistry, genetics)
  • RNA, Viral (chemistry, genetics)
  • Viroids (physiology)

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