cells

Review
Melatonin and Indole-3-Acetic Acid Synergistically Regulate
Plant Growth and Stress Resistance
Min Zhang 1,† , Chunxue Gao 2,† , Ling Xu 3,† , Hui Niu 1 , Qian Liu 1 , Yixiao Huang 1 , Guoshuai Lv 2 ,
Hengshan Yang 2, * and Minhui Li 1,2,3,4,5,6, *

1 Inner Mongolia Key Laboratory of Characteristic Geoherbs Resources Protection and Utilization,
College of Pharmacy, BaoTou Medical College, Baotou 014040, China
2 University Engineering Research Center of Chinese (Mongolia) Ecological Planting Medicinal
Materials (Nurture) in Inner Mongolia Autonomous Region, College of agronomy,
Inner Mongolia Minzu University, Tongliao 028000, China
3 School of Life Sciences, Inner Mongolia University, Hohhot 010021, China
4 College of Pharmacy, Inner Mongolia Medical University, Hohhot 010110, China
5 Inner Mongolia Hospital of Traditional Chinese Medicine, Hohhot 010020, China
6 Inner Mongolia Traditional Chinese & Mongolian Medical Research Institute, Hohhot 010010, China
* Correspondence: yanghengshan@imun.edu.cn (H.Y.);
prof_liminhui@yeah.net or 102008164@btmc.edu.cn (M.L.)
† These authors contributed equally to this work.

Abstract: Plant growth and development exhibit plasticity, and plants can adapt to environmental
changes and stress. Various phytohormones interact synergistically or antagonistically to regulate
these responses. Melatonin and indole-3-acetic acid (IAA) are widespread across plant kingdom.
Melatonin, an important member of the neuroendocrine immune regulatory network, can confer
autoimmunity and protect against viral invasion. Melatonin functions as a plant growth regulator
Citation: Zhang, M.; Gao, C.; Xu, L.; and biostimulant, with an important role in enhancing plant stress tolerance. IAA has a highly
Niu, H.; Liu, Q.; Huang, Y.; Lv, G.; complex stress response mechanism, which participates in a series of stress induced physiological
Yang, H.; Li, M. Melatonin and changes. This article reviews studies on the signaling pathways of melatonin and IAA, focusing
Indole-3-Acetic Acid Synergistically on specific regulatory mechanisms. We discuss how these hormones coordinate plant growth and
Regulate Plant Growth and Stress development and stress responses. Furthermore, the interactions between melatonin and IAA and
Resistance. Cells 2022, 11, 3250. their upstream and downstream transcriptional regulation are discussed from the perspective of
https://doi.org/10.3390/
modulating plant development and stress adaptation. The reviewed studies suggest that, at low
cells11203250
concentrations, melatonin promotes IAA synthesis, whereas at high levels it reduces IAA levels.
Academic Editors: Julien Pirrello and Similarly to IAA, melatonin promotes plant growth and development. IAA suppresses the melatonin
Panagiotis Kalaitzis induced inhibition of germination. IAA signaling plays an important role in plant growth and
development, whereas melatonin signaling plays an important role in stress responses.
Received: 10 August 2022
Accepted: 14 October 2022
Published: 16 October 2022
Keywords: phytohormone; stress response; indole-3-acetic acid; melatonin; gene regulatory network;
auxins
Publisher’s Note: MDPI stays neutral
with regard to jurisdictional claims in
published maps and institutional affil-
iations. 1. Introduction
During their growth and development, plants must adapt to various physiological
responses triggered by environmental stress. These stress responses and the development
Copyright: © 2022 by the authors. of stress tolerance are driven essentially by plant hormones and their intricate crosstalk [1].
Licensee MDPI, Basel, Switzerland. Plant hormones, small organic molecules with remarkable physiological effects even at
This article is an open access article very low concentrations, are universally involved in plant biological processes and regulate
distributed under the terms and cell signal transduction to balance growth and stress responses [2]. Abscisic acid (ABA)
conditions of the Creative Commons is the most studied plant hormone involved in stress responses; however, other phyto-
Attribution (CC BY) license (https:// hormones such as auxin, cytokinin, melatonin, and brassinosteriod are also involved in
creativecommons.org/licenses/by/ plant responses to environmental stress. Synergistic or antagonistic interactions between
4.0/).

Cells 2022, 11, 3250. https://doi.org/10.3390/cells11203250 https://www.mdpi.com/journal/cells

Cells 2022, 11, 3250 2 of 18

different plant hormones can play a crucial role in multiple processes associated with plant
responses to stress as well as other conditions.
Auxin, the first plant hormone discovered, affects multiple stages of plant growth
and development. Auxins are generally considered to promote differentiation, as they
initiate plant development and regulate the realization of organ morphology. However, the
growth-promoting effects of auxins on plant stems, buds, and roots vary with concentration.
Melatonin was first discovered in plants in 1995 and is now considered a regulator of plant
growth. Similar to its observed effects in animals, melatonin has several specific functions in
plant physiology, including regulating plant growth and conferring resistance to biological
and abiotic stresses.
Accumulating evidence from recent studies indicates that the interaction between
indole-3-acetic acid (IAA) and melatonin plays an important role in plant growth and
adaptation to stress responses. Genetic and biochemical studies have revealed the regula-
tory mechanisms of the IAA and melatonin signaling pathways. The identities of related
genes have mainly been reported in Arabidopsis thaliana. At a certain low concentration,
melatonin works in parallel with IAA and functions as its analog in promoting the induc-
tion, growth, and development of lateral and adventitious roots. At a high concentration,
melatonin either reduces the IAA content, or maintains it at a constant level [3]. However,
under semi-arid conditions, exogenous melatonin increases the content of endogenous
IAA in maize, thus ensuring plant survival under stress and significantly improving maize
yield [4]. However, the interaction between melatonin and IAA remains controversial, and
the specific roles of these hormones have not been elucidated. Here, we review the research
progress on crosstalk networks between melatonin and IAA, focusing on the upstream
and downstream transcriptional regulation of key components between the two hormones.
We also elucidate the signal exchange between melatonin and IAA which guides plant
development and stress adaptation.

2. Biosynthesis of Melatonin and IAA

At present, an increasing number of studies are investigating the metabolic pathways
of melatonin and IAA. We found that tryptophan is the common precursor of both mela-
tonin and IAA. We have summarized the synthesis of these hormones in the tryptophan
metabolic pathway (Figure 1). Melatonin biosynthesis begins in a wide array of plant
species through tryptophan. L-tryptophan decarboxylase (PSID) catalyzes the conversion
of aromatic-L-amino-acid (DDC) into tryptamine, as shown in Figure 1; next, tryptamine
5-hydroxylase (CYP71P1) catalyzes tryptamine in serotonin, which is transformed by two
steps to melatonin. Among some other species such as Hypericum perforatum, tryptophan,
is catalyzed by tryptophan 5-hydroxylase (TPH) in 5-hydroxytryptophan, and then DDC
transforms 5-hydroxytryptophan to serotonin. The same metabolic pathway is described
for synthesis of melatonin in animals. The serotonin is converted to N-acetyl-serotonin
in the next two steps, using serotonin N-acetyltransferase (SNAT)/arylalkylamine N-
acetyltransferase (AANAT), and then catalysis of acetylserotonin O-methyltransferase
(ASMT)/caffeic acid 3-O-methyltransferase (COMT) N-acetyl-serotonin results in mela-
tonin. Tryptophan-is not only a provider of melatonin but also an indole-3-acetic pre-
cursor acid (IAA), perhaps implying that melatonin has a multifunctional role in plants.
NADPH:oxygen oxidoreductase (N-hydroxylating) (E 1.14.1.13.-) catalyzes the conversion
of tryptamine into N-hydroxytryptamine, which is then converted to indole-3-acetaldoxime
under the action of N-hydroxyl-tryptamine oxidoreductase (E 2.5.1.-).
Cells 2022,11,
Cells2022, 11,3250
x FOR PEER REVIEW 33 of 18
18

Figure 1.
Figure 1. Synthesis
Synthesis pathways
pathways of
of IAA
IAA and
and melatonin
melatonin in
in tryptophan
tryptophan metabolism.
metabolism.

Tryptophan
TryptophanN-monooxygenase
N-monooxygenase(CYP79B1_2)/CYP79B3
(CYP79B1_2)/CYP79B3 is used to directly directly transform
transform
tryptophan
tryptophaninto indole-3-acetaldoxim.
into indole-3-acetaldoxim. Next, Next,
aromatic aldoximealdoxime
aromatic N-monooxygenase (CYP83B1)
N-monooxygenase
converts
(CYP83B1) indole-3-acetaldoxim to S-(indolylaceto-hydroximoyl)-L-cysteine.
converts indole-3-acetaldoxim S-(Indolylaceto-
to S-(indolylaceto-hydroximoyl)-L-cysteine. S-
Hydroximoyl)-L-cysteine is converted to indole-3-thiohydroximate
(Indolylaceto-Hydroximoyl)-L-cysteine by S-alkyl-thiohydroximate
is converted to indole-3-thiohydroximate by S-al-
lyase (SUR1), indole-3-thiohydroximate
kyl-thiohydroximate synthesizes lindolylme thyldesulfoglucosinolate
lyase (SUR1), indole-3-thiohydroximate synthesizes lindolylme
through N-hydroxythioamide S-beta-glucosyltransferase
thyldesulfoglucosinolate through N-hydroxythioamide (UGT74B1), Then, lindolylme
S-beta-glucosyltransferase
thyldesulfoglucosinolate synthesizes glucobrassicin through aromatic
(UGT74B1), Then, lindolylme thyldesulfoglucosinolate synthesizes glucobrassicin desulfoglucosino-
late sulfotransferase
through (ST5A), and glucobrassicin
aromatic desulfoglucosinolate is synthesized
sulfotransferase into glucobrassicin
(ST5A), and indole-3-acetonitrile
is syn-
catalyzed
thesized intoby myrosinase (E 3.2.1.147)
indole-3-acetonitrile and indoleacetaldoxime
catalyzed by myrosinase (E dehydratase
3.2.1.147) and(CYP71A13).
indolea-
Indole-3-acetaldoxim
cetaldoxime dehydratase is directly transformed
(CYP71A13). into indole-3-acetonitrile
Indole-3-acetaldoxim through
is directly CYP71A13
transformed into
and aldehyde dehydrogenase (ALDH)/aldehyde dehydrogenase
indole-3-acetonitrile through CYP71A13 and aldehyde dehydrogenase (ALDH)/aldehyde family 7 member A1
(ALDH7A1)/aldehyde dehydrogenase family 9 member A1
dehydrogenase family 7 member A1 (ALDH7A1)/aldehyde dehydrogenase family 9(ALDH9A1). Finally, indole-3-
acetonitrile
member A1is(ALDH9A1).
transformedFinally,
into indole-3-acetic acid (IAA)
indole-3-acetonitrile through nitrilase
is transformed (E 3.5.5.1).
into indole-3-acetic
Tryptophan is synthesized
acid (IAA) through nitrilase (E 3.5.5.1).by tryptophan 2-monooxygenase (iaaM) into indole-3-
acetamide, and indole-3-acetonitrile
Tryptophan can also be
is synthesized by tryptophan converted into (iaaM)
2-monooxygenase indole-3-acetamide by
into indole-3-ac-
nitrile hydratase subunit alpha (nthA)/nitrile hydratase subunit beta (nthB).
etamide, and indole-3-acetonitrile can also be converted into indole-3-acetamide by nitrile Amidase (E
3.5.1.4) and indoleacetamide hydrolase (iaaH) catalyze the conversion of indole-3-acetamide
hydratase subunit alpha (nthA)/nitrile hydratase subunit beta (nthB). Amidase (E 3.5.1.4)
to
andIAA. Another IAA synthesis
indoleacetamide hydrolasepathway is tryptophan
(iaaH) catalyze catalyzed by
the conversion L-amino-acid oxidase
of indole-3-acetamide to
(IL4L1)/tryptophan aminotransferase (Tam1)/aromatic amino acid aminotransferase I
IAA. Another IAA synthesis pathway is tryptophan catalyzed by L-amino-acid oxidase
(ARO8)/L-tryptophan—pyruvate aminotransferase (TAA1) to synthesize indole pyru-
(IL4L1)/tryptophan aminotransferase (Tam1)/aromatic amino acid aminotransferase I
vate, and then by indole-3-acetaldehyde oxidase (AAO1_2)/benzaldehyde dehydrogenase
(ARO8)/L-tryptophan---pyruvate aminotransferase (TAA1) to synthesize indole py-
(AAO4) and indole-3-pyruvate monooxygenase (YUCCA) to synthesize IAA from indole
ruvate, and then by indole-3-acetaldehyde oxidase (AAO1_2)/benzaldehyde dehydrogen-
pyruvate. It can be seen from the above that the upstream regulation of melatonin and IAA
ase (AAO4) and indole-3-pyruvate monooxygenase (YUCCA) to synthesize IAA from in-
has been thoroughly studied at the present stage, and there is indeed a certain relationship
dole pyruvate. It can be seen from the above that the upstream regulation of melatonin
between IAA and melatonin, which lays a foundation for our in-depth discussion on the
and IAA has been thoroughly studied at the present stage, and there is indeed a certain
signal transduction mechanism of melatonin and IAA.
relationship between IAA and melatonin, which lays a foundation for our in-depth dis-
cussion
3. on theSignaling
Melatonin signal transduction
in Plants mechanism of melatonin and IAA.
Owing to the increasing achievement of interesting results in melatonin-related re-
search in animals, researchers have turned their attention to the functions of melatonin
Cells 2022, 11, x FOR PEER REVIEW 4 of 18

