Biotechnology Advances 31 (2013) 11531171

Contents lists available at ScienceDirect

Biotechnology Advances
journal homepage: www.elsevier.com/locate/biotechadv

Research review paper

Phytohormones and microRNAs as sensors and regulators

of leaf senescence: Assigning macro roles to small molecules
Maryam Sarwat a,, Afsar Raza Naqvi b, Parvaiz Ahmad c,, Muhammad Ashraf d, Nudrat Aisha Akram e
a
Pharmaceutical Biotechnology, Amity Institute of Pharmacy, Amity University, Uttar Pradesh (AUUP), NOIDA, India
b
School of Dentistry, Department of Periodontics, University of North Carolina at Chapel, Hill, USA
c
Department of Botany, A. S. College, Srinagar, 190008, University of Kashmir, Srinagar, India
d
Department of Botany, University of Agriculture, Faisalabad, Pakistan
e
Department of Botany, GC University, Faisalabad, Pakistan

a r t i c l e i n f o a b s t r a c t

Article history: Ageing or senescence is an intricate and highly synchronized developmental phase in the life of plant parts includ-
Received 28 May 2012 ing leaf. Senescence not only means death of a plant part, but during this process, different macromolecules under-
Received in revised form 26 January 2013 go degradation and the resulting components are transported to other parts of the plant. During the period from
Accepted 2 February 2013 when a leaf is young and green to the stage when it senesces, a multitude of factors such as hormones, environ-
Available online 20 February 2013
mental factors and senescence associated genes (SAGs) are involved. Plant hormones including salicylic acid,
Keywords:
abscisic acid, jasmonic acid and ethylene advance leaf senescence, whereas others like cytokinins, gibberellins,
Leaf senescence and auxins delay this process. The environmental factors which generally affect plant development and growth,
Senescence associated genes (SAGs) can hasten senescence, the examples being nutrient dearth, water stress, pathogen attack, radiations, high temper-
Cytokinins ature and light intensity, waterlogging, and air, water or soil contamination. Other important inuences include
Ethylene carbohydrate accumulation and high carbon/nitrogen level. To date, although several genes involved in this com-
Auxins plex process have been identied, still not much information exists in the literature on the signalling mechanism of
PCD leaf senescence. Now, the Arabidopsis mutants have paved our way and opened new vistas to elucidate the signal-
MicroRNA
ling mechanism of leaf senescence for which various mutants are being utilized. Recent studies demonstrating the
Arabidopsis mutants
role of microRNAs in leaf senescence have reinforced our knowledge of this intricate process. This review provides
a comprehensive and critical analysis of the information gained particularly on the roles of several plant growth
regulators and microRNAs in regulation of leaf senescence.
2013 Elsevier Inc. All rights reserved.

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1154
1.1. Senescence, its roles and mechanism . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1154
2. Regulation of leaf senescence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1155
2.1. Gene expression during senescence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1156
2.2. Perception and transduction of senescence signals . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1156
2.3. Transcription regulation of gene expression . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1157
3. Progression of senescence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1157
3.1. Chloroplast degradation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1157
3.2. Role of mitochondria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1158
3.3. Loss of membrane integrity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1158
4. Senescence and Programmed Cell Death (PCD) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1158
5. Involvement of phytohormones in leaf senescence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1159
5.1. Cytokinins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1159
5.2. Ethylene . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1159
5.3. ABA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1159
5.4. Jasmonic acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1163
5.5. Salicylic acid . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1163

Corresponding authors.
E-mail addresses: maryam21_7@yahoo.com (M. Sarwat), parvaizbot@yahoo.com (P. Ahmad).

0734-9750/$ see front matter 2013 Elsevier Inc. All rights reserved.
http://dx.doi.org/10.1016/j.biotechadv.2013.02.003
1154 M. Sarwat et al. / Biotechnology Advances 31 (2013) 11531171

5.6. Brassinosteroids (BRs) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1163

5.7. Auxins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1163
5.8. Gibberellins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1164
5.9. Polyamines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1164
6. Stress and senescence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1164
6.1. Role of transcription factors in regulation of leaf senescence under stress conditions . . . . . . . . . . . . . . . . . . . . . . . . 1165
6.2. Role of reactive oxygen species . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1165
7. Micro RNA and senescence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1165
7.1. miR164 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1165
7.2. miR319 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1166
8. Conclusion and future prospects . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1166
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1166

