Curr Genet (2016) 62:283–286
DOI 10.1007/s00294-015-0540-8
REVIEW
Environmental pH adaption and morphological transitions
in Candida albicans
Han Du1 · Guanghua Huang1
Received: 3 November 2015 / Revised: 4 November 2015 / Accepted: 5 November 2015 / Published online: 18 November 2015
© Springer-Verlag Berlin Heidelberg 2015
Abstract The human fungal pathogen Candida albicans
encounters a wide range of pH stresses during its com-
mensal and pathogenic lifestyles. It has been well studied
that environmental pH regulates the yeast–filamentous
growth transition in this fungus. White–opaque switching is
another type of phenotypic transitions in C. albicans. White
and opaque cells are two morphologically and functionally
distinct cell types, which differ in many aspects including
global gene expression profiles, virulence, mating compe-
tency, and susceptibility to antifungals. The switch between
white and opaque cell types is heritable and epigenetically
regulated. In a recently study, Sun et al. (Eukaryot Cell
14:1127–1134, 2015) reported that pH plays a critical role
in the regulation of the white–opaque phenotypic switch
and sexual mating in C. albicans via both the conserved
Rim101-mediated pH sensing and cAMP signaling path-
ways. The effect of pH on the two biological processes may
represent a balancing act between host environmental adap-
tation and sexual reproduction in this pathogenic fungus.
Keywords Candida albicans · pH regulation ·
Morphological transitions · Epigenetic regulation · Sexual
development · Rim101 · cAMP signaling pathway
Communicated by M. Kupiec.
* Guanghua Huang
huanggh@im.ac.cn
1
State Key Laboratory of Mycology, Institute of Microbiology,
Chinese Academy of Sciences, Beijing 100101, China
Introduction
Candida albicans is a common member of the microbiota
in the human gastrointestinal tract, mouth, and genital tract.
It causes superficial infections in both immune-competent
and immune-compromised individuals and life-threatening
invasive diseases in immune-compromised patients (Brown
et al. 2012). To survive and proliferate, C. albicans must be
able to sense and respond rapidly to changes of the environ-
ment in the diverse niches of the human host. The change
of ambient pH is one of the major challenges often encoun-
tered by this pathogen. C. albicans can virtually colonize
any organs of humans with a wide range of pH conditions
(from pHs of <2 to pHs of >10) (Davis 2003; Kararli 1995;
Selvig and Alspaugh 2011). pH homeostasis and adapta-
tion to its changes is critical for the growth and virulence of
C. albicans. It has been well investigated that pH controls
yeast–filamentous growth transitions and pathogenesis in
C. albicans [reviewed in (Biswas et al. 2007; Davis 2003,
2009; Huang 2012)]. In a recent study, we reported that
pH regulates white–opaque transitions, another phenotypic
system, and sexual mating in C. albicans (Sun et al. 2015).
pH‑regulated filamentation and white–opaque
switching in C. albicans
Phenotypic plasticity is one of the most striking features
of C. albicans biology. The transitions between differ-
ent morphological phenotypes represent a response of C.
albicans to environmental changes (Biswas et al. 2007;
Sudbery 2011; Whiteway and Bachewich 2007). Different
cellular morphologies play different roles in the processes
of infection and adaptation to different host niches. Acidic
pH represses the yeast-to-filamentous growth transition,
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while neutral and alkaline pH conditions promote filamen-
tation (Biswas et al. 2007; Davis 2003). Since the pH of
the human blood and tissues is neutral or weakly alkaline,
filamentation under these pH conditions would facilitate
C. albicans cells to invade and colonize host tissues. C.
albicans can also actively modify the environmental pH
through metabolism of available nutrients. For example,
in the presence of glucose, the growth of C. albicans can
acidify the medium by glycolysis. Alternatively, under the
conditions of carbon deprivation, C. albicans uses amino
acids as the carbon source and alkalinizes the extracellular
environment by the production of ammonia. This alteration
of environmental pH affects yeast–filamentous growth tran-
sitions and may play a role in colonizing the host and caus-
ing infections (Stewart et al. 1989; Vylkova et al. 2011).