Cells 2022, 11, 3250

3. Melatonin Signaling in Plants 4 of 18
Owing to the increasing achievement of interesting results in melatonin-related re-
search in animals, researchers have turned their attention to the functions of melatonin in
plants. Melatonin plays an important role in plant development and resistance to stress
in plants. Melatonin plays an important role in plant development and resistance to
[5–8]. Its biosynthesis is induced by abiotic stressors such as cold, drought, and heavy
stress [5–8]. Its biosynthesis is induced by abiotic stressors such as cold, drought, and
metals, activating
heavy metals, particular
activating stress response
particular factors. Itfactors.
stress response participates in and regulates
It participates in and plant
regu-
growth, promotes root growth after seed germination [9], affects the
lates plant growth, promotes root growth after seed germination [9], affects the flowering flowering time [10],
and
timeregulates sugar metabolism
[10], and regulates in plantsin[11].
sugar metabolism The[11].
plants signal
Thepathway of melatonin
signal pathway in plants
of melatonin in
has also been gradually elucidated. The first phytomelatonin receptor
plants has also been gradually elucidated. The first phytomelatonin receptor in Arabidopsis in Arabidopsis thali-
ana, candidate
thaliana, candidateG-protein
G-proteincoupled
coupledreceptor
receptor 2 (CAND2),
2 (CAND2),a amembrane
membraneprotein protein that
that binds
binds
readily with melatonin, was identified in 2018 [12]. Treatment with
readily with melatonin, was identified in 2018 [12]. Treatment with exogenous melatonin exogenous melatonin
at
at aa 50
50 µM
μM concentration
concentrationupregulated
upregulatedthe theRNA
RNApolymerase
polymerase genes
genes RPOTm
RPOTm andand RPOTmp
RPOTmp by
by promoting the CAND2 receptor and heterotrimeric G protein
promoting the CAND2 receptor and heterotrimeric G protein α subunit (GPA1) coupled α subunit (GPA1) cou-
pled to CAND2
to CAND2 [13].[13]. In Arabidopsis,
In Arabidopsis, Cand2/pmrt1
Cand2/pmrt1 is is locatedon
located onthe
theplasma
plasma membrane
membrane (PM), (PM),
interacts
interacts with
with GPA1,
GPA1, and and regulates
regulates stomatal
stomatal movement
movement via via the
the reactive
reactive oxygen
oxygen species
species
(ROS) signaling
(ROS) signaling pathway
pathway mediated
mediated by by NADPH
NADPH oxidase
oxidase [14].
[14].
Treatment
Treatment with 1 µM melatonin activated MAPK3 and MAPK6
with 1 μM melatonin activated MAPK3 and MAPK6 via via upstream
upstream MAPKMAPK
kinases (MKKs), including MAPKK4, MAPKK5, MAPKK7, and and MAPKK9 [15].
MAPKK9 [15]. However,
However,
this activation
activationwas wasnotnotassociated
associatedwithwithGG protein
protein signaling,
signaling,as as
thethelevels of Gofprotein
levels β did
G protein β
not
did change in a in
not change knockout
a knockout mutant (agb1)
mutant of Arabidopsis.
(agb1) of Arabidopsis.ThisThis
indicated
indicatedthatthat
melatonin
melatonin in-
duced
induced innate
innateimmunity
immunitywas wastriggered
triggeredby byMAPK
MAPK signals
signals from multiple
multiple MAPKK/MAPK
MAPKK/MAPK
components. MEKK MEKKactivation
activationisisconsidered
consideredtoto bebemediated
mediated viaviamelatonin
melatonin receptors
receptorsor
receptor
or receptor kinases thatthat
kinases recognize pathogen-derived
recognize pathogen-derived molecular patterns
molecular (PAMPs)
patterns and effec-
(PAMPs) and
tors, leading
effectors, to the
leading activation
to the of PAMP-triggered
activation of PAMP-triggered immunity
immunity(PTI)(PTI)andandeffector-triggered
effector-triggered
immunity (ETI), respectively [16]. Melatonin Melatonin can can induce
induce plant responses by activating
MAPKKK3 and oxidative signal-inducible 1 (OXI1) [17] (Figure (Figure 2).2).

Figure 2. Melatonin
Melatonin signaling
signalingpathway
pathwayfrom
frommelatonin
melatoninperception to action.
perception Melatonin
to action. promotes
Melatonin the
promotes
the production
production of itsofreceptor
its receptor protein,
protein, enabling
enabling upstream
upstream MKK toMKK to activate
activate MAPK3MAPK3 and MAPK6,
and MAPK6, thereby
thereby activating
activating melatonin melatonin
signal. signal.

peroxide (H
Hydrogen peroxide (H22OO2)2 )and
andnitric
nitric oxide
oxide (NO)
(NO) cancan regulate
regulate plant
plant responses
responses to
to en-
environmental stress. The suppression of NADPH oxidase (RBOH)
vironmental stress. The suppression of NADPH oxidase (RBOH) activity or the activity or the removal
O22blocks
of H22O blocksthe
theexpression
expressionof ofstress-related
stress-relatedgenes,
genes,such
suchas CDPK1, MAPK1,
asCDPK1, MAPK1, ERF4, and
ERD15, further weakening melatonin induced defense responses [18]. NO and melatonin
can interact to regulate glutathione (GSH) levels and glutathione reductase (GR) activity
and maintain low levels of H2O22,,thus thus regulating
regulating resistance
resistance to
to salt
salt stress
stress [19].
[19].
Cells 2022, 11, 3250 5 of 18

4. IAA Signaling in Plants

Auxin, which at low concentrations promotes plant growth, is critical for plant growth
and stress resistance [20,21]. Its reported responses have become increasingly difficult to
integrate into the accepted canonical auxin signaling pathways, and there is abundant
literature on the noncanonical auxin signal transduction pathways. The auxin receptor
transport inhibitor response 1 (TIR1) is central to both mechanistic pathways: its canonical
pathway is localized in the nucleus and its noncanonical pathway in the cyto-plasmic matrix
and plasma membrane [22]. The tir1-1 mutant of Arabidopsis thaliana shows defective root
growth when heat shock protein 90 (HSP90) is inhibited. HSP90 forms a complex with
the auxin receptor transport inhibitory response 1 (TIR1) and, when inhibited, impairs the
nuclear localization of TIR1 and eliminates plant responses to auxins [23].
The IAA signal transduction pathway has been described in detail. Canonical auxin
signal transduction mainly depends on the following three protein families: the F-box
transport inhibitor response 1/auxin signaling F-box protein (TIR1/AFB) auxin co-receptors;
the auxin/indole-3-acetic acid (Aux/IAA) transcriptional repressors; and the auxin response
factor (ARF) transcription factors [24,25]. The accumulation of IAA leads to faster hy-
drolysis of the Aux/IAA protein and alleviates the inhibition of auxin response genes by
allowing the formation of ARF dimers. Although this proposed model cannot explain
the function of the ARF subtype, it is considered as an inhibitor. The TIR1/ABF protein
binds to the Aux/IAA transcription inhibitor and participates in its polyubiquitination and
subsequent proteasome-based degradation. This pathway rapidly induces auxin response
genes—including Aux/IAAs and the auxin-responsive Gretchen Hagen3 (GH3) family of
auxin homeostasis regulators—and subsequently triggers a negative feedback loop. How-
ever, when the IAA level is low, the Aux/IAA transcription inhibitors interact with ARFs
and inhibit their activity [26].
The interaction between the Aux/IAA protein and the SCFTIR1 complex is regulated by
auxin, which simultaneously affects TIR1 and related proteins. This pathway demonstrates
the unique mode of action of SCF ligase; namely, auxin promotes the interaction between
the TIR1 and Aux/IAA proteins by binding to TIR1 [27–29]. ARFs participate in a prominent
noncanonical pathway. The N-terminal domains of ARFs also function as dimerization
domains. Most ARFs have a conserved C-terminal Phox/Bem1p box (PB1) domain that is
responsible for binding AUX/IAA proteins via the canonical auxin pathway. The ARF3
variant ETTIN (ETT) participates in noncanonical auxin ETT-signaling, regulating growth
and tissue patterning in an IAA-dependent mechanism.
Noncanonical auxin signaling is associated with various protein kinases, including D6
protein kinases, PINOID, mitogen-activated protein kinases [MAPKs], and PM-associated
kinases. These contribute to the action of auxin by moderating PINFORMED (PIN) protein
localization (phosphorylation) and activity [22]. It remains unclear whether auxin binding
protein 1 (ABP1) functions as an auxin receptor. In Arabidopsis, ABP1 is secreted and
binds specifically to the native auxin IAA at the typical acidic pH. These findings indicate
that ABP1 is the auxin receptor for TMK1-based cell surface signaling, which mediates
the global phospho-response and auxin canalization [30,31]. Both IAA and melatonin,
therefore, play important roles in plant growth and resilience by interacting with MAPK
kinases (Figure 3).
 x FOR PEER REVIEW
Cells 2022, 11, 3250 66of
of 18

Figure 3. IAA signaling pathway from IAA perception to action. After receiving the IAA signal, the
interaction between the
interaction between thethree
threeprotein
proteinfamilies
familiesofofauxin
auxinandand key
key components
components promotes
promotes thethe produc-
production
tion of up-regulated
of up-regulated genegene encoding
encoding GH3 GH3 protein
protein to activate
to activate the downstream
the downstream IAA signal.
IAA signal.

