Journal of Plant Physiology 258–259 (2021) 153391

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Journal of Plant Physiology

journal homepage: www.elsevier.com/locate/jplph

Epigenetic regulation and intercellular communication during male

gametophyte development
Xiaorong Huang, Meng-Xiang Sun *
College of Life Science, State Key Laboratory of Hybrid Rice, Wuhan University, Wuhan 430072, China

A R T I C L E I N F O A B S T R A C T

Keywords The male gametophyte of angiosperms has long been recognized as an ideal system for the study of the molecular
Male gametophyte mechanisms regulating cell fate determination. Recent findings on histone variants in two cell lineages,
Epigenetic regulation vegetative-cell-derived small interfering RNA and transposable element expression provide new power for
Cell-to-cell communication
relevant investigations.

The haploid microspore undergoes mitosis twice to produce a highly
simplified male gametophyte with smaller male germline cells
embedded within a larger vegetative cell, forming a “cell within a cell”
construction (Hackenberg and Twell, 2019). Male gametophyte devel­
opment is recognized as a model for studying cell polarity, cell cycle, cell
fate determination, and cell signaling. In past decades, numerous studies
have indicated that this short and simple developmental process is
actually governed by a very complex mechanism. Liu and Wang (2021)
summarized relevant advances in this field and give a clear account of
the functional specialization of two cell lineages. They highlight the
functions of temporary callose walls, dynamic nuclear pore density,
transcription repression, and histone variants, and also discuss the
intercellular movement of vegetative-cell-derived small interfering RNA
(siRNA) and the roles of transposable element (TE) silence or expression
in male gametogenesis.
As chromatin is highly diffuse in vegetative cells but condensed in
sperm cells, the relationship between cell fate and its epigenetic char­
acteristics in male gametophyte has drawn great attention. In their re­
view, Liu and Wang (2021) summarize the dynamics of linker histone
H1 during male gametophyte development. H1 is retained in male
germline cells, but lost in vegetative cells, which is significant for
maintaining transcriptional homeostasis in two cell lineages (Liu and
Wang, 2021). H1 depletion facilitates heterochromatin relaxation and
contributes to TE activation in vegetative cells (He et al., 2019).
Furthermore, H3 variants and histone posttranslational modifications
(PTMs) act in the regulation of male gametophyte development, but the
types and relative abundance of histone markers show diversity between
two cell lineages of pollen from different plants (Borg and Berger, 2015).
In A. thaliana, the heterochromatin of vegetative cell decondenses,
which is accompanied by a unidirectional progressive loss of the
centromeric H3 variant CenH3 (HTR12) and the repressive mark
H3K9me2. The ectopic H3K9me2 speckle formation and chromatin
compaction in vegetative cells occurs owing to loss of SET DOMAIN
GROUP2-mediated H3K4me3 deposition, indicating that both H3K4me3
and H3K9me2 are crucial for chromatin remodeling in male gameto­
phytes (Pinon et al., 2017). In contrast, CenH3, HTR10 (AtMGH3), and
H3K9me2 specifically accumulate in sperm cells, highlighting their
distinct fate and ongoing state of proliferation compared to vegetative
cells (Okada et al., 2005; Schoft et al., 2009). Male germline cells have
lower metabolic and transcriptional activities than vegetative cells. The
loading of the male germline cell-specific HTR10 in the sperm chromatin
combines with the silencing of H3K27me3 writers and activity of
H3K27me3 erasers, suggesting that male germline cells seem to have
developed specific epigenetic mechanisms to regulate the expression of
genes required for germline specialization (Borg et al., 2020), although
sperm chromatin is highly compacted. However, the absence of specific
histone variants and abnormal histone PTMs in two cell lineages of male
gametophyte do not disturb cell identities (Okada et al., 2005; Pinon
et al., 2017; Borg et al., 2020). Thus, further studies are needed to reveal
the roles of epigenetic regulation in cell fate determination during male
gametophyte development.
As introduced by Liu and Wang (2021), male gametogenesis occurs
in a relatively closed microenvironment created by vegetative cells.
Germline cells are quarantined from vegetative cells by plasma mem­
brane, and even have a real cell wall in some species. Microspore
asymmetric cell division determines vegetative cell and male germline
identities and leads to the gene expression dimorphism (Hackenberg and
Twell, 2019). The genetic mechanisms of regulating pollen