1. Introduction senescence, e.g., SEN1 gene observed in Arabidopsis, contribute signi-

Normally, senescence is a synchronized series of events occurring in
plants during the course of development, but a stress of any type may
bring about senescence in plants much earlier than usual (Lim et al.,
2007; Lobell et al., 2012). The changes with time that fall under the gen-
eral term ageing are not necessarily deteriorative, although in the long
run errors will accumulate and living tissues will show signs of wear
and tear. According to some proposed models, senescence is an acceler-
ated form of ageing. Ageing is a term, like senescence that has become
associated with deterioration referring to changes that occur with
time (Thomas et al., 2003). Premature senescence can reduce the crop
productivity up to 50% and is one of the primary causes of crop losses
worldwide (Navabpour et al., 2003).
In senescence, a series of metabolic events take place. For example,
leaf ageing is a synchronized and sequential process in which the chlo-
roplast rst undergoes degeneration, followed by hydrolysis and
remobilization of macromolecules to other parts. Finally, disintegration
of nucleus as well as mitochondrion takes place (Buchanan-Wollaston
et al., 2003, 2005; Wang, 2012). Furthermore, several genes have so
far been identied which are up- or down-regulated during senescence.
For example, a study using ATH1 microarray taken from Arabidopsis ge-
nome sequence has shown up-regulation of 303 and down-regulation
of 81 transcription factors during dark-induced senescence (Lin and
Wu, 2004). Recently, a crucial role of some other genes such as NAC
family, WRKY53 TFs, WRKY6 and AtNAP, has been observed in leaf senes-
cence (Besseau et al., 2012; Sharabi-Schwager et al., 2010).
Other factors inuencing leaf senescence include plant hormones.
Hormones like, jasmonates (He et al., 2002), ethylene (Grbic and
Bleecker, 1995), salicylic acid (Morris et al., 2000) and ABA (Zeevaart
and Creelman, 1988) promote leaf senescence, whereas others like cy-
tokinins (Jung et al., 2006), gibberellins (Mutui et al., 2006) and auxins
(Kim et al., 2011a) hinder senescence. Overall, all the environmental
factors which cause deleterious effects on plant growth (Ahmad et al.,
2008, 2010a, 2010b; Koyro et al., 2011) can speed up senescence
(Lers, 2007). Furthermore, carbohydrate accumulation and accessibility
of high carbon to low nitrogen level represent other factors that can reg-
ulate leaf ageing (Wingler et al., 2006).
Studies on the elucidation of the mechanism of senescence in several
aerobic organisms have led to an intriguing stress resistance theory of
ageing which indicated that improvement against stresses could delay
senescence thereby expanding the life span of plants (Pardon, 2007).
This is further supported by a study on Arabidopsis, where delayed-
leaf-senescence mutants including gigantea (gi-3), ore9, ore3 and ore1
have been found markedly tolerant to oxidative disorders (Cao et al.,
2006; Woo et al., 2004). Furthermore, a possible coupling between leaf
senescence and stress responses has been described by Lim et al.
(2007). Chen et al. (2002) showed an overlapping role of about 28 senes-
cence induced genes in tolerance to various stresses. This phenomenon
has also been observed in the case of other genes have a key role in
cantly to leaf senescence during stresses (Schenk et al., 2005). Recently,
it has been shown that over-expression of the genes involved in cold ac-
climation like CBF2 and CBF3 delayed leaf senescence (Sharabi-Schwager
et al., 2010).
From all the above-mentioned reports it is amply clear that the se-
nescence phenomenon is very complex, which is regulated by a series
of metabolic events as well as genes. Thus, in the present review all
factors responsible for causing or accelerating senescence have been
comprehensively discussed. How far different phytohormones and
microRNAs participated signicantly to control senescence under
non-stress and stress conditions have also been explicitly described
in the review (Fig. 1).
1.1. Senescence, its roles and mechanism
Senescence is an intricate but much regulated metabolic process. It
goes on throughout the life of the plant. It involves synchronized mobi-
lization of nutrients like nitrogen, carbon and minerals from senescing
tissues to other plant parts (Milla et al., 2007; Wang, 2012). Loss of chlo-
rophyll and decrease in the total RNA and protein contents also take
place during this deteriorative process (Diaz et al., 2005). Although
the kinds of senescence reported are leaf, ower and fruit senescence,
in this review we have emphasized mainly on leaf senescence.
An interesting question arises here that how leaf senescence is initi-
ated and how the plant recognizes a particular developmental stage to
initiate the senescence program. Some studies provide a clue that
sugar concentration plays a considerable role in the coordination and
regulation of leaf senescence which specically depends on leaf photo-
synthetic capacity (Jang et al., 1997; Wingler et al., 2006). Others
suggest reduced metabolic rate as a trigger during age-dependent se-
nescence, but not in dark- or hormone-induced senescence. For exam-
ple, in Arabidopsis mutants oresara 4-1 (ore4-1), a lesion in chloroplast
functions resulted in reduced metabolic rate (Woo et al., 2002).
The overall structure of the leaf cells changes as the senescence pro-
cess starts in them. However, the vascular system of the plant is
maintained until the very late stages as it is required for the transporta-
tion of nutrients from the senescing tissues into young tissues as well as
reproductive organs like owers and seeds (Cholewa and Grifth, 2004;
Fukuda, 1996; Wang, 2012).
Senescence effectively inuences the expression of certain genes
involved in degradation of proteins into amino acids, RNA to low molec-
ular weight nitrogen compounds, and membrane lipids to fatty acids
and sugars (Buchanan-Wollaston, 1997; Zimmermann and Zentgraf,
2005). However, the DNA content remains unchanged till the very
late stages of senescence and the DNA fragmentation is recorded at
the end phase (Campisi and Fagagna, 2007; Huna et al., 2011; Orzaez
and Granell, 1997). These are the inherent changes which take place
during senescence. Furthermore, there are a number of factors which
cause premature senescence, including environmental factors like
 M. Sarwat et al. / Biotechnology Advances 31 (2013) 11531171
Endogenous factors
Phytohormones
Reproduction
Plant age & size
Meristem indeterminacy
Onset of Leaf Senescence
Initiation phase
[Shift in gene expression e.g. Senescence Associated Genes (SAGs); transduction of
senescence signals; over -accumulation of reactive oxygen species]
Re-organization phase
(Chlorophyll degradation; degradation of proteins, lipids and other macromolecules; nutrient
translocation by the phloem)
Terminal phase
(Cell deathdisruption of organelles)
Leaf abscision (Not always)
Fig. 1. Different factors involved in senescence (partially adopted from Munne-Bosch, 2008 with slight modication with permission).
abiotic e.g., nutrients deciency, drought, salinity, waterlogging, heat
stress and oxidative disorders caused by different stresses and biotic
factors particularly pathogen attack to plants.
Worldwide, extensive research work on molecular biology of leaf
senescence is being conducted on the model plant Arabidopsis
thaliana because of its short life cycle, readily distinguishable devel-
opmental changes and reproducible senescence program, but such
work in other plant species is not much yet.
2. Regulation of leaf senescence
Leaf senescence is a well-regulated phenomenon which takes place
through different genetically programmed events. It involves a myriad
of physiological changes such as the increase in number of senescing
leaves especially during owering and seed development. The aging
leaves generally serve as a source of nutrients during reproduction, and
the nutrients which are stored in the seeds, serve as the reserve food
for the next generation. Furthermore, age of the leaf and subsequently
age of the plant is the most important endogenous factor which regulates
leaf senescence. As the plant gets older, its photosynthetic activity drops,
and this can be contemplated as a senescence-inducing signal (Guo and
Gan, 2012; Mohapatra et al., 2010; Smart, 1994). In annual plants, photo-
synthetic rate of leaf shows a consistent decline after the full expansion
of the leaf (Hensel et al., 1993), however, it varies from species to species.
For example, in A. thaliana the leaves undergo fast-aging. A decline of al-
most 50% in its photosynthetic capacity has been observed within one
week preceding full expansion of its leaves if kept under continuous
light (Hensel et al., 1993). It is now evident that high amount of sugar
down-regulates photosynthesis related genes (Bilgin et al., 2010;
Chaves et al., 2009; Rolland et al., 2002). In fact, a close relationship has
been reported between the autophagic degradation of chloroplasts
through rubisco-containing bodies (RCBs) and the carbon content of
1155
Environmental factors
Photoperiod
Nutrient deficiencies
Salinity
Heat or cold
Light stress
Water stress
Ozone
leaves (Izumi and Ishida, 2011). As the leaf grows, expands and pro-
gresses towards senescence, the RCB production generally increases,
whereas in the later stages of senescence, the RCB production drops
and a large amount of carbohydrates accumulates, primarily glucose
and fructose.
Chlorophyll degradation is a promising process during leaf senes-
cence, which involves degradation of thylakoid membranes as well as
various chlorophyllprotein complexes. Plant mutants which retain
greenness by improving chlorophyll during leaf senescence are known
as stay-green mutants. It has been reported that non-functional
stay-green mutants, defective in chlorophyll degradation, can effective-
ly maintain greenness but not leaf functionality during senescence
(Morita et al., 2009). A protein termed as STAYGREEN PROTEIN (SGR)
is known to play a variety of physiological roles in plants particularly
during leaf senescence. For example, this protein can disassemble chlo-
rophyllprotein complexes in photosystem II and works together with
photosystem II proteins (Aubry et al., 2008). In rice, the SGR has been
reported to be absent from mature leaves (Zhou et al., 2011). Rice mu-
tants lacking SGR have long-lived chloroplasts and SGR over-expressing
lines show rapid degradation of chloroplasts in the developing leaves
(Park et al., 1998). The transcript level of SGR is specically induced
during leaf senescence (Park et al., 1998). SGR is a homolog of SGN1
in A. thaliana. Arabidopsis mutants acd2 (accelerated cell death 2) and
acd1 (accelerated cell death 1) (Mach et al., 2001; Pruinsk et al.,
2003) lack SGN1 activity.
The permeabilization of chloroplast membrane is a common event
that takes place during progression of senescence, but its mechanism is
still unknown. In animal system, arachidonic-type 15-lipoxygenases
have been reported to be involved in the permeabilization of the outer
membranes of mitochondria, peroxisomes and ER particularly during
differentiation of reticulocytes and keratinocytes (van Leyen et al.,
1998). Similarly, we can assume the role of 13-LOX enzymes present in
1156 M. Sarwat et al. / Biotechnology Advances 31 (2013) 11531171
chloroplasts (Bell et al., 1995; Feussner et al., 1995; Nemchenko et al.,
2006; Rangel et al., 2002; Seltmann et al., 2010) during plant senescence.
Furthermore, destruction of chloroplast may take place due to mem-
brane fatty acid peroxidation (Blokhina et al., 2003; Kunz et al., 2009).
Senescence is naturally an irreversible process, which takes place
due to decrease in photosynthesis due to inactivation of rubisco,
while activation of proteinase, lipid peroxidation and membrane
leakiness (Navabpour et al., 2003; Palma et al., 2002; Sato et al.,
2007). Generally, leaf protein and chlorophyll levels are considered
as metabolic markers of leaf senescence. Expression of genes involved
in protein synthesis and photosynthesis declines with an increase in
that of senescence-associated genes (SAGs) during senescence. More-
over, Navabpour et al. (2003) reported that SAGs express in response
to enhanced production of ROS, because cellular oxidation is a key
phenomenon during leaf senescence (Chen and Dickman, 2004;
Navabpour et al., 2003; Zentgraf and Hemleben, 2008). Generally,
the naturally rmly controlled balance between generation and
counteracting of ROS has been reported to be perturbed during the
progression of senescence in different cellular compartments either
by reduction in antioxidants or excess generation of ROS (Dai et al.,
2012; Zentgraf and Hemleben, 2008).
Recent genetic evidence suggests that ROS act as signals that acti-
vate genetically programmed pathways of gene expression that regu-
late cell suicide processes (Foyer and Noctor, 2005; Zentgraf and
Hemleben, 2008). Plants use ROS as second messengers in many signal
transduction cascades from mitosis to PCD (Bhattacharjee, in press;
Foyer and Noctor, 2005; Vanacker et al., 2006), and thus ROS accumula-
tion is essential to plant development as well as defence. Similarly, the
plant antioxidant defence network in the form of antioxidant com-
pounds and antioxidant enzymes, is necessary in controlling the
life-span of the ROS signals as well as in preventing uncontrolled cellu-
lar oxidation.
2.1. Gene expression during senescence
As mentioned earlier, leaf senescence is under strict genetic control,
and it requires several genes which express/repress in response to differ-
ent metabolic processes, signalling pathways and transcription factor ac-
tivities (Gadjev et al., 2008; He et al., 2001; Lim et al., 2003; Tenea et al.,
2011). All these trigger different biochemical/physiological processes.
Cell senescence is also associated with activation of the TOR (target of
rapamycin) nutrient- and mitogen-sensing pathway, which promotes
cell growth, even though cell cycle is blocked (Blagosklonny, 2011). Up
till now, many senescence-associated genes (SAGs) have been identied
in different plant species. Gepstein et al. (2003) identied 70 genes stim-
ulating macromolecule degradation, detoxication of oxidative metabo-
lites, induction of defense mechanisms, and signalling and other
regulatory events during senescence. Li et al. (2012) developed a curated
database of genes potentially associated with leaf senescence, the Leaf
Senescence Database (LSD) and gene networks to identify common reg-
ulators of leaf senescence in A. thaliana. They found that phytohormones
play a critical role in regulating leaf senescence. Functional analysis of
candidate SAGs in LSD revealed that a WRKY transcription factor
WRKY75 and a Cys2/His2-type transcription factor AZF2 are positive reg-
ulators of leaf senescence and loss-of-function of WRKY75 or AZF2
delays leaf senescence. During another study, Espinoza et al. (2007)
suggested that different virus-induced genes are also expressed at ele-
vated levels during natural senescence in vigorous plants. They showed
the role of SAGs during compatible viral interactions in A. thaliana and
grapevine, and reported that SAGs encode proteins such as proteases, li-
pases, and proteins involved in the mobilization of nutrients and min-
erals, transporters, transcription factors, proteins related to translation,
and antioxidant enzymes.
Recently, an SAG SPA15 has been identied in sweet potato
(Ipomoea batatas) but its function is still unknown. To elucidate the
role of SPA15 in leaf senescence, an orthologue of SPA15 referred to
as ARABIDOPSIS A-FIFTEEN (AAF) has been recently identied by
Chen et al. (2012). The overexpression of AAF in Arabidopsis has
been shown to promote leaf senescence. Products of these SAGs
cause degradation of a variety of macromolecules and remobilization
of nutrients thereby speeding up the process of senescence. The com-
plex regulatory network of senescence involves an array of various
metabolic pathways (Lim et al., 2007). One of those networks was
recorded in Arabidopsis (He et al., 2001). A few years ago, while work-
ing with senescence leaves of Arabidopsis, Guo et al. (2004) have
shown the involvement of 2500 genes as is evident from single pass
sequencing of its cDNA library. Another study entailing the microar-
ray analyses of Arabidopsis cDNAs has shown the change in the
expression of about 20% of the genes during leaf senescence
(Buchanan-Wollaston et al., 2003; Zentgraf et al., 2004). However, in-
formation on the regulation of these genes is not much in the litera-
ture. Recently, TOR (target of rapamycin) is reported as a negative
regulator of leaf senescence (Dobrenel et al., 2011). It has been ob-
served that leaf and root biomass increases in plants overexpressing
TOR. These plants produce more seeds and are more resistant to dif-
ferent stresses (Dobrenel et al., 2011). Targeted silencing of TOR by
RNAi (RNA interference) approach leads to a reduced organ growth
and early senescence accompanied by high accumulation of sugars
and amino acids (Dobrenel et al., 2011). Recently, Liu et al. (2011)
have generated a leaf senescence database (LSD, http://www.
eplantsenescence.org/) which comprises 1145 SAGs from 21 plant
species. They have categorized the SAGs based on their role in leaf se-
nescence. This site provides rich information to researchers working
to elucidate the function and expression of different SAGs as well as
explore the signalling network, and the origin and evolution of
SAGs. It is expected that this database will expand considerably in
the future as a number of groups are actively pursuing research on
uncovering unidentied SAGs in different plant species.
2.2. Perception and transduction of senescence signals
Various factors including sugars, environmental stresses, and plant
growth regulators are involved in the perception and signalling of se-
nescence. The role of some of the phytohormones on signal transduc-
tion in senescence are well studied, like ethylene (Klee, 2004), while
the research on other phytohormones is still in infancy.
Various studies have revealed receptor kinases to serve as receivers
and/or transducers of intrinsic or extrinsic signals. For example, leaf se-
nescence is reported to induce receptor like kinase (RLK) in tomato
(SARK) (Hajouj et al., 2000), lecRK a1 (Riou et al., 2002), SIRK
(Robatzek and Somssich, 2002), Paul27 in Populus tremula (Bhalerao et
al., 2003) and At5g48380 in Arabidopsis (Gepstein et al., 2003). In
Arabidopsis alone, 610 RLK genes constitute a large gene family (Shiu
and Bleecker, 2003). Of them, 44 are found in senescing leaves (Guo et
al., 2004). To have a better detail of signal perception of leaf senescence
functional study of these RLK genes is essential. Involvement of MAPK
signalling cascade in senescence was rst postulated by Guo et al.
(2004) with the representation of 3 MAPKs, 3 MAPKKs, 9 MAPKKKs,
and 1 MAPKKKK in one of the senescent leaf EST databases. MAPKs are
also reported from other plant species, e.g., Paul31 from P. tremula
MAPK, EDR1 from Arabidopsis and zmMPK5 from maize. Paul31 is the
most strongly expressed gene specic to autumn senescing in Populus
leaves (Bhalerao et al., 2003). Li et al. (2006) cloned and characterized
a receptor-like kinase (RLK) gene, GmSARK (Glycine max senescence-
associated receptor-like kinase) in soybean. This gene is reported to be
involved in regulating leaf senescence. ABA is considered as an
up-regulator of leaf senescence. While elucidating the role of ABA in
senescence signalling in Arabidopsis, Lee et al. (2011) have examined
the role of an ABA-induced gene, RPK1, which induces leucine-rich
repeat-containing receptor protein kinase 1. Furthermore, the authors
observed that rpk1 mutants showed decreased sensitivity to
ABA-induced senescence, but they did not alter JA-or ethylene-induced
 M. Sarwat et al. / Biotechnology Advances 31 (2013) 11531171
senescence, conrming considerable responsiveness of RPK1 to ABA. The
expression of RPK1 at different phases of growth was found to be
variable, being maximal at the adult stage, the stage at which senescence
process is generally accelerated. Tang and Innes (2002) reported
ethylene-induced senescence in Arabidopsis plants by over-expressing
kinase decient EDR1 gene. Some studies revealed involvement of calci-
um dependent protein kinases, calcium binding proteins, G proteins,
protein phosphatases, and 14,3,3 proteins in leaf senescence (de Silva
et al., 2011; Gepstein et al., 2003; Guo et al., 2004; Guterman et al.,
2003). Thus, there is an indication of association of other signal transduc-
tion pathways such as calcium signalling, G protein mediated signalling,
phosphatases, and 14,3,3 proteins with leaf senescence.
Furthermore, nitric oxide (NO) and calcium (Ca 2+) also show their
vital role in plant senescence signalling cascade. Calcium (Ca 2+) can hin-
der senescence, especially in detached leaves (Ma and Berkowitz, 2011;
Poovaiah and Leopold, 1973). It has been observed that Ca2+ can defer
methyl jasmonate (MeJA)-induced leaf senescence in rice (Ma and
Berkowitz, 2011). During an investigation, overexpression of a bacterial
NO dioxygenase (NOD), which has a key role in detoxifying NO in trans-
genic Arabidopsis, resulted in premature leaf senescence. However, exog-
enous treatment of NO delayed early senescence occurring in transgenic
Arabidopsis NOD plants (Mishina et al., 2007). Recently, Ma and
Berkowitz (2011) have shown that Arabidopsis cyclic nucleotide gated
channel 2 (CNGC2) has an important role in senescence signalling in-
duced by calcium inux. The CNGC2 loss-of-function mutant dnd1
resulted in decreased Ca 2+ uptake and early senescence have been ob-
served as compared to the wild type. In addition, endogenous NO con-
tent in dnd1 leaves was less than that in the leaves of non-transgenic
lines. Thus, the present study suggests that NO is a negative regulator
and Ca2+ as a positive regulator in senescence signalling.
2.3. Transcription regulation of gene expression
Transcription is regulated by transcription factors, connected to def-
inite cis-elements on the target gene promoters, which, in turn, cause
differential gene expression by activation and/or suppression of their
respective genes. About 1500 transcription factor genes were reported
in the Arabidopsis genome, which altogether constitute more than 30
gene families (Riechmann et al., 2000). Interestingly, more than 130
of them were reported from leaf senescence EST collection of
Arabidopsis (Guo et al., 2004). These senescence transcription factors
belong to the families of NAC, WRKY, AP2/EREBP, C2H2, C3H, C2C2, zinc
nger proteins, MYB, HB, bZIP, and bHLH, etc. (Guo et al., 2004). Previ-
ously, Chen et al. (2002) also showed senescence induced expression of
43 genes out of 402 transcription factor genes on a microarray.
Among the best studied leaf senescence associated transcription fac-
tors, WRKYs are the promising ones. The WRKY family is characterized of
possessing a 60 amino acid motif, the WRKY domain (Eulgem et al.,
2000). Of these, two WRKY proteins including WRKY6 and WRKY53,
have been found to be up-regulated during leaf senescence (Eulgem et
al., 2000; Hinderhofer and Zentgraf, 2001). The AtWRKY6 exerts its effect
by modulating calmodulin-response genes and other senescence-
induced kinases, known as SARK and SIRK (Robatzek and Somssich,
2002). SARK and SIRK show similarity in structure as they consist of a
SerThr kinase domain, a trans-membrane domain as well as an extra-
cellular leucine-rich domain (Yoshida, 2003). Further, Robatzek and
Somssich (2001, 2002) worked on Arabidopsis WRKY6 and SIRK expres-
sion during leaf senescence. It is well known that SIRK is a receptor
kinase, so WRKY6 binds to the promoter of SIRK and control its expres-
sion. The WRKY6 positively regulates PR1 (a defense related gene that
is also expressed during leaf senescence), possibly through its transcrip-
tion factor NPR1 (Zhou et al., 2000). Hinderhofer and Zentgraf (2001)
have shown induction of WRKY53 at early stages of leaf senescence. Se-
nescence is reported to get speeded up if WRKY53 is over-expressed in
Arabidopsis plants and delayed when RNAi or insertional mutation
suppressed this gene (Miao et al., 2004). The downstream target genes
1157
of WRKY53 can be stress related genes, defense-related genes, and
some SAGs (Miao et al., 2004; Zhang et al., 2011; Zhou et al., 2011).
Arabidopsis WRKY transcription factors have been reported to partic-
ipate in the gene expression during trichome development, wounding
and pathogen defense along with senescence (Zhou et al., 2011). Tran-
scription of WRKY22 was reported to be promoted by darkness and de-
creased by light. Over-expression of AtWRKY22 (dark-treated) and
knockout lines indicated phenotypes with varying response to senes-
cence (Zhou et al., 2011). Furthermore, AtNAP is a NAC family transcrip-
tion factor gene that contributes signicantly in leaf senescence, but its
mechanism needs to be elucidated (Zhang et al., 2011).
TCP is another transcription factor family involved in regulating leaf
senescence (Guo et al., 2010; Pulido and Laufs, 2010). TCPs can be
grouped into class 1 and class 2 (Cubas et al., 1999; Kosugi and
Ohashi, 2002). Interestingly, both differ in their effect on senescence
regulation. Class 1 TCPs (e.g. TCP20) are positive regulators of growth,
while in contrast, class 2 TCPs including TB1 and CYCDICH are negative
regulators (Kosugi and Ohashi, 2002; Li et al., 2005; Uberti-Manassero
et al., 2012). Balance of these classes is necessary for proper growth of
organs. The class 2 TCP genes include CINCINNATA (CIN) and JAW-D
(Palatnik et al., 2003). The gene CIN controls cell division arrest in
leaves. Thus, it is involved in senescence response. TCPs also exert
