Abstract
The fertilization of flowering plants is a critical event in their reproduction. ZnF1, a C2H2-like zinc finger protein, is highly expressed in the reproductive organs during Brassica oleracea self-pollination. However, BoZnF1’s functions during these biological processes have not been well characterized. Genetically modified mutants were employed to investigate the potential roles of ZnF1 in Arabidopsis fertilization. A Promoter expression analysis indicated that BoZnF1 is primarily expressed in root and reproductive organs, especially in filaments and pistils. When transformed into Arabidopsis protoplasts, the green fluorescent protein–BoZnF1 fusion fluoresced in the nuclei. A T-DNA insert (znf1) in Arabidopsis disrupted inflorescence development, including elongation of the gynophore and papilla cells on the stigma and the failure of anther sac cracking, resulting in defective pollen morphology. By hybrid pollination between wild-type and znf1, we found that wild-type pollen grains grew slowly on the znf1 stigma, leading to a reduced seed-setting rate. In addition, in vivo yeast two-hybrid screening and co-immunoprecipitation were performed to identify the binding protein of ZnF1. A TEOSINTE BRANCHED 1-CYCLOIDEA-PCF family transcription factor, BoTCP4, was identified as a ZnF1-interacting partner. We found that znf1 plays a role in the auxin pathway. When znf1 plants were treated with N-1-naphthylphthalamic acid, an auxin-transport inhibitor, the defects were exacerbated, leading to the ectopic formation of the gynophore in the ovarian tissue. Thus, ZnF1 may play a negative regulatory role and influence reproduction through the auxin pathway in Brassica.
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Acknowledgements
This work was funded by the National Natural Science Foundation of China (Grant No. 31572127), a special foundation of central institution basic research (Grant No. XDJK2017C032).
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XPL, HCZ and LQZ designed the experiments. XPL, HCZ, QYL and THZ performed the experiments. XPL, ZJ and XJB analyzed the data. YKW, QYL, YZZ and THZ contributed analyses tools. XPL and HCZ wrote the manuscript. XPL, LQZ and RLC revised the manuscript. All the authors have read and agreed to the submitted version of the manuscript.
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Lian, X., Zhang, H., Zeng, J. et al. C2H2-like zinc finger protein 1 causes pollen and pistil malformation through the auxin pathway. Plant Growth Regul 90, 505–518 (2020). https://doi.org/10.1007/s10725-019-00568-1
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DOI: https://doi.org/10.1007/s10725-019-00568-1