The Circadian Clock Regulates the Photoperiodic

Response of Hypocotyl Elongation through a

Coincidence Mechanism in Arabidopsis thaliana
Aims
• Understand the molecular mechanisms in which the circadian clock
regulates hypocotyl elongation in light/dark cycles

• Previous studies done in constant light/dark, so don’t know the

impact of photoperiod on hypocotyl elongation

• Implications for how plants adapt to seasonal changes

 Overview
Mutant knockouts
and overexpressors qRT-PCR Hypocotyl measurements

GA enriched
media

Figure 6A
Photoperiodic response of hypocotyl elongation

Hypothesis: Hypocotyl elongation is photoperiod dependent

Background:
Other processes, like flowering (Suarez-Lopez et al., 2001), are photoperiod dependent
 Photoperiodic response of hypocotyl elongation

Grew A. thaliana on plates in different

photoperiods and measured the hypocotyl
length.

Hypocotyl elongation is short-day dependent.

Figure 1
 Photoperiodic response of hypocotyl elongation

Figure 1
 Photoperiodic response of hypocotyl elongation

Hypothesis: Hypocotyl elongation is

regulated by total light intensity per
day

Different light intensities with the

same photoperiod had the same
hypocotyl elongation.

Figure 2
 Photoperiodic response of hypocotyl elongation

Figure 1 Figure S1
 The PHYB and GA mediated
pathways

Hypothesis: The photoperiodic response of hypocotyl elongation can be explained by PHYB

and GA control of PIF4/5

Background:
PHYB senses light and directly interacts with PIF4/5 (Huq and Quail, 2002).
Gibberellin has a light-mediated impact on hypocotyl elongation (Nozue and Maloof, 2006).
 The PHYB and GA mediated
pathways

Linear trend in mutants, implying

involvement in the regulation of
hypocotyl elongation.

Figure 2
 The PHYB and GA mediated
pathways
Positive regulators of hypocotyl
elongation.

Constitutive expression causes

elongation linear to dark period.

Figure 2
 The PHYB and GA mediated
pathways

Increased hypocotyl elongation but

is still photoperiod-dependent.

Is addition of GA different to
expression within the plant?

Figure 2
 The PHYB and GA mediated
pathways
phyB mediated hypocotyl
elongation is only partly
dependent on PIF4/5.

GA mediated hypocotyl
elongation is dependent on
PIF4/5.

“This light dependent degradation of PIF5 in

plants was confirmed in this study with PIF5-
HAox seedling”

Figure 6
 The Circadian Clock

Hypothesis: The circadian clock is the mechanism behind the non-linear hypocotyl
elongation in different photoperiods.

Background:
Hypocotyl elongation happens diurnally at predawn (Nozue et al., 2007) and the circadian
clock is known to control PIF expression (Fujimori et al., 2004).
 The Circadian Clock

toc1 – elongated hypocotyl in long

days

cca1 lhy - needs even shorter days

for hypocotyl elongation

Figure 3
 The Circadian Clock

Overexpression mutants show the

opposite

Figure 3
 The Circadian Clock

Photoperiod-dependent hypocotyl
elongation.

No hypocotyl elongation in LL.

Figure 3
 The Circadian Clock

Hypothesis: the circadian clock modulates

the expression of PIF4/5

PIF4/5 are expressed diurnally (Fujimori et

al. 2004).

Hypocotyl elongation in clock mutants is

photoperiod dependent.

Figure 4
 The Circadian Clock

Hypothesis: the circadian clock modulates

the expression of PIF4/5

PIF4/5 are expressed diurnally (Fujimori et

al. 2004).

Hypocotyl elongation in clock mutants is

photoperiod dependent.

Figure 4
 The Circadian Clock

Hypothesis: the circadian clock modulates

the expression of PIF4/5

PIF4/5 are expressed diurnally (Fujimori et

al. 2004).

Hypocotyl elongation in clock mutants is

photoperiod dependent.

Figure 4
 The Circadian Clock

Tested PIF expression in

remaining clock genes.

Decrease in PIF accumulation,

so shorter hypocotyls.

cca1 lhy was grown at

19D/5L, all other mutants at
14D/10L.

Figure 8
 The Circadian Clock
Hypothesis: The circadian clock modulates the expression of PIF4/5 in different
photoperiods

Figure 5
 The Circadian Clock

toc1 PIF expression only measured at 1 photoperiod.

Figure 5
 The Circadian Clock
Hypothesis: The circadian clock modulates the expression of PIF4/5 in different
photoperiods

Figure 5
 The Circadian Clock
prr7 prr5 phyB triple mutant
is additive, so these pathways
are independent.

Prr7 prr5 phyB +GA cancels

out all the pathways leading
to very elongated hypocotyls.

No Col +GA or PIF5ox +GA.

Figure 6
 The Circadian Clock

WT and mutants grown at

different photoperiods for 25
days.

Mature plants showed similar

trends to seedlings.

Figure 9
 Clock mutations – dark period dependent
hypocotyl elongation

WT has photoperiod dependent hypocotyl

elongation

pif4/5 cause reduced hypocotyl elongation

(implying factor X)

Figure 7
Conclusions

• PHYB and GA signals are integrated into a PIF4/5 response which is

gated by the circadian clock. PIF4/5 then promote hypocotyl
elongation under short day conditions.

• Significant support for the involvement of PHYB, GA and the circadian

clock, but didn’t do any experiments involving DELLAs.
Research since ?
• https://www.cell.com/current-biology/pdf/S0960-9822(14)01418-3.p
df

• https://www.tandfonline.com/doi/full/10.4161/psb.22863

• https://academic.oup.com/pcp/article/53/11/1965/1865349?login=fa
lse
References
Huq 2002 - https://www.embopress.org/doi/full/10.1093/emboj/21.10.2441
Suarez lopez 2001 –
https://www.nature.com/articles/35074138

Nozue and Maloof 2006

https://onlinelibrary.wiley.com/doi/full/10.1111/j.1365-3040.2005.01489.x
Nozue et al 2007
https://www.nature.com/articles/nature05946

Fujimori et al 2007
https://academic.oup.com/pcp/article/45/8/1078/1837026?login=false