Developmental phase change in plants

Plants progress through several developmental phases before reaching maturity; after germination,
shoots undergo stages of juvenile and adult vegetative growth before transitioning into a reproductive
phase. The term “vegetative phase change” (VPC) has been used differently throughout the literature
on this topic, in this essay I will use the term to refer to the transition from juvenile to adult stages of
vegetative growth. VPC is controlled by the master regulator miR156, an evolutionarily conserved
microRNA, that influences the expression of several other factors in order to coordinate progression
to the adult vegetative phase. miR156 expression is age-dependent but it is still largely unknown how
this is regulated; the measuring of “age” in plants is a topic still surrounded by mystery so the
investigation of factors such as miR156 is of great importance. In this essay, I will discuss how
different morphological changes are coordinated during VPC as well as how the timing of these
events may be measured.

miR156 and its targets

The regulation of vegetative phase change by miR156 has been studied extensively in the model
organism Arabidopsis, as well as in maize and rice. In Arbabidopsis, this was discovered by
investigating loss-of-function mutations of HASTY as this gene was identified as being responsible for
accelerating vegetative phase change (Bollman et al., 2003). Further analysis revealed that miR156
was greatly reduced in HASTY mutants and importantly, was expressed at significantly higher levels
in young shoots than in older ones (Wu and Poethig, 2006). The gradual, age-dependent decline of
miR156 is central to the transition into the adult vegetative phase.

miR156 works by inhibiting its targets, the SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE

(SPL) transcription factors (Hyun, Richter and Coupland, 2016). 11 of the 17 SPL genes in
Arabidopsis are repressed my miR156 with SPL3/4/5 and SPL9/15, which represent two classes of
SPLs, having the most significant effects (Poethig, 2013). SPLs promote the adult vegetative phase,
so by repressing them, miR156 inhibits development to this stage. miR156 expression is highest at the
seedling stage; as this level reduces, SPLs are repressed less so are able to promote the pathways that
enable development of adult characteristics (Wu et al., 2009). As shown in figure 1, SPLs have
several targets, and it is highly likely that there are yet more to be discovered. A key part of this
regulatory pathway is the activation of another microRNA, miR172, by SPL9/15. miR172 was
initially identified through random sequencing of miRNAs known to accumulate during plant
development, the same authors also identified APETALA2 (AP2) as a target of miR172 by
identifying genes with regions of homology (Park et al., 2002). As miR172 is promoted by SPL9/15,
the decline of miR156 (suppressing SPLs) causes an increase in miR172, enabling it to carry out
functions such as inhibiting AP2-like floral repressor genes to allow progression of the shoot to later
phases where flowering can occur. The relative abundances of miR156 and miR172 are a key factor in
determining the timing of vegetative phase change (Chuck et al., 2007).

Development of adult leaf morphology

In Arabidopsis, vegetative phase change is characterised by the development of adult leaf

morphology. This involves the leaves growing to be larger, more elongated and serrated (Figure 1).
Trichomes, hair like extensions of epidermal cells, are present on the adaxial (top) side of juvenile
leaves, abaxial trichomes appear only in the adult phase (Wang et al., 2021). This transition is
coordinated by the decline of miR156 and the subsequent increase in expression of SPLs 3/4/5 and
9/15.

Wu et al. (2009) found that SPL9/15 affects all aspects of leaf morphology while SPL3/4/5 affects
only the epidermal identity (i.e. abaxial trichome growth) as double mutants of SPL9 and SPL15
presented late growth of abaxial trichomes with smaller, rounder leaves. Plants grown with miR156
resistant SPL3/4/5 (so would not be downregulated) only showed accelerated abaxial trichome
growth, this is explained by the fact that SPL3/4/5 do not seem to regulate miR172 so are likely to
coordinate adult epidermal identity in a different way. When SPL9/15 is no longer repressed my
miR156, it promotes the expression of miR172; this is because SPL9 is a direct transcriptional
activator of the major precursor gene miR172b (Wu et al., 2009). miR172 targets AP2-like
transcription factors such as TOE1 and TOE2, these genes were first identified for their role in
repressing floral development however it was later shown that they are also repressed in vegetative
phase change to enable abaxial trichome growth (Aukerman and Sakai, 2003; Wu et al., 2009).

Figure 1. (A) Leaf of Arabidopsis plant showing adaxial

and abaxial trichomes. (B) Stages of leaf development in
Arabidopsis; adult leaves are larger, more elongated and
serrated, unlike the smaller, rounded juvenile leaves.
(Adapted from Wang et al. 2021 and Huijser and Schmid,
2011).
Development of competence to flower

“Competence to flower” is acquired by adult shoots as they undergo vegetative phase change, it
describes the ability of shoots to flower and subsequently reproduce; shoots with competence to
flower are capable of flowering in response to environmental stimuli such as photoperiod and
temperature (Hyun, Richter and Coupland, 2016). In Arabidopsis, once shoots have acquired
competence to flower, exposure to longer daylight hours triggers flowering through activating the
CONSTANS (CO) then FLOWERING LOCUS T (FT) genes (Suárez-López et al., 2001). SPLs
upregulate FT, so miR156 prevents early shoots from responding to photoperiod cues by inhibiting
SPL expression; as miR156 levels drop over time, the shoot eventually develops competence to
flower as SPL levels reach high enough levels to promote FT expression (Wang, Czech and Weigel,
2009).

Shoot Apical Meristem growth

The spatial and temporal distribution of miR156

Control of the timing of miR156 expression

References

Aukerman, M. and Sakai, H., 2003. Regulation of Flowering Time and Floral Organ Identity by a
MicroRNA and Its APETALA2-Like Target Genes. The Plant Cell, 15(11), pp.2730-2741.

Bollman, K., Aukerman, M., Park, M., Hunter, C., Berardini, T. and Poethig, R., 2003. HASTY, the
Arabidopsis ortholog of exportin 5/MSN5, regulates phase change and morphogenesis. Development,
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Chuck, G., Cigan, A., Saeteurn, K. and Hake, S., 2007. The heterochronic maize mutant Corngrass1
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Huijser, P. and Schmid, M., 2011. The control of developmental phase transitions in plants.
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Hyun, Y., Richter, R. and Coupland, G., 2016. Competence to Flower: Age-Controlled Sensitivity to
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Park, W., Li, J., Song, R., Messing, J. and Chen, X., 2002. CARPEL FACTORY, a Dicer Homolog,
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Suárez-López, P., Wheatley, K., Robson, F., Onouchi, H., Valverde, F. and Coupland, G., 2001.
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Wang, X., Shen, C., Meng, P., Tan, G. and Lv, L., 2021. Analysis and review of trichomes in plants.
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