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Cloning and characterization of four differentially expressed cDNAs encoding NFYA homologs involved in responses to ABA during somatic embryogenesis in Japanese larch (Larix leptolepis)

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Abstract

In this study, four nuclear transcription factor Y subunit alpha (NFYA) homologous genes, LaNFYA1, LaNFYA2, LaNFYA3, and LaNFYA4, were isolated from Japanese larch (Larix leptolepis). The deduced amino acid sequences of the LaNFYAs harbored a highly conserved core region consisting of two functional domains: a subunit association domain and a DNA-binding homeodomain. Further analysis showed that miR169 target sequences were present in the 3′ end of all four LaNFYA mRNA sequences. A transient expression analysis of the LaNFYA-GFP fusion proteins in tobacco leaf epidermal cells revealed that the LaNFYA proteins were localized in the nucleus. An analysis of tissue expression levels of the LaNFYAs showed that expression was highest in the stems and lowest in the needles, demonstrating a pattern of temporal- and spatial-specific expression. Their mRNA accumulation patterns during somatic embryogenesis, and the effects of abscisic acid (ABA) on their transcript levels and somatic embryo maturation were investigated. The results showed that the transcripts of the four genes gradually increased and peaked at 7 days, followed by a decline as the somatic embryos matured, and that the removal of the ABA treatment did not affect the mRNA abundance of LaNFYA1, LaNFYA2, and LaNFYA3 during the first 14 days, but up-regulated the abundance of LaNFYA4 mRNA. We concluded that LaNFYAs may be involved with the early determination and maturation of the somatic embryo, which is regulated by responses to ABA and miR169. These results provide new insights into the regulatory mechanisms of somatic embryogenesis.

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Acknowledgments

This work was supported by the National Natural Science Foundation of China (31330017) and the National High Technology Research and Development Program of China (2013AA102704, 2011AA100203).

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Authors and Affiliations

  1. Laboratory of Cell Biology, Research Institute of Forestry, Chinese Academy of Forestry, Beijing, 100091, China

    Li-feng Zhang, Wan-feng Li, Hai-yan Xu & Li-wang Qi

  2. Research Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry, Beijing, 100091, China

    Su-ying Han

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  1. Li-feng Zhang
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Correspondence to Su-ying Han.

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Zhang, Lf., Li, Wf., Xu, Hy. et al. Cloning and characterization of four differentially expressed cDNAs encoding NFYA homologs involved in responses to ABA during somatic embryogenesis in Japanese larch (Larix leptolepis). Plant Cell Tiss Organ Cult 117, 293–304 (2014). https://doi.org/10.1007/s11240-014-0440-5

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