White and opaque cells are two distinct cell types
(Anderson and Soll 1987; Slutsky et al. 1987). White cells
are small and relatively round, while opaque cells are
large and elongated. Under scanning microscopy, the sur-
face of the cell wall of mature opaque cells is rough and
exhibits a “pimpled” pattern, while that of white cells is
smooth. Opaque cells constitutively produce several types
of secreted aspartyl proteinases (Saps), which cause deg-
radation of host tissues. Therefore, opaque cells are better
at cutaneous infections than white cells. White cells exhibit
high virulence in mouse systemic infection systems pos-
sibly due to their stronger abilities of filamentation and
stress resistance in response to diverse host environments
(Kvaal et al. 1999; Xie et al. 2013). The switching between
white and opaque cell types is bi-stable, heritable, and
epigenetically regulated (Lohse and Johnson 2009; Slut-
sky et al. 1987; Soll 2009). Each cell type can maintain its
original cell type for many generations and can also spon-
taneously switch to the other phenotype. Both white and
opaque cells are single-celled yeast, but differ in a number
of biological aspects, including colony morphologies, cel-
lular appearances, global gene expression profiles, meta-
bolic profiles, susceptibility to environmental stresses and
the host immune cells, ability of biofilm development, and
mating competency (Huang 2012; Mathé and Van Dijck
2013; Prasad and Singh 2013; Soll 2009). The environ-
mental and genetic regulation of white–opaque transitions
occurs in a bi-directional or “yin-yang” fashion. A pleth-
ora of host-related environmental factors is involved in
this epigenetic regulation (Alby and Bennett 2009; Hnisz
et al. 2009; Huang 2012; Huang et al. 2009, 2010; Ram-
irez-Zavala et al. 2008; Soll 2009; Sun et al. 2015; Fig. 1).
For example, CO2 and N-acetylglucosamine (GlcNAc),
two abundant molecules in the host gut, induce white-to-
opaque switching (Huang et al. 2009, 2010). The com-
bined use of these two environmental cues can even induce
white-to-opaque switching in a subset of MTL heterozy-
gous (a/α) strains (Xie et al. 2013), which were thought to
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Curr Genet (2016) 62:283–286
Fig. 1  Environmental regulation of white–opaque switching in C.
albicans. UV ultraviolet, Wh white cell type, Op opaque cell type.
Cells (of GH1349, a derivative of WO-1) were grown on agar plates
of Lee’s medium containing 5 μg/ml phloxine B, a red dye exclu-
sively staining opaque cells
be opaque incompetent (Lockhart et al. 2002; Miller and
Johnson 2002). CO2 promotes the opaque phenotype pri-
marily through the cAMP signaling pathway and some
unidentified mechanisms, which converge on the transcrip-
tion factor Flo8 (Du et al. 2012). Flo8 finally activates the
expression of the master regulator gene of white–opaque
switching, WOR1 (Du et al. 2012; Huang et al. 2006; Sri-
kantha et al. 2006; Zordan et al. 2006). Similar to its role in
filamentation, GlcNAc induces white-to-opaque switching
majorly through the cAMP signaling pathway (Huang et al.
2010). High temperatures (>27 °C) promote opaque-to-
white switching when glucose is used as the carbon source,
but induce white-to-opaque switching when GlcNAc is
used as the carbon source (Huang et al. 2010). In the recent
study, Sun et al. found that acidic pH has an inducing effect
on white-to-opaque switching in several clinical isolates
of C. albicans (Sun et al. 2015). Unlike CO2 and GlcNAc,
the inducing effect of acidic pH is highly dependent on the
genetic background of strains. pH conditions affect both
CO2- and GlcNAc-induced opaque cell formation. Under
acidic pH conditions, both CO2 and GlcNAc cause a higher
rate of induction of the opaque phenotype (Sun et al. 2015).
CO2 and pH are tightly linked in environmental and bio-
logical systems. The dissociation of CO2 leads to acidifica-
tion of the intracellular environment. It remains to be inves-
tigated whether CO2-induced opaque cell formation is due
to acidification of the intracellular environment.
pH‑regulated sexual mating in C. albicans
Sexual reproduction is common in eukaryotic organisms.
White–opaque switching controls sexual mating in C.
albicans (Miller and Johnson 2002). To mate, white cells
must first switch to the opaque cell type since only opaque
cells can mate efficiently. Therefore, the inducing effect of
acidic pH on the opaque phenotype would be expected to
facilitate sexual mating in C. albicans. However, we found
that opaque cells mate much less efficiently at acidic pHs
Curr Genet (2016) 62:283–286
than at neutral or weakly alkaline pHs (Sun et al. 2015). In
Saccharomyces cerevisiae, pre-treatment of the MATa and
MATα cells in acidic pH media significantly increases its
mating efficiency (Soellick and Uhrig 2001). The induc-
tion of sexual agglutinability by pheromone in Hansenula
wingei is also highly dependent on pH (Yanagishima and
Fujimura 1981). Sexual reproduction is the engine of
genetic diversity, but it increases “costs” compared to asex-
ual reproduction (Barton and Charlesworth 1998; Ni et al.