their effect by regulating WRKY53, and in turn onset of early senescence
gene expression. ORE1 is another transcription factor which may bind
to the promoters of SAGs and cause their activation. ORE1 is regulated
by ethylene-insensitive 2 (EIN2) and others in an age-dependent
manner.
Another transcription factor RAV1 (RAV family) plays a key func-
tion during initial events of leaf senescence (Woo et al., 2010; Zhao
et al., 2008). In a study, the role of RAV1 has been found to be closely
interlinked with leaf senescence. The RAV1 mRNA increased with age
and reached to a maximum level on the initiation of senescence, but
interestingly declined again at the nal stage of senescence which
suggested that RAV1 is a positive regulator of senescence and its ex-
pression is sufcient to cause leaf senescence (Woo et al., 2010).
Similarly SAG113, a gene encoding a Golgi-localized PP2C family
protein phosphatase, regulates ABA-induced stomatal opening/closing
and dehydration, especially during leaf senescence (Zhang et al.,
2011). Recently, Zhang et al. (2011) investigated that two genes namely
SAG113 and AtNAP were over-expressed during ABA accumulation and
leaf senescence. Overall, chemical induction of AtNAP also induces ex-
pression of SAG113. However, when AtNAP was knocked out, the ABA-
and senescence-induced expression of SAG113 also decreased.
From all the above reports it is amply clear that a variety of tran-
scription factors exist in the leaves, which contributed signicantly
in the regulation of leaf senescence. However, different types of tran-
scription factors regulate the transcription phenomenon to a varying
extent thereby regulating leaf senescence variably. With further ad-
vancement in knowledge on the interactive role of all such types of
transcription factors in regulating leaf senescence it would be possi-
ble to enhance crop yield by delaying stress-induced senescence.
3. Progression of senescence
3.1. Chloroplast degradation
Senescence starts with the degradation of chloroplast (Hrtensteiner
and Krutler, 2011). It can be observed in the form of disorientation of
granum stacks and swelling of the thylakoids resulting into plastoglobuli
enlargement (Gan and Amasino, 1997; Smart, 1994; Thomas and
Stoddart, 1980; Zentgraf and Hemleben, 2008; Zimmermann and
Zentgraf, 2005). A variety of intrinsic as well as extrinsic factors like
ageing and different types of stresses may trigger the senescence
program by causing loss in photosynthetic capacity or membrane
integrity (Azad et al., 2008; Franklin et al., 2012; Quirino et al., 2000).
The most important and widely occurring chloroplast protein,
1158 M. Sarwat et al. / Biotechnology Advances 31 (2013) 11531171
ribulose-1,5-bisphosphate carboxylase/oxygenase (rubisco) is rapidly
and selectively degraded during senescence, however, the mechanism
is still not fully known (Khanna-Chopra, 2011). During senescence, a
non-yellowing gene (NYE1) has been reported to be a key regulator of
leaf chlorosis (Ren et al., 2007; Wei et al., 2011). Similarly, in rice a
novel protein RLS1 was found to be up-regulated and caused chloroplast
degradation during dark-induced leaf senescence (Wang, 2012). The
degrading chloroplast is termed as gerontoplast by some researchers
(Matile et al., 1999; van Doorn and Yoshimoto, 2010). The chlorophyll
and degraded products are transferred to the cell vacuole (Borrmann et
al., 2009; Matile et al., 1999; Pruinsk et al., 2007), probably to detoxify
the cell from this potentially phototoxic pigment (Christ et al., 2012;
Hrtensteiner and Krutler, 2011). The mobilization of chloroplasts to
the vacuole is done by rubisco-containing bodies (RCBs), which are gen-
erally spherical in shape (Izumi and Ishida, 2011).
Despite the above changes taking place in the chloroplast during se-
nescence, the thylakoid membrane particularly its uidity and compo-
sition remains unaltered, but the membrane-bound electron transport
system I and II stied greatly during senescence (Falqueto et al., 2009;
Hutter et al., 2004). Thus, the measurement of loss of photochemical ca-
pacity (Fv/Fm; which reects the functionality of photosystem II) at a
particular growth stage is one of the most promising criteria to detect
senescence at that particular stage of a plant (John et al., 1995; Lim et
al., 2007, 2010; Oh et al., 1996, 1997; Zimmermann and Zentgraf, 2005).
From all these reports it is amply clear that the degradation of
chloroplast is the rst promising intracellular symptom of senescence
taking place in a leaf. Not only ultrastructure of a chloroplast is dam-
aged during senescence but several metabolites occurring in the chlo-
roplast are also affected.
3.2. Role of mitochondria
In animal systems, mitochondrion is the main organelle which plays
an important role in the regulation of programmed cell death (PCD),
whereas in contrast, in plant cells, this commission is carried out by
the chloroplast. In fact, the mitochondrial functions are maintained dur-
ing the whole breakdown process of the cell so they provide metabolic
energy until the late phase of senescence (Keech, 2011; Thomson and
Plat-Aloia, 1987; Zapata et al., 2005; Zimmermann and Zentgraf,
2005). From animal mitochondria, discharging of cytochrome c is a
key event during apoptosis (similar to PCD in plants) (Balk and
Leaver, 2001). This can be supported by a study in which it has been
shown that an impediment in the release of mitochondrial cytochrome
c caused inhibition in apoptosis in animal cells which is closely involved
with the activity of mitochondrial hexokinase (Eckardt, 2006). Further-
more, Kim et al. (2006) found that mitochondrial hexokinase regulates
PCD in plants, e.g., in Nicotiana benthamiana, virus-induced gene silenc-
ing (VIGS) of Hxk1 induced PCD. Likewise, overexpression of mitochon-
drial hexokinases in Arabidopsis enhanced survival under PCD-inducing
conditions (Eckardt, 2006). However, in plants in addition to mitochon-
dria, chloroplasts play a considerable role in controlling PCD due to
stress-induced over-production of ROS (Doyle et al., 2010). It is evident
from a study with Arabidopsis that antioxidant treatment enhanced PCD
induction, revealing that there is a composite interaction among chloro-
plasts, nuclear protein synthesis, light and ROS during plant PCD (Doyle
et al., 2010).
3.3. Loss of membrane integrity
The loss of membrane integrity is an important aspect of the senes-
cence program. Due to this phenomenon mobilization of membrane
lipids takes place which is contemplated as crucial for providing energy
for senescence to take place (Lim et al., 2007; Thompson et al., 1998).
The release of lipids in this way results in increased membrane perme-
ability. Long ago, Dhindsa et al. (1981) reported that leaf senescence in
Nicotiana tabacum could be due to the effect of cumulative membrane
deterioration caused by ROS-induced lipid peroxidation. Merzlyak and
Hendry (1994) reported that membrane lipid peroxidation is one of
the promising effects of senescence in plant leaves. While assessing
the inuence of salt stress on senescence-related attributes in rice seed-
lings Lutts et al. (1996) reported that increased membrane permeability
is one of the foremost symptoms of leaf senescence. The authors suc-
cessfully discriminated different rice genotypes for their tolerance to
salt stress using membrane permeability as a very efcient selection
criterion.
4. Senescence and Programmed Cell Death (PCD)
First experimental study on programmed nature of PCD was
carried-out in 1980s and 1990s (Bozhkov and Lam, 2011; Walbot et al.,
1983), although 20002010 is the most promising decade for the study
of PCD (Bozhkov and Lam, 2011). Many external and internal factors
are known to cause PCD in plants, which depend on the expression of
various genes involved in this process. For example, two independent
stresses (heat treatment and by allowing the cultures to senesce)
have been reported to induce PCD in plants which occur due to
up-regulation of certain genes during both processes (Feliciano et al., in
press; Guo and Gan, 2012). Such genes were considered to be involved
in all types of PCD and they may serve as markers for this process
(Elmore, 2007; Palavan-Unsal et al., 2005). Examples are oxidative
stress-related genes (CSD1, CSD3, and GPX) cysteine proteinases, some
transcription factors, and hypersensitivity response (HR)-related genes
(Al-Taweel et al., 2011; Gill and Tuteja, 2010; Swidzinski et al., 2004).
In the case of leaf senescence, PCD is regulated by several active genetic
programs. Of the different types of cells in a leaf, the mesophyll cells are
the rst that are affected during leaf senescence which later proliferate to
other cell types and eventually to the whole leaf tissue (Desvoyes et al.,
2006). In this way, leaf senescence is different from other types of PCD
(van Doorn and Woltering, 2004), as the former involves whole tissues
and the latter is limited to specic tissues and cells. Moreover, senes-
cence in the case of leaves occurs in patches and progresses slowly in
comparison with other PCDs known.
This slow progression in senescence ensures effective transport of
nutrients to other parts. Different studies have reported the typical
PCD like symptoms in the late phases of senescing leaves of Arabidopsis,
tobacco, etc. (Cao et al., 2003; Simeonova et al., 2000; Uzelac et al.,
2008). Some of these effects are DNA laddering, chromatin condensa-
tion and vacuolar collapse. A study on barley has also revealed disinte-
gration of nuclear DNA (nDNA) and condensation of chromatin as parts
of PCD (Kolodziejek et al., 2007). A similar order of events, rst frag-
mentation and then chromatin condensation, has been reported during
senescence (Kolodziejek et al., 2007; Tripathi and Tuteja, 2007). Unlike
senescence, PCD is not coordinated even in the neighbouring cells and
does not spread at the constant rate. It has been observed that in the ini-
tial stage of senescence, the concentration of NO reaches to ceiling in the
cytoplasm of mesophyll cells and during later stages this concentration
declines, suggesting a vital role of NO signalling in induction of PCD (da
Silva et al., 2011; de Michele et al., 2009; Kolodziejek et al., 2007).
Although senescence is a type of PCD (van Doorn and Woltering,
2004), its properties differ from those of PCD (Thomas et al., 2003). As
stated earlier, senescence involves degradation of macromolecules like
DNA, RNA and proteins and mobilization of carbon, nitrogen, phospho-
rus and other minerals within the plant body (Fischer, 2007). Senes-
cence has been reported to have a close link with PCD because of the
reason that the key hydrolytic enzymes (proteases and nucleases) in-
volved in the breakdown of macromolecules are encoded by different
senescence associated genes (SAGs) (Farage-Barhom et al., 2011). Var-
ious studies conrm the up-regulation of these endonucleases during
the course of senescence (like BFN1) (Farage-Barhom et al., 2008,
2011) and PCD (like ZEN1) (Ito and Fukuda, 2002). Moreover, the pro-
tein sequences of senescence associated nuclease BFN1 are highly sim-
ilar to ZEN1, a PCD-related nuclease (Ito and Fukuda, 2002). Both can be
 M. Sarwat et al. / Biotechnology Advances 31 (2013) 11531171
grouped into type I nuclease family. These are single-strand-specic
endonucleases, like S1 nuclease which can degrade both RNA
and single-stranded DNA (Desai and Shankar, 2003). Different environ-
mental factors can induce endonucleases (Muramoto et al., 1999;
Saumonneau et al., 2012; Yupsanis et al., 2001). Enhanced nuclease ac-
tivity has been reported during different phases of leaf senescence
(Aleksandrushkina et al., 2008; Farage-Barhom et al., 2011). A variety
of other PCD processes like hypersensitive response (HR), aleurone
cell death, endosperm development, and tracheary element (TE) differ-
entiation have also been reported to be controlled due to up-regulation
of nucleases (Dominguez et al., 2004; Farage-Barhom et al., 2008, 2011).
Although involvement of mitochondria has not been conrmed in
the programmed cell death (PCD) of leaf senescence, Swidzinski et al.
(2004) have reported down-regulation of the mitochondrial adenine
nucleotide transporter, suggesting an early event in the occurrence of
plant PCD. Recently, in a comprehensive review, Bozhkov and Lam
(2011) stated comparative analyses of cell death mechanisms in
plants, animals and fungi will provide an evolutionary framework
for understanding PCD function and control in various biological
systems as well as improvement in biomass and wood production,
disease and stress resistance. Signicant results in this eld will pro-
vide great impact on agriculture and forestry to cope with the
ever-increasing demand for food production.
5. Involvement of phytohormones in leaf senescence
Hormonal regulation of senescence is an interesting topic in its
own right as it involves a complex interplay of signalling molecules
at various stages of leaf senescence. Global expression proling and
availability of genetic mutants have paved the way to unravel the
mystery of these remarkable biomolecules and their role in senes-
cence (Table 1).
5.1. Cytokinins
Of the various plant hormones studied for their role in senescence,
cytokinins receive the maximum attention, because of their consider-
able role in delaying leaf-senescence (Kim et al., 2006; Lara et al.,
2004; Richmond and Lang, 1957; Xu and Huang, 2009). Thus, they
are of great benet for the economically important plants. The levels
of cytokinins within a cell gradually decline as leaf senescence pro-
gresses, and there is a general consensus that high levels of cellular
cytokinins delay leaf senescence (Kim et al., 2006; Lara et al., 2004;
McCabe et al., 2001). To further reinforce this observation, certain
studies (Frebort et al., 2011; Hirose et al., 2008; Xu et al., 2010)
have demonstrated the down-regulation of cytokinin biosynthesis
genes such as adenosine phosphate isopentenyl-transferase (IPT)
and cytokinin synthase as well as up-regulation of cytokinin oxida-
tion genes (cytokinin oxidase) during leaf senescence (Table 1).
The gain-of-function analysis of Arabidopsis ore12-1 mutant shows
insight into the molecular mechanism of cytokinin-mediated delayed
senescence response (Kim et al., 2006; Lim et al., 2007, 2010). Its
loss-of-function mutant AHK3 (Arabidopsis Histidine Kinase 3) causes
reduced sensitivity to cytokinins thereby prompting senescence even
in the presence of cytokinins. The gene AHK3 possibly exerts its effect
by phosphorylating the Arabidopsis response regulator 2 (ARR2)
(Horak et al., 2008; Kudryakova et al., 2008).
The exact mechanism how cytokinins prevent senescence in
the plant is not known. However, a correlation between cytokinins
and the enzyme extracellular invertase has been reported (Lara et
al., 2004). With the inhibition of extracellular invertase activity,
cytokinin-regulated postponement in leaf senescence is also repressed,
which suggests the vital role of this enzyme in cytokinin-induced delay
in leaf senescence. Furthermore, invertase activity enhances during
carbohydrate partitioning, which acts as a trigger for leaf senescence.
In contrast, another hexokinase AtHXK1, an intracellular enzyme,
1159
accelerates leaf senescence. Isopentenyl transferase (IPT) is a key en-
zyme in the biosynthesis of cytokinins, which also plays a signicant
role in delaying leaf senescence. But, the double transgenic tomato
plants overexpressing both AtHXK1 and IPT show early senescence
phenotype (Swartzberg et al., 2011). This indicates the dominant nature
of AtHXK1 gene and so dominance of intracellular sugar sensing via
AtHXK1 over extracellular sugar sensing mediated by IPT (Swartzberg
et al., 2011). This indicates the complexities of hormonal signalling dur-
ing senescence.
Cytokinins also mediate their action by modulating a number of
transcription factor genes like GATA22, HAT4, HAT22 and bHLH64
(Kllmer et al., 2011; Wang, 2012). For example, overexpression of
HAT22 has been shown to cause reduction in chlorophyll content of
seedlings and an early onset of leaf senescence in these plants
(Kllmer et al., 2011). Further studies on the various transcription
factor genes listed here or discovered in future may elucidate the
mechanism of leaf senescence.
5.2. Ethylene
While cytokinins suppress senescence, ethylene is known since
long for hastening leaf senescence (Abeles et al., 1988). The underly-
ing genetic mechanism is beginning to be unravelled with studies fo-
cusing on genes of the ethylene biosynthetic pathway. The three
major genes encoding ACC synthase, ACC oxidase and nitrilase are
reported to be up-regulated in many plant species during leaf senes-
cence (Hirose et al., 2008; Van der Graaff et al., 2006). Further conr-
mation came from some Arabidopsis mutant studies. For example,
two mutants, ethylene-insensitive 2 (ein2) and ethylene-resistant 1
(etr1), being decient in ethylene signal transduction, delayed leaf se-
nescence (Lim et al., 2007). It was observed that leaf senescence did
not appear earlier in transgenic plants constitutively overproducing
ethylene (Lim et al., 2007; Yang et al., 2008). This suggests that ethyl-
ene alone cannot drive leaf aging.
Certain age-dependent factors are found to be a part of ethylene-
regulated leaf senescence. In Arabidopsis the old1 (onset of leaf death
1) mutant shows an earlier-onset of senescence (Jing et al., 2002). In
combination with ethylene, these mutants exhibit further accelera-
tion of senescence. In the old1etr1 double mutant where ETR1 gene
is also mutated, ethylene perception was impaired (Gan, 2003;
Lim et al., 2007). Thus, despite showing age-dependent early onset
of leaf senescence, there is no acceleration when treated with
ethylene.
5.3. ABA
The role of ABA in plant development as well as leaf senescence
is well known (Table 1). Exogenous application of ABA has long
been known to promote leaf abscission and senescence (Zeevaart
and Creelman, 1988). The level of ABA certainly increases in
senescing leaves and the exogenous application of ABA induces ex-
pression of several SAGs (Choi and Hwang, 2011; Lim et al., 2007;
Weaver et al., 1998). The correlation between ABA and senescence
is further conrmed by the up-regulation of both ABA and leaf se-
nescence during environmental stresses (drought, high salinity
and low temperature) (Tuteja, 2007). For gaining insight into the
cellular mediators of ABA-induced senescence, ABA inducible
receptor-like kinase gene of Arabidopsis, RPK1 has been comprehen-
sively studied (Lee et al., 2011). Its expression increases during the
progression of leaf senescence. Loss-of-function mutants in RPK1
showed delayed natural senescence. Interestingly, these mutants
show reduced sensitivity to ABA-induced senescence, but little
change to JA- or ethylene-induced senescence (Lee et al., 2011).
The role of RPK1 in leaf senescence depends upon developmental
stage of a leaf. Over-expression of RPK1 at an early stage causes
growth retardation but not senescence, while during adult stages
1160
Table 1
Role of different phytohormones in regulation of plant senescence.
Phytohormones Plant species
Cytokinins Rose (Rosa spp.)
Tobbaco (Nicotiana tabacum)
Tobbaco (Nicotiana tabacum)
Petunia (Petunia hybrida
Carnation (Dianthus
caryophyllus)
Tomato (Lycopersicon
esculentum)
Creeping bentgrass
(Agrostis stolonifera)
Agrostis stolonifera
Arabidopsis thaliana
Arabidopsis thaliana
Tomato
(Lycopersicon esculentum)
Arabidopsis thaliana
Auxins Arabidopsis thaliana
Tobacco (Nicotiana tabacum)
Tropaeolum majus
Arabidopsis thaliana
Arabidopsis thaliana
Arabidopsis thaliana
Arabidopsis thaliana
Arabidopsis thaliana
Ethylene Watercress (Nasturtium spp.)
Nicotiana sylvestris
Arabidopsis thaliana
M. Sarwat et al. / Biotechnology Advances 31 (2013) 11531171
Effect Reference
Application of the cytokinin (N6-henzyladenine) prolonged existence of fresh or aged Mayak and Halevy, 1970
owers of a short-lived rose variety, which shows that cytokinins take part in the
endogenous regulation of senescence in rose petals
In cytokinin-induced transgenic tobacco plants the production of cytokinins in Wingler et al., 1998
senescing tissue suppressed the rubisco, hydroxypyruvate reductase (HPR), and
some other enzymes participated in photosynthetic processes, which caused
delay in senescence, which might be due to reduction in photosynthesis-involved
proteins
Exogenous application of cytokinins increased its endogenous concentration which delayed Lara et al., 2004
senescence and caused nutrient mobilization
Overproduction of cytokinins in petunia owers transformed with PSAG12-IPT delayed Chang et al., 2003
corolla senescence and reduced sensitivity to ethylene
Stem and leaf tissues of carnation (Dianthus caryophyllus) plants appeared to contain a Eisinger, 1977
natural antisenescence factor (kinetin), and removal of these tissues from carnation
owers promoted senescence. However, exogenous application of kinetin (510 g/ml)
considerably delayed senescence of owers on removal of stem and leaf tissues. In
addition, the life span of cut-owers was increased with kinetin treatment
Over-expression of two (SAG12 and SAG13) senescence-associated Arabidopsis genes in Swartzberg et al., 2011
tomato plants expressing isopentenyl transferase (IPT) causes delay in leaf senescence
and promotes owering
Creeping bentgrass transformed with SAG12-ipt showed greater tiller as well as root Xu and Huang, 2009
formation and restricted leaf senescence under high temperature stress
The transformation of ipt (adenine isopentenyl transferase gene) regulating CK synthesis in Xu et al., 2010
a C3 perennial grass species, Agrostis stolonifera resulted in delayed senescence due to
change in protein contents and energy metabolism
Loss-of-function mutant AHK3 (Arabidopsis Histidine Kinase 3) caused reduced Horak et al., 2008
sensitivity to cytokinins thereby prompting senescence even in the presence of
cytokinins. The gene AHK3 possibly exerted its effect by phosphorylating the
Arabidopsis response regulator 2 (ARR2)
Expression of four different cytokinin-regulated transcription factor genes, GATA22, Kllmer et al., 2011
HAT4, HAT22 and bHLH64 has shown that of all the genes, overexpression of HAT22
lowered the seedling chlorophyll content and induced premature leaf senescence
The double transgenic tomato plants over-expressing both AtHXK1 and IPT appeared as Swartzberg et al., 2011
an early senescence phenotype
ARR5-gene expression in natural leaf senescence and detached leaf senescence in the Kudryakova et al., 2008
dark using Arabidopsis thaliana plants overexpressing PARR5-GUS gene showed that
natural leaf senescence was dependent on decreased GUS-activity
Auxin response factors (ARFs) known as transcription factors regulate plant responses Ellis et al., 2005
to auxins. It has been found that arf2 mutant plants delayed rosette plant aging,
senescence and owering
Transgenic lines showed greater lateral branches as well as prolonged preservation of Khodakovskaya et al., 2006
chlorophyll under dark incubation
Roles of naphthalene acetic acid (NAA), indole butyric acid (IBA) and indole acetic acid Karatas et al., 2010
(IAA) were investigated on some physiological processes under dark-induced
senescence in Tropaeolum majus. IAA accelerated chlorophyll and carotenoid loss, while
improved protein contents
Two allelic mutations, ore14-1 and ore14-2 induced a signicant delay in different Lim et al., 2010
senescence related attributes. Overexpression of ORE14 showed that it encodes ARF2, a
member of the auxin response factor (ARF), which mediated early auxin-induced gene
expression thereby positively regulating the leaf senescence in Arabidopsis
Auxins cause down-regulation of over a half of the genes (IAA-TPs and AUX/IAA) of Van der Graaff et al., 2006
auxin transport which may result in uneven distribution of auxins
Characterization of three arf2 mutants such as arf2-7, arf2-6 and arf2-8 causes delayed Okushima et al., 2005
owering, dark green rosette leaves, long and thick inorescence and sterility in early
appeared owers as well as delayed senescence and abscission, as compared to
wild-type plants
Arabidopsis plants over-expressing YUCCA6 (a family of avin monooxygenase proteins) Kim et al., 2011a
displayed dramatic longevity, exhibited delayed dark-and hormone-induced senescence
Transgenic plants overexpressing GmSARK, led to early leaf senescence, chloroplast Xu et al., 2011
degradation and alteration in ower morphology
While observing the efcacy of various inhibitors of ethylene synthesis such as Philosoph-Hadas et al., 1994
aminoethoxyvinylglycine (AVG), CO2 and Ag+ in retarding senescence processes it was
observed that application of CO2 (11%) to watercress bunches signicantly retarded the
senescence process. In addition, exogenous treatment of AVG (0.1 mM) or Ag+ (30 M)
to detached leaves signicantly reduced chlorophyll contents as compared to that of
proteolysis which showed that the delayed senescence cannot be associated only to its
action as an anti-ethylene agent. Therefore, it can be suggested that all
senescence-associated processes are not regulated by ethylene
Transgenic N. sylvestris plants expressing Arabidopsis mutant ethylene receptor ETR1-1 Yang et al., 2008
or the tomato EIN3-like (LeEIL1) gene show time-course leaf senescence in transgenic
lines as compared to wild type plants. However, petal necrosis was delayed, These
observations support a role for ethylene in regulating a spectrum of developmental
events associated with organ senescence and tissue necrosis
The association between age and ethylene to induce leaf senescence was observed in Jing et al., 2005
different accessions and mutants of Arabidopsis. Prolonged exposure of ethylene
 M. Sarwat et al. / Biotechnology Advances 31 (2013) 11531171 1161

