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Background.  We have previously performed an analysis of the cold-responsive transcriptome in the mature leaves of tiger lily ( Lilium lancifolium ) by gene co-expression network identification. The results has revealed that a  ZFHD  gene, notated as encoding zinc finger homeodomain protein, may play an essential regulating role in tiger lily response to cold stress.    Methods.  A further investigation of the  ZFHD  gene (termed as  LlZFHD4 ) responding to osmotic stresses, including cold, salt, water stresses, and abscisic acid (ABA) was performed in this study. Based on the transcriptome sequences, the coding region and 5′ promoter region of  LlZFHD4  were cloned from mature tiger lily leaves. Stress response analysis was performed under continuous 4 °C, NaCl, PEG, and ABA treatments. Functional characterization of  LlZFHD4  was conducted in transgenic  Arabidopsis , tobacco, and yeast.    Results.   LlZFHD4  encodes a nuclear-localized protein consisting of 180 amino acids. The N-terminal region of LlZFHD4 has transcriptional activation activity in yeast. The 4 °C, NaCl, PEG, and ABA treatments induced the expression of  LlZFHD4 . Several stress- or hormone-responsive cis-acting regulatory elements (T-Box,  Box I. and ARF) and binding sites of transcription factors (MYC, DRE and W-box) were found in the core promoter region (789 bp) of  LlZFHD4 . Also, the  GUS  gene driven by  LlZFHD4  promoter was up-regulated by cold, NaCl, water stresses, and ABA in  Arabidopsis . Overexpression of  LlZFHD4  improved cold and drought tolerance in transgenic  Arabidopsis ; higher survival rate and better osmotic adjustment capacity were observed in  LlZFHD4  transgenic plants compared to wild type (WT) plants under 4 °C and PEG conditions. However,  LlZFHD4  transgenic plants were less tolerant to salinity and more hypersensitive to ABA compared to WT plants. The transcript levels of stress- and ABA-responsive genes were much more up-regulated in  LlZFHD4  transgenic  Arabidopsis  than WT. These results indicate  LlZFHD4  is involved in ABA signaling pathway and plays a crucial role in regulating the response of tiger lily to cold, salt and water stresses.

peerj

Functional characterization of <i>Lilium lancifolium</i> cold-responsive Zinc Finger Homeodomain (<i>ZFHD</i>) gene in abscisic acid and osmotic stress tolerance

Functional characterization of Lilium lancifolium cold-responsive Zinc Finger Homeodomain (ZFHD) gene in abscisic acid and osmotic stress tolerance