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PtFCA from precocious trifoliate orange is regulated by alternative splicing and affects flowering time and root development in transgenic Arabidopsis

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Abstract

The transition to flowering is a major developmental switch in flowering plants. The nuclear RNA-binding protein FCA responds to seasonal signals and abscisic acid (ABA), which can control the flowering time via ambient temperature and autonomous pathways. Citrus FCA ortholog (PtFCA) has been isolated and characterized from precocious trifoliate orange (Poncirus trifoliata L. Raf). Three alternatively spliced transcripts of PtFCA (PtFCA1, PtFCA2, and PtFCA3) were isolated. The expression pattern of PtFCA indicated that it may be involved in phase transition in precocious trifoliate orange. A functional complementation experiment of PtFCA indicated that PtFCA1 partially rescued the late-flowering phenotype of the fca-1 mutant. There was no influence on flowering time of transgenic Arabidopsis by PtFCA3 as compared with PtFCA2, which exhibits delayed flowering time in a fca-1 background. Meanwhile, these three transcripts also showed different abilities to regulate root development in the fca-1 background. The study of protein–protein interactions suggested that PtFCA may form higher order complexes with PtFY and PtNF-YA7 to regulate timing of the transition from the vegetative to reproductive phase in precocious trifoliate orange. ABA and ambient temperature treatments changed the expression of PtFCA and interaction protein. These findings reveal that PtFCA may play important roles in flowering time and root development of precocious trifoliate orange through the formation of multiple protein complexes.

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Acknowledgments

We are grateful to Prof. Jia-Ling Yao for her helpful discussion in this manuscript. This research was supported financially by the National Natural Science Foundation of China (grant nos. 31130046, 31471863, 31360469, 31372046, and 31521092) and Fundamental Research Funds for the Central Universities (Program No. 2662016PY037).

Author contributions

X-YA carried out the yeast two-hybrid, bimolecular fluorescence complementation, and PCR experiments; X-YA and T-JL prepared the plant material and obtained the transgenic plants; J-ZZ and C-GH designed the experiments and the study; X-YA and J-ZZ wrote the paper. All authors discussed the data obtained. All authors reviewed and provided comments upon preparation of the manuscript.

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  1. Xiao-Yan Ai

    Present address: Institute of Pomology and Tea, Hubei Academy of Agricultural Sciences, Wuhan, 430064, China

Authors and Affiliations

  1. Key Laboratory of Horticultural Plant Biology (Ministry of Education), College of Horticulture and Forestry Science, Huazhong Agricultural University, Wuhan, 430070, China

    Xiao-Yan Ai, Jin-Zhi Zhang, Tian-Jia Liu & Chun -Gen Hu

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  1. Xiao-Yan Ai
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Correspondence to Chun -Gen Hu.

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The authors declare that they have no competing interests.

Data archiving statement

The PtFCA1, PtFCA2, and PtFCA3 have been deposited in GenBank under accession nos. KX440390, KX440391, and KX440392, respectively.

Additional information

Communicated by W.-W. Guo

Xiao-Yan Ai and Jin-Zhi Zhang contributed equally to this work.

Electronic supplementary material

Figure S1

Gene sequences, structures, and phylogenetic relationship of PtFCA. A: Sequence alignment between PtFCA2 and PtFCA3. The gray box represents the sequence differences between PtFCA2 and PtFCA3. B: Alternatively spliced transcripts were validated in precocious trifoliate orange by RT-PCR, M: Marker (DL2000). Lanes 1–4: PtFCA1 in trifoliate orange, 5–7: PtFCA2/3 in precocious trifoliate orange. C: Phylogenic tree of FCA showing evolutionary relationships. The sequences with the highest percent sequence similarity are grouped together. The accession numbers of FCA proteins used in this analysis are HvFCA (FJ188402.2), VvFCA (GU300763.1), RcFCA (XM_002519206.1), GmFCA-like (XM_003529468.1), PsFCAgamma (AY805329.1), MtFCA (XM_003609755.1), FCAalpha (Z82993.1), FCAbeta (Z82991.1), FCAdelta (Z82990.1), FCAgamma (Z82989.1), OsFCA-1 (AY274928.1), OsFCA-2 (AY311344.1), OsFCA-3 (AY311343.1), OsFCA-4 (AY331574.1). (JPEG 4617 kb)

Table S1

List of primer sequences used in this study. (XLS 25 kb)

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Ai, XY., Zhang, JZ., Liu, TJ. et al. PtFCA from precocious trifoliate orange is regulated by alternative splicing and affects flowering time and root development in transgenic Arabidopsis . Tree Genetics & Genomes 12, 85 (2016). https://doi.org/10.1007/s11295-016-1035-6

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