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Wheat microRNA Member TaMIR444a Is Nitrogen Deprivation-Responsive and Involves Plant Adaptation to the Nitrogen-Starvation Stress

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Abstract

microRNAs (miRNAs) are involved in regulating various plant developmental processes and mediating plant-adaptive responses to nutrient deprivation. In this study, the characterization of a wheat miRNA member TaMIR444a and the role of this miRNA in mediating plant tolerance to the N-starvation stress were investigated. Results indicated that the expression levels of TaMIR444a and NtMIR444a, the homologue of TaMIR444a in tobacco, were upregulated in roots and leaves under N deprivation, whereas the transcription of their target genes showed reverse expression patterns in above tissues. These results suggest that miR444a is conserved across plant species of dicots and monocots and can possibly establish the miRNA/target modules for mediating plant response to N deficiency. Overexpression of TaMIR444a in tobacco improved the plant growth feature, biomass, N content, photosynthetic parameters, and antioxidant enzymatic activities under N deprivation. Based on microarray analyses, a large number of genes were identified to be differentially expressed in the TaMIR444a-overexpressing plants; these differential genes are categorized into functional groups of signal perception and transduction, transcription regulation, primary and secondary metabolism, phytohormone response, cellular protection and defensive responsiveness, etc. qPCR analyses revealed that the nitrate transporter (NRT) genes NtNRT1.1-s, NtNET1.1-t, and NtNRT2.1 and the antioxidant enzyme genes (AEEs) NtCAT1;1, NtPOD1;3, and NtPOD4 were significantly upregulated by TaMIR444a, suggesting that the altered transcription of these NRT and AEE genes is associated with the improvement of the N acquisition and the cellular ROS detoxification in the N-deprived transgenic plants. Together, our findings demonstrate that miR444a acts as one critical regulator in mediating plant tolerance to the N-starvation stress through modulation of the regulatory networks associated with N acquisition, cellular ROS homeostasis, and carbon assimilation. Our findings have provided insights into the mechanisms of plant tolerance to N deficiency mediated by the distinct miRNA pathways.

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    Authors and Affiliations

    1. College of Agronomy, Agricultural University of Hebei, Baoding, 071001, China

      Si Gao, Chengjin Guo, Yongsheng Zhang, Feifei Zhang & Kai Xiao

    2. College of Life Sciences, Agricultural University of Hebei, Baoding, 071001, China

      Xiaoming Du & Juntao Gu

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    1. Si Gao
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    2. Chengjin Guo
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    3. Yongsheng Zhang
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    4. Feifei Zhang
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    5. Xiaoming Du
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    7. Kai Xiao
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    Corresponding author

    Correspondence to Kai Xiao.

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    The authors declare that they have no conflict of interest.

    Funding

    This study was funded by the National Natural Science Foundation of China (nos. 31371618 and 31571664), the National Transgenic Major Program of China (no. 2011ZX08008), and the Key Laboratory of Crop Growth Regulation of Hebei Province.

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    Si Gao and Chengjin Guo contributed equally to this work.

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    Fig. S1

    Alignment of precursor sequences of TaMIR444a and its tobacco homologue NtMIR444a (DOC 120 kb)

    Fig. S2

    Expression patterns of TaMIR444 and NtMIR444a in roots and leaves of wheat and tobacco under the N-sufficient and -deficient conditions detected by semiquantitative RT-PCR (DOC 225 kb)

    Fig. S3

    Expression patterns of TaMIR444a and NtMIR444a in response to longer N deprivation treatment (DOC 28 kb)

    Fig. S4

    Molecular characterization of the transgenic tobacco plants (DOC 146 kb)

    Fig. S5

    Expression patterns of the randomly selected upregulated and downregulated genes detected by microarray analyses (DOC 880 kb)

    Table S1

    Primers used for gene expression analysis of TaMIR444a and NtMIR444a, their interacting target genes, the internal standard tubulin genes as well as for the selection gene bar (DOC 37 kb)

    Table S2

    Primers used for gene expression analysis of the randomly selected differential genes in the microarray analysis (DOC 44 kb)

    Table S3

    Primers used for expression analyses of the tobacco NRT, SOD, CAT, and POD genes (DOC 48 kb)

    Table S4

    The differentially upregulated genes by TaMIR444a (XLS 286 kb)

    Table S5

    The differentially downregulated genes by TaMIR444a (XLS 291 kb)

    Table S6

    Functional groups of the differentially upregulated genes by TaMIR444a (XLS 58 kb)

    Table S7

    Functional groups of the differentially downregulated genes by TaMIR444a (XLS 58 kb)

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    Gao, S., Guo, C., Zhang, Y. et al. Wheat microRNA Member TaMIR444a Is Nitrogen Deprivation-Responsive and Involves Plant Adaptation to the Nitrogen-Starvation Stress. Plant Mol Biol Rep 34, 931–946 (2016). https://doi.org/10.1007/s11105-016-0973-3

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