Planta

, Volume 232, Issue 6, pp 1281–1288

Specific down-regulation of PAL genes by artificial microRNAs in Populus trichocarpa

  • Rui Shi
  • Chenmin Yang
  • Shanfa Lu
  • Ronald Sederoff
  • Vincent L. Chiang
Original Paper

DOI: 10.1007/s00425-010-1253-3

Cite this article as:
Shi, R., Yang, C., Lu, S. et al. Planta (2010) 232: 1281. doi:10.1007/s00425-010-1253-3

Abstract

Artificial microRNAs (amiRNAs) are similar to microRNAs (miRNAs) in that they are able to reduce the abundance of specific transcripts in plants by RNA-Induced Silencing Complex (RISC)-mediated cleavage and degradation, but differ in that they are designed for specific targets. The long generation times of forest trees have limited the discovery of mutations by conventional genetics. AmiRNAs can create gene-specific transcript reduction in transgenic trees in a single generation and may have broad application for functional genomics of trees. In this paper, we describe the specific down-regulation of multiple genes in the phenylalanine ammonia-lyase (PAL) gene family of Populus trichocarpa using amiRNA sequences incorporated in a P. trichocarpa miRNA-producing precursor, ptc-MIR408. Two different amiRNA constructs were designed to specifically down-regulate two different subsets of PAL genes, revealing differential regulation within the gene family. Down-regulation of subset A (PAL2, PAL4 and PAL5) by amiRNA-palA led to an increase in transcript abundance of subset B (PAL1 and PAL3). The reciprocal effect was not observed.

Keywords

Artificial microRNA Phenylalanine ammonia-lyase Populus trichocarpa transformation Down-regulation PolyA tailing-based real time RT-PCR Quantitation 

Supplementary material

425_2010_1253_MOESM1_ESM.pdf (64 kb)
Supplementary material 1 (PDF 63 kb)

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Rui Shi
    • 1
  • Chenmin Yang
    • 1
  • Shanfa Lu
    • 1
    • 2
  • Ronald Sederoff
    • 1
  • Vincent L. Chiang
    • 1
    • 3
  1. 1.Forest Biotechnology Group, Department of Forestry and Environmental ResourcesNorth Carolina State UniversityRaleighUSA
  2. 2.Medicinal Plant Cultivation Research Center, Institute of Medicinal Plant DevelopmentChinese Academy of Medical Sciences and Peking Union Medical CollegeBeijingPeople’s Republic of China
  3. 3.Department of Forest BiomaterialsNorth Carolina State UniversityRaleighUSA

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