5. Relationships between the Melatonin and IAA Pathways

Melatonin and IAA have well documented synergistic effects on plant growth and
development.
development. Auxins Auxins and
and melatonin
melatonin cooperatively
cooperatively induce
induce adventitious
adventitious rootroot formation.
formation.
The growth promoting
The growth promoting activity
activity of
of melatonin
melatonin is one of
is one of its
its auxin-like
auxin-like effects.
effects. Treatment
Treatment with with
exogenous melatonin
exogenous melatonin has been shown
has been shown to to increase
increase thethe content
content of of auxin
auxin signal
signal transduction
transduction
genes (IAA19
genes (IAA19 and IAA24) in
and IAA24) in tomato
tomato seedlings.
seedlings. NO NO acts
acts as
as downstream
downstream signalsignal ofof melatonin
melatonin
to enhance the auxin signal and induce adventitious roots in tomato
to enhance the auxin signal and induce adventitious roots in tomato explants [32]. There- explants [32]. There-
fore, exogenous melatonin can activate the auxin signaling pathway.
fore, exogenous melatonin can activate the auxin signaling pathway. However, melatonin However, melatonin
signaling also
signaling also antagonizes
antagonizes IAA
IAA biosynthesis.
biosynthesis. In In transgenic
transgenic plants
plants with
with melatonin
melatonin overex-
overex-
pression, the decreased expression levels of key auxin biosynthesis
pression, the decreased expression levels of key auxin biosynthesis genes (including genes (including YUC1,
YUC2,
YUC1, YUC5,
YUC2, YUC6,
YUC5, andYUC6,TAR2),
and as well as
TAR2), asthe
welleffects
as the of effects
melatonin on auxin transport,
of melatonin on auxin
significantly reduce the level of IAA in plant roots. These findings
transport, significantly reduce the level of IAA in plant roots. These findings indicate that melatonin
indicate that
negatively negatively
melatonin regulates auxin biosynthesis
regulates [33].
auxin biosynthesis [33].
IAA and
IAA andmelatonin
melatoninsignals
signalscancanalso be positively
also be positively cross-regulated.
cross-regulated. Transcriptome
Transcriptome anal-
ysis has revealed similar expression patterns of melatonin and auxin,
analysis has revealed similar expression patterns of melatonin and auxin, confirming a confirming a partial
overlap between the regulatory pathways of these hormones. The melatonin-mediated
partial overlap between the regulatory pathways of these hormones. The melatonin-me-
effects of the auxin pathway on root elongation were further investigated in plants that
diated effects of the auxin pathway on root elongation were further investigated in plants
were auxin deletion mutants yucQ (yc3, yuc5, yuc7, yuc8, or yuc9 mutations) or had been
that were auxin deletion mutants yucQ (yc3, yuc5, yuc7, yuc8, or yuc9 mutations) or had
treated with inhibitors of auxin synthesis (L-AOPP) or auxin transport (TIBA). When auxin
been treated with inhibitors of auxin synthesis (L-AOPP) or auxin transport (TIBA). When
synthesis was completely inhibited, there was no obvious effect of melatonin on root elon-
auxin synthesis was completely inhibited, there was no obvious effect of melatonin on
gation, suggesting that this effect is preconditioned on the presence of auxin [34]. Optimal
root elongation, suggesting that this effect is preconditioned on the presence of auxin [34].
concentrations of IAA (0–0.2 mg/L) and melatonin (0–0.2 mmol/L) in liquid MS medium
Optimal concentrations of IAA (0–0.2 mg/L) and melatonin (0–0.2 mmol/L) in liquid MS
promoted the growth of hairy roots in Isatis indigotica, whereas higher concentrations
medium promoted the growth of hairy roots in Isatis indigotica, whereas higher concen-
caused the de-differentiation of hairy roots. Treatment with 0.2 mg/L IAA and 0.2 mmoL/L
trations caused the de-differentiation of hairy roots. Treatment with 0.2 mg/L IAA and 0.2
melatonin increased the activities of enzymes (superoxide dismutase [SOD], peroxidase
mmoL/L melatonin increased the activities of enzymes (superoxide dismutase [SOD], pe-
[POD], and catalase [CAT]) and the indirubin content of Isatis tinctoria hairy roots under
roxidase
salt stress,[POD], and catalase
and melatonin had[CAT])
a greater and the indirubin
effect than IAA content of Isatis tinctoria
[35]. In Helianthus hairy
tuberosus L.,roots
IAA
under salt stress, and melatonin had a greater effect than IAA [35].
improved the growth index to a greater extent, whereas melatonin had greater effects on In Helianthus tuberosus
L., IAA improved
photosynthesis andthe
the growth index
antioxidant to a greater
system. Moreover,extent, whereas
melatonin melatonin
promoted roothad greater
elongation,
effects on photosynthesis and the antioxidant system. Moreover, melatonin
dependent on the presence of IAA. These findings indicate that the combination of these promoted root
elongation, dependent on the presence of IAA.
enzymes can greatly improve stress resistance in plants [36]. These findings indicate that the combina-
tion of these enzymes can greatly improve stress resistance in plants [36].
Cells 2022, 11, 3250 7 of 18

5.1. IAA Signaling Regulation by Melatonin

Melatonin shares similarities with IAA in terms of its chemical structure and biosyn-
thetic pathway, suggesting a potential link in signal transduction [37,38]. Melatonin treat-
ment can increase IAA levels, and melatonin exhibits similar activity to IAA [39], with
similar effects on plant growth and development. For example, melatonin promotes plant
flowering, photosynthesis, senescence, and seed development [37,40,41]. These phyto-
hormones co-regulate plant growth and development. When seeds were treated with
melatonin and IAA simultaneously, IAA inhibited the inhibitory effect of melatonin on
seed germination, indicating that IAA antagonizes melatonin in the regulation of seed
germination [42]. Using the IAA-responsive marker construct DR5::GUS, it was demon-
strated that the effects of melatonin on Arabidopsis roots are independent of IAA signal
transduction [36]. However, other researchers have reported that melatonin has both posi-
tive and negative effects on the production of endogenous growth hormones. For example,
in Arabidopsis, several IAA-related transcription factors (Including NAC019, TCH4, FLA8,
and PIN5) were up- or down-regulated after treatment with melatonin [43]. In another
root growth study on Arabidopsis, melatonin was shown to regulate IAA distribution by
regulating IAA transport, thereby enhancing lateral root development. This indicated
that melatonin and IAA synergistically promote lateral root development in wild-type
Arabidopsis thaliana [44]. Of the 16 IAA-related genes identified, 12 genes (AXR3, At1G29500,
SAUR68, TT5, At4G38860, At4G00880, TT4, PIN5, At2G21050, CYP83A1, WAG1, and SHY2)
were found to be down-regulated by melatonin and 4 genes (ACS8, At3G12830, AtGSTU1,
and GH3.3) were up-regulated, suggesting a potential role of melatonin in regulating IAA
transport [44]. A comprehensive analysis of IAA synthesis, the PIN protein, and IAA
reaction markers in Arabidopsis also showed that melatonin regulated the root meristem by
inhibiting IAA synthesis and polar IAA transport [33].
Melatonin plays a protective role against oxidative stress, and oxidative stress can
increase plant melatonin levels [45–47]. Melatonin promotes tolerance to low temperatures
and osmotic stress, reduces physiological damage from stress, and improves salt tolerance
and resistance to fungal diseases in plants [48,49]. Most studies on the roles of auxins
in abiotic stress have focused on water-related (particularly drought), salt, and heavy
metal stress [50]. However, IAA and melatonin also interact with each other to jointly
promote plant resistance to biotic and abiotic stresses. A study identified 51 genes related
to IAA response and signaling (including MDC12, PBS3, ATGSTU1, HAI1, AT1G63840,
ACS8, AT1G63720, TCH4, BT2, AT3G51660, CNI1, MPK11, and GH3.3) that were altered by
melatonin. Among these, 29 genes (including MDC12, MPK11, GH3.3, ZF2, NAC019, PMZ,
and SYP122) were up-regulated and 23 genes (including AT2G41820, PG2, Prx37, FLA8,
AT1G31710, UGT74B1, WAG1, TT4, PIN5, and AUX1) were down-regulated in response to
melatonin. The study also found that most of the IAA-responsive genes down-regulated in
response to melatonin were involved in IAA transport and homeostasis [43]. In addition,
one of the up-regulated genes encodes GH3 protein, an IAA-amino synthase that combines
amino acids with IAA to inactivate it [51]. This suggests that Arabidopsis seedlings respond
to excess auxin in response to high levels of melatonin. However, none of the known genes
on the IAA biosynthetic pathway (including TAA, YUC, CYP79B, CYP450, TDC, AMI, and
TIR) showed significantly altered expression in response to melatonin [52].
The YUCCA (YUC) proteins, tryptophan aminotransferase of Arabidopsis (TAA) family,
and TAA-related 1 and 2 proteins play important roles in the biosynthesis of IAA. Wang
et al. [33] explored the effects of different concentrations of melatonin on the transcription
levels of these genes. The transcription levels of YUC1, YUC2, YUC5, YUC6, and TAR2 de-
creased significantly after treatment with 600 µM melatonin, whereas those of YUC3, YUC4,
YUC7, and YUC8 increased after treatment. The relative expression levels of YUC3 and
YUC8 in roots were 1.5 times lower after treatment than those in the control. The YUCCA
gene family contains key enzymes that catalyze IAA biosynthesis. A study analyzed the
transcription levels of the YUCCA gene family after treatment with low concentrations of
IAA and melatonin and found that both YUCCA5 and YUCCA8 were down-regulated. This
Cells 2022, 11, 3250 8 of 18

