Plant Molecular Biology

, Volume 77, Issue 6, pp 619–629 | Cite as

Regulation of auxin response by miR393-targeted transport inhibitor response protein 1 is involved in normal development in Arabidopsis

  • Zhe-Hao Chen
  • Mao-Lin Bao
  • Yu-Zhe Sun
  • Yan-Jun Yang
  • Xiao-Hong Xu
  • Jun-Hui Wang
  • Ning Han
  • Hong-Wu Bian
  • Mu-Yuan Zhu


miR393, which is encoded by MIR393a and MIR393b in Arabidopsis, post-transcriptionally regulates mRNAs for the F-box auxin receptors TIR1 (Transport Inhibitor Response Protein 1), AFB1 (Auxin Signaling F-box Protein 1), AFB2 and AFB3. However, biological functions of the miR393-TIR1/AFBs module in auxin response and plant development is not fully understood. In the study herein, we demonstrate that miR393 accumulated in response to exogenous IAA treatment, and its induction was due to enhanced MIR393b transcription but not MIR393a. Overexpression of a miR393-resistant form of TIR1 (mTIR1) enhanced auxin sensitivity and led to pleiotropic effects on plant development including inhibition of primary root growth, overproduction of lateral roots, altered leave phenotype and delayed flowering. Furthermore, miR393 level was increased in 35S:mTIR1 plant, suggesting that TIR1 promoted the expression of miR393 by a feedback loop. The interaction between miR393 and its target indicates a fine adjustment to the roles of the miR393-TIR1 module, which is required for auxin responses in plant development.


microRNAs miR393 Transport inhibitor response 1 (TIR1Auxin response Development Arabidopsis thaliana 



We sincerely thank Dr. Mark Estelle (Section of Cell and Developmental Biology, UCSD) for kindly providing the tir1-1 and tir1-1 afb1-1 afb2-1 afb3-1 quadruple mutants and Dr. C-Y Huang (The University of Adelaide, Australia) for helpful discussions and critical reading the manuscript. This work was supported by the National Science Foundation of China (Grant No. 30571197, No. 30972016), National High Technology Research and Development Program of China (863 Program) (No. 2007AA10Z141) and Zhejiang Provincial Natural Science Foundation of China (Grant No. Y3080323).

Supplementary material

11103_2011_9838_MOESM1_ESM.tif (1.9 mb)
Supplementary material 1 (TIFF 1991 kb). Supplemental Figure 1. Determination of endogenous TIR1 in wild-type, 35S:mTIR1 and 35S:TIR1 transgenic lines. Endogenous and miR393-resistant TIR1 (mTIR1) transcripts were amplified by RT-PCR and distinguished by digestion with the restriction enzyme NsbI, which cuts only mTIR1
11103_2011_9838_MOESM2_ESM.tif (3.2 mb)
Supplementary material 2 (TIFF 3279 kb). Supplemental Figure 2. Rosette leaves of 4-week-old 35S:MIR393 plants. a The first leaf with serrate margins was slightly delayed in 35S:MIR393 plants. Leaves from 4-week-old wild-type, 35S:MIR393a and 35S:MIR393b transgenic plants were cut and arranged in the order of appearance. Scale bar = 1 cm. b Rosette leaves of 4-week-old 35S:MIR393 plants displayed no remarkable differences in size or numbers. Wild-type, 35S:MIR393a and 35S:MIR393b transgenic plants grown under short-day conditions were photographed with side view (left) and top view (right)
11103_2011_9838_MOESM3_ESM.tif (2 mb)
Supplementary material 3 (TIFF 2053 kb). Supplemental Figure 3. Relative expression levels of TIR1 in Col-0 and tir1-1 plants. TIR1 level in tir1-1 plants was not significantly different from the level in wild-type. Relative transcript levels were normalized using UBQ5 as a standard. The relative expression levels in wild-type plants were set to 1.0. Error bars represent SD from three PCR results, and similar results were obtained in three independent experiments
11103_2011_9838_MOESM4_ESM.tif (3.3 mb)
Supplementary material 4 (TIFF 3373 kb). Supplemental Figure 4. pTIR1:mTIR1-tir1-1 plants display pleiotropic effects similar to 35S:mTIR1. a 5-day-old pTIR1:mTIR1-tir1-1 transgenic plants displayed downward cotyledons and upward petioles. b 10-day-old pTIR1:mTIR1-tir1-1 plants displayed downward curly true leaves with long and twisted petioles. cd The narrow and bended rosette leaves from 4-week-old pTIR1:mTIR1-tir1-1 were much smaller in size and fewer in numbers. Transgenic plants grown under short-day conditions were photographed with side view (c) and top view (d)
11103_2011_9838_MOESM5_ESM.doc (52 kb)
Supplementary material 5 (DOC 55 kb)
11103_2011_9838_MOESM6_ESM.doc (46 kb)
Supplementary material 6 (DOC 51 kb)
11103_2011_9838_MOESM7_ESM.xls (688 kb)
Supplementary material 7 (XLS 688 kb)


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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Zhe-Hao Chen
    • 1
  • Mao-Lin Bao
    • 1
  • Yu-Zhe Sun
    • 1
  • Yan-Jun Yang
    • 1
  • Xiao-Hong Xu
    • 1
  • Jun-Hui Wang
    • 1
  • Ning Han
    • 1
  • Hong-Wu Bian
    • 1
  • Mu-Yuan Zhu
    • 1
  1. 1.State Key Laboratory of Plant Physiology and Biochemistry, College of Life SciencesZhejiang UniversityHangzhouChina

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