Discussion
Light regulates plant
photomorphogenesis.
With a large number of genes which are involved in such
photomorphogenesis processes were identified as light receptors
(Datta et al., 2006;
Kircher et al., 2002;
Peter H. Quail, 2002), signal
transduction factors (Gangappa et al.,
2013; Osterlund et al., 2000)or
degradation proteins (Crocco, Holm,
Yanovsky, & Botto, 2010; Crocco et al.,
2015; Delker et al., 2014), one of the
immediate questions is how these genes act in a network to mediate
various light-related phenotypes. It has been proved that there were
multiple pathways that are interlinked to form a gene network of
photomorphogenesis (Lau & Deng, 2012;
H.-J. Lee, Park, Ha, Baldwin, & Park,
2017). Among these pathways, it is worth mentioning that the ones
formed by a subset of family genes termed COL genes
(Cheng & Wang, 2005). These family genes
played multiple roles in plant development
(Datta et al., 2006;
Graeff et al., 2016;
Muntha et al., 2018;
Tripathi et al., 2017;
Wang et al., 2014). As an effort toward
COLs networking, we investigated the relationship between COL3and COL13 , and provided evidence that these two COLs and HY5 were
connected together to form an HY5-COL3-COL13 regulatory chain that
controls hypocotyl elongation in Arabidopsis(Fig.
7b). In addition, we also proposed a possible
COP1-dependent
COL3-COL13 feedback pathway, to optimize this regulatory pathway (Fig.
7b).
Hypocotyl elongation is a genetically well-controlled process to
response the light. In Arabidopsis , several key genes are
required for hypocotyl growth. Among these, COP1 is a negative regulator
(McNellis, von Arnim, & Deng, 1994),
while HY5 and COL3 are considered to be positive ones
(Datta et al., 2006;
Hardtke et al., 2000). A previous study
showed that COL3 played roles in flowering and hypocotyl elongation
(Datta et al., 2006), and COL3 is known to
interact with B-BOX32 to regulate
flowering(Tripathi et al., 2017).
However, there has no research on how COL3 regulates hypocotyl
elongation. To explore how the COL family genes, COL3 in particular,
function in the regulation of hypocotyl elongation will be facilitated
by identifying the downstream genes. In this study, we demonstrated thatCOL13 , whose RNA accumulated to high level in hypocotyl (Fig.1),
was one more positive regulator in the regulation of hypocotyl
elongation under red light conditions. For example, overexpression ofCOL13 or knock down its transcript resulted in a shorter or
longer hypocotyl, respectively (Fig. 2). To further define and
characterize COL13 , we analyzed genetic interactions betweencol13 and col3. Seedlings of the col13 andcol3 mutants showed reduced inhibition of hypocotyl elongation in
red light (Fig.3). Analysis of col3 col13 double mutants and
COL13 transgenic plants revealed that COL3 is epistatic toCOL13 with respect to hypocotyl elongation (Fig. 3). Given thatcol3 hy5 double mutants behaved like the hy5 mutation
(Datta et al., 2006), we hypothesized that
there is a HY5-COL3-COL13 regulatory chain for controlling hypocotyl
growth. As expected, our data proved that HY5 directly targeted
the promoter of COL3 and COL3 directly bound to the promoter ofCOL13 (Fig. 4a-c), indicating that HY5, COL3 and COL13 constitute
a hypocotyl regulatory pathway.
CONSTANS-LIKE genes belong to BBX family. Given that
BBX
family members are commonly involved in photomorphogenesis and that they
can interact with other BBX proteins to regulate plant
growth(Tripathi et al., 2017;
Wang et al., 2014), it is possible that
COL3 interacts with other BBX proteins (eg. COL13/B-BOX11) to regulate
plant development in light. Indeed, we provided evidence that COL13 can
interact with COL3 (Fig. 6). Furthermore, we found that the expression
of COL13 promoted the interaction between COP1 and COL3 (Fig. 7a). To
our knowledge, COP1 is responsible for degradation of several positive
transcription factors such as COL3 in the dark
(Datta et al., 2006;
Dornan et al., 2004;
Duek et al., 2004;
Lau & Deng, 2012;
Osterlund et al., 2000;
Seo et al., 2004;
Seo et al., 2003). Increasing the bind
activity of COP1 and COL3 would lead to the degradation of COL3. As a
result, there would be less COL3 to active the expression of COL13 (Fig.
7b). The COP1-dependent COL3-COL13 feedback pathway could enrich the
regulation network in hypocotyl elongation.