Table 1 (continued)
Phytohormones Plant species Effect Reference

enhanced the yellowing of leaves suggesting that leaf senescence also depends on the
duration of ethylene application
Petunia (Petunia spp.) Ethylene caused premature senescence of leaves and owers Jones and Pasian, 2011
Arabidopsis thaliana Senescence-associated receptor-like kinase (SARK), a leucine-rich repeat-receptor-like Xu et al., 2011
protein kinase, obtained from Glycine max acts as a positive regulator of leaf senescence.
Transgenic A. thaliana plants for inducible overexpression of GmSARK, led to early leaf
senescence, chloroplast destruction, and abnormal ower morphology. Inhibition of
ethylene biosynthesis alleviated the senescence-induced by GmSARK
Nicotiana mutabilis Up-regulation of 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) transcripts Macnisha et al., 2010
elevated the rates of ethylene production at the onset of petal wilting
Hibiscus rosa-sinensis Flowers treated with 1-aminocyclopropane-1-carboxylic acid (ACC), a precursor of Trivellini et al., 2011
ethylene biosynthesis, enhanced ower senescence
Arabidopsis thaliana Interaction between leaf age and ethylene was assessed in isolated mutants with altered Jing et al., 2002
leaf senescence referred to as onset oeafdeath (old) mutants. The old1 is represented by
three recessive alleles and displayed earlier senescence both in air and upon ethylene
exposure. Analyses of the chlorophyll degradation, ion leakage and SAG expression showed
that leaf senescence was enhanced in ethylene-treated plants
Arabidopsis thaliana Ethylene-insensitive mutant etr2-1 shows leaf expansion, etiolated seedlings and leaf Sakai et al., 1998
senescence
Arabidopsis thaliana Ethylene insensitivity was conferred by ETR1 gene which promoted ethylene signal Chang et al., 1993
transduction pathway of Arabidopsis thaliana
Tomato (Lycopersicon Ethylene-induced the triple response in etiolated hypocotyls of various tomato Lanahan et al., 1994
esculentum) mutants except for Never ripe (Nr). Lack of ripening in this mutant was found to be
caused by ethylene insensitivity. In addition, overexpression of
1-aminocyclopropane-1-carboxylate (ACC) synthase in plants indicated that
they are phenotypically opposites of Nr plants
Nemesia strumosa Over-expression of ethylene receptor gene Cm-ETR1/H69A declined ethylene sensitivity Cui et al., 2004
in Nemesia strumosa. All the transgenic plants showed enhanced ower longevity as
compared to that of the wild types
Salicylic acid Arabidopsis thaliana The atg mutant phenotypes promote PCD. It was found that the cell death in the atg Yoshimoto et al., 2009
(SA) mutants is associated with SA signal
Arabidopsis thaliana Senescence-induced expression of the cysteine protease gene SAG12 was dependent on Morris et al., 2000
the presence of SA. In addition, delayed yellowing and declined necrosis in the mutant
plants defective in SA suggest a key role of SA in plant cell death
Arabidopsis thaliana Transgenic Arabidopsis overexpressing ACBP3 (ACBP3-OEs) displayed accelerated leaf Xiao et al., 2010
senescence. Furthermore, enhanced senescence in ACBP3-OEs transgenic plants depends
on SA, but not on jasmonates
Arabidopsis thaliana WRKY53 and WRKY70 were previously implicated as positive and negative regulators of Besseau et al., 2012
senescence, respectively. Involvement of two additional WRKY TFs, WRKY 54 and
WRKY30, has also been suggested in this process. Expression in wild-type and salicylic
acid-decient mutants suggests a common but not exclusive role for SA in induction of
WRKY30, 53, 54, and 70 during senescence which showed that WRKY53, WRKY54, and
WRKY70 may participate in a regulatory network that integrates internal and
environmental factors to modulate the onset and the progression of leaf senescence,
possibly through an interaction with WRKY30
ABA Arabidopsis thaliana Arabidopsis mutant oresara9 (ore9) leaf showed prolonged longevity during natural Woo et al., 2001
senescence by delaying the commencement of different senescence symptoms
Arabidopsis thaliana It has been believed that a senescing leaf loses water greater as compared to a Zhang et al., 2011
non-senescing leaf, and ABA has a prominent role in enhancing leaf senescence. They
identied a leaf senescence associated gene, SAG113, which is usually expressed in
senescing leaves and is induced by ABA. However, over-expression of SAG113 was
declined in aba2 and abi4 mutants
Arabidopsis thaliana Leaf senescence mutants [ore1 (oresara1), ore3 and ore9] show delayed senescence Kim et al., 2011b
phenotypes in dark-induced and age-dependent senescence. So, it is suggested that
ORE1, ORE3 and ORE9 can be used for the appropriate progression of leaf senescence
mediated by ABA as well as MeJA and ethylene
Cotton (Gossypium spp.) Leaf senescence is closely associated with reduced accumulation of zeatin and its Dong et al., 2008
riboside (Z + ZR) and isopentenyl and its adenine (iP + iPA) rather than enhanced ABA
or dihydrozeatin and its riboside (DHZ + DHZR) in cotton leaves
Arabidopsis thaliana Abscisic acid (ABA)-responsive NAC transcription factor VND-INTERACTING2 (VNI2) Yang et al., 2011
integrates ABA-induced stress signals into leaf aging by regulating RESPONSIVE TO
DEHYDRATION (RD) and COLD-REGULATED (COR) genes
Arabidopsis thaliana Age-dependent senescence was signicantly delayed in RPK1 decient mutants. Lee et al., 2011
However, overexpression of RPK1 at the reproductive stage signicantly enhanced the
senescence as well as cell death. Furthermore, induction of RPK1 at an early growth
stage reduced growth without generating senescence symptoms, which indicates that
the role of RPK1 in senescence is age dependent
Jasmonic acid Rice (Oryza sativa) A new zinc nger protein, OsDOS delays leaf senescence in rice. It is observed that Kong et al., 2006
(JA) expression of OsDOS was down-regulated during leaf senescence, pollination as well as
panicle development. RNAi knockdown of OsDOS increased age-dependent leaf
senescence, while its over-expression caused a considerable delay in leaf senescence,
which indicates that it acts as a negative regulator of leaf senescence
Cucurbita pepo Foliar-applied 100 M methyl jasmonate (MeJA) reduced chlorophyll content, Ananieva et al., 2007
chloroplast transcriptional activity and photosynthetic rate more than did by
darkness. This indicates that MeJA is a more potent inducer of senescence in
C. pepo seedlings

(continued on next page)

1162
Table 1 (continued)
Phytohormones Plant species
Jasmonic acid Arabidopsis thaliana
(JA)
Arabidopsis thaliana
Arabidopsis thaliana
Arabidopsis thaliana
Arabidopsis thaliana
Arabidopsis thaliana
Brassinosteroids Arabidopsis thaliana
(BRs)
Arabidopsis thaliana
Jujube (Ziziphus jujuba)
Wheat (Triticum aestivum)
Tomato (Lycopersicon
esculentum)
Gibberellins Paris polyphylla
Paris polyphylla
Zoysiagrass (Zoysia japonica)
Tomato (Lycopersicon
esculentum)
Taraxacum megallorrhizon,
Rumex pulcber
and Tropaeolum majus
Pea (Pisum sativum)
Pea (Pisum sativum)
Polyamines Oat (Avena sativa)
Rosa bourboniana and
Rosa damascena
Tobacco (Nicotiana tabacum)
Barley (Hordeum vulgare)
Lactuca sativa
Peach (Prunus persica)
Apple (Malus domestica)
M. Sarwat et al. / Biotechnology Advances 31 (2013) 11531171
Effect Reference
JA induced premature senescence in detached as well as attached leaves in wild-type He et al., 2002
Arabidopsis indicating that JA-signalling is essential for JA to induce leaf senescence
Exogenous application of JA enhanced leaf senescence in both drought sensitive and Yun-xia et al., 2010
drought tolerant genotypes and it was signicantly attributed to chlorophyll reduction
as well as increased malondialdehyde (MDA) contents
TCP (TEOSINTE BRANCHED/CYCLOIDEA/PCF) transcription factor genes modulate Schommer et al., 2008
hormone responses including biosynthesis of JA. It was shown that miR319-controlled
TCP transcription factors coordinated two sequential processes in leaf development as
well as leaf growth, which was in fact down-regulated, while, in contrast, leaf
senescence was positively regulated
Senescence-associated genes were identied in which the reporter gene GUS is He and Gan, 2003
up-regulated by both senescence and JA. Consistent with the GUS expression data,
another gene OPR1 was indeed up-regulated by both senescence and JA, which
suggests that leaf senescence and JA may share a common molecular mechanism
for modulating OPR1
Exogenous application of MeJA caused increased expression of SAGs such as SEN4, SEN5, Xiao et al., 2004
and VPE and loss of chlorophyll content and photochemical efciency in Arabidopsis.
Mutant studies have further conrmed the role of JA in leaf senescence as JA-induced
senescence is absent in JA-insensitive mutant
The 35 coronatine insensitive 1 (COI1)-dependent JA-regulated proteins were identied Shan et al., 2011
in Arabidopsis. Of them, Rubisco activase (RCA) was a COI1-dependent JA-repressed
protein. Furthermore, loss of RCA caused premature senescence
Mutants decient in BR biosynthesis (det2) or the BR receptor BRI1 exhibited delayed Yang et al., 2011
senescence, indicating that BRs are positive regulator of senescence
BRI1-associated kinase 1 (BAK1) has a key role in BR signalling as well as Jeong et al., 2010
BR-independent plant defense response. Transgenic Arabidopsis plants expressing
BAK7 showed growth reduction and early senescence
Exogenous application of BRs (5 M) effectively enhanced the activities of superoxide Zhu et al., 2010
dismutase (SOD), phenylalanine ammonia-lyase, polyphenoloxidase as well as catalase
and delayed fruit senescence and maintained fruit quality by reducing ethylene
production
Exogenously-applied epibrassinolide (eBL) at the rate of 10 and 0.1 M accelerated leaf Salam-a, 2007
senescence in wheat plants due to increased peroxidase activity and decreased protease
activity and chlorophyll contents
Exogenous application of BRs to pericarp discs resulted in enhanced lycopene contents Vardhini and Rao, 2002
and decreased chlorophyll pigments. In addition, it decreased ascorbic acid, while
increased carbohydrate contents. BR-induced fruit ripening was associated with
increased ethylene production, resulting in acceleration of fruit-senescence
Application of gibberellic acid (GA3) considerably enhanced the endogenous GAs Li et al., 2010
(GA4 + GA7) level and slowed down the senescence of shoots, and the degradation of
proteins and chlorophyll. While activities of chlorophyllase, Mg-dechelation and
peroxidase increased more in control plants than in those treated with GA3
Foliar-applied gibberellin A3 (GA3) showed antagonistic effects of GA and ABA on Yu et al., 2009
senescence
The seedlings treated with GA3 delayed leaf senescence induced by darkness Cheng et al., 2009
When leaf discs of tomato were treated with solutions containing GA3 (4.33 106 M), Parmar and Hammond, 1971
senescence was retarded as compared to that in non-treated plants due to delay in
degradation of chlorophyll, and improved levels of total protein, RNA, and DNA
ABA reduced the senescence retarding effect of GA on excised leaf discs of the three plant Back and Richmond, 1971
species more than that of cytokinins. In addition, GA did not reverse the effects of ABA
GA3 application in vitro resulted in rapid accumulation of GDA-1 mRNA in LD-grown G2 Li et al., 1998
pea apical buds, which is compatible with its delaying effect on apical senescence
The G2 line of pea exhibited apical senescence-delaying phenotype under short-day
(SD) conditions. The initiation of cell death signals in the terminal oral meristem was
involved in the regulation of apical senescence in pea plants
Foliar-applied diaminopropane, spermidine and spermine enhanced the chlorophyll Borrell et al., 1997
contents, while decreased malondialdehyde (MDA) levels, lipoxygenase activity as
well as enzyme-protein levels, which induced anti-senescence effects
Polyamine biosynthetic inhibitors diuoromethylornithine (DFMO), Sood and Nagar, 2008
diuoromethylarginine (DFMA) and methylglyoxal-bis(guanylhydrazone) (MGBG)
enhanced senescence, however, exogenously-applied spermine considerably delayed
this phenomenon
In excised owers, foliar-applied spermine caused an increase in free and acid-soluble Serani-Fracassini et al., 2002
conjugated PAs levels, resulting in delayed programmed cell death as well as senescence
The kinetin conserved low TGase levels/activity during leaf senescence which showed a Sobieszczuk-Nowicka et al.,
key component of the kinetin action in retarding leaf senescence 2009
Spermidine declined chlorophyll degradation and promoted endogenous TGase activity. Serani-Fracassini et al., 2010
However, exogenously applied TGase enhanced chlorophyll synthesis three-fold. The
senescence-delaying effects of spermidine were found to be regulated by TGase
Field applications of spermidine (1 mM) on peach fruit showed reduced ethylene level Torrigiani et al., 2012
and esh softening. The endogenous concentration of free as well as insoluble
conjugated PAs increased 0.32.6-fold, while it caused reduction in non-treated plants
which showed that spermidine impedes fruit development as well as fruit ripening by
interacting with multiple hormonal pathways
Methylglyoxal bis-(guanylhydrazone) (MGBG)-treated fruit had higher ethylene Pang et al., 2010
production as compared to that by non-treated fruits. The expression of apple
 M. Sarwat et al. / Biotechnology Advances 31 (2013) 11531171 1163

Table 1 (continued)
Phytohormones Plant species Effect Reference

SAM synthase (MdSAMS) in MGBG-treated fruit was slightly higher than that in control.
In addition, considerable changes in free polyamine contents indicated a relationship
between polyamine and ethylene pathways
Tomato (Lycopersicon Over-expression of ySpdSyn (yeast spermidine synthase) in tomato enhanced Nambeesan et al., 2010
esculentum) intracellular
level of spermidine in the leaf as well as in the fruits. Furthermore, ySpdSyn transgenic
fruits had a longer shelf life, reduced shriveling and delayed senescence as compared to
non-transgenic plant fruits