indicated that the promoting of root length in wild-type Arabidopsis at low concentrations
of melatonin was closely related to IAA [34]. In addition, the effects of melatonin treat-
ment on PIN proteins (especially PIN1, PIN3, and PIN7) were also explored. The authors
found that treatment with 600 µM melatonin significantly reduced the levels of PIN1 in
pre-culture and of PIN3 and PIN7 in the root cap and pre-culture area. This indicated that
melatonin treatment inhibited the expression of PIN1, PIN3, and PIN7. Experiments on root
development also showed that PIN1/3/7 played important roles in the melatonin mediated
inhibition of the root meristem.
The biosynthetic pathways of melatonin and IAA share the same substrate, trypto-
phan. Low concentrations of exogenous IAA increase melatonin production, whereas high
concentrations of melatonin reduce IAA production and the levels of PIN1/3/7 in Arabidopsis
roots [44]. These dual actions of exogenous melatonin also lead to high levels of IAA in the
cytoplasm and an increase in lateral root growth. At present, it is unclear how cytoplasmic
IAA regulates lateral root growth. One possibility is that cytoplasmic IAA increases the
level of cytosolic Ca2+ ions, and that increased calcium ion signaling ultimately leads to
lateral root development [53]. Transcriptome analysis of the expression patterns of low
concentration melatonin and IAA revealed many similarities between the two; for example,
the genes regulated by melatonin and IAA were enriched in several common pathways in
Arabidopsis thaliana [34]. The promotion of root growth by low concentrations of melatonin
depends on the presence of IAA, and when the transport or synthesis of IAA is severely
inhibited, melatonin cannot promote root growth.
Studies on the interaction between melatonin and IAA have typically lacked a di-
rect comparison between melatonin and IAA treatments under the same experimental
conditions. In a recent study, DR5::GFP and AOX1a::LUC were used as markers for IAA
response and mitochondrial retrograde signaling, respectively. These were used together
with melatonin or IAA-treated Arabidopsis rosette leaves, and transcriptome analysis
was conducted to investigate the potential molecular crosstalk between melatonin and
IAA [54]. The results showed that melatonin treatment did not affect the expression of
the IAA-responsive DR5::GFP gene in Arabidopsis seedlings. Unlike IAA, melatonin also
did not affect mitochondrial retrograde signal transduction, but affected the expression of
photosynthesis related genes. These results may indicate a trade-off between growth and
defense. The small IAA up-regulating RNA (SAUR) family is one of three gene families
known to be significantly elevated and transiently induced upon treatment with IAA [55].
The findings of several studies suggest that SAURs play an important role in IAA-mediated
tissue root elongation in Arabidopsis and negatively affect IAA biosynthesis and polar IAA
transport in rice [56]. IAA causes swelling of the cell wall [57], and melatonin may reduce
the transcriptional level of genes in the SAUR family, thus reducing cell wall expansion
and limiting the potential for pathogen invasion.
The aforementioned findings suggest that, in Arabidopsis, melatonin can regulate
lateral and adventitious root induction in a parallel manner to IAA [58]. Melatonin can
also interfere with IAA action by altering the IAA carrier, thereby altering the local IAA
gradient [33,43,58]. Some researchers have proposed a working model to explain the roles of
melatonin as an IAA-like regulator [59]. The signaling molecule NO is involved in various
physiological processes during plant growth and development and is also an important
regulator of stress responses and pathophysiological processes [60,61]. NO regulates plant
roots, adventitious roots, lateral roots, root hair formation, and root geotropism [62–65].
The application of a NO donor can also mimic the effects of IAA. In Arabidopsis thaliana,
auxin induces NO production in roots, mediated by nitrate reductase and the induction
of S-nitrosothiols from proteins, regulating the activation of cell division and subsequent
adventitious or lateral root formation, suggesting that NO plays a very important role
in the induction of IAA [66]. Furthermore, NO mediates the IAA response, leading to
the formation of dominant roots, and NO acts downstream of auxin [67]. One study
reported that PINFORMED (PIN) proteins, specifically PIN1, PIN3 and PIN7, are directly
involved in auxin transport in plant roots, Melatonin treatment induced changes in apical
Cells 2022, 11, x FOR PEER REVIEW 9 of 18

Cells 2022, 11, 3250 9 of 18

that PINFORMED (PIN) proteins, specifically PIN1, PIN3 and PIN7, are directly involved
in auxin transport in plant roots, Melatonin treatment induced changes in apical IAA traf-
IAA trafficking
ficking by up-regulating
by up-regulating several
several PIN PIN proteins
proteins (PIN1, PIN3, andPIN3,
(PIN1, andPIN7)
PIN7)and and IAA
IAA signaling
signaling
genes (IAA19genes
and(IAA19
IAA24).andIn IAA24).
contrast,Inthecontrast, the of
expression expression
these same ofproteins
these same proteins
(PIN1, PIN3,
(PIN1, PIN3, and PIN7) is inhibited in Arabidopsis, thereby affecting root growth
and PIN7) is inhibited in Arabidopsis, thereby affecting root growth [33]. The proposed [33]. The
proposed model suggests that the dual control of NO levels by melatonin
model suggests that the dual control of NO levels by melatonin and IAA may fine tuneand IAA may fine
tune plant responses during growth, rooting, and tropicalization through the
plant responses during growth, rooting, and tropicalization through the IAA carrier pro- IAA carrier
protein
tein [48][48] (Figure
(Figure 4). 4).

Figure 4.4. Regulation

Figure Regulation of
of IAA
IAA signaling
signaling by
by melatonin
melatonin include
include key
key components
components and
and transcriptional
transcriptional
regulation.
regulation.

5.2.
5.2. IAA-Regulated
IAA-Regulated Melatonin
Melatonin Signaling
Signaling
In
In view
view of of the
the similarity
similarity in in chemical
chemical structure
structure andand biosynthetic
biosynthetic pathway
pathway between
between
IAA
IAA andand melatonin,
melatonin, researchers
researchers have have been
been exploring
exploring their
their effects
effects onon plant
plant growth
growth and and
development.
development. A virus induced gene silencing silencing (VIGS)
(VIGS) technique
technique waswas used
used toto determine
determine
that
that inhibition
inhibition of of DREB1α
DREB1α and and IAA3
IAA3 significantly
significantly reduced
reduced melatonin
melatonin induced
induced saline–alkali
saline–alkali
tolerance Solanum lycopersicum
tolerance in Solanum lycopersicum (tomato). Furthermore, the physiological function
in (tomato). Furthermore, the physiological function of of
DREB1α and IAA3 mediated melatonin in improving saline–alkali
DREB1α and IAA3 mediated melatonin in improving saline–alkali tolerance of tomato tolerance of tomato was
further investigated.
was further It wasIt found
investigated. that inhibiting
was found DREB1α
that inhibiting and IAA3
DREB1α significantly
and IAA3 reduced
significantly re-
the
ducedexpression of genes
the expression of related to ion to
genes related transport, organic
ion transport, acid accumulation,
organic acid accumulation, stomatal
sto-
movement,
matal movement, water retention and antioxidant
water retention enzymesenzymes
and antioxidant in roots induced by melatonin.
in roots induced They
by melato-
found that DREB1α and IAA3 are key downstream genes of melatonin
nin. They found that DREB1α and IAA3 are key downstream genes of melatonin induced induced saline–alkali
tolerance
saline–alkaliin tomato,
tolerance and the melatonin
in tomato, and the-DREB1α-IAA3 cascade signaling
melatonin -DREB1α-IAA3 cascadenetwork
signaling plays
net-
multiple roles in regulating tomato growth and stress tolerance balance
work plays multiple roles in regulating tomato growth and stress tolerance balance [68]. [68]. The effects of
exogenous 2,4-D (an auxin analogue) on endogenous hormones were
The effects of exogenous 2,4-D (an auxin analogue) on endogenous hormones were de- detected in embryonic
callus
tectedof inEriobotrya
embryonic prinoides
callus ofvar dadunens.prinoides
Eriobotrya The results showed changes
var dadunens. in melatonin
The results showed changesrelated
to culture time, first decreasing and then increasing after 30 days [69]. In Arabidopsis
in melatonin related to culture time, first decreasing and then increasing after 30 days [69]. thaliana,
the addition of auxin polar transport inhibitor TIBA indicated that the regulation of primary
In Arabidopsis thaliana, the addition of auxin polar transport inhibitor TIBA indicated that
root growth by melatonin and cytokinin was dependent on auxin transport. This indicates
the regulation of primary root growth by melatonin and cytokinin was dependent on
that the polar transport of auxin plays an important role in this process [70]. However, the
auxin transport. This indicates that the polar transport of auxin plays an important role in
results indicate that the expected molecular regulatory mechanisms of auxin and melatonin
this process [70]. However, the results indicate that the expected molecular regulatory
are less well studied. This is a very promising area of future exploration. In addition, only
mechanisms of auxin and melatonin are less well studied. This is a very promising area
a few genes are involved in their temporal regulation, which hinders research on their
of future exploration. In addition, only a few genes are involved in their temporal regula-
participation in plant stress. Gene expression analysis of auxin and melatonin regulation
tion, which hinders research on their participation in plant stress. Gene expression analy-
in response to various abiotic stresses is expected to elucidate the regulatory pathways of
sis of auxin and melatonin regulation in response to various abiotic stresses is expected to
these hormones and the underlying molecular mechanisms.
Cells 2022, 11, 3250 10 of 18

5.3. Interactions between Melatonin and IAA Levels to Balance Growth and Adaptation to Stress
Melatonin, an important stress response hormone in plants, accumulates under ad-
verse conditions such as drought, cold, flood, and heavy metal stress. This inhibits the
production of auxin, as both auxin and melatonin are synthesized with tryptophan as the
starting material. When IAA is the dominant signal in Arabidopsis thaliana, it is inhibited
by the zinc finger protein (ZAT6); in contrast, ZAT6 knockout plants are not sensitive to
melatonin regulated auxin biosynthesis [71]. Most studies on the interaction between mela-
tonin and auxin focus on plant roots. Although some studies have reported that melatonin
induced changes in plant root development may not be related to auxin signaling [39], stud-
ies on rice have shown that melatonin directly or indirectly activates the auxin signaling
pathway to shape root structure [72]. In addition, the promoting effect of melatonin on
lateral root development disappeared in knockout mutants of auxin transport (including
pin5, wag1, tt4, and tt5) in Arabidopsis thaliana. This indicated that melatonin and auxin
synergistically promoted lateral root development in wild-type Arabidopsis thaliana [44].
IAA plays an important role in mediating root changes during abiotic stress [73]. The
production of local maximum IAA inhibits cell elongation locally, thus preventing the
emergence of lateral roots. In Arabidopsis thaliana, the local minimum IAA was found to
trigger a transition from cell division to cell differentiation [74]. In the typical IAA signaling
pathway, the binding of IAA to its receptor activates the E3 ubiquitin ligase complex SCF,
resulting in the degradation of the Aux/IAA transcriptional repressor. This degradation
allows the ARF to regulate the expression of IAA response genes [75,76]. In Arabidopsis,
typical IAA perception includes six receptors that lead to the activation of target genes TIR1
and five AFB proteins (AFB1–AFB5) [77]. The accumulation and differential perception
of IAA in roots modulates several types of abiotic stresses [78–80]. A study on salt stress
showed that different treatment times lead to responses from different genes. For example,
TIR1 was up-regulated after six hours of salt stress treatment [78], whereas TIR1 and AFB2
were down-regulated at the protein level after four hours of salt stress treatment [81].
Interestingly, the overexpression of TIR1 (an anti-degradation form) endowed plants with a
salt tolerant phenotype and increased the seed germination rate, lateral root density, and
Na+ exclusion, among other parameters [78]. AFB3 has been identified to play a key role
under limited nitrate conditions, and its downstream signal components have also been
identified [82]. For example, the NAC4 transcription factor has been described as a signal
component downstream of AFB3 in response to nitrate [83]. Recent studies also show
that SZF1 is a key transcription factor in the IAA-dependent response to salt stress, as it
regulates NAC4 [84]. A study on drought stress revealed that, in Arabidopsis, some drought
stress genes are regulated by IAA. Osmotic stress inhibits cell division during the growth
of Arabidopsis leaves by inducing plant ARFs, at least in part [85]. In addition, other related
genes have been found to play important roles in the abiotic stress response of Arabidopsis.
Drought and salt stress significantly reduced the expression of TSB1, resulting in decreased
levels of tryptophan and IAA [86]. The same study also reported that a large amount of
H2 O2 produced by plants under drought or salt stress could sulfenylate the cysteine at
position 308 of the TSB1 protein. This inhibited the tryptophan synthase activity of TSB1,
thereby reducing the levels of tryptophan and IAA.
High temperature stress is known to have an important impact on the growth, devel-
opment, and geographical distribution of plants. In Arabidopsis, high temperature stress
can significantly induce the expression of a key transcription factor (PIF4) and an IAA
synthesis-related gene (YUC8) in the light signaling pathway. In a study on heavy metal
stress, researchers reported that aluminum stress can induce the specific ectopic expression
of key IAA synthesis-related genes (TAA1 and YUCCA) in the root tip transformation
region. This caused the excessive accumulation of IAA in this region, thus inhibiting the
elongation of main roots. This indicates that aluminum toxicity affects the normal growth
of plant roots by regulating local IAA synthesis, polar transport, and corresponding signal
transduction [87]. In addition, cadmium toxicity can also affect the growth and develop-
ment of plants by disturbing the internal homeostasis of IAA [88]. Auxins can directly
Cells 2022, 11, 3250 11 of 18