* Corresponding author.
E-mail address: mxsun@whu.edu.cn (M.-X. Sun).

https://doi.org/10.1016/j.jplph.2021.153391
Received 10 February 2021; Accepted 12 February 2021
Available online 17 February 2021
0176-1617/© 2021 Published by Elsevier GmbH.
X. Huang and M.-X. Sun
compartmentation are focused, with a more comprehensive under­
standing of the relationship between asymmetric segregation and cell
fate determination. The significance of the callose wall during micro­
sporogenesis and male gametogenesis is emphasized. Callose plays
crucial roles in the multiple stages of male gametophyte development in
flowering plants, and its spatiotemporal deposition or degradation is
often the sign for cell fate transition (Liu and Wang, 2021). Interestingly,
a current work provides strong support for this viewpoint. STICKY
GENERATIVE CELL (SGC) prevents callose deposition in the nascent
generative cell, thereby allowing the generative cell to fully enter into
the vegetative cell and then correctly proliferate and differentiate into
male gametes in A. thaliana. In sgc pollen, the generative cell is immo­
bilized at the pollen wall because of ectopic callose accumulation that
disturbs its well-timed internalization and differentiation. Meanwhile,
the vegetative cell expresses the germline-specific marker and exhibits a
mixed cell fate (Oh et al., 2021). This seems to indicate a critical role of
closer contact between two cell lineages in germline establishment. In
fact, the intercellular communication between vegetative cells and male
germline cells does exist. In addition to providing nutrients, the vege­
tative cell also sacrifices the specific ABA-hypersensitive germination3
transcripts and the TE-derived siRNAs to sperm cells (Slotkin et al.,
2009; Jiang et al., 2015; Martínez et al., 2016). Most plant TEs have lost
the ability to transpose: however, TEs exhibit strong spatiotemporal
expression in certain gametophytic cells. Liu and Wang (2021) review in
detail TE reactivation and its regulatory mechanisms in vegetative cells.
The A. thaliana vegetative cell performs developmental TE reactivation,
and the TE-derived siRNAs are transported into sperm cells to reinforce
cognate TE silencing and maintain genome stability (Slotkin et al., 2009;
Martínez et al., 2016). A recent study of the tomato male germline cells
indicates that sperm cells and generative cells have highly similar 5-mC
DNA methylomes, and suggests that the strengthening of TE silencing by
siRNA may be pre-established in generative cells (Lu et al., 2020).
Although some crucial questions remain unanswered, e.g., whether
there are unknown siRNA importers localized in the plasma membrane
of male germline cells and whether the cytoplasmic projections are
involved in signaling, siRNA delivery between vegetative and male
germline cells is discussed and three putative models of intercellular
transfer of siRNA are proposed (Liu and Wang, 2021). This provides
ideas for follow-up research. On the other hand, unlike in A. thaliana, the
suppression of TE expression may be relaxed in the sperm cells of plant
species with large genomes. Appropriate TE expression and trans­
position in germline cells could facilitate the production and inheritance
of certain beneficial mutations to adapt to the changing environment
(Liu and Wang, 2021). Moreover, in contrast to receiving transcripts and
siRNAs, might germline cells also send messages to vegetative cells?
Although findings show that the absence of sperm cells does not seem to
affect the maturation and function of vegetative cells (Zhang et al.,
Journal of Plant Physiology 258–259 (2021) 153391
2017), the mechanisms of cell-to-cell communication in male gameto­
phytes are worthy to be further elucidated.
Declaration of Competing Interest
The authors report no declarations of interest.
Acknowledgements
The authors’ work is funded by the National Natural Science Foun­
dation of China (31991201 and 31800265).
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