senescence is accelerated thereby leading to cell death (Lee et al.,
2011; Osakabe et al., 2010). Similar response is noticed for exoge-
nous application of ABA, which does not have an effect on young
leaves but causes senescence in older leaves (Lee et al., 2011; Park
et al., 1998). This indicates that ABA function in senescence is medi-
ated by RPK1.
5.4. Jasmonic acid
The positive role of methyl jasmonate (MeJA) and its precursor
jasmonic acid (JA) in leaf senescence has been well documented in a
number of plants like Arabidopsis, oat, rice, etc. (He et al., 2002; Kim et
al., 2009; Kong et al., 2006; Shan et al., 2011; Ueda and Kato, 1980).
Exogenous application of MeJA causes increased expression of SAGs
such as SEN4, SEN5, and -VPE and loss of chlorophyll content and pho-
tochemical efciency in Arabidopsis. Mutant studies have further con-
rmed the role of JA in leaf senescence as JA-induced senescence is
absent in the JA-insensitive mutant coronatine insensitive 1 (coi1)
(Shan et al., 2011; Xiao et al., 2004). JA shows its role in leaf senescence
by repressing the protein rubisco activase (RCA), and this repression is
coil dependent. The genetic and physiological studies show that this re-
pression of RCA associated with JA-regulated leaf senescence, and that
loss of RCA causes general senescence-linked characteristics (Shan et
al., 2011). The JA-induced leaf senescence was further elaborated by
the observation of OsDOS) gene, which is a negative regulator for leaf
senescence (Kong et al., 2006; Lim et al., 2007). Studies on the OsDOS
overexpressing plants and RNAi knockdown plants have revealed that
many of the JA signalling-dependent genes were up-regulated in the
RNAi transgenic lines but down-regulated in the over-expressing trans-
genic lines (Shan et al., 2011). These observations validate the
co-regulation of JA signalling pathway with senescence.
5.5. Salicylic acid
Although the role of salicylic acid (SA) has been widely studied in
disease resistance, it also shows involvement in leaf senescence
(Table 1). Similar to JA the endogenous level of SA is four times higher
in the leaves undergoing senescence (Lim et al., 2003, 2007).
Arabidopsis mutants like npr1 and pad4, defective in SA signalling, ex-
hibit reduced expression of a number of SAGs such as PR1a, chitinase,
and SAG12 (Buchanan-Wollaston et al., 2003; Lim et al., 2007; Quirino
et al., 2000; Rao et al., 2002). Transcriptome proling reveals high
level of similarities between SA pathway and senescence pathway in
gene modulation (Van der Graaff et al., 2006). Morris et al. (2000) and
Buchanan-Wollaston et al. (2005) also postulated the involvement of
SA in the regulation of gene expression during developmental
senescence.
As SA is also involved in pathogen mediated response and cell death,
its involvement in senescence-associated cell death has been proposed
(Coll et al., 2011; Feys et al., 2005). There is an evidence which indicates
that SA might be involved in senescence-associated cell death (Coll et
al., 2011; Feys et al., 2005; Rao et al., 2002). The evidence came from
analysis of Arabidopsis pad4 mutants, defective in the SA signalling path-
way, also showed impaired cell death (Brosch and Kangasjrvi, 2012;
Yoshimoto et al., 2009).
5.6. Brassinosteroids (BRs)
BRs are generally known as positive regulators of senescence. For
example, mutants decient in BR biosynthesis (det2) or the BR receptor
BRI1 exhibit delayed senescence (Yang et al., 2011). However, contrari-
ly, the microarray expression proling has shown no effect on the ex-
pression of BR biosynthesis genes during senescence (Van der Graaff
et al., 2006). AtBRL3 is a BR receptor that functions in vascular differen-
tiation (Cano-Delgado et al., 2004). The signicant up-regulation of the
gene of this receptor during senescence supports the notion that BRs
might function in controlling the vascular system for efcient transport
of metabolites during senescence (Cano-Delgado et al., 2004; Wang and
Chory, 2006; Yang et al., 2011). However, the molecular mechanism of
BR action on leaf senescence is not fully known yet (He et al., 2001; Lim
et al., 2007). The effects of exogenous application of BRs (5 M) on se-
nescence were assessed in jujube fruit (Zhu et al., 2010). BRs effectively
enhanced the activities of superoxide dismutase (SOD), phenylalanine
ammonia-lyase, polyphenoloxidase as well as catalase and delayed
fruit senescence and maintained fruit quality by reducing ethylene pro-
duction (Zhu et al., 2010). A few years ago, Salam-a (2007) found
that exogenous application of epibrassinolide (eBL) at 10 and 0.1 M
accelerated leaf senescence in wheat plants due to increased peroxidase
activity and decreased protease activity and chlorophyll contents
(Salam-a, 2007). However, in contrast, it has also been observed
that BRs are also involved in delaying leaf senescence by improving
chlorophyll pigments. Chlorophyll breakdown is one of the vital indica-
tors for the measurement of leaf senescence and BRs are reported to be
involved in adversely affecting the photosynthetic apparatus of plant
organs (Rao et al., 2002; Salam-a, 2007; Vardhini and Rao, 2002).
As there is no much information available in the literature on the role
of BRs in leaf senescence so the mechanism how BRs regulate leaf senes-
cence is not fully known.
5.7. Auxins
Auxins, referred to as generally plant development hormones, play a
signicant role in leaf senescence (Kim et al., 2011a; Lim et al., 2007,
2010; Sexton and Roberts, 1982). Evidence came from up-regulation of
some key auxin biosynthetic genes (Van der Graaff et al., 2006), like
tryptophan synthase (TSA1), IAAld oxidase (AO1), and nitrilases
(NIT1-3), and consequently increased auxin concentration in the
senescing leaves. Further, when auxins are applied exogenously they
are reported to cause down-regulation of some SAGs (Hong et al.,
2000; Jones et al., 2010; Kim et al., 2011a; Noh and Amasino, 1999).
Thus, auxins are contemplated as negative regulators of leaf senescence.
Another interesting observation to note is that auxin concentration
changes considerably during senescence (Addicott et al., 1955; Ellis et
al., 2005; Lim et al., 2007; Van der Graaff et al., 2006). They cause
down-regulation of over a half of the genes of auxin transport (Van der
Graaff et al., 2006) which may result in uneven distribution of auxins.
Analysis of genetic mutants defective in auxin signalling further rein-
forces their role in leaf senescence (Ellis et al., 2005; Lim et al., 2007,
2010; Okushima et al., 2005). For example, a transcription repressor of
auxin signalling pathway, AUXIN RESPONSE FACTOR 2 (ARF2), is
up-regulated during leaf senescence (Ellis et al., 2005; Lim et al., 2010).
1164 M. Sarwat et al. / Biotechnology Advances 31 (2013) 11531171
It has been reported that the T-DNA mutants of ARF2 cause delay in leaf
senescence, suggesting their role as positive regulators in the process
(Lim et al., 2007; Xu et al., 2011). YUCCA6, which is a member of avin
monooxygenase family of proteins, catalyses a rate-limiting step in
auxin biosynthesis (Kim et al., 2011a; Mashiguchi et al., 2011) and
Arabidopsis plants over-expressing YUCCA6, e.g., the 35S:YUC6 transgen-
ic plants and yuc6-1D activation mutant, exhibit high-auxin phenotypes
and show markedly increased longevity (Kim et al., 2011a). All these re-
ports clearly depict the effective role of auxins in regulation of leaf
senescence.
5.8. Gibberellins
Like other plant hormones, gibberellins also have a signicant im-
pact on leaf senescence (Table 1). As chlorophyll degradation is one of
the potential indicators of leaf senescence, so effect of any agent on
chlorophyll degradation can be directly related to leaf senescence. Chlo-
rophyll breakdown was considered to be associated with endogenous
gibberellin content in shoots of Paris polyphylla during senescence (Li
et al., 2010; Pandey et al., 2007). Delayed leaf senescence with exoge-
nous application of gibberellins has been reported in various plant spe-
cies. For example, plant species in which GA3 delays senescence include
lettuce (Aharoni and Richmond, 1978), Rumex crispus and R. obtusifolius
(Leopold, 1971), Catharanthus roseus (Pandey et al., 2007), nasturtium
(Tropaeolum majus) (Beevers, 1966), and Dioscorea rotundata (Ile et
al., 2006).
Recently, a study with perennial plant P. polyphylla (Li et al., 2010)
has shown that exogenous application with gibberellic acid (GA3) con-
siderably enhanced the level of internal GAs (GA4 +GA7) resulting in re-
tardation of shoot senescence, which was attributed primarily to the
suppression of degradation of proteins and chlorophyll as well as
lipoxygenase activity (Li et al., 2010). A few years ago, it has been con-
rmed that both GA and ABA act antagonistically to regulate different de-
velopmental processes. This has been revealed after determining
physiological changes taking place in aging leaves associated with senes-
cence; in particular the changes related to the hormonal and oxidative
metabolisms have showed that they signicantly hamper senescence
of aerial parts and slow the decline of pigments and total soluble
proteins. However, when plants were treated with paclobutrazol
(PCB), an inhibitor of GA-synthesis, concentration of ABA rose while
that of GAs declined, resulting in accelerated senescence, indicating an
antagonistic role of GA and ABA in the leaf senescence of P. polyphylla
(Yu et al., 2009). In zoysiagrass, seedlings treated with GA3 delayed the
darkness-induced leaf senescence. The rst genome-wide trial of
senescence was carried-out in a species of turfgrass Zoysiagrass (Zoysia
japonica), during which the expression of genes responsive to staying
green were identied which showed differences between GA3-treated
and non-treated plants under dark conditions. Expression of ve genes,
i.e., An (diphthine synthase gene), Met, V-ATPase, SAM and Cry
(Cryptochrome gene), was investigated. Both real-time PCR and
RT-PCR indicated the differential expression of these genes, which can
be associated with delayed senescence due to GA treatment (Cheng et
al., 2009). However, the effects of gibberellin on natural senescence
and the relationship between gibberellins and senescence as well as
their interaction with other plant hormones, are not fully understood.
5.9. Polyamines
Polyamines (PAs) include spermine (Spm), putrescine (Put),
spermidine (Spd) and cadaverine (Cad) which are low molecular
weight cations synthesized in almost all biological systems including
plants (Altman et al., 1977; Kaur-Sawhney et al., 2003; Nambeesan et
al., 2010; Pandey et al., 2000). In general, PAs levels have been
reported to be high in actively growing tissues (Bachrach, 1973;
Galston and Kaur-Sawhney, 1980; Mattoo et al., 2010) and low in
senescing tissues (Altman and Bachrach, 1981; Nambeesan et al.,
2010; Serani-Fracassini et al., 1980). Natural PAs are actively in-
volved in events related to PCD (Kusano et al., 2008). As suppression
in the levels of endogenous polyamines has been observed in
senescing/aging leaves, so it is believed that they have a key role in
controlling leaf senescence (Chen and Kao, 1991). So far it is evident
that PAs are involved in many plant developmental processes such
as oral initiation and development, cell division, embryogenesis,
root growth, reproductive organ development, tuberization, fruit de-
velopment, ripening, and leaf senescence (Galston et al., 1997;
Nambeesan et al., 2010; Tiburcio et al., 2002).
A number of reports available show the role of PAs in delaying senes-
cence of leaf, ower as well as fruit (Pandey et al., 2000). For example, in
barley, Sobieszczuk-Nowicka et al. (2009) investigated the changes in
the levels of PAs bound to thylakoids in senescing leaves and found
that Put and Spd increased as senescence progressed, whereas Spm de-
creased. Furthermore, PAs associate with numerous molecules by the
binding action of different types of proteins (Serani-Fracassini and del
Duca, 2008). In chloroplasts, covalent binding of PAs to proteins might
play a considerable role in the post-translational modications of struc-
tural enzymes/proteins (Dondini et al., 2003; Sobieszczuk-Nowicka et
al., 2007, 2008). Transglutaminases (TGases), a protein family, as well
as Pas are involved in reactions leading to senescence/PCD in animals
(Kuehn and Phillips, 2005; Serani-Fracassini and Del Duca, 2008). How-
ever, no information is available on the role of these biomolecules in se-
nescence as well as PCD in plants (Lorand and Graham, 2003; Seiler and
Raul, 2005).
Biogenic, growth-promoting and anti-senescence PAs are key factors
in seed production, fruit ripening and senescence, owering, embryo de-
velopment, and fruit set (Nambeesan et al., 2008). In different studies, ex-
pression of a yeast SAMdc in tomato indicated that S-adenosylmethionine
(SAM) availability was not limiting for ethylene biosynthesis. The conse-
quences of ripening-specic expression of ySAMdc in tomato include
prolonged vine life, high amount of lycopene and enhanced fruit quality
(Mattoo and Handa, 2008). In another investigation, expression of yeast
spermidine synthase (ySpdSyn) was found to be associated with in-
creased intracellular levels of Spd in the leaf and fruit development
(Nambeesan et al., 2010). Overall, senescence in the transgenic plants
was delayed as compared to that in the WT plants. So, it can be suggested
that PAs, particularly Spd, are involved in increasing fruit shelf life, prob-
ably by reducing post-harvest decay and senescence (Mehta et al., 2002;
Nambeesan et al., 2010).
Overall, of various hormones discussed above, cytokinins receive a
maximum attention in relation to their signicant role in leaf senescence.
Cytokinins and auxins are considered as negative regulators of senes-
cence in plants. In contrast, ethylene and ABA promote leaf senescence.
The afrmative role of JA and MeJA in leaf senescence has been well doc-
umented in a number of plants. In addition, BRs are involved in delaying
leaf senescence. A number of reports available show the role of PAs and
gibberellins in delaying senescence of leaf, ower as well as fruit. Cur-
rently, efforts are being made to elucidate how far different plant hor-
mones can regulate leaf senescence. Unravelling of mechanism of
signal transduction in leaf senescence would provide an insight to con-
trol this phenomenon in different plants including crops for attaining
maximum crop production.
6. Stress and senescence
Stresses (both biotic and abiotic) can induce leaf senescence, as re-
vealed by the up-regulation of SAGs under these conditions (Ahmad
et al., 2011a, 2011b, 2011c; Koyro et al., 2011). Many defense related
(DR) and pathogenesis related (PR) genes also are up-regulated dur-
ing senescence (Quirino et al., 2000). Genomic studies on A. thaliana
(Gepstein et al., 2003; Guo et al., 2004; Lin and Wu, 2004) and
P. tremula (Andersson et al., 2004; Bhalerao et al., 2003) further con-
rm the role of DR genes in leaf senescence.
 M. Sarwat et al. / Biotechnology Advances 31 (2013) 11531171
6.1. Role of transcription factors in regulation of leaf senescence under
stress conditions
There is now a growing body of evidence that stressful conditions/
agents can regulate transcription factors involved in the regulation of
genes which induce leaf senescence (Breeze et al., 2011; Lee et al.,
2012; Woo et al., 2010). For example, a recently identied abscisic
acid (ABA)-responsive NAC transcription factor VND-INTERACTING2
(VNI2) has been reported to negatively regulate leaf senescence
(Yang et al., 2011). VNI2 over-expressing plants show delayed senes-
cence, whereas it is accelerated in a VNI2-decient mutant. VNI2 tran-
scription factor binds to the promoters of the COR (COLD-REGULATED)
and RD (RESPONSIVE TO DEHYDRATION) genes. The COR and RD
over-expressing plants exhibit prolonged leaf longevity (Nakashima
et al., 2011; Yang et al., 2011). Thus, VNI2 transcription factor provided
a molecular link which assimilated plant responses to different stresses
and delays leaf senescence (Yang et al., 2011). Recently, new roles are
identied for the acyl-CoA-binding proteins (ACBPs) which are impor-
tant constituents of phospholipid metabolism (Xiao and Chye, 2011).
These proteins have been reported to modulate plant development
and delay leaf senescence by generating freezing tolerance, oxidative
stress tolerance, heavy metal resistance and pathogen resistance
(Xiao and Chye, 2011). Efforts are currently underway at different in-
stitutions to identify specic transcription factors and elucidate their
role in regulation of genes which induce leaf senescence. This advanced
knowledge will certainly help control leaf senescence.
6.2. Role of reactive oxygen species
Reactive oxygen species (ROS) are also known to play a signicant
role in stress signalling and senescence (Bhattacharjee, 2005; Jimnez
et al., 1998; Lee et al., 2011; Lin and Kao, 1998; Munne-Bosch and
Alegre, 2002). ROS generation is one of the earliest responses of plant
cells under abiotic stresses (Ahmad et al., 2008, 2010a) and senescence
(Khanna-Chopra, 2011; Lee et al., 2012). The disturbance in balance be-
tween production and scavenging of ROS under different environmen-
tal stresses resulted in increased intracellular ROS levels (Ahmad et al.,
2010a, 2010b, 2011a, 2011b, 2011c; Koyro et al., 2011). ROS are delete-
rious to biomolecules, like proteins, lipids, carbohydrates, DNA and ulti-
mately result in cell death (Ahmad et al., 2008, 2010a, 2010b, 2011a,
2011b, 2011c; Koyro et al., 2011). Environmental stresses cause
ROS-linked damage to chloroplasts. A similar scene is observed in
normal senescence as ROS-scavenging ability declined on ageing.
ROS function by initiation of protein degradation through the pro-
teolytic enzymes cysteine and serine proteases is also evident
(Khanna-Chopra, 2011).
Research on the catalase (a H2O2-scavenging enzyme) throws light