and positively regulate the resistance of plants to cadmium toxicity. This regulation is
mainly achieved by reducing the transport efficiency of Cd2+ from root to aboveground
parts and inducing the activities of some antioxidant enzymes, such as SOD, CAT, and
POD. These findings indicate that some key genes related to auxin synthesis, metabolism,
polar transport, and signal transduction are directly regulated by stress. However, the
regulation mechanisms of auxin at the molecular level under conditions of abiotic stress
need to be further examined in future studies. In addition, the molecular mechanisms of
synergistic or antagonistic interactions between phytohormones under stressful conditions
should also be investigated.
Melatonin has been shown to play an important role in plant responses to abiotic
stress. A study of drought stress revealed that melatonin conferred resistance to oxida-
tion, regulated infiltration, and promoted plant growth, thus slowing down the negative
effects of drought stress on plants and helping plants adapt to the drought environment.
Treatment with exogenous melatonin also increases the levels of proline, soluble sugar,
and soluble protein in plants [89] and plays a role in the osmotic protection of cells [90],
thus effectively improving the root water absorption capacity of seedlings under drought
stress. In Arabidopsis, the ectopic expression of MzSNAT5 can improve melatonin synthesis
in the mitochondria, reduce oxidative damage, and improve the drought tolerance of
plants [91]. Under salt stress treatment, melatonin can alleviate the damage caused by
salt stress. The main physiological mechanisms of this include the following: inhibiting
chlorophyll decomposition and increasing the photosynthetic rate [92]; enhancing the
activities of antioxidant enzymes and reducing the ROS content [93]; and regulating the
absorption and transport of Na+ , Cl– , and K+ ions and regulating ion homeostasis [94].
Heat stress can cause changes in a series of metabolic processes, leading to the excessive
production of ROS, photoinhibition, protein denaturation, damage to biofilm structure
and function, and inhibition of protein synthesis [95]. However, melatonin significantly
up-regulates the expression of heat shock factor HSFA1 to activate several heat response
genes—including HSFA2, HSA32, HSP90, and HSP101—thus improving the heat tolerance
of Arabidopsis plants [96]. In addition, treatment with exogenous melatonin promotes the
activity of antioxidant enzymes and improves the germination of Arabidopsis seeds under
heat stress [97]. Endogenous melatonin also alleviates the toxicity of heavy metals, and
the application of exogenous melatonin effectively alleviates heavy metal stress in plants.
Taken together, these results indicate that plants can increase the production of endogenous
melatonin in response to adverse environments, and that melatonin plays a crucial role in
Cells 2022, 11, x FOR PEER REVIEW regulating a series of physiological processes (such as plant responses to abiotic
12 of 18 stress) [98]
(Figure 5).

Figure
Figure5.5.Regulation of melatonin
Regulation signaling
of melatonin by IAA
signaling byinclude key components
IAA include and transcriptional
key components and transcriptional
regulation.
regulation.
When environmental stress conditions are alleviated and appropriate levels of water
and temperature are provided, the endogenous melatonin level decreases, whereas IAA
synthesis increases. The plant shifts from a poorly defended state and resumes growth.
The interaction between the two hormone signals helps regulate the balance of key en-
zymes and substances, thus modulating plant growth and development and the transi-
Cells 2022, 11, 3250 12 of 18

When environmental stress conditions are alleviated and appropriate levels of water
and temperature are provided, the endogenous melatonin level decreases, whereas IAA
synthesis increases. The plant shifts from a poorly defended state and resumes growth. The
interaction between the two hormone signals helps regulate the balance of key enzymes
and substances, thus modulating plant growth and development and the transitions related
to stress response.

5.4. Crosstalk between Melatonin and IAA in Other Plant Species

Melatonin and auxin have been shown to regulate growth and stress responses not
only in Arabidopsis, but also in other plant species. Plant resistance to various stresses
is associated with improved antioxidant capacity. In Zoysia japonica, melatonin not only
increases the antioxidant capacity, but also affects the expression of IAA related genes,
which play an important role in seed germination [99]. The combined use of melatonin
and IAA can increase the levels of chlorophyll, total soluble protein, total soluble sugar,
and ascorbic acid in wheat under stress, and can also enhance the antioxidant capacity
of the plants [100]. Maize yield is limited in arid areas, and the application of exogenous
melatonin can improve plant tolerance, thereby increasing maize yield. Corn seeds soaked
with melatonin exhibit increased levels of zeatin + zeatin riboside, IAA, and gibberellic
acid, which promote plant growth [4]. Both melatonin and IAA enhance plant tolerance to
salt stress [101].
IAA and melatonin are important indole compounds in plants. Arnao et al. studied
the growth of lupin plants [102] and various monocots [103] to examine these two indole
compounds and reported their similarities. Since then, several other studies have demon-
strated that melatonin can induce vegetative growth in a similar way to IAA, but through
other means [59]. For instance, growth depends on melatonin concentration and tissue
type. Therefore, roots are more sensitive to growth than aerial tissues (such as the talus and
leaves). Growth is inhibited at high concentrations of melatonin, similar to IAA. Recent
studies have also shown that low concentrations of IAA activate melatonin biosynthesis,
whereas high concentrations of melatonin inhibit IAA biosynthesis. However, more studies
have shown that melatonin regulates signaling elements such as TIR1, Aux/IAA, and SAURs
to mediate the activation of growth processes [104].
Melatonin treatment has been shown to induce a slight (1.4–2.0-fold) increase in
endogenous IAA, compared to untreated Brassica napus [105] and tomato [32] plants. How-
ever, in transgenic melatonin overproducing plants, IAA levels have been reported to drop
significantly. Tomato plants overexpressing sheep SNA HIOMT and Arabidopsis plants
overexpressing apple HIOMT had endogenous melatonin levels up to 6-fold higher than
those of wild-type plants, whereas endogenous IAA levels decreased 7-fold in tomato and
1.4-fold in Arabidopsis. The phenotypes of these transgenic plants clearly resembled those of
an auxin-like response, including the promotion of root growth and rooting, reduced apical
dominance, and the induction of root primordia by melatonin independent of the IAA
signaling pathway. IAA, but not melatonin, activated the auxin-inducible gene expression
marker DR5::GUS in Arabidopsis [39,106]. These data suggest that melatonin can act in
parallel with IAA in the induction of lateral and adventitious roots [103]. In cucumber,
melatonin has been shown to up- or down-regulate approximately 320 genes related to root
development. Some of these transcription factors and ethylene transcription factors can
negatively regulate root-related genes, thus inhibiting root formation. Liang et al. found
that IAA-related genes were significantly activated under melatonin treatment. Several
transcription factors and candidate cis-regulatory elements involved in root growth and
development and auxin-related processes are encoded by co-differentially expressed genes.
This suggests that melatonin-mediated root growth occurs in an auxin signaling pathway-
dependent manner. In addition, gravity response analysis has been used to identify the
process by which melatonin affects auxin regulation in rice roots [72]. Melatonin regulates
IAA distribution by regulating auxin transport, thereby promoting lateral root develop-
ment [107]. Chen et al. found that the application of 0.1 mM melatonin promoted root
Cells 2022, 11, 3250 13 of 18

growth, whereas 100 mM melatonin inhibited root growth. Furthermore, this stimulatory
effect was only detected in 2-day-old seedlings, and older seedlings (4-day-old) appeared
to be less sensitive to both the stimulating and inhibitory effects of melatonin. Exogenous
melatonin (0.1 mM) also increased the levels of endogenous free IAA in roots, whereas
higher concentrations of melatonin had no significant effect on endogenous IAA levels [105].
As a signal molecule, melatonin can regulate leaf senescence. Endogenous melatonin is
also associated with the overexpression of CsASMT (a key gene for melatonin synthesis),
which can delay the senescence of dark-stimulated leaves. A greenhouse study on cu-
cumbers reported that melatonin inhibited leaf senescence by reducing ABA biosynthesis
and inhibiting its signaling pathway, while promoting IAA biosynthesis and its signaling
pathway [108]. In apple, melatonin promotes the formation of adventitious roots, mainly
by promoting IAA levels and up-regulating MdWOX11 in the stress induction stage [109].

6. Conclusions and Perspective

IAA is involved in almost all aspects of plant growth and development, including
responses to external stimuli (such as biotic and abiotic stresses). Melatonin also plays an
important role in plant growth and stress response. This paper summarizes the known
literature on interactions between melatonin and IAA in regulating plant growth and
development. First, melatonin regulates IAA levels and promotes IAA biosynthesis at low
concentrations. Specifically, melatonin enhances core signal transduction genes related to
IAA (such as IAA19 and IAA24), several PIN proteins (PIN1, PIN3, and PIN7) and key genes
related to IAA biosynthesis (YUC1, YUC2, YUC5, YUC6, and TAR2), thereby enhancing
lateral root development. However, melatonin reduces IAA levels at high concentrations.
The exogenous application of low concentrations of IAA also increases melatonin produc-
tion. Second, melatonin has an IAA-like effect on plant growth and development, and can
promote plant flowering, photosynthesis, senescence, and seed development. IAA can
also modulate the antagonistic effects of melatonin on seed germination. When seeds are
treated with both melatonin and IAA, IAA suppresses the inhibitory effects of melatonin
on seed germination. Third, NO signaling molecules are involved in various physiological
processes during plant growth and development and are also important regulators of stress
response and pathophysiological processes. The dual control of NO levels by melatonin
and IAA may regulate plant growth and environmental response through IAA carrier
proteins. Finally, the combination of melatonin and IAA can improve the stress resistance
of plants.
Both melatonin and IAA can regulate each other’s levels; however, the mechanisms
of how plants regulate melatonin from the perspective of IAA need further exploration.
In the future, systems biology approaches—especially combinations of different “omics”
approaches with CRISPR/Cas9 technology—should be used to accelerate the identification
of key genes related to the biosynthesis, degradation, and signaling pathways of melatonin
and IAA. Analyzing the changes in melatonin content and signaling in mutant lines for IAA
biosynthesis, degradation, and signal transduction will also help reveal the relationship be-
tween melatonin and IAA. Both of these hormones are known to communicate extensively
with other hormone signaling pathways by regulating their biosynthesis or signal trans-
duction. Therefore, uncovering the crosstalk between IAA and melatonin would contribute
to our understanding of more complex networks of crosstalk among plant hormones.

