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Balanced activity of microRNA166/165 and its target transcripts from the class III homeodomain-leucine zipper family regulates root growth in Arabidopsis thaliana

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Abstract

Key message

Overexpression of miR166/165 down-regulates target HD - ZIP IIIs and promotes root growth by enhancing cell division and meristematic activity, whereas overexpression of HD - ZIP IIIs inhibits root growth in Arabidopsis thaliana.

Abstract

Post-embryonic growth of higher plants is maintained by active meristems harbouring undifferentiated cells. Shoot and root apical meristems (SAM and RAM) utilize both similar and distinct signalling mechanisms for their maintenance in Arabidopsis thaliana. An important regulatory role in this context has the interaction of microRNAs with their target mRNAs, mostly encoding transcription factors. One class of microRNA166/165 (miR166/165) has been implicated in the maintenance of SAM and vascular patterning. Here, we show that miR166/165 plays an important role in root growth also by negatively regulating its target transcripts, HD-ZIP IIIs, in the RAM. While overexpression of miR166 promotes RAM activity, overexpression of its targets reduces RAM activity. These results reveal a conserved role of miR166/165 in the maintenance of SAM and RAM activity in A. thaliana.

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Abbreviations

ARF:

AUXIN RESPONSE FACTOR

ATHB:

Arabidopsis thaliana HOMEOBOX

dag:

Days after germination

eTM:

Endogenous target mimic

HD-ZIP III:

CLASS III HOMEODOMAIN-LEUCINE ZIPPER

miR166/165 :

microRNA166/165

PHB:

PHABULOSA

PHV:

PHAVOLUTA

PLT:

PLETHORA

QC:

Quiescent centre

RAM:

Root apical meristem

REV:

REVOLUTA

SAM:

Shoot apical meristem

SCR:

SCARECROW

SHR:

SHORT ROOT

WOX5:

WUSCHEL-RELATED HOMEOBOX 5

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Acknowledgments

We thank DBT (Department of Biotechnology, Government of India) for a Ramalingaswami fellowship (BT/HRD/35/02/06/2008) grant to AKS, CSIR (Council for Scientific and Industrial Research, India) for a fellowship to AS and SS, and NIPGR (National Institute of Plant Genome Research, India) for internal funding and facility to carry out this work. We thank ABRC for Arabidopsis seeds and Prof. X. Wang (Institute of Genetics and Developmental Biology, China) for eTM-miR166.

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi, 110067, India

    Archita Singh, Sharmila Singh & Ananda K. Sarkar

  2. National Institute of Science Education and Research, Bhubaneswar, 751005, India

    Kishore C. S. Panigrahi

  3. Plant Biotechnology, Faculty of Biology, University of Freiburg, Schänzlestr 1, 79104, Freiburg, Germany

    Ralf Reski

  4. BIOSS, Centre for Biological Signalling Research, 79104, Freiburg, Germany

    Ralf Reski

  5. FRIAS, Freiburg Institute for Advanced Studies, 79104, Freiburg, Germany

    Ralf Reski

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  1. Archita Singh
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  2. Sharmila Singh
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  3. Kishore C. S. Panigrahi
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  4. Ralf Reski
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  5. Ananda K. Sarkar
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Corresponding authors

Correspondence to Ralf Reski or Ananda K. Sarkar.

Additional information

Communicated by P. Kumar.

Electronic supplementary material

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299_2014_1573_MOESM1_ESM.tif

Supplementary material 1 Suppl. Fig. 1 Down-regulation of HD-ZIP IIIs by miR166/165 overexpression enhances root growth in Arabidopsis. a Real-time quantitative RT-PCR analysis showing relative expression level of different HD-ZIP III in miR166-Oe (in comparison to wild type). b–c Confocal images of roots showing RAM size (the distance between two arrows) of miR166a-Oe #45 in comparison to wild type (n = 10 for each genotype). Quantification of RAM size in miR166a-Oe #45 and wild type (n = 6 or more for each genotype). e–f Confocal images of roots showing RAM size (the distance between two arrows) of miR166a-Oe #46 and wild type (n = 10 for each genotype). g Quantification of RAM size in miR166a-Oe #46 and wild type (n = 9 or more for each genotype). h–i Quantification of cortical cell number in RAM of miR166-Oe #45 (n = 9) and #46 (n = 9) root in comparison to wild type. ‘n’ represents number of plants. At least two biological replicates were used. Error bars represent SD (standard deviation). Asterisks indicate significant difference at P < 0.05 (ANOVA). Scale bars represent 100 μm (b, c, e and f) (TIFF 2107 kb)

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Singh, A., Singh, S., Panigrahi, K.C.S. et al. Balanced activity of microRNA166/165 and its target transcripts from the class III homeodomain-leucine zipper family regulates root growth in Arabidopsis thaliana . Plant Cell Rep 33, 945–953 (2014). https://doi.org/10.1007/s00299-014-1573-z

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