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Ectopic expression of the apple Md-miR172e gene alters flowering time and floral organ identity in Arabidopsis

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Abstract

The miRNA miR172 is involved in the regulation of flowering time and floral organ development by specifically restricting the transcripts of target gene APETELA2 (AP2) transcription factors. In our study, Md-miR172e and its target genes, MdAP2 and MdAP2-1 to MdAP2-7, were isolated from the apple cultivar Royal Gala (Malus × domestica). Phylogenetic tree analysis revealed that eight MdAP2 genes were similar to the AtAP2 subfamily and were putative targets of miR172. qRT-PCR and western blotting analyses indicated Md-miR172e regulation of the expression of the target gene MdAP2 at the translation level. Next, an over-expression construct 35S::Md-miR172e was generated and transformed into Arabidopsis. qRT-PCR showed that Md-miR72e expression and mature miR172e accumulation increased, and transgenic plants exhibited early flowering (20–30 days early in flowering) under long days and floral defects compared with wild-type. Taken together, these results suggest that miR172 and its target AP2-like genes are involved in flower developmental processes, particularly with regards to flowering time and floral organ development, and miR172 mediates a conserved regulatory pathway in apple and Arabidopsis.

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Abbreviations

ARF:

Auxin response factor

AP2:

APETELA2

SPL:

Squamosa promoter binding protein-like protein

RT:

Reverse transcriptase

EST:

Expressed sequence tag

CaMV:

Cauliflower mosaic virus

WT:

Wild type

DCL1:

DICER-LIKE 1

TOE1:

TARGET Of EAT 1

SMZ:

SCHLAFMUTZE

SNZ:

SCHNARCHZAPFEN

qRT-PCR:

Quantificational real-time polymerase chain reaction

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Acknowledgments

This work was supported by NSFC (31171946), PCSIRT (IRT1155) and 948 Project from Ministry of Agriculture of China (2011-G21).

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Authors and Affiliations

  1. National Key Laboratory of Crop Biology, Shandong Agricultural University, Tai-an, 271018, Shandong, China

    Qiang Zhao, Chao Sun, Dan-Dan Liu, Yu-Jin Hao & Chun-Xiang You

  2. National Research Center for Apple Engineering and Technology, Shandong Agricultural University, Tai-an, 271018, Shandong, China

    Qiang Zhao, Chao Sun, Dan-Dan Liu, Yu-Jin Hao & Chun-Xiang You

  3. College of Horticulture Science and Engineering, Shandong Agricultural University, Tai-an, 271018, Shandong, China

    Qiang Zhao, Chao Sun, Dan-Dan Liu, Yu-Jin Hao & Chun-Xiang You

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  1. Qiang Zhao
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  2. Chao Sun
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  3. Dan-Dan Liu
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  4. Yu-Jin Hao
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  5. Chun-Xiang You
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Correspondence to Chun-Xiang You.

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Zhao, Q., Sun, C., Liu, DD. et al. Ectopic expression of the apple Md-miR172e gene alters flowering time and floral organ identity in Arabidopsis . Plant Cell Tiss Organ Cult 123, 535–546 (2015). https://doi.org/10.1007/s11240-015-0857-5

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  • DOI: https://doi.org/10.1007/s11240-015-0857-5

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