Author Contributions: Conceptualization, M.L. and M.Z.; writing—original draft preparation, M.Z.;
C.G., L.X. and H.N.; writing—review and editing, M.Z.; C.G., L.X., H.Y. and H.N.; visualization,
M.Z.; C.G., L.X., Q.L., Y.H. and G.L. All authors have read and agreed to the published version of
the manuscript.
Funding: This research was funded by China Agriculture Research System of MOF and MARA, grant
number CARS-21; Scientific Research and Innovation Project of postgraduates in Inner Mongolia
Autonomous region in 2021, grant number S20210168Z; The Key Project at Central Government
Level: The Ability Establishment of Sustainable Use for Valuable Chinese Medicine Resources, grant
Cells 2022, 11, 3250 14 of 18

number 2060302; Open Fund Project of Key Laboratory of Chinese and Mongolian Medicine of Inner
Mongolia Autonomous Region, grant number MYX2022-K05.
Data Availability Statement: Not applicable.
Conflicts of Interest: The authors declare no conflict of interest.

Abbreviations
Indole-3-acetic acid (IAA), indole-3-pyruvate monooxygenase (YUCCA), abscisic acid (ABA),
reactive oxygen species (ROS), nitric oxide (NO), hydrogen peroxide (H2O2), transport inhibitory
response 1 (TIR1), auxin (Aux), superoxide dismutase (SOD), peroxidase (POD), catalase (CAT).

References
1. Salvi, P.; Manna, M.; Kaur, H.; Thakur, T.; Gandass, N.; Bhatt, D.; Muthamilarasan, M. Phytohormone signaling and crosstalk in
regulating drought stress response in plants. Plant Cell Rep. 2021, 40, 1305–1329. [CrossRef]
2. Wang, P.; Zhao, Y.; Li, Z.; Hsu, C.C.; Liu, X.; Fu, L.; Hou, Y.J.; Du, Y.; Xie, S.; Zhang, C.; et al. Reciprocal regulation of the TOR
kinase and ABA receptor balances plant growth and stress response. Mol. Cell 2018, 69, 100–112.e6. [CrossRef] [PubMed]
3. Yang, L.; You, J.; Li, J.; Wang, Y.; Chan, Z. Melatonin promotes Arabidopsis primary root growth in an IAA-dependent manner.
J. Exp. Bot. 2021, 72, 5599–5611. [CrossRef]
4. Ahmad, S.; Kamran, M.; Zhou, X.; Ahmad, I.; Meng, X.; Javed, T.; Iqbal, A.; Wang, G.; Su, W.; Wu, X.; et al. Melatonin improves the
seed filling rate and endogenous hormonal mechanism in grains of summer maize. Physiol. Plant 2021, 172, 1059–1072. [CrossRef]
5. Zhao, Y.Q.; Zhang, Z.W.; Chen, Y.E.; Ding, C.B.; Yuan, S.; Reiter, R.J.; Yuan, M. Melatonin: A Potential Agent in Delaying Leaf
Senescence. Crit. Rev. Plant Sci. 2021, 40, 1–22. [CrossRef]
6. Debnath, B.; Islam, W.; Li, M.; Sun, Y.; Lu, X.; Mitra, S.; Hussain, M.; Liu, S.; Qiu, D. Melatonin mediates enhancement of stress
tolerance in plants. Int. J. Mol. Sci. 2019, 20, 1040. [CrossRef]
7. Korkmaz, A.; Değer, Ö.; Szafrańska, K.; Köklü, Ş.; Karaca, A.; Yakupoğlu, G.; Kocacinar, F. Melatonin effects in enhancing chilling
stress tolerance of pepper. Sci. Hortic. 2021, 289, 110434. [CrossRef]
8. Gu, Q.; Wang, C.; Xiao, Q.; Chen, Z.; Han, Y. Melatonin confers plant cadmium tolerance: An update. Int. J. Mol. Sci. 2021,
22, 11704. [CrossRef]
9. Park, S.; Back, K. Melatonin promotes seminal root elongation and root growth in transgenic rice after germination. J. Pineal Res.
2012, 53, 385–389. [CrossRef] [PubMed]
10. Lee, H.Y.; Lee, K.; Back, K. Knockout of Arabidopsis serotonin N-Acetyltransferase-2 reduces melatonin levels and delays
flowering. Biomolecules 2019, 9, 712. [CrossRef]
11. Zhao, H.; Su, T.; Huo, L.; Wei, H.; Jiang, Y.; Xu, L.; Ma, F. Unveiling the mechanism of melatonin impacts on maize seedling
growth: Sugar metabolism as a case. J. Pineal Res. 2015, 59, 255–266. [CrossRef]
12. Wei, J.; Li, D.; Zhang, J.; Shan, C.; Rengel, Z.; Song, Z.; Chen, Q. Phytomelatonin receptor PMTR1-mediated signaling regulates
stomatal closure in Arabidopsis thaliana. J. Pineal Res. 2018, 65, e12500. [CrossRef] [PubMed]
13. Bychkov, I.A.; Kudryakova, N.V.; Shugaev, A.G.; Kuznetsov, V.V.; Kusnetsov, V.V. The melatonin receptor CAND2/PMTR1 is
involved in the regulation of mitochondrial gene expression under photooxidative stress. Dokl. Biochem. Biophys. 2022, 502, 15–20.
[CrossRef] [PubMed]
14. Li, D.; Wei, J.; Peng, Z.; Ma, W.; Yang, Q.; Song, Z.; Sun, W.; Yang, W.; Yuan, L.; Xu, X.; et al. Daily rhythms of phytomelatonin
signaling modulate diurnal stomatal closure via regulating reactive oxygen species dynamics in Arabidopsis. J. Pineal Res. 2020,
68, e12640. [CrossRef]
15. Lee, H.Y.; Back, K. Mitogen-activated protein kinase pathways are required for melatonin-mediated defense responses in plants.
J. Pineal Res. 2016, 60, 327–335. [CrossRef]
16. Chisholm, S.T.; Coaker, G.; Day, B.; Staskawicz, B.J. Host-microbe interactions: Shaping the evolution of the plant immune
response. Cell 2006, 124, 803–814. [CrossRef] [PubMed]
17. Lee, H.Y.; Back, K. Melatonin is required for H2 O2 - and NO-mediated defense signaling through MAPKKK3 and OXI1 in
Arabidopsis thaliana. J. Pineal Res. 2017, 62, e12379. [CrossRef] [PubMed]
18. Gong, B.; Yan, Y.; Wen, D.; Shi, Q. Hydrogen peroxide produced by NADPH oxidase: A novel downstream signaling pathway in
melatonin-induced stress tolerance in Solanum lycopersicum. Physiol. Plant. 2017, 160, 396–409. [CrossRef] [PubMed]
19. Kaur, H.; Bhatla, S.C. Melatonin and nitric oxide modulate glutathione content and glutathione reductase activity in sunflower
seedling cotyledons accompanying salt stress. Nitric Oxide. 2016, 59, 42–53. [CrossRef] [PubMed]
20. Naser, V.; Shani, E. Auxin response under osmotic stress. Plant Mol. Biol. 2016, 91, 661–672. [CrossRef] [PubMed]
21. Wang, D.; Chen, Q.; Chen, W.; Guo, Q.; Xia, Y.; Wang, S.; Jing, D.; Liang, G. Physiological and transcription analyses reveal the
regulatory mechanism of melatonin in inducing drought resistance in loquat (Eriobotrya japonica Lindl.) seedlings. Environ. Exp.
Bot. 2021, 181, 104291. [CrossRef]
22. Kubeš, M.F.; Napier, R.M. Non-canonical auxin signalling: Fast and curious. J. Exp. Bot. 2019, 70, 2609–2614. [CrossRef] [PubMed]
Cells 2022, 11, 3250 15 of 18