on the involvement of H2O2 in leaf senescence. Catalase regulates the
intracellular H2O2 (Ahmad et al., 2008, 2010a, 2010b; Koyro et al.,
2011). During leaf senescence CATALASE2 (CAT2) expression is
down-regulated and is postulated to be involved in the increase of
H2O2 level (Smykowski et al., 2010). A DNA-binding protein, G-Box
Binding Factor1 (GBF1), reported as the CAT2 promoter has further in-
creased our knowledge of involvement of CAT2 in senescence
(Smykowski et al., 2010). At the onset of senescence in gbf1 mutant
plants, the level of CAT2 decreases; H2O2 does not remain at elevated
level and there is delayed expression of SAGs (Miao et al., 2004;
Smykowski et al., 2010). All this clearly indicates the involvement of
H2O2 in leaf senescence possibly mediated by GBF1. Another recently
identied H2O2 responsive gene is the transcription factor ORS1, a
paralog of ORE1/ANAC092/AtNAC2, a previously observed regulator
of leaf senescence (Balazadeh et al., 2011; Kim et al., 2009). The pro-
moter sequences of both the genes are highly conserved in family
Brassicaceae. ORS1, similar to ORE1, is a positive regulator of senescence
(Balazadeh et al., 2011; Kjaersgaard et al., 2011). Global expression
1165
analyses show its overlapping role in salt and H2O2 signalling pathways
(Balazadeh et al., 2011).
Not only transcription factors and ROS are involved in leaf senes-
cence under stress conditions, there are several other metabolites
which play an active role in causing leaf senescence under stress con-
ditions. Thus, the role all such metabolites need to be fully elucidated.
7. Micro RNA and senescence
MicroRNAs are single stranded, genome encoded, and small RNA
molecules that regulate key levels of target transcript either by cleav-
age or translation suppression (Naqvi et al., 2009, 2012). There are
>1000 miRNAs reported from various plants and this list is still in-
creasing day by day. MicroRNAs have been associated with diverse
developmental and physiological processes in plants (Branscheid et
al., 2010; Naqvi et al., 2010, 2011a, 2011b; Yoon et al., 2010). Howev-
er, very few reports demonstrate a clear evidence of their role in plant
aging. Recently, two independent studies highlight the contribution
of at least two miRNAs viz., miR164 and miR319 in regulating plant
aging (Pulido and Laufs, 2010; Schommer et al., 2008). We have
discussed below the role of these miRNAs in senescence.
7.1. miR164
Recently, Kim et al. (2009) identied ORE1 gene that is a positive
regulator of senescence in Arabidopsis. The ORE1 mutants exhibit de-
layed aging symptoms which include high photochemical activity, re-
duced chlorophyll loss, reduction in dying cells and induction of aging
associated genes, etc. (Kim et al., 2009; Lim et al., 2007; Schippers et
al., 2008). Genetic analysis has shown that the corresponding loci en-
code for a NAC2 protein. Previously, it was demonstrated that the ex-
pression of this particular transcript increases with aging (Guo and
Gan, 2006; Guo et al., 2004). Being a transcription factor, the NAC2
was observed to be localized in nucleus and it probably regulates
transcription of various downstream senescence related genes.
In plants, majority of transcription factors are known to be under di-
rect control of microRNAs (Rajagopal et al., 2006). Therefore, ORE1 was
searched for potential miRNA binding sites on the encoded transcript.
Interestingly, it was found that miR164, an evolutionary conserved
miRNA among plant lineage that regulates CUC/NAC family of transcrip-
tion factors, binds to ORE1 (Mishra and Mukherjee, 2007; Zhu et al.,
2012). This was further conrmed that miR164 levels increase with
aging, an expected pattern exhibited by most of the bonade
microRNAtarget pairs (Kim et al., 2009; Pulido and Laufs, 2010).
MiR164 family, in Arabidopsis, has three different members viz.,
miR164a, miR164b and miR164c and all of them target CUC/NAC tran-
scription family genes. These genes have well dened roles in leaf mor-
phology where they control organ boundary and serration (Nikovics et
al., 2006).
The levels of ORE1 were checked in miR164 triple mutants and in
miR164 over-expressing lines of Arabidopsis (Kim et al., 2009). It was
observed that ORE1 expressed at higher levels in mutants while signif-
icant reduction was noticed in case of miR164 over-expressing lines
(Kim et al., 2009). Similarly, accumulation of ORE1 was noticed in a
transgenic line of Arabidopsis expressing ORE1 resistant to miR164
binding. Taken together, these results clearly suggest that miR164 fam-
ily members regulate the levels of ORE1 transcripts. Further, miR164 tri-
ple mutant plants have high levels of ORE1 at early stages of infection
but no signicant changes were noticed at later stages. This indicated
that miR164 controls aging at early stages of plant growth. Thus,
miR164 gene family acts as a negative regulator of plant aging.
Now the question arises as to how miR164 levels decline with aging
in plants? To address this, the miR164 levels were analysed in other ORE
mutants. Intriguingly, it was found that miR164 levels remained
unchanged in ORE3-1 mutant. The ORE3-1 locus encodes EIN-2 tran-
scription factor which mediates age-dependent down-regulation of
1166 M. Sarwat et al. / Biotechnology Advances 31 (2013) 11531171
miR164 (Chakrabarti, 2010; Kim et al., 2009). Moreover, the ORE1
mRNA accumulated at high levels in EIN2 mutant. Thus, while EIN2 in-
duces ORE1 transcription, the same TF leads to reduction in miR164
transcript levels. In agreement to this, Arabidopsis mutants for both
EIN2 and ORE1 exhibit much delayed expression of senescence related
genes like SAG12 and consequently delayed loss of Fv/Fm, when com-
pared with either of the single mutants (Lim et al., 2007) which indicat-
ed that ORE has a denite role in delaying aging.
7.2. miR319
MiR319 is one the earliest miRNAs identied and is highly con-
served across the plant kingdom. It has been extensively studied in
terms of the targets it regulates which include well known transcrip-
tion factors (TF) TEOSINTE BRANCHED/CYCLOIDEA/PCF (TCP).
MiR319 targets ve different TCP members and any alteration in
this control leads to leaf deformation, in both Arabidopsis and tomato
(Ori et al., 2007; Palatnik et al., 2003). These TCP TFs control the cell
division and thus maintain the leaf morphology. Jasmonic acid (JA)
is one of the plant hormones that are well recognized in controlling
leaf senescence (Schommer et al., 2008; Yun-xia et al., 2010). Several
miRNAs are known to directly target genes involved in biosynthesis/
transport of hormones (Lafos et al., 2011). This study provides yet an-
other, albeit indirect, mode of regulation of miRNA-mediated regula-
tion of plant hormones. The study by Schommer et al. (2008)
proposes a new role to miR319-targeted TCP TF in leaf senescence
by affecting the JA biosynthesis.
In global transcriptome proling, it was noticed that a large num-
ber of genes were induced in miR319 resistant TCP4 over-expressing
plants, including those involved in oxylipin biosynthesis (Pulido and
Laufs, 2010; Schommer et al., 2008). The promoter analysis revealed
that these 8 of the 19 oxylipin biosynthesis pathway genes shared a
GGACCA motif which is a core element in the promoters of
TCP-response genes (Schommer et al., 2008). This was further vali-
dated by performing EMSA based in vitro interaction studies. Impor-
tantly, the in planta experiments clearly show that treatment of the
wild type and jaw-D plant with exogenous MeJA could restore the se-
nescence delay (Schommer et al., 2008). This evidence demarcates
that jaw-D senescence phenotype is a manifestation of JA biosynthe-
sis but not the hormonal response. Collectively, these data show
that while miR319 negatively regulates the TCP TFs, the expression
of jasmonic acid biosynthesis genes is positively regulated by TCP TF
(Koyama et al., 2010; Mitsuda and Ohme-Takagi, 2009). By corollary,
high levels of miR319 may lead to delay in senescence. This study
provides a new paradigm of miRNA control of plant hormones by
maintaining the biosynthesis and not just the hormonal signalling.
8. Conclusion and future prospects
Senescence is a complex but well orchestrated process in the life of a
plant. A plethora of internal as well as external factors control the cas-
cade events going on during aging. To date, although several mecha-
nisms perturbed in this complex process have been identied, still not
much information exists in the literature on the signalling mechanism
of leaf senescence. In addition to physiology and molecular biology, bio-
informatic studies will prove to be more benecial in understanding the
leaf senescence phenomenon. Besides SAGs, several regulatory ele-
ments involved in leaf ageing have been recently identied and others
yet need to be discovered. Arabidopsis senescence mutants have
contributed signicantly to identifying novel genes controlling senes-
cence. In addition to T-DNA insertion mutants, chemical mutants of
Arabidopsis like ORE1/AHK3 have also proved useful. Global expression
proling of these mutants will be very helpful in understanding the
complex regulatory pathways of senescence. The latest entrants re-
sponsible for regulating these gene regulatory networks are miRNAs
and ta-siRNAs. These classes of small non-coding RNAs regulate the
expression of several genes from leaf morphogenesis to leaf senescence.
miR164 and miR319 target ORE1 (NAC) and TCP transcription factors,
respectively, mediate pathways converging at senescence. However, it
is still a matter of discussion how miRNAs exhibit coordination in
these well diversied levels. An organized approach in studying
miRNAs controlling leaf senescence should be the immediate need of
the time.
Stress-induced disturbance in balance between accumulation and
scavenging of ROS is believed to result in enhanced intracellular ROS
levels, thereby signicantly stimulating the process of senescence.
However, identication and then silencing/expressing defense and
pathogenesis-related genes using advanced molecular techniques
could be used to delay leaf senescence.
Hormonal regulation of senescence is very important as it involves
a complex interplay of signalling molecules at various stages of senes-
cence. Global expression proling and availability of genetic mutants
have unravelled the mystery of these marvellous molecules. Howev-
er, the exact mechanism how hormones delay or hasten the process
of plant senescence is not yet well known. Therefore, there is still a
dire need to study the role of hormones in senescence at the physio-
logical and molecular levels. It could be helpful in preventing losses
during the storage of fruits, legumes, owers, and crops. Particularly,
by modulating the hormones involved in the senescence pathway,
the shelf-life of these economically important plants can be increased.
Therefore, the introgression of genes as well as identication of QTLs
related to different hormones synthesis involved directly in the pro-
cess of senescence could be further studied. Certainly, future research
on leaf senescence will be an amalgamation of bioinformatical predic-
tions, hormonal regulation of senescence, miRNA regulations of genes
and microarray based genetic proling of Arabidopsis mutants that
will unravel the mechanism underlying leaf aging.
References
Abeles FB, Dunn LJ, Morgens P, Callahans A, Dinterman RE, Schmidit J. Induction of
33-kD and 60-kD peroxidases during ethylene-induced senescence of cucumber
cotyledons. Plant Physiol 1988;87:60915.
Addicott FT, Lynch RS, Carns HR. Auxin gradient theory of abscission regulation.
Science 1955;121:6445.
Aharoni N, Richmond E. Endogenous gibberellin and abscisic acid content as related to
senescence of detached lettuce leaves. Plant Physiol 1978;62:2248.
Ahmad P, Ashraf M, Younis M, Hu X, Kumar A, Akram NA, et al. Role of transgenic
plants in agriculture and biopharming. Biotechnol Adv 2011a;30:52440.
Ahmad P, Jaleel CA, Salem MA, Nabi G, Sharma S. Roles of enzymatic and
non-enzymatic antioxidants in plants during abiotic stress. Crit Rev Biotechnol
2010a;30:16175.
Ahmad P, Jeleel CA, Azooz MM. Free radical production, oxidative damage and antiox-
idant defense mechanisms in plants under abiotic stress. In: Ahmad P, Umar S, ed-
itors. Oxidative stress: Role of antioxidants in plants. New Delhi, India: Studium
Press Pvt. Ltd.; 2011b. p. 1953.
Ahmad P, Nabi G, Ashraf M. Cadmium-induced oxidative damage in mustard [Brassica
juncea (L.) Czern. & Coss.] plants can be alleviated by salicylic acid. S Afr J Bot
2011c;77:3644.
Ahmad P, Sarwat M, Sharma S. Reactive oxygen species, antioxidants and signaling in
plants. J Plant Biol 2008;51:16773.
Ahmad P, Umar S, Sharma S. Mechanism of free radical scavenging and role of phytohor-
mones in plants under abiotic stresses. In: Ashraf M, Ozturk M, Ahmad MSA, editors.
Plant Adaptation and Phytoremediation. Dordrecht/Heidelberg/London/NY: Springer;
2010b. p. 99-108.
Aleksandrushkina NI, Kof EM, Seredina AV, Borzov AA, Vanyushin BF. Degradation of
DNA and endonuclease activity associated with senescence in the leaves of pea
of normal and aphyllous genotypes. Russ J Plant Physiol 2008;55:2332.
Al-Taweel K, Fernando WGD, Brl-Babel AL. Construction and characterization of a
cDNA library from wheat infected with Fusarium graminearum Fg 2. Int J Mol Sci
2011;12:61326.
Altman A, Bachrach U. Involvement of polyamines in plant growth and senescence. In:
Caldarera CM, Zappia V, Bachrach U, editors. Advances in Polyamine Research. New
York: Raven Press; 1981. p. 36575.
Altman A, Kaur-Sawhney R, Galston AW. Stabilization of oat leaf protoplasts through
polyamine-mediated inhibition of senescence. Plant Physiol 1977;60:5704.
Ananieva K, Ananiev ED, Mishev K, Georgieva K, Malbeck J, Kamnek M, et al. Methyl
jasmonate is a more effective senescence-promoting factor in Cucurbita pepo
(zucchini) cotyledons when compared with darkness at the early stage of senes-
cence. J Plant Physiol 2007;164:117987.
Andersson A, Keskitalo J, Sjodin A, Bhalerao R, Sterky F, Wissel K, et al. A transcriptional
timetable of autumn senescence. Genome Biol 2004;5:R24.
 M. Sarwat et al. / Biotechnology Advances 31 (2013) 11531171
Aubry S, Mani J, Hrtensteiner S. Stay-green protein, defective in Mendel's green coty-
ledon mutant, acts independent and upstream of pheophorbide a oxygenase in the
chlorophyll catabolic pathway. Plant Mol Biol 2008;67:24356.
Azad AK, Ishikawa T, Ishikawa T, Sawa Y, Shibata H. Intracellular energy depletion trig-
gers programmed cell death during petal senescence in tulip. J Exp Bot 2008;59:
208595.
Bachrach U. Function of Naturally Occurring Polyamines. New York: Academic Press;
1973.
Back A, Richmond AE. Interrelations between gibberellic acid, cytokinins and abscisic
acid in retarding leaf senescence. Physiol Plant 1971;24:769.
Balazadeh S, Kwasniewski M, Caldana C, Mehrnia M, Zanor MI, Xue GP, et al. ORS1, an
H2O2-responsive NAC transcription factor, controls senescence in Arabidopsis
thaliana. Mol Plant 2011;4:34660.
Balk J, Leaver CJ. The PET1-CMS mitochondrial mutation in sunower is associated with
premature programmed cell death and cytochrome c release. Plant Cell 2001;13:
180318.
Beevers L. Effect of gibberellic acid on the senescence of leaf discs of nasturtium
(Tropaeolum majus). Plant Physiol 1966;41:10746.
Bell E, Creelman RA, Mullet JE. A chloroplast lipoxygenase is required for wound-induced
jasmonic acid accumulation in Arabidopsis. Proc Natl Acad Sci U S A 1995;92:86759.
Besseau S, Li J, Palva ET. WRKY54 and WRKY70 co-operate as negative regulators of leaf
senescence in Arabidopsis thaliana. J Exp Bot 2012;63(7):266779.
Bhalerao R, Keskitalo J, Sterky F, Erlandsson R, Bjorkbacka H, Birve SJ, et al. Gene ex-
pression in autumn leaves. Plant Physiol 2003;131:43042.
Bhattacharjee S. Reactive oxygen species and oxidative burst: roles in stress, senes-
cence and signal transduction in plants. Curr Sci 2005;89:111321.
Bhattacharjee S. The language of reactive oxygen species signaling in plants. J Bot in press.
Bilgin DD, Zavala JA, Zhu J, Clough SJ, Orti DR, Delucia EH. Biotic stress globally down
regulates photosynthesis genes. Plant Cell Environ 2010;33:1597613.
Blagosklonny MV. Cell cycle arrest is not senescence. Ageing 2011;3:94-101.
Blokhina O, Virolainen E, Fagerstedt K. Antioxidants, oxidative damage and oxygen
deprivation stress: a review. Ann Bot 2003;91:17994.
Borrell LC, Farris R, Piug-Parellada P, Tiburcio AF. Polyamines inhibit lipid peroxidation
in senescing oat leaves. Physiol Plant 1997;99:38590.
Borrmann D, Junqueira RM, Sinnecker P, Gomes MSO, Castro IA, Marquez UML. Chem-
ical and biochemical characterization of soybean produced under drought stress.
Cinc Tecnol Aliment Campinas 2009;29:67681.
Bozhkov PV, Lam E. Green death: revealing programmed cell death in plants. Cell Death
Differ 2011;18:123940.
Branscheid A, Sieh D, Pant BD, May P, Devers EA, Elkrog A, et al. Expression pattern suggests
a role of MiR399 in the regulation of the cellular response to local Pi increase during
arbuscular mycorrhizal symbiosis. Mol Plant Microbe Interact 2010;23:91526.
Breeze E, Harrison E, McHattie S, Hughes L, Hickman R, Hill C. High-resolution temporal
proling of transcripts during Arabidopsis leaf senescence reveals a distinct chro-
nology of processes and regulation. Plant Cell 2011;23:87394.
Brosch M, Kangasjrvi J. Low antioxidant concentrations impact on multiple signalling
pathways in Arabidopsis thaliana partly through NPR1. J Exp Bot 2012;63(5):
184961.
Buchanan-Wollaston V, Earl S, Harrison E, Mathas E, Navabpour S, Page T, et al. The molec-
ular analysis of leaf senescence: a genomics approach. Plant Biotechnol J 2003;1:3-22.
Buchanan-Wollaston V, Page T, Harrison E, Breeze E, Lim PO. Comparative
transcriptome analysis reveals signicant differences in gene expression and sig-
naling pathways between developmental and dark/starvation-induced senescence
