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Stage and tissue-specific modulation of ten conserved miRNAs and their targets during somatic embryogenesis of Valencia sweet orange

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Abstract

Somatic embryogenesis (SE) is a remarkable process of plant somatic cells developing into an embryo capable of forming a complete plant. MiRNAs play important roles in plant development by regulating expression of their target genes, but its function in SE has rarely been studied. Herein, ten conserved miRNAs with critical functions in plant development are detected by stem-loop qRT-PCR in the SE system of Valencia sweet orange. Sixteen unigenes from citrus are predicted to be targeted by six of the miRNAs. Cleavage sites on 15 of these target mRNAs are mapped by 5′RACE, of which ten are reported in this study. Transcript abundances of the 16 target unigenes are detected by qRT-PCR during SE process. Stage and tissue-specific expressions of miRNAs and their targets suggest their possible modulation on SE of citrus callus: miR156, 168 and 171 exert regulatory function during somatic embryo induction process; miR159, 164, 390 and 397 are related to globular-shaped embryo formation; miR166, 167 and 398 are required for cotyledon-shaped embryo morphogenesis; in addition, target genes of miR164, 166 and 397 are associated with SE disability of nonembryogenic callus. Exploration of miRNA-mediated modulation on SE is expected to provide new insights into plant cell totipotency, as well as how to maintain and enhance the embryogenic capacity of somatic cells.

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Abbreviations

miRNA:

MicroRNA

SE:

Somatic embryogenesis

NEC:

Nonembryogenic callus

EC:

Embryogenic callus

E1/2/3/4:

Embryogenic callus induced for 1/2/3/4 weeks

GE:

Globular-shaped somatic embryo

CE:

Cotyledon-shaped somatic embryo

qRT-PCR:

Quantitative reverse transcription-polymerase chain reaction

5′RACE:

5′ Rapid amplification of cDNA ends

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Acknowledgments

This research was financially supported by the National Basic Research Program of China (2011CB100600), the National Natural Science Foundation (No. 30921002), and the Hubei provincial Natural Science Foundation (No. 2008CDA069).

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Authors and Affiliations

  1. Key Laboratory of Horticultural Plant Biology (Ministry of Education), National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan, 430070, China

    Xiao-Meng Wu, Mei-Ya Liu, Xiao-Xia Ge, Qiang Xu & Wen-Wu Guo

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  1. Xiao-Meng Wu
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  2. Mei-Ya Liu
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  3. Xiao-Xia Ge
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Correspondence to Wen-Wu Guo.

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Wu, XM., Liu, MY., Ge, XX. et al. Stage and tissue-specific modulation of ten conserved miRNAs and their targets during somatic embryogenesis of Valencia sweet orange. Planta 233, 495–505 (2011). https://doi.org/10.1007/s00425-010-1312-9

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