23. Watanabe, E.; Mano, S.; Hara-Nishimura, I.; Nishimura, M.; Yamada, K. HSP90 stabilizes auxin receptor TIR1 and ensures
plasticity of auxin responses. Plant Signal. Behav. 2017, 12, e1311439. [CrossRef] [PubMed]
24. Salehin, M.; Bagchi, R.; Estelle, M. SCFTIR1/AFB-based auxin perception: Mechanism and role in plant growth and development.
Plant Cell. 2015, 27, 9–19. [PubMed]
25. Wang, R.; Estelle, M. Diversity and specificity: Auxin perception and signaling through the TIR1/AFB pathway. Curr. Opin. Plant
Biol. 2014, 21, 51–58. [CrossRef]
26. Benjamins, R.; Scheres, B.; Benjamins, R.; Scheres, B. Auxin: The looping star in plant development. Annu. Rev. Plant Biol. 2008,
59, 443–465. [CrossRef]
27. Dharmasiri, N.; Dharmasiri, S.; Estelle, M. The F-box protein TIR1 is an auxin receptor. Nature 2005, 435, 441–445. [CrossRef]
28. Kepinski, S.; Leyser, O. The Arabidopsis F-box protein TIR1 is an auxin receptor. Nature 2005, 435, 446–451. [CrossRef]
29. Kepinski, S.; Leyser, O. Auxin-induced SCFTIR1-Aux/IAA interaction involves stable modification of the SCFTIR1 complex. Proc.
Natl. Acad. Sci. USA 2004, 101, 12381–12386. [CrossRef]
30. Friml, J.; Gallei, M.; Gelová, Z.; Johnson, A.; Mazur, E.; Monzer, A.; Rodriguez, L.; Roosjen, M.; Verstraeten, I.; Živanović, B.D.;
et al. ABP1-TMK auxin perception for global phosphorylation and auxin canalization. Nature. 2022, 609, 575–581. [CrossRef]
31. Kou, X.H.; Zhao, X.Y.; Wu, B.D.; Wang, C.; Wu, C.; Yang, S.; Zhou, J.Q.; Xue, Z.H. Auxin Response Factors Are Ubiquitous in Plant
Growth and Development, and Involved in Crosstalk between Plant Hormones: A Review. Appl. Sci. 2022, 12, 1360. [CrossRef]
32. Wen, D.; Gong, B.; Sun, S.; Liu, S.; Wang, X.; Wei, M.; Yang, F.; Li, Y.; Shi, Q. Promoting roles of melatonin in adventitious root
development of Solanum lycopersicum L. by regulating auxin and nitric oxide signaling. Front. Plant Sci. 2016, 7, 718. [PubMed]
33. Wang, Q.; An, B.; Wei, Y.; Reiter, R.J.; Shi, H.; Luo, H.; He, C. Melatonin regulates root meristem by repressing auxin synthesis and
polar auxin transport in Arabidopsis. Front. Plant Sci. 2016, 7, 1882. [CrossRef] [PubMed]
34. Yang, L. Differential Transcriptome Induced by Melatonin and Auxin and Functional Analysis of 14-3-3 Gene Families in
Arabidopsis thaliana and Brachypium Discus. Ph.D. Thesis, Wuhan Botanical Garden, Chinese Academy of Sciences, Wuhan,
China, 2017. (In Chinese)
35. Zhang, X.Q. Induction and Effect of Exogenous Melatonin on Hairy Roots of Isatis Indigotica. Master’s Thesis, Northwest
University, Xi’an, China, 2014. (In Chinese)
36. Zhang, Y. Regulation of Indole-Acetic Acid and Melatonin on Aluminum Toxicity of Helianthus tuberosus L. Master’s Thesis,
Zhejiang Normal University, Jinhua, China, 2021. (In Chinese).
37. Shi, H.; Reiter, R.J.; Tan, D.X.; Chan, Z. Indole-3-acetic acid Inducible 17 positively modulates natural leaf senescence through
melatonin-mediated pathway in Arabidopsis. J. Pineal Res. 2015, 58, 26–33. [CrossRef]
38. Park, W.J. Melatonin as an endogenous plant regulatory signal: Debates and perspectives. J. Plant Biol. 2011, 54, 143–149.
[CrossRef]
39. Pelagio-Flores, R.; Muñoz-Parra, E.; Ortiz-Castro, R.; López-Bucio, J. Melatonin regulates Arabidopsis root system architecture
likely acting independently of auxin signaling. J. Pineal Res. 2012, 53, 279–288. [CrossRef]
40. Byeon, Y.; Park, S.; Kim, Y.S.; Park, D.H.; Lee, S.; Back, K. Light-regulated melatonin biosynthesis in rice during the senescence
process in detached leaves. J. Pineal Res. 2012, 53, 107–111. [CrossRef]
41. Lei, X.Y.; Zhu, R.Y.; Zhang, G.Y.; Dai, Y.R. Attenuation of cold-induced apoptosis by exogenous melatonin in carrot suspension
cells: The possible involvement of polyamines. J. Pineal Res. 2004, 36, 126–131. [CrossRef] [PubMed]
42. Lv, Y.; Pan, J.; Wang, H.; Reiter, R.J.; Li, X.; Mou, Z.; Zhang, J.; Yao, Z.; Zhao, D.; Yu, D. Melatonin inhibits seed germination by
crosstalk with abscisic acid, gibberellin, and auxin in Arabidopsis. J. Pineal Res. 2021, 70, e12736. [CrossRef]
43. Weeda, S.; Zhang, N.; Zhao, X.; Ndip, G.; Guo, Y.; Buck, G.A.; Fu, C.; Ren, S. Arabidopsis transcriptome analysis reveals key roles
of melatonin in plant defense systems. PLoS ONE 2014, 9, e93462. [CrossRef]
44. Ren, S.; Rutto, L.; Katuuramu, D. Melatonin acts synergistically with auxin to promote lateral root development through fine
tuning auxin transport in Arabidopsis thaliana. PLoS ONE 2019, 14, e0221687. [CrossRef] [PubMed]
45. Arnao, M.B.; Hernández-Ruiz, J. Chemical stress by different agents affects the melatonin content of barley roots. J. Pineal Res.
2009, 46, 295–299. [CrossRef]
46. Arnao, M.B.; Hernández-Ruiz, J. Growth conditions influence the melatonin content of tomato plants. Food Chem. 2013, 138,
1212–1214. [CrossRef] [PubMed]
47. Arnao, M.B.; Hernández-Ruiz, J. Growth conditions determine different melatonin levels in Lupinus albus L. J. Pineal Res. 2013, 55,
149–155. [CrossRef] [PubMed]
48. Li, C.; Wang, P.; Wei, Z.; Liang, D.; Liu, C.; Yin, L.; Jia, D.; Fu, M.; Ma, F. The mitigation effects of exogenous melatonin on
salinity-induced stress in Malus hupehensis. J. Pineal Res. 2012, 53, 298–306. [CrossRef] [PubMed]
49. Yin, L.; Wang, P.; Li, M.; Ke, X.; Li, C.; Liang, D.; Wu, S.; Ma, X.; Li, C.; Zou, Y.; et al. Exogenous melatonin improves Malus
resistance to Marssonina apple blotch. J. Pineal Res. 2013, 54, 426–434. [CrossRef] [PubMed]
50. Blakeslee, J.J.; Spatola Rossi, T.; Kriechbaumer, V. Auxin biosynthesis: Spatial regulation and adaptation to stress. J. Exp. Bot. 2019,
70, 5041–5049. [CrossRef] [PubMed]
51. Ludwig-Müller, J. Auxin conjugates: Their role for plant development and in the evolution of land plants. J. Exp. Bot. 2011, 62,
1757–1773. [CrossRef] [PubMed]
52. Mano, Y.; Nemoto, K. The pathway of auxin biosynthesis in plants. J. Exp. Bot. 2012, 63, 2853–2872. [CrossRef] [PubMed]
Cells 2022, 11, 3250 16 of 18

53. Dindas, J.; Scherzer, S.; Roelfsema, M.R.G.; von Meyer, K.; Müller, H.M.; Al-Rasheid, K.A.S.; Palme, K.; Dietrich, P.; Becker, D.;
Bennett, M.J.; et al. AUX1-mediated root hair auxin influx governs SCFTIR1/AFB-type Ca2+ signaling. Nat. Commun. 2018,
9, 1174. [CrossRef] [PubMed]
54. Zia, S.F.; Berkowitz, O.; Bedon, F.; Whelan, J.; Franks, A.E.; Plummer, K.M. Direct comparison of Arabidopsis gene expression
reveals different responses to melatonin versus auxin. BMC Plant Biol. 2019, 19, 567. [CrossRef]
55. Hagen, G.; Guilfoyle, T. Auxin-responsive gene expression: Genes, promoters and regulatory factors. Plant Mol. Biol. 2002, 49,
373–385. [CrossRef]
56. Kant, S.; Bi, Y.M.; Zhu, T.; Rothstein, S.J. SAUR39, a small auxin-up RNA gene, acts as a negative regulator of auxin synthesis and
transport in rice. Plant Physiol. 2009, 151, 691–701. [CrossRef]
57. Majda, M.; Robert, S. The role of auxin in cell wall expansion. Int. J. Mol. Sci. 2018, 19, 951. [CrossRef]
58. Arnao, M.B.; Hernández-Ruiz, J. Melatonin: Plant growth regulator and/or biostimulator during stress? Trends Plant Sci. 2014, 19,
789–797. [CrossRef]
59. Arnao, M.B.; Hernández-Ruiz, J. Growth activity, rooting capacity, and tropism: Three auxinic precepts fulfilled by melatonin.
Acta Physiol. Plant. 2017, 39, 127. [CrossRef]
60. Di, D.W.; Zhang, C.; Guo, G.Q. Involvement of secondary messengers and small organic molecules in auxin perception and
signaling. Plant Cell Rep. 2015, 34, 895–904. [CrossRef]
61. Hussain, A.; Mun, B.G.; Imran, Q.M.; Lee, S.U.; Adamu, T.A.; Shahid, M.; Kim, K.M.; Yun, B.W. Nitric oxide mediated
transcriptome profiling reveals activation of multiple regulatory pathways in Arabidopsis thaliana. Front. Plant Sci. 2016, 7, 975.
[CrossRef]
62. Pagnussat, G.C.; Simontacchi, M.; Puntarulo, S.; Lamattina, L. Nitric oxide is required for root organogenesis. Plant Physiol. 2002,
129, 954–956. [CrossRef]
63. Correa-Aragunde, N.; Graziano, M.; Lamattina, L. Nitric oxide plays a central role in determining lateral root development in
tomato. Planta 2004, 218, 900–905. [CrossRef] [PubMed]
64. Hu, X.; Neill, S.J.; Tang, Z.; Cai, W. Nitric oxide mediates gravitropic bending in soybean roots. Plant Physiol. 2005, 137, 663–670.
[CrossRef] [PubMed]
65. Lombardo, M.C.; Graziano, M.; Polacco, J.C.; Lamattina, L. Nitric oxide functions as a positive regulator of root hair development.
Plant Signal Behav. 2006, 1, 28–33. [CrossRef] [PubMed]
66. Chen, Y.H.; Kao, C.H. Calcium is involved in nitric oxide- and auxin-induced lateral root formation in rice. Protoplasma 2012, 249,
187–195. [CrossRef] [PubMed]
67. Correa-Aragunde, N.; Cejudo, F.J.; Lamattina, L. Nitric oxide is required for the auxin-induced activation of NADPH-dependent
thioredoxin reductase and protein denitrosylation during root growth responses in Arabidopsis. Ann. Bot. 2015, 116, 695–702.
[CrossRef] [PubMed]
68. Yan, Y.Y. Regulation Mechanism of Melatonin and Nitric Oxidc on Abiotic Stress Tolerance and Branch Growth in Tomato. Ph.D.
Thesis, Shandong Agricultural University, Taian, China, 2021.
69. Guo, Y.Q. Effect of Exogenous 2,4-D on Proliferation of Embryogenic Callus of Daduhe Loquat. Master’s Thesis, Southwest
University, Chongqing, China, 2020.
70. Li, J.Z. Mechanism of Interactions between Melatonin and Cytokinin in Regulating Arabidopsis Primary Root Growth. Master’s
Thesis, University of Chinese Academy of Sciences, Beijing, China, 2017.
71. Shi, H.; Zhang, S.; Lin, D.; Wei, Y.; Yan, Y.; Liu, G.; Reiter, R.J.; Chan, Z. Zinc finger of Arabidopsis thaliana 6 is involved in
melatonin-mediated auxin signaling through interacting indeterminate domain 15 and indole-3-acetic acid 17. J. Pineal Res. 2018,
65, e12494. [CrossRef] [PubMed]
72. Liang, C.; Li, A.; Yu, H.; Li, W.; Liang, C.; Guo, S.; Zhang, R.; Chu, C. Melatonin Regulates Root Architecture by Modulating
Auxin Response in Rice. Front. Plant Sci. 2017, 8, 134. [CrossRef] [PubMed]
73. Korver, R.A.; Koevoets, I.T.; Testerink, C. Out of shape during stress: A Key role for auxin. Trends Plant Sci. 2018, 23, 783–793.
[CrossRef]
74. Di Mambro, R.; De Ruvo, M.; Pacifici, E.; Salvi, E.; Sozzani, R.; Benfey, P.N.; Busch, W.; Novak, O.; Ljung, K.; Di Paola, L.; et al.
Auxin minimum triggers the developmental switch from cell division to cell differentiation in the Arabidopsis root. Proc. Natl.
Acad. Sci. USA 2017, 114, E7641–E7649. [CrossRef]
75. Woodward, A.W.; Bartel, B. Auxin: Regulation, action, and interaction. Ann. Bot. 2005, 95, 707–735. [CrossRef]
76. Maraschin Fdos, S.; Memelink, J.; Offringa, R. Auxin-induced, SCF(TIR1)-mediated poly-ubiquitination marks AUX/IAA proteins
for degradation. Plant J. 2009, 59, 100–109. [CrossRef]
77. Prigge, M.J.; Platre, M.; Kadakia, N.; Zhang, Y.; Greenham, K.; Szutu, W.; Pandey, B.K.; Bhosale, R.A.; Bennett, M.J.; Busch, W.; et al.
Genetic analysis of the Arabidopsis TIR1/AFB auxin receptors reveals both overlapping and specialized functions. eLife 2020, 9,
e54740. [CrossRef] [PubMed]
78. Chen, Z.; Hu, L.; Han, N.; Hu, J.; Yang, Y.; Xiang, T.; Zhang, X.; Wang, L. Overexpression of a miR393-resistant form of transport
inhibitor response protein 1 (mTIR1) enhances salt tolerance by increased osmoregulation and Na+ exclusion in Arabidopsis
thaliana. Plant Cell Physiol. 2015, 56, 73–83. [CrossRef] [PubMed]
Cells 2022, 11, 3250 17 of 18

