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In fruit trees, flowering is a key event followed by fruit development and seed production.  Gigentea  ( GI ), a clock-associated gene, is known to contribute to photoperiodic flowering and circadian clock control in  Arabidopsis thaliana . However, its functions in woody fruit trees remain unclear. In this study, a 2000 bp promoter fragment of the longan ( Dimocarpous longan )  DlGI  gene was isolated from the genomic DNA of longan ‘Honghezi’ by polymerase chain reaction amplification. The  DlGI  promoter contained two main types of potential  cis -acting elements: light-responsive and hormone-responsive elements. The promoter was fused with the β-glucuronidase ( GUS ) reporter gene of pBI121 to generate the pDlGI:GUS construct.  GUS  histochemical staining of transgenic  A. thaliana  revealed that  DlGI  might play a role in different developmental phases of longan. Exposure of transgenic  A. thaliana  to varying light intensities showed that the  GUS  activity increases with increased light intensity. Transient expression of pDlGI::GUS in  Nicotiana benthamiana  showed that the  GUS  activity was higher and reached peak a few hours earlier under short-day (SD) than long-day conditions. Exposure to different hormonal treatments revealed that the transcript level of  GUS  was activated by gibberellin (GA 3 ) and indoleacetic acid (IAA) but suppressed by abscisic acid and methyl jasmonate treatment. In addition,  N. benthamiana  transient assay and dual-luciferase assay revealed that the presence of  early flowering 4  ( ELF4 ) homologs of longan ( DlELF4-1  and  DlELF4-2 ) significantly activated the  DlGI  promoter. The positive response of  DlGI  promoter to high light-intensity, SD photoperiod, GA 3  and IAA signals, and  DlELF4  transcription factor suggest that  DlGI  may function as a circadian clock and play a role in responding to SD conditions and other signals in flower initiation of longan.

Identification and Characterization of DlGI Promoter Involved in Photoperiod, Light Intensity, Hormone, and DlELF4 Response from Longan