in Arabidopsis. Plant J 2005;42:56785.
Buchanan-Wollaston V. The molecular biology of leaf senescence. J Exp Bot 1997;48:18199.
Campisi J, Fagagna F. Cellular senescence: when bad things happen to good cells. Nat
Rev Mol Cell Biol 2007;8:72940.
Cano-Delgado A, Yin Y, Yu C, Vafeados D, Mora-Garcia S, Cheng JC, et al. BRL1 and BRL3
are novel brassinosteroid receptors that function in vascular differentiation in
Arabidopsis. Development 2004;131:534151.
Cao J, Jiang F, Sodmergen, Cui K. Time-course of programmed cell death during leaf se-
nescence in Eucommia ulmoides. J Plant Res 2003;162:7-12.
Cao L, Kim S, Xiao C, Wang RH, Coumoul X, Wang X, et al. ATM-Chk2-p53 activation
prevents tumorigenesis at an expense of organ homeostasis upon Brca1 deciency.
EMBO J 2006;25:216777.
Chakrabarti M. Evolutionary perspective of nicotine to nornicotine conversion, its reg-
ulation and characterization of ein2 mediated ethylene signaling in tobacco. Doc-
toral Dissertations. Paper 88; 2010.
Chang C, Kwok SF, Bleecker AB, Meyerowitz EM. Arabidopsis ethylene-response gene
ETR1: similarity of product to two-component regulators. Science 1993;262:
53944.
Chang H, Jones ML, Banowetz GM, Clark DG. Overproduction of cytokinins in petunia
owers transformed with PSAG12-IPT delays corolla senescence and decreases
sensitivity to ethylene. Plant Physiol 2003;132:217483.
Chaves MM, Flexas J, Pinheiro C. Photosynthesis under drought and salt stress: regula-
tion mechanisms from whole plant to cell. Ann Bot 2009;103:55160.
Chen C, Dickman MB. Bcl-2 family members localize to tobacco chloroplasts and inhibit
programmed cell death induced by chloroplast-targeted herbicides. J Exp Bot
2004;55:261723.
Chen CT, Kao CH. Senescence of rice leaves XXIX. Ethylene production, polyamine level
and polyamine biosynthetic enzyme activity during senescence. Plant Sci 1991;78:
1938.
Chen HJ, Wu SD, Lin ZW, Huang GJ, Lin YH. Cloning and characterization of a sweet potato
calmodulin SPCAM that participates in ethephon-mediated leaf senescence, H2O2
elevation and senescence-associated gene expression. J Plant Physiol 2012;169:
52941.
1167
Chen W, Provart NJ, Glazebrook J, Katagiri F, Chang HS, Eulgem T, et al. Expression pro-
le matrix of Arabidopsis transcription factor genes suggests their putative func-
tions in response to environmental stresses. Plant Cell 2002;14:55974.
Cheng X, Dai X, Zeng H, Li Y, Tang W, Han L. Gene expression involved in dark-induced leaf
senescence in zoysiagrass (Zoysia japonica). Plant Biotechnol Rep 2009;3:28592.
Choi DS, Hwang BK. Proteomics and functional analyses of pepper abscisic acid-
responsive 1 (ABR1), which is involved in cell death and defense signaling. Plant
Cell 2011;23:82342.
Cholewa E, Grifth M. The unusual vascular structure of the corm of 405 Eriophorum
vaginatum: implications for efcient retranslocation of nutrients. J Exp Bot
2004;55:73141.
Christ B, Schelbert S, Aubry S, Sssenbacher I, Mller T, Krutler B, et al. MES16, a mem-
ber of the methylesterase protein family, specically demethylates uorescent
chlorophyll catabolites during chlorophyll breakdown in Arabidopsis. Plant Physiol
2012;158:62841.
Coll NS, Epple P, Dangl JL. Programmed cell death in the plant immune system. Cell
Death Differ 2011;18:1-10.
Cubas P, Lauter N, Doebley J, Coen E. The TCP domain: a motif found in proteins regu-
lating plant growth and development. Plant J 1999;18:21522.
Cui M, Takada K, Ma B, Ezura H. Overexpression of a mutated melon
ethylenereceptorgeneCm-ETR1/H69A confers reduced ethylene sensitivity in a
heterologous plant, Nemesia strumose. Plant Sci 2004;167:22538.
da Silva D, Lachaud C, Cotelle V, Brire C, Grat S, Mazars C, et al. Nitric oxide production
is not required for dihydrosphingosine-induced cell death in tobacco BY-2 cells.
Plant Signal Behav 2011;6:7369.
Dai H, Shan C, Wei Y, Liang JG, Yang T, Sa WQ, et al. Sub-cellular localization of cadmi-
um in hyperaccumulator Populus canescens. Afr J Biotechnol 2012;11:377987.
de Michele R, Vurro E, Rigo C, Costa A, Elviri L, di Valentin M, et al. Nitric oxide is in-
volved in cadmium-induced programmed cell death in Arabidopsis suspension cul-
tures. Plant Physiol 2009;150:21728.
de Silva K, Laska B, Brown C, Sederoff HW, Khodakovskaya M. Arabidopsis thaliana
calcium-dependent lipid-binding protein (AtCLB): a novel repressor of abiotic
stress response. J Exp Bot 2011;62:267989.
Desai NA, Shankar V. Single-strand-specic nucleases. FEMS Microbiol Rev 2003;26:
45791.
Desvoyes B, Ramirez-Parra E, Xie Q, Chua NH, Gutierrez C. Cell type-specic role of the
retinoblastoma/E2F pathway during Arabidopsis leaf development. Plant Physiol
2006;140:6780.
Dhindsa RS, Plumb-Dhindsa P, Thorpe TA. Leaf senescence: correlated with increased
levels of membrane permeability and lipid peroxidation, and decreased levels of
superoxide dismutase and catalase. J Exp Bot 1981;32:93-101.
Diaz C, Purdy S, Christ A, Morot-Gaudry JF, Wingler A, Masclaux-Daubresse C. Charac-
terization of markers to determine the extent and variability of leaf senescence in
Arabidopsis. A metabolic proling approach. Plant Physiol 2005;138:898908.
Dobrenel T, Marchive C, Sormani R, Moreau M, Mozzo M, Montan MH, et al. Regula-
tion of plant growth and metabolism by the TOR kinase. Biochem Soc Trans
2011;39:47781.
Dominguez F, Moreno J, Cejudo FJ. A gibberellin-induced nuclease is localized in the
nucleus of wheat aleurone cells undergoing programmed cell death. J Biol Chem
2004;279:115306.
Dondini L, Del Duca S, Dall'Agata L, Bassi R, Gastaldelli M, Della Mea M, et al.
Suborganellar localisation and effect of light on Helianthus tuberosus chloroplast
transglutaminases and their substrates. Planta 2003;217:8495.
Dong H, Niu Y, Li W, Zhang D. Effects of cotton rootstock on endogenous cytokinins and
abscisic acid in xylem sap and leaves in relation to leaf senescence. J Exp Bot
2008;59:1295304.
Doyle SM, Diamond M, McCabe PF. Chloroplast and reactive oxygen species involve-
ment in apoptotic-like programmed cell death in Arabidopsis suspension cultures.
J Exp Bot 2010;61:47382.
Eckardt NA. Programmed cell death in plants: a role for mitochondrial-associated
hexokinases. Plant Cell 2006;18:20979.
Eisinger W. Role of cytokinins in carnation ower senescence. Plant Physiol 1977;59:
7079.
Ellis CM, Nagpal P, Young JC, Hagen G, Guilfoyle TJ, Reed JW. AUXIN RESPONSE
FACTOR1 and AUXIN RESPONSE FACTOR2 regulate senescence and oral organ ab-
scission in Arabidopsis thaliana. Development 2005;132:456374.
Elmore S. Apoptosis: a review of programmed cell death. Toxicol Pathol 2007;35:495516.
Espinoza C, Medina C, Somerville S, Arce-Johnson P. Senescence-associated genes in-
duced during compatible viral interactions with grapevine and Arabidopsis. J Exp
Bot 2007;58:3197212.
Eulgem T, Rushton PJ, Robatzek S, Somssich IE. The WRKY superfamily of plant tran-
scription factors. Trends Plant Sci 2000;5:199206.
Falqueto AR, Cassol D, Magalhes Junior AM, Oliveira AC, Bacarin MA. Physiological
analysis of leaf senescence of two rice cultivars with different yield potential.
Pesqui Agropecu Bras 2009;44:695700.
Farage-Barhom S, Burd S, Sonego L, Mett A, Belausov E, Gidoni D, et al. Localization of
the Arabidopsis senescence and cell death-associated BFN1 nuclease: from the ER
to fragmented nuclei. Mol Plant 2011;4:106273.
Farage-Barhom S, Burd S, Sonego L, Perl-Treves R, Lers A. Expression analysis of the
BFN1 nuclease gene promoter during senescence, abscission, and programmed
cell death-related processes. J Exp Bot 2008;59:324758.
Feliciano A, Snchez-Sendra B, Kondoh H, LLeonart ME. MicroRNAs regulate key effec-
tor pathways of senescence. J Aging Res in press.
Feussner I, Hause B, Vrs K, Parthier B, Wasternack C. Jasmonate-induced lipoxygenase
forms are localized in chloroplasts of barley leaves (Hordeum vulgare cv Salome).
Plant J 1995;7:94957.
1168 M. Sarwat et al. / Biotechnology Advances 31 (2013) 11531171
Feys BJ, Wiermer M, Bhat RA, Moisan LJ, Medina-Escobar N, Neu C, et al. Arabidopsis
senescence-associated gene101 stabilizes and signals within an enhanced disease
susceptibility1 complex in plant innate immunity. Plant Cell 2005;17:260113.
Fischer AM. Nutrient remobilization during leaf senescence. In: Gan S, editor. Senes-
cence Processes in Plants. Oxford: Blackwell Publishing; 2007. p. 87-107.
Foyer CH, Noctor G. Oxidant and antioxidant signalling in plants: a re-evaluation of the
concept of oxidative stress in a physiological context. Plant Cell Environ 2005;28:
105671.
Franklin DJ, Airs RL, Fernandes M, Bell TG, Bongaerts RJ, Berges JA, et al. Identication of
senescence and death in Emiliania huxleyi and Thalassiosira pseudonana: cell stain-
ing, chlorophyll alterations, and dimethylsulfoniopropionate (DMSP) metabolism.
Limnol Oceanogr 2012;57:30517.
Frebort I, Kowalska M, Hluska T, Frbortov J, Galuszka P. Evolution of cytokinin bio-
synthesis and degradation. J Exp Bot 2011;62:243152.
Fukuda H. Xylogenesis: initiation, progression, and cell death. Annu Rev Plant Physiol
Plant Mol Biol 1996;47:299325.
Gadjev I, Stone JM, Gechev TS. Programmed cell death in plants: new insights into
redox regulation and the role of hydrogen peroxide. Int Rev Cell Mol Biol
2008;270:87-144.
Galston AW, Kaur-Sawhney R, Altabella T, Tiburcio AF. Plant polyamines in reproduc-
tive activity and responseto abiotic stress. Bot Acta 1997;110:197207.
Galston AW, Kaur-Sawhney R. Polyamines and plant cells. Plant Physiol 1980;11:58.
Gan S, Amasino R. Making sense of senescence: molecular genetic regulation and ma-
nipulation of leaf senescence. Plant Physiol 1997;113:3139.
Gan S. Mitotic and postmitotic senescence in plants. Sci SAGE KE 2003:re7.
Gepstein S, Sabehi G, Carp MJ, Hajouj T, Nesher MFO, Yariv I, et al. Large scale identi-
cation of leaf senescence-associated genes. Plant J 2003;36:62942.
Gill SS, Tuteja N. Reactive oxygen species and antioxidant machinery in abiotic stress
tolerance in crop plants. Plant Physiol Biochem 2010;48:90930.
Grbic V, Bleecker AB. Ethylene regulates the timing of leaf senescence in Arabidopsis.
Plant J 1995;8:595602.
Guo S, Zhang X, Gan L, Zhu C, Gan Y. Effect of poly (ethylene oxide)-poly (propylene
oxide)-poly (ethylene oxide) micelles on pharmacokinetics and intestinal toxicity
of irinotecan hydrochloride: potential involvement of breast cancer resistance pro-
tein (ABCG2). J Pharm Pharmacol 2010;62:97384.
Guo Y, Cai Z, Gan S. Transcriptome of Arabidopsis leaf senescence. Plant Cell Environ
2004;27:52149.
Guo Y, Gan S. AtNAP, a NAC family transcription factor, has an important role in leaf se-
nescence. Plant J 2006;46:60112.
Guo Y, Gan S. Convergence and divergence in gene expression proles induced by leaf
senescence and 27 senescence-promoting hormonal, pathological and environ-
mental stress treatments. Plant Cell Environ 2012;35:64455.
Guterman A, Hajouj T, Gepstein S. Senescence-associated mRNAs that may participate
in signal transduction and protein tracking. Physiol Plant 2003;118:43946.
Hajouj T, Michelis R, Gepstein S. Cloning and characterization of a receptor-like protein
kinase gene associated with senescence. Plant Physiol 2000;124:130514.
He Y, Fukushige H, Hildebrand DF, Gan S. Evidence supporting a role of jasmonic acid in
Arabidopsis leaf senescence. Plant Physiol 2002;128:87684.
He Y, Gan S. Molecular Characteristics of Leaf Senescence. Recent Research Develop-
ments in Plant Molecular Biology. Kerala, India: Research Signpost; 2003. p. 1-17.
He Y, Tang W, Swain JD, Green AL, Jack TP, Gan S. Networking senescence regulating
pathways by using Arabidopsis enhancer trap lines. Plant Physiol 2001;126:
70716.
Hensel L, Grbic V, Baumgarten DA, Bleecker AB. Developmental and age-related pro-
cesses that inuence the longevity and senescence of photosynthetic tissues in
Arabidopsis. Plant Cell 1993;5:55364.
Hinderhofer K, Zentgraf U. Identication of a transcription factor specically expressed
at the onset of leaf senescence. Planta 2001;213:46973.
Hirose N, Takei K, Kuroha T, Kamada-Nobusada T, Hayashi H, Sakakibara H. Regulation
of cytokinin biosynthesis, compartmentalization and translocation. J Exp Bot
2008;59:7583.
Hong SB, Sexton R, Tucker ML. Analysis of gene promoters for two tomato
polygalacturonases expressed in abscission zones and the stigma. Plant Physiol
2000;123:86981.
Horak J, Grefen C, Berendzen KW, Hahn A, Stierhof YD, Stadelhofer B, et al. The Arabidopsis
thaliana response regulator ARR22 is a putative AHP phospho-histidine phosphatase
expressed in the chalaza of developing seeds. BMC Plant Biol 2008;8:77.
Hrtensteiner S, Krutler B. Chlorophyll breakdown in higher plants. Biochim Biophys
Acta 2011;1807:97788.
Huna A, Salmina K, Jascenko E, Duburs G, Inashkina I, Erenpreisa J. Self-renewal signal-
ling in presenescent tetraploid IMR90 Cells. J Aging Res 2011;14:103253.
Hutter E, Renner K, Pster G, Stockl P, Jansen-Durr P, Gnaiger E. Senescence-associated
changes in respiration and oxidative phosphorylation in primary human bro-
blasts. Biochem J 2004;380:91928.
Ile EI, Craufurd PQ, Battey NH, Asiedu R. Phases of dormancy in yam tubers (Dioscorea
rotundata). Ann Bot 2006;97:497504.
Ito J, Fukuda H. ZEN1 is a key enzyme in the degradation of nuclear DNA during
programmed cell death of tracheary elements. Plant Cell 2002;14:320111.
Izumi M, Ishida H. The changes of leaf carbohydrate contents as a regulator of autoph-
agic degradation of chloroplasts via rubisco-containing bodies during leaf senes-
cence. Plant Signal Behav 2011;6:6857.
Jang JC, Leon P, Zhou L, Sheen J. Hexokinase as a sugar sensor in higher plants. Plant Cell
1997;9:5-19.
Jeong YJ, Shang Y, Kim BH, Kim SY, Song JH, Lee JS, et al. BAK7 displays unequal genetic
redundancy with BAK1 in brassinosteroid signaling and early senescence in
Arabidopsis. Mol Cells 2010;29:25966.
Jimnez A, Hernndez JA, Pastori GM, del Ro LA, Sevilla F. Role of the ascorbategluta-
thione cycle of mitochondria and peroxisomes in the senescence of pea leaves.
Plant Physiol 1998;118:132735.
Jing HC, Schippers JH, Hille J, Dijkwel PP. Ethylene-induced leaf senescence depends on
age-related changes and OLD genes in Arabidopsis. J Exp Bot 2005;56:291523.
Jing HC, Sturre MJG, Hille J, Dijkwel PP. Arabidopsis onset of leaf death mutants identify
a regulatory pathway controlling leaf senescence. Plant J 2002;32:5163.
John I, Drake R, Farrell A, Cooper W, Lee P, Horton P, et al. Delayed leaf senescence in
ethylene-decient ACC-oxidase antisense tomato plants: molecular and physiolog-
ical analysis. Plant J 1995;7:48390.
Jones B, Gunners SA, Petersson SW, Tarkowski P, Graham N, May S, et al. Cytokinin
regulation of auxin synthesis in Arabidopsis involves a homeostatic feedback loop
regulated via auxin and cytokinin signal transduction. Plant Cell 2010;22:295669.
Jones ML, Pasian C. Managing Postproduction Shrink in the Greenhouse Industry. In:
Buckeye, editor. Columbus, OH: Ohio Nursery and Landscape Association; 2011.
[22:29].
Jung KH, Han MJ, Lee DY, Lee YS, Schreiber L, Franke R, et al. Wax-decient anther is
involved in cuticle and wax production in rice anther walls and is required for pol-
len development. Plant Cell 2006;18:301532.
Karatas I, Oztrk L, Ersahin Y, Okatan Y. Effects of auxin on photosynthetic pigments
and some enzyme activities during dark-induced senescence of tropaeolum leaves.
Pak J Bot 2010;42:18818.
Kaur-Sawhney R, Tiburcio AF, Altabella TA, Galston AW. Polyamines in plants: an over-
view. J Cell Mol Biol 2003;2:1-12.
Keech O. The conserved mobility of mitochondria during leaf senescence reects differ-
ential regulation of the cytoskeletal components in Arabidopsis thaliana. Plant Sig-
nal Behav 2011;6(1):14750.
Khanna-Chopra R. Leaf senescence and abiotic stresses share reactive oxygen
species-mediated chloroplast degradation. Protoplasma 2011;249:46981.
Khodakovskaya M, Zhao D, Smith W, Li Y, McAvoy R. Expression of ipt gene controlled
by an ethylene and auxin responsive fragment of the LEACO1 promoter increases
ower number in transgenic Nicotiana tabacum. Plant Cell Rep 2006;25:118192.
Kim HJ, Ryu H, Hong SH, Woo HR, Lim PO, Lee IC, et al. Cytokinin-mediated control of
leaf longevity by AHK3 through phosphorylation of ARR2 in Arabidopsis. Proc Natl
Acad Sci U S A 2006;103:8149.
Kim JH, Woo HR, Kim J, Lim PO, Lee IC, Choi SH, et al. Trifurcate feed-forward regulation
of age-dependent cell death involving miR164 in Arabidopsis. Science 2009;323:
10537.
Kim JI, Murphy AS, Baek D, Lee SW, Yun DJ, Bressan RA, et al. YUCCA6 over-expression
demonstrates auxin function in delaying leaf senescence in Arabidopsis thaliana.
J Exp Bot 2011a;62:398192.
Kim JH, Chung KM, Woo HR. Three positive regulators of leaf senescence in Arabidopsis,
ORE1, ORE3 and ORE9, play roles in crosstalk among multiple hormone-mediated
senescence pathways. Genes Genomics 2011b;33:37381.
Kjaersgaard T, Jensen MK, Christiansen MW, Gregersen P, Kragelund BB, Skriver K.
Senescence-associated barley NAC (NAM, ATAF1,2, CUC) transcription factor inter-
acts with radical-induced cell death through a disordered regulatory domain. J Biol
Chem 2011;28:3541829.
Klee HJ. Ethylene signal transduction. Moving beyond Arabidopsis. Plant Physiol
2004;135:6607.
Kllmer I, Werner T, Schmlling T. Ectopic expression of different cytokinin-regulated
transcription factor genes of Arabidopsis thaliana alters plant growth and develop-
ment. J Plant Physiol 2011;168:13207.
Kolodziejek I, Koziol-Lipinska J, Waleza M, Korczynski J, Mostowska A. Aspects of
programmed cell death during early senescence of barley leaves: possible role of
nitric oxide. Protoplasma 2007;232:97-108.
Kong Z, Li M, Yang W, Xu W, Xue Y. A novel nuclear-localized CCCH-type zinc nger
protein, OsDOS, is involved in delaying leaf senescence in rice (Oryza sativa L.).
Plant Physiol 2006;141:137688.
Kosugi S, Ohashi Y. DNA binding and dimerization specicity and potential targets for
the TCP protein family. Plant J 2002;30:33748.