79. Iglesias, M.J.; Terrile, M.C.; Bartoli, C.G.; D’Ippólito, S.; Casalongué, C.A. Auxin signaling participates in the adaptative response
against oxidative stress and salinity by interacting with redox metabolism in Arabidopsis. Plant Mol. Biol. 2010, 74, 215–222.
[CrossRef] [PubMed]
80. Park, J.E.; Park, J.Y.; Kim, Y.S.; Staswick, P.E.; Jeon, J.; Yun, J.; Kim, S.Y.; Kim, J.; Lee, Y.H.; Park, C.M. GH3-mediated auxin
homeostasis links growth regulation with stress adaptation response in Arabidopsis. J. Biol. Chem. 2007, 282, 10036–10046.
[CrossRef] [PubMed]
81. Iglesias, M.J.; Terrile, M.C.; Windels, D.; Lombardo, M.C.; Bartoli, C.G.; Vazquez, F.; Estelle, M.; Casalongué, C.A. MiR393
regulation of auxin signaling and redox-related components during acclimation to salinity in Arabidopsis. PLoS ONE 2014,
9, e107678.
82. Vidal, E.A.; Moyano, T.C.; Riveras, E.; Contreras-López, O.; Gutiérrez, R.A. Systems approaches map regulatory networks
downstream of the auxin receptor AFB3 in the nitrate response of Arabidopsis thaliana roots. Proc. Natl. Acad. Sci. USA 2013, 110,
12840–12845. [CrossRef]
83. Vidal, E.A.; Álvarez, J.M.; Gutiérrez, R.A. Nitrate regulation of AFB3 and NAC4 gene expression in Arabidopsis roots depends on
NRT1.1 nitrate transport function. Plant Signal. Behav. 2014, 9, e28501. [CrossRef]
84. Garrido-Vargas, F.; Godoy, T.; Tejos, R.; O’Brien, J.A. Overexpression of the Auxin Receptor AFB3 in Arabidopsis Results in Salt
Stress Resistance and the Modulation of NAC4 and SZF1. Int. J. Mol. Sci. 2020, 21, 9528. [CrossRef]
85. Kalve, S.; Sizani, B.L.; Markakis, M.N.; Helsmoortel, C.; Vandeweyer, G.; Laukens, K.; Sommen, M.; Naulaerts, S.; Vissenberg, K.;
Prinsen, E.; et al. Osmotic stress inhibits leaf growth of Arabidopsis thaliana by enhancing ARF-mediated auxin responses. New
Phytol. 2020, 226, 1766–1780. [CrossRef]
86. Liu, W.C.; Song, R.F.; Zheng, S.Q.; Li, T.T.; Zhang, B.L.; Gao, X.; Lu, Y.T. Coordination of plant growth and abiotic stress responses
by tryptophan synthase subunit 1 through modulation of tryptophan and ABA homeostasis in Arabidopsis. Mol. Plant. 2022, 15,
973–990. [CrossRef]
87. Yang, Z.B.; Geng, X.; He, C.; Zhang, F.; Wang, R.; Horst, W.J.; Ding, Z. TAA1-regulated local auxin biosynthesis in the root-
apex transition zone mediates the aluminum-induced inhibition of root growth in Arabidopsis. Plant Cell 2014, 26, 2889–2904.
[PubMed]
88. Hu, Y.F.; Zhou, G.; Na, X.F.; Yang, L.; Nan, W.B.; Liu, X.; Zhang, Y.Q.; Li, J.L.; Bi, Y.R. Cadmium interferes with maintenance of
auxin homeostasis in Arabidopsis seedlings. J. Plant Physiol. 2013, 170, 965–975. [CrossRef] [PubMed]
89. Li, J.J.; Zeng, L.; Cheng, Y.; Lu, G.Y.; Fu, G.P.; Ma, H.Q.; Liu, Q.Y.; Zhang, X.K.; Zou, X.L.; Li, C.S. Exogenous melatonin alleviates
damage from drought stress in Brassica napus L. (rapeseed) seedlings. Acta Physiol. Plant. 2018, 40, 43. [CrossRef]
90. Antoniou, C.; Chatzimichail, G.; Xenofontos, R.; Pavlou, J.J.; Panagiotou, E.; Christou, A.; Fotopoulos, V. Melatonin systemically
ameliorates drought stress-induced damage in Medicago sativa plants by modulating nitro-oxidative homeostasis and proline
metabolism. J. Pineal Res. 2017, 62, e12401. [CrossRef]
91. Wang, L.; Feng, C.; Zheng, X.; Guo, Y.; Zhou, F.; Shan, D.; Liu, X.; Kong, J. Plant mitochondria synthesize melatonin and enhance
the tolerance of plants to drought stress. J. Pineal Res. 2017, 63, e12429. [CrossRef]
92. Wang, L.Y.; Liu, J.L.; Wang, W.X.; Sun, Y. Exogenous melatonin improves growth and photosynthetic capacity of cucumber under
salinity-induced stress. Photosynthetica 2016, 54, 19–27. [CrossRef]
93. Chen, Y.E.; Mao, J.J.; Sun, L.Q.; Huang, B.; Ding, C.B.; Gu, Y.; Liao, J.Q.; Hu, C.; Zhang, Z.W.; Yuan, S.; et al. Exogenous melatonin
enhances salt stress tolerance in maize seedlings by improving antioxidant and photosynthetic capacity. Physiol. Plant. 2018, 164,
349–363. [CrossRef]
94. Jiang, C.; Cui, Q.; Feng, K.; Xu, D.; Li, C.; Zheng, Q. Melatonin improves antioxidant capacity and ion homeostasis and enhances
salt tolerance in maize seedlings. Acta Physiol. Plant. 2016, 38, 82. [CrossRef]
95. Xu, W.; Cai, S.Y.; Zhang, Y.; Wang, Y.; Ahammed, G.J.; Xia, X.J.; Shi, K.; Zhou, Y.H.; Yu, J.Q.; Reiter, R.J.; et al. Melatonin enhances
thermotolerance by promoting cellular protein protection in tomato plants. J. Pineal Res. 2016, 61, 457–469. [CrossRef]
96. Shi, H.; Tan, D.X.; Reiter, R.J.; Ye, T.; Yang, F.; Chan, Z. Melatonin induces class A1 heat-shock factors (HSFA1s) and their possible
involvement of thermotolerance in Arabidopsis. J. Pineal Res. 2015, 58, 335–342. [CrossRef]
97. Hernández, I.G.; Gomez, F.J.; Cerutti, S.; Arana, M.V.; Silva, M.F. Melatonin in Arabidopsis thaliana acts as plant growth regulator
at low concentrations and preserves seed viability at high concentrations. Plant Physiol. Biochem. 2015, 94, 191–196. [CrossRef]
[PubMed]
98. Hardeland, R. Melatonin in Plants—Diversity of Levels and Multiplicity of Functions. Front. Plant Sci. 2016, 7, 198. [CrossRef]
[PubMed]
99. Dong, D.; Wang, M.; Li, Y.; Liu, Z.; Li, S.; Chao, Y.; Han, L. Melatonin influences the early growth stage in Zoysia japonica Steud.
by regulating plant oxidation and genes of hormones. Sci. Rep. 2021, 11, 12381. [CrossRef] [PubMed]
100. Zafar, S.; Akhtar, M.; Perveen, S.; Hasnain, Z.; Khalil, A. Attenuating the adverse aspects of water stress on wheat genotypes by
foliar spray of melatonin and indole-3-acetic acid. Physiol. Mol. Biol. Plants 2020, 26, 1751–1762. [CrossRef]
101. Li, J.; Liu, J.; Zhu, T.; Zhao, C.; Li, L.; Chen, M. The role of melatonin in salt stress responses. Int. J. Mol. Sci. 2019, 20, 1735.
[CrossRef]
102. Hernández-Ruiz, J.; Cano, A.; Arnao, M.B. Melatonin: Growth-stimulating compound present in lupin tissues. Planta 2004, 220,
140–144. [CrossRef]
Cells 2022, 11, 3250 18 of 18

103. Hernández-Ruiz, J.; Cano, A.; Arnao, M.B. Melatonin acts as a growth-stimulating compound in some monocot species. J. Pineal
Res. 2005, 39, 137–142. [CrossRef]
104. Tan, X.; Long, W.; Zeng, L.; Ding, X.; Cheng, Y.; Zhang, X.; Zou, X. Melatonin-induced transcriptome variation of rapeseed
seedlings under salt stress. Int. J. Mol. Sci. 2019, 20, 5355. [CrossRef]
105. Chen, Q.; Qi, W.B.; Reiter, R.J.; Wei, W.; Wang, B.M. Exogenously applied melatonin stimulates root growth and raises endogenous
indoleacetic acid in roots of etiolated seedlings of Brassica juncea. J. Plant Physiol. 2009, 166, 324–328. [CrossRef]
106. Koyama, F.C.; Carvalho, T.L.G.; Alves, E.; da Silva, H.B.; de Azevedo, M.F.; Hemerly, A.S.; Garcia, C.R. The structurally related
auxin and melatonin tryptophan-derivatives and their roles in Arabidopsis thaliana and in the human malaria parasite Plasmodium
falciparum. J. Eukaryot. Microbiol. 2013, 60, 646–651. [CrossRef]
107. Lee, K.; Back, K. Melatonin-deficient rice plants show a common semidwarf phenotype either dependent or independent of
brassinosteroid biosynthesis. J. Pineal Res. 2019, 66, e12537. [CrossRef] [PubMed]
108. Jing, T.; Liu, K.; Wang, Y.; Ai, X.; Bi, H. Melatonin positively regulates both dark- and age-induced leaf senescence by reducing
ROS accumulation and modulating abscisic acid and auxin biosynthesis in cucumber plants. Int. J. Mol. Sci. 2022, 23, 3576.
[CrossRef]
109. Mao, J.; Niu, C.; Li, K.; Chen, S.; Tahir, M.M.; Han, M.; Zhang, D. Melatonin promotes adventitious root formation in apple by
promoting the function of MdWOX11. BMC Plant Biol. 2020, 20, 536. [CrossRef]