Initial embedding of TRANSPARENT TESTA GLABRA 1 in the Arabidopsis thaliana flowering time regulatory pathway

Botanical Institute, Department of Biology, University of Cologne, Cologne, Germany
DOI
10.7287/peerj.preprints.27974v1
Subject Areas
Developmental Biology, Plant Science
Keywords
TTG1, flowering time, PRR, circadian clock, FT, FLC, bHLH92, Arabidopsis thaliana
Copyright
© 2019 Paffendorf et al.
Licence
This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ Preprints) and either DOI or URL of the article must be cited.
Cite this article
Paffendorf BAM, Qassrawi R, Meys AM, Trimborn L, Schrader A. 2019. Initial embedding of TRANSPARENT TESTA GLABRA 1 in the Arabidopsis thaliana flowering time regulatory pathway. PeerJ Preprints 7:e27974v1

Abstract

Pleiotropic regulatory factors mediate concerted responses of the plant’s trait network to endogenous and exogenous cues. TRANSPARENT TESTA GLABRA 1 (TTG1) is a pleiotropic regulator that has been predominantly described in its role as a regulator of early accessible developmental traits. Although its closest homologs LIGHT-REGULATED WD1 (LWD1) and LWD2 are regulators of photoperiodic flowering, a role of TTG1 in flowering time regulation has not been reported.

Here we reveal that TTG1 is a regulator of flowering time in Arabidopsis thaliana and changes transcription levels of different targets within the flowering time regulatory pathway. TTG1 mutants flower early and TTG1 overexpression lines flower late at long-day conditions. Consistently, TTG1 can suppress the transcript levels of the floral integrators FLOWERING LOCUS T and SUPPRESSOR OF OVEREXPRESSION OF CO1 and can act as an activator of circadian clock components. Moreover, TTG1 might form feedback loops at the protein level. The TTG1 protein interacts with PSEUDO RESPONSE REGULATOR (PRR)s and basic HELIX-LOOP-HELIX 92 (bHLH92) in yeast. In planta, the respective pairs exhibit interesting patterns of localization including a recruitment of TTG1 by PRR5 to subnuclear foci. This mechanism proposes additional layers of regulation by TTG1 and might aid to specify the function of bHLH92.

Within another branch of the pathway, TTG1 can elevate FLOWERING LOCUS C (FLC) transcript levels. FLC mediates signals from the vernalization, ambient temperature and autonomous pathway and the circadian clock is pivotal for the plant to synchronize with diurnal cycles of environmental stimuli like light and temperature. Our results suggest an unexpected positioning of TTG1 upstream of FLC and upstream of the circadian clock. In this light, this points to an adaptive value of the role of TTG1 in respect to flowering time regulation.

Author Comment

This is a submission to PeerJ for review.

Supplemental Information

Characterization of ttg1-21 and ttg1-22

DOI: 10.7287/peerj.preprints.27974v1/supp-1

Alignment of TTG1, LWD1 and LWD2 CDS containing the primer binding sited for “TTG1 both” and “TTG1 no LWD” qRT-PCR experiments

DOI: 10.7287/peerj.preprints.27974v1/supp-2

Additional blots and analysis of protein levels in TTG1 overexpressor lines

DOI: 10.7287/peerj.preprints.27974v1/supp-3

Circadian profiles of endogenous and overexpressed TTG1 transcript levels

DOI: 10.7287/peerj.preprints.27974v1/supp-4

Similar to CO, GI transcript levels can not explain the significant reduction of FT transcript levels in TTG1 overexpression lines

DOI: 10.7287/peerj.preprints.27974v1/supp-5

Flowering time analysis of TTG1 gene regulatory network bHLH factor mutants and respective overexpression lines in A. thaliana Col-0 background at warm long day conditions

DOI: 10.7287/peerj.preprints.27974v1/supp-6

PRRs can re-localize TTG1 to different distinct (sub-)nuclear localizations and TTG1 can prevent the nuclear enrichment of CFP-bHLH92 as revealed by CLSM of nuclei in epidermal cells of infiltrated tobacco leaves

DOI: 10.7287/peerj.preprints.27974v1/supp-8

Epidermal areas of infiltrated tobacco leaves revealing the nuclear enrichment of RFP-TTG1 upon co-expression with PRRs and the abolished enrichment of CFP-bHLH92 by co-expression of RFP-TTG1

DOI: 10.7287/peerj.preprints.27974v1/supp-9

Datalogger results from growth condition measurements

DOI: 10.7287/peerj.preprints.27974v1/supp-10

Results of phenotyping experiments and statistics

DOI: 10.7287/peerj.preprints.27974v1/supp-11

Results of qRT-PCR experiments for circadian TTG1, CO, FT, SOC1 transcript levels in overexpression lines and statistics

DOI: 10.7287/peerj.preprints.27974v1/supp-12

Qualitative analysis of YFP-fluorescence in the overexpression lines as seen and classified by eye at a fluorescence stereo microscope

DOI: 10.7287/peerj.preprints.27974v1/supp-13

Results of circadian qRT-PCR experiments and statistics

DOI: 10.7287/peerj.preprints.27974v1/supp-14