ORANGE negatively regulates flowering time in Arabidopsisthaliana.

Floral transition is an important process in plant development, which is regulated by at least four flowering pathways: the photoperiod, vernalization, autonomous, and gibberellin (GA)-dependent pathways. The DnaJ-like zinc finger domain-containing protein ORANGE (OR) was originally cloned from the cauliflower or mutant, which has distinct phenotypes of the carotenoid-accumulating curd, the elongated petioles, and the delayed-flowering time. OR has been demonstrated to interact with phytoene synthase for carotenoid biosynthesis in plastids and with eukaryotic release factor 1-2 (eRF1-2) in the nucleus for the first two phenotypes, respectively. In this study, we showed that overexpression of OR in Arabidopsis thaliana resulted in a delayed-flowering phenotype resembling the cauliflower or mutant. Our results indicated that OR negatively regulates the expression of the flowering integrator genes FLOWERING LOCUS T (FT) and SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (SOC1). Both GA3 and vernalization treatments could not rescue the delayed-flowering phenotype of the OR-overexpressing seedlings, suggesting the repression of floral transition by OR does not depend on SOC1-mediated vernalization or GA-dependent pathways. Moreover, our analysis revealed that transcripts of OR and FT fluctuated in opposite directions diurnally, and the overexpression of OR repressed the accumulation of CONSTANS (CO), FT, and SOC1 transcripts in a 16 h/8 h light/dark long-day cycle. Our results indicated the possibility that OR represses flowering through the CO-FT-SOC1-mediated photoperiodic flowering pathway.

An intein-split transactivator for intersectional neural imaging and optogenetic manipulation.

The cell-type-specific recording and manipulation is instrumental to disentangle causal neural mechanisms in physiology and behavior and increasingly requires intersectional control; however, current approaches are largely limited by the number of intersectional features, incompatibility of common effectors and insufficient gene expression. Here, we utilized the protein-splicing technique mediated by intervening sequences (intein) and devised an intein-based intersectional synthesis of transactivator (IBIST) to selectively control gene expression of common effectors in multiple-feature defined cell types in mice. We validated the specificity and sufficiency of IBIST to control fluorophores, optogenetic opsins and Ca2+ indicators in various intersectional conditions. The IBIST-based Ca2+ imaging showed that the IBIST can intersect five features and that hippocampal neurons tune differently to distinct emotional stimuli depending on the pattern of projection targets. Collectively, the IBIST multiplexes the capability to intersect cell-type features and controls common effectors to effectively regulate gene expression, monitor and manipulate neural activities.