Koyama T, Mitsuda N, Seki M, Shinozaki K, Ohme-Takagi M. TCP transcription factors
regulate the activities of asymmetric leaves and miR164, as well as the auxin re-
sponse, during differentiation of leaves in Arabidopsis. Plant Cell 2010;22:357488.
Koyro HW, Ahmad P, Geissler N. Abiotic Stress Responses in Plants: An Overview. In:
Ahmad P, Prasad MNV, editors. Environmental Adaptations and Stress Tolerance of
Plants in the Era of Climate Change. NY, USA: Springer, Science + Business Media;
2011. p. 1-28.
Kudryakova NV, Kusnetsov VV, Shtratnikova VY, Kulaeva ON. Effects of cytokinin and
senescence-inducing factors on expression of PARR5-GUS gene construct during
leaf senescence in transgenic Arabidopsis thaliana plants. Plant Growth Regul
2008;56:2130.
Kuehn GD, Phillips GC. Roles of polyamines in apoptosis and other recent advances in
plant polyamines. Crit Rev Plant Sci 2005;24:12330.
Kunz HH, Scharnewski M, Feussner K, Feussner I, Flgge UI, Fulda M, et al. The ABC
transporter PXA1 and peroxisomal -oxidation are vital for metabolism in mature
leaves of Arabidopsis during extended darkness. Plant Cell 2009;21:273349.
Kusano T, Berberich T, Tateda C, Takahashi Y. Polyamines: essential factors for growth
and survival. Planta 2008;228:36781.
Lafos M, Kroll P, Hohenstatt MLH, Thorpe FL, Clarenz O, Schubert D. Dynamic regulation of
H3K27 trimethylation during Arabidopsis differentiation. PLoS Genet 2011;7:e1002040.
Lanahan MB, Yen HC, Giovannoni JJ, Klee HJ. The never ripe mutation blocks ethylene
perception in tomato. Plant Cell 1994;6:52130.
Lara MEB, Garcia MCG, Fatima T, Ehness R, Lee TK, Proels R, et al. Extracellular invertase
is an essential component of cytokinin mediated delay of senescence. Plant Cell
2004;16:127687.
 M. Sarwat et al. / Biotechnology Advances 31 (2013) 11531171
Lee IC, Hong SW, Whang SS, Lim PO, Nam HG, Koo JC. Age-dependent action of an
ABA-inducible receptor kinase, RPK1, as a positive regulator of senescence in
Arabidopsis leaves. Plant Cell Physiol 2011;52:65162.
Lee S, Seo PJ, Lee H, Park C. A NAC transcription factor NTL4 promotes reactive oxygen
species production during drought-induced leaf senescence in Arabidopsis. Plant J
2012;70:83144.
Leopold AC. Antagonism of some gibberellin actions by a substituted pyrimidine. Plant
Physiol 1971;48:53740.
Lers A. Environmental regulation of leaf senescence. In: Gan S, editor. Senescence Pro-
cesses in Plants. Oxford: Blackwell; 2007. p. 10944.
Li C, Potuschak T, Colon-Carmona A, Gutierrez RA, Doerner P. Arabidopsis TCP20 links
regulation of growth and cell division control pathways. Proc Natl Acad Sci U S A
2005;102:1297883.
Li HY, Guo ZF, Zhu YX. Molecular cloning and analysis of a pea cDNA that is expressed in
darkness and very rapidly induced by gibberellic acid. Mol Gen Genet 1998;259:3937.
Li JR, Yu K, Wei JR, Ma Q, Wang BQ, Yu D. Gibberellin retards chlorophyll degradation
during senescence of Paris polyphylla. Biol Plant 2010;54:3959.
Li Y, Lee KK, Walsh S, Smith C, Hadingham S, Sorefan K. Establishing glucose- and
ABA-regulated transcription networks in Arabidopsis by microarray analysis and
promoter classication using a Relevance Vector Machine. Genome Res 2006;16:
41427.
Li Z, Peng J, Wen X, Guo H. Gene network analysis and functional studies of
senescence-associated genes reveal novel regulators of Arabidopsis leaf senes-
cence. J Integr Plant Biol 2012;54:52639.
Lim PO, Kim HJ, Nam HG. Leaf senescence. Annu Rev Plant Biol 2007;58:11536.
Lim PO, Lee IC, Kim J, Kim HJ, Ryu JS, Woo HR, et al. Auxin response factor 2 (ARF2)
plays a major role in regulating auxin-mediated leaf longevity. J Exp Bot
2010;61:141930.
Lim PO, Woo HR, Nam HG. Molecular genetics of leaf senescence in Arabidopsis. Trends
Plant Sci 2003;8:2728.
Lin JN, Kao CH. Effect of oxidative stress caused by hydrogen peroxide on senescence of
rice leaves. Bot Bull Acad Sin 1998;39:1615.
Lin JF, Wu SH. Molecular events in senescing Arabidopsis leaves. Plant J 2004;39:61228.
Liu X, Li Z, Jiang Z, Zhao Y, Peng J, Jin J, et al. LSD: a leaf senescence database. Nucleic
Acids Res 2011;39:D11037.
Lobell DB, Sibley A, Ortiz-Monasterio JI. Extreme heat effects on wheat senescence in
India. Nat Clim Change 2012;2:1869.
Lorand L, Graham R. Transglutaminases: crosslinking enzymes with pleiotropic func-
tions. Nat Rev Mol Cell Biol 2003;4:14056.
Lutts S, Kinet J, Bouharmont J. NaCl induced senescence in leaves of rice (Oryza sativa)
cultivars differing in salinity resistance. Ann Bot 1996;78:38998.
Ma W, Berkowitz GA. Cyclic nucleotide gated channel and Ca2+-mediated signal trans-
duction during plant senescence signalling. Plant Signal Behav 2011;6:4135.
Mach JM, Castillo AR, Hoogstraten R, Greenberg JT. The Arabidopsis accelerated cell
death gene ACD2 encodes red chlorophyll catabolite reductase and suppresses
the spread of disease symptoms. Proc Natl Acad Sci U S A 2001;98:7716.
Macnisha AJ, Jiang C, Negre-Zakharova F, Reida MS. Physiological and molecular
changes during opening and senescence of Nicotiana mutabilis owers. Plant Sci
2010;179:26772.
Mashiguchi K, Tanaka K, Sakai T, Sugawara S, Kawaide H, Natsume M, et al. The main auxin
biosynthesis pathway in Arabidopsis. Proc Natl Acad Sci U S A 2011;108:185127.
Matile P, Hortensteiner S, Thomas H. Chlorophyll degradation. Annu Rev Plant Physiol
Plant Mol Biol 1999;50:6795.
Mattoo AK, Minocha SC, Minocha R, Handa AK. Polyamines and cellular metabolism in
plants: transgenic approaches reveal different responses to diamine putrescine versus
higher polyamines spermidine and spermine. Amino Acids 2010;38:40513.
Mattoo Al, Handa AK. Higher polyamines restore and enhance metabolic memory in
ripening fruit. Plant Sci 2008;174:38693.
Mayak S, Halevy AH. Cytokinin activity in rose petals and its relation to senescence.
Plant Physiol 1970;46:4979.
McCabe MS, Garratt LC, Schepers F, Jordi WJRM, Stoopen GM, Davelaar E, et al. Effects
of PSAG12-IPT gene expression on development and senescence in transgenic let-
tuce. Plant Physiol 2001;127:50516.
Mehta RA, Cassol T, Li N, Ali N, Handa AK, Mattoo AK. Engineered polyamine accumu-
lation in tomato enhances phytonutrient content, juice quality and vine life. Nat
Biotechnol 2002;20:6138.
Merzlyak M, Hendry G. Free radical metabolism, pigment degradation and lipid perox-
idation in leaves during senescence. In: Crawford R, Hendry G, Goodman B, editors.
Oxygen and Environmental Stress in Plants, 102 B. Proc. Royal Soc., Edinburgh;
1994. p. 45971.
Miao Y, Laun T, Zimmermann P, Zentgraf U. Targets of theWRKY53transcription factor
and its role during leaf senescence in Arabidopsis. Plant Mol Biol 2004;55:85367.
Milla R, Palacio S, Maestro-Martnez M, Montserrat-Mart G. Leaf exchange in a
Mediterranean shrub: water, nutrient, non-structural carbohydrate and osmolyte
dynamics. Tree Physiol 2007;27:95160.
Mishina TE, Lamb C, Zeier J. Expression of a NO degrading enzyme induces a senes-
cence programme in Arabidopsis. Plant Cell Environ 2007;30:3952.
Mishra NS, Mukherjee SK. A peep into the plant miRNA world. Plant Sci J 2007;1:19.
Mitsuda N, Ohme-Takagi M. Functional analysis of transcription factors in Arabidopsis.
Plant Cell Physiol 2009;50:123248.
Mohapatra PK, Patro L, Raval MK, Ramaswamy NK, Biswal UC, Biswal B. Senescence-
induced loss in photosynthesis enhances cell wall -glucosidase activity. Physiol
Plant 2010;138:34655.
Morita R, Sato Y, Masuda Y, Nishimura M, Kusaba M. Defect in non-yellow coloring 3,
an a/b hydrolase-fold family protein, causes a stay-green phenotype during leaf se-
nescence in rice. Plant J 2009;59:94052.
1169
Morris K, MacKerness SA, Page T, John CF, Murphy AM, Carr JP, et al. Salicylic acid has a
role in regulating gene expression during leaf senescence. Plant J 2000;23:67785.
Munne-Bosch S. Do perennials really senesce? Trends Plant Sci 2008;13:21620.
Munne-Bosch S, Alegre L. Plant aging increases oxidative stress in chloroplasts. Planta
2002;214:60815.
Muramoto Y, Watanabe A, Nakamura T, Takabe T. Enhanced expression of a nuclease
gene in leaves of barley plants under salt stress. Gene 1999;234:31521.
Mutui TM, Emongor VE, Hutchinson MJ. The effects of gibberellin on the vase life
and ower quality of Alstroemeria cut owers. Plant Growth Regul 2006;48:
20714.
Nakashima K, Takasakia H, Mizoib J, Shinozakic K, Yamaguchi-Shinozaki K. NAC transcrip-
tion factors in plant abiotic stress responses. Biochim Biophys Acta 2011;1819:97-103.
Nambeesan S, Datsenka T, Ferruzzi MG, Malladi A, Mattoo AK, Handa AK.
Overexpression of yeast spermidine synthase impacts ripening, senescence and
decay symptoms in tomato. Plant J 2010;63:83647.
Nambeesan S, Mattoo AK, Handa AK. Polyamines and regulation of ripening and
senescence. In: Paliyath G, Murr DP, Handa AK, Lurie S, editors. Postharvest Biology
and Technology of Fruits, Vegetables, and Flowers. Ames, IA: Wiley-Blackwell
Publishing; 2008. p. 31940.
Naqvi AR, Choudhury NR, Haq QMR. Small RNA-mediated defensive and adaptive re-
sponses in plants. Sust Agric Rev 2011a;7:12960.
Naqvi AR, Choudhury NR, Mukherjee SK, Haq QMR. In silico identication of tomato
microRNA/microRNA sequences that exhibit propensity to bind ToLCV associated
genomes and most of their encoded ORFs. Plant Physiol Biochem 2011b;49:137.
Naqvi AR, Haq QM, Mukherjee SK. MicroRNA proling of Tomato leaf curl New Delhi
virus (ToLCNDV) infected tomato leaves indicates that miR159/319 and miR172
might be linked with leaf curl disease. Virol J 2010;7:281.
Naqvi AR, Islam MN, Choudhury NR, Haq QM. The fascinating world of RNA interfer-
ence. Int J Biol Sci 2009;5:97-117.
Naqvi AR, Sarwat M, Hasan S, Choudhury NR. Biogenesis, functions and fate of plant
microRNAs. J Cell Physiol 2012;227:31638.
Navabpour S, Morris K, Allen R, Harrison E, A-H-Mackerness S, Buchanan-Wollaston V.
Expression of senescence-enhanced genes in response to oxidative stress. J Exp Bot
2003;54:228592.
Nemchenko A, Kunze S, Feussner I, Kolomiets M. Duplicate maize 13-lipoxygenase
genes are differentially regulated by circadian rhythm, cold stress, wounding,
pathogen infection, and hormonal treatments. J Exp Bot 2006;57:376779.
Nikovics K, Blein T, Peaucelle A, Ishida T, Morin H, Aida M, et al. The balance between
the miR164A and CUC2 genes controls leaf margin serration in Arabidopsis. Plant
Cell 2006;18:292945.
Noh YS, Amasino R. Identication of a promoter region responsible for the senescence-
specic expression of SAG12. Plant Mol Biol 1999;41:18194.
Oh SA, Lee SY, Chung IK, Lee CH, Nam HG. A senescence associated gene of Arabidopsis
thaliana is distinctively regulated during natural and articially induced leaf senes-
cence. Plant Mol Biol 1996;30:73954.
Oh SA, Park JH, Lee GI, Paek KH, Park SK, Nam HG. Identication of three genetic loci
controlling leaf senescence in Arabidopsis thaliana. Plant J 1997;12:52735.
Okushima Y, Mitina I, Quach HL, Theologis A. AUXIN RESPONSE FACTOR 2 (ARF2): a
pleiotropic developmental regulator. Plant J 2005;43:2946.
Ori N, Cohen AR, Etzioni A, Brand A, Yanai O, Shleizer S, et al. Regulation of
LANCEOLATE by miR319 is required for compound-leaf development in tomato.
Nat Genet 2007;39:78791.
Orzaez D, Granell A. DNA fragmentation is regulated by ethylene during petal senes-
cence in Pisum sativum. Plant J 1997;11:13744.
Osakabe Y, Mizuno S, Tanaka H, Maruyama K, Osakabe K, Todaka D. Overproduction of
the membrane-bound receptor-like protein kinase 1, RPK1, enhances abiotic stress
tolerance in Arabidopsis. J Biol Chem 2010;285:9190201.
Palatnik JF, Allen E, Wu X, Schommer C, Schwab R, Carrington JC, et al. Control of leaf
morphogenesis by microRNAs. Nature 2003;425:25763.
Palavan-Unsal N, Buyuktuncer E, Tufekci MA. Programmed cell death in plants. J Cell
Mol Biol 2005;24:9-23.
Palma JM, Sandalio LM, Corpas FJ, Romero-Puertas MC, McCarthy I, del Ro LA. Plant
proteases, protein degradation, and oxidative stress: role of peroxisomes. Plant
Physiol Biochem 2002;40:52130.
Pandey S, Gupta K, Mukherjee AK. Impact of cadmium and lead on Catharanthus roseus:
a phytoremediation study. J Environ Biol 2007;28:65562.
Pandey S, Ranade SA, Nagar PK, Kumar N. Role of polyamines and ethylene as modula-
tors of plant senescence. J Biosci 2000;25:2919.
Pang XM, Nada K, Kurosawa T, Ban Y, Moriguchi T. Effect of methylglyoxal
bis-(guanylhydrazone) on polyamine and ethylene biosynthesis of apple fruit
after harvest. Acta Physiol Plant 2010;32:100510.
Pardon MC. Stress and ageing interactions: a paradox in the context of shared etiolog-
ical and physiopathological processes. Brain Res Rev 2007;54:25173.
Park JH, Oh SA, Kim YH, Woo HR, Nam HG. Differential expression of senescence-
associated mRNAs during leaf senescence induced by different senescence induc-
ing factors in Arabidopsis. Plant Mol Biol 1998;37:44554.
Parmar AS, Hammond HD. The effect of boron and gibberellic acid on the senescence of
leaf discs from Lycopersicon esculentum cv. Rutgers. Physiol Plant 1971;25:16977.
Philosoph-Hadas S, Meir S, Akiri B, Kanner J. Oxidative defense systems in leaves of
three edible herb species in relation to their senescence rates. J Agric Food Chem
1994;42:233176.
Poovaiah BW, Leopold AC. Deferral of leaf senescence with calcium. Plant Physiol
1973;52:2369.
Pruinsk A, Anders I, Aubry S, Schenk N, Tapernoux-Lthi E, Mller T, et al. In vivo par-
ticipation of red chlorophyll catabolite reductase in chlorophyll breakdown. Plant
Cell 2007;19:36987.
1170 M. Sarwat et al. / Biotechnology Advances 31 (2013) 11531171
Pruinsk A, Tanner G, Anders I, Roca M, H rtensteiner S. Chlorophyll breakdown:
pheophorbide a oxygenase is a Rieske-type iron-sulfur protein, encoded by the ac-
celerated cell death 1 gene. Proc Natl Acad Sci U S A 2003;100:15259526.
Pulido A, Laufs P. Co-ordination of developmental processes by small RNAs during leaf
development. J Exp Bot 2010;61:127791.
Quirino BF, Noh YS, Himelblau E, Amasino RM. Molecular aspects of leaf senescence.
Trends Plant Sci 2000;5:27882.
Rajagopal R, Vaucheret H, Trejo J, Bartel DP. A diverse and evolutionarily uid set of
microRNAs in Arabidopsis thaliana. Genes Dev 2006;20:340725.
Rangel M, Machado OLT, Cunha M, Jacinto T. Accumulation of chloroplast-targeted
lipoxygenase in passion fruit leaves in response to methyl jasmonate. Phytochem-
istry 2002;60:61925.
Rao MV, Lee HI, Davis KR. Ozone-induced ethylene production is dependent on salicylic
acid, and both salicylic acid and ethylene act in concert to regulate ozone-induced
cell death. Plant J 2002;32:44756.
Ren G, An K, Liao Y, Zhou X, Cao Y, Zhao H, et al. Identication of a novel chloroplast
protein AtNYE1 regulating chlorophyll degradation during leaf senescence in
Arabidopsis. Plant Physiol 2007;144:142941.
Richmond AE, Lang A. Effect of kinetin on protein content and survival of detached
Xanthium leaves. Science 1957;125:6501.
Riechmann JL, Heard J, Martin G, Reuber L, Jiang C, Keddie J, et al. Arabidopsis transcrip-
tion factors: genomewide comparative analysis among eukaryotes. Science
2000;290:210510.
Riou C, Herve C, Pacquit V, Dabos P, Lescure B. Expression of an Arabidopsis lectin kinase
receptor gene, lecRK-a1, is induced during senescence, wounding and in response
to oligogalacturonic acids. Plant Physiol Biochem 2002;40:4318.
Robatzek S, Somssich IE. A new member of the Arabidopsis WRKY transcription factor
family, AtWRKY6, is associated with both senescence- and defense-related pro-
cesses. Plant J 2001;28:12333.
Robatzek S, Somssich IE. Targets of AtWRKY6 regulation during plant senescence and
pathogen defense. Genes Dev 2002;16:113949.
Rolland F, Moore B, Sheen J. Sugar sensing and signaling in plants. Plant Cell 2002;14:
S185205.
Salam-a S. The effect of epibrassinolide on senescence in wheat leaves. Biotechnol Tripathi SK, Tuteja N. Integrated signaling in ower senescence. Plant Signal Behav
Biotechnol EQ 2007;21:635.
Sakai H, Hua J, Chen QG, Chang C, Medrano LJ, Bleecker AB, et al. ETR2 is an ETR1-like gene
involved in ethylene signaling in Arabidopsis. Proc Natl Acad Sci U S A 1998;95:
58127.
Sato Y, Morita R, Nishimura M, Yamaguchi H, Kusaba M. Mendel's green cotyledon
gene encodes a positive regulator of the chlorophyll-degrading pathway. Proc
Natl Acad Sci U S A 2007;104:1416974.
Saumonneau A, Laloi M, Lallemand M, Rabot A, Atanassova R. Dissection of the
transcriptional regulation of grape ASR and response to glucose and abscisic acid.
J Exp Bot 2012;63:1495510.
Schenk PM, Kazan K, Rusu AG, Manners JM, Maclean DJ. The SEN1 gene of Arabidopsis is
regulated by signals that link plant defence responses and senescence. Plant Phys-
iol Biochem 2005;43:997-1005.
Schippers JHM, Nunes-Nesi A, Apetrei R, Hille J, Fernie AR, Dijkwela PP. The Arabidopsis
onset of leaf death mutation of quinolinate synthase affects nicotinamide adenine
dinucleotide biosynthesis and causes early ageing. Plant Cell 2008;20:290925.
Schommer C, Palatnik JF, Aggarwal P, Chtelat A, Cubas P, Farmer EE, et al. Control of
jasmonate biosynthesis and senescence by miR319 targets. PLoS Biol 2008;6:230.
Seiler N, Raul F. Polyamines and apoptosis. J Cell Mol Med 2005;9:62342.
Seltmann MA, Stingl NE, Lautenschlaeger JK, Krischke M, Mueller MJ, Berger S. Differ-
ential impact of lipoxygenase 2 and jasmonates on natural and stress-induced se-
nescence in Arabidopsis. Plant Physiol 2010;152:194050.
Serani-Fracassini D, Bagni N, Cionini PG, Benini A. Polyamines and nucleic acids dur-
ing the rst cell cycle of Helianthus tuberosus tissue after the dormancy break.
Planta 1980;148:3327.
Serani-Fracassini D, Del Duca S, Monti F, Poli F, Sacchetti G, Bregoli AM, et al.
Transglutaminase activity during senescence and programmed cell death in the co-
rolla of tobacco (Nicotiana tabacum) owers. Cell Death Differ 2002;9:30921.
Serani-Fracassini D, Del Duca S. Transglutaminases: widespread cross-linking en-
zymes in plants. Ann Bot 2008;102:14552.
Serani-Fracassini D, Sandro AD, Duca SD. Spermine delays leaf senescence in Lactuca
sativa and prevents the decay of chloroplast photosystems. Plant