2011). The increased mating efficiency due to phenotypic
switching in C. albicans could be offset by the repressing
effect of acidic pH conditions on mating. White–opaque
switching involves rapid or short-term adaptations to the
environmental changes, while sexual mating is critical for
genetic variations and long-term adaptation. pH could play
a balancing act on these two biological processes. Sun et al.
(2015) further demonstrated that acidic pH represses sex-
ual mating in C. albicans through the pheromone response
pathway (Sun et al. 2015). In contrast to the neutral or
weak alkaline pH conditions, the expression of a-phero-
mone and the α-pheromone receptor gene STE2 in “MTLa”
cells could not be efficiently induced by α-pheromone or
opaque α cells under acidic pH conditions. Mutation assays
indicate that the Ste2-MAPK pheromone response pathway
is essential for this induction.
The Rim101‑pH sensing pathway
The conserved PacC/Rim101 pathway plays a critical role
in pH sensing in fungi (Maeda 2012). As mentioned ear-
lier, Rim101 regulates pH-controlled morphological transi-
tions and virulence in C. albicans (Davis 2009). Deletion
of RIM101 suppresses alkaline pH-induced filamentation
(Davis et al. 2000). Phr1 and Phr2 are two genes, encoding
beta-1,3- and beta-1, 6-glucan crosslinking glycosidases
in C. albicans, differentially regulated by Rim101 under
different pH conditions (Davis et al. 2000; Fonzi 1999).
PHR1 is expressed at pHs >5.5, while PHR2 is exclusively
expressed at acidic pHs (<5.0). The phr1/phr1 mutant
exhibits growth defects under alkaline pH conditions, while
the phr2/phr2 mutant grows poorly under acidic pH condi-
tions. Deletion of RIM101 and PHR1 had no obvious effect
on opaque-to-white switching under different pH condi-
tions, while deletion of PHR2 resulted in a mass conversion
to the white phenotype under acidic pH conditions (Sun
et al. 2015). It has been suggested that Rim101 is processed
to different versions of proteins under acidic and alkaline
pH conditions (Li et al. 2004). Under alkaline pH condi-
tions, Rim101 is processed to a 74-kD active form, while
under acidic conditions, it is processed to a 65-kD protein
with unknown function (Li et al. 2004). This 65-kD protein
could be an active form under acidic pH conditions in C.
285
albicans. Consistently, deletion of RIM101 led to the loss
of acidic pH-induced opaque phenotype, suggesting that
Rim101 plays a major role in this regulation (Sun et al.
2015).
The cAMP signaling pathway
The cAMP signaling pathway is involved in the regulation
of many biological aspects in C. albicans (Wang 2013).
Mutants of this pathway (including ras1/ras1, cyr1/cyr1,
and pde2/pde2 mutants) undergo the white-to-opaque
switch at a rate similar to that of the wild-type control (Sun
et al. 2015). However, white and opaque cells of the cyr1/
cyr1 mutant (mutant of the adenylyl cyclase gene) exhibit
a distinct growth defects under different pH conditions.
White cells of the cyr1/cyr1 mutant show growth defects
under alkaline pH conditions, while opaque cells of the
mutant grow worse under acidic pH conditions. Therefore,
the cAMP signaling pathway could regulate the two cell
populations indirectly through the control of growth rate.
Conclusion
Humans are natural reservoirs for C. albicans. In the long
history of association with humans, this fungus has evolved
and well adapted to diverse niches of the host. The involve-
ment of pH regulation in the unique phenotypic switch and
sexual reproduction could represent one of these adaption
mechanisms to the fluctuating environment. The balancing
role of pH in these two biological processes would not only
contribute to its rapid environmental adaptation ability, but
also the increase of genetic diversity in the context of long-
term evolution in C. albicans.
Acknowledgments The authors thank Drs. Mira Edgerton and Yue
Wang for their insightful comments on the manuscript. The work in
the Huang lab was supported by Grants from the Chinese National
Natural Science Foundation (31370175, 31170086, and 81322026 to
G.H.) and the “100 Talent Program” Grant from the Chinese Acad-
emy of Sciences (to G.H.).
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