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RNA interference-mediated knock-down of α1,3-fucose and β1,2-xylose in rice N-glycans

  • Joon-Soo Sim
  • Yeon-Hee Lee
  • Ha Young Chung
  • Yong-Hwan Kim
  • Bum-Soo HahnEmail author
SHORT COMMUNICATION

Abstract

RNA interference (RNAi) was performed for the down-regulated expression of two endogeneous core α1,3-fucosyltransferase (FucT) and β1,2-xylosyltransferase (XylT) genes in rice (Oryza sativa). We constructed three RNAi vectors for the suppression of two glycosyltransferase transcriptions and transformed them into the rice plants by Agrobacterium. The transgenic plants were selected by PCR and down-regulated expressions of FucT and XylT in the mRNA level were verified by RT-PCR and qRT-PCR. In addition, structural properties of N-glycans from glycoproteins in transgenic rice leaves were analyzed by MALDI-TOF MS and were compared with those of wild-type rice leaves. The compositional change of plant-specific N-glycans in total endogenous rice leaf glycoproteins was induced as consequence of the knock-down of two N-glycan maturation glycosyltransferases by using RNAi. Compositional change of N-glycans in three selected RNAi suppression plants was observed to decrease the ratio of three major hybrid, hybrid bisected and paucimanosidic-type N-glycans: GlcNAc3Man3Xyl1Fuc1, GlcNAc4Man3Xyl1-Fuc1 and GlcNAc2Man3Xyl1Fuc1.

Keywords

Rice N-glycan β1,2-xylosyltransferase α1,3-fucosyltransferase RNA interference 

Abbreviations

PCR

Polymerase chain reaction

mRNA

Messenger RNA

RT

Reverse transcription

qRT-PCR

Quantitative real-time PCR

MALDI-TOF

Matrix assisted laser desorption ionization-time of flight mass spectrometry

GlcNAc

N-Acetylglucosamine

Man

Mannose

Xyl

Xylose

Fuc

Fucose

cDNA

Complementary DNA

NCBI

National center for biotechnology information

Blast

Basic local alignment search tool

Notes

Acknowledgments

This project was supported by grant (to B.S.H.) from National Academy of Agricultural Science (PJ008541). This study was supported by 2014 Postdoctoral Fellowship Program (to H.Y.C.) of National Academy of Agricultural Science, RDA, Korea.

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Copyright information

© Society for Plant Biochemistry and Biotechnology 2014

Authors and Affiliations

  • Joon-Soo Sim
    • 1
  • Yeon-Hee Lee
    • 1
  • Ha Young Chung
    • 1
  • Yong-Hwan Kim
    • 1
    • 2
  • Bum-Soo Hahn
    • 1
    Email author
  1. 1.National Academy of Agricultural ScienceSuwonSouth Korea
  2. 2.Chonbuk National University R&D CenterJeonju-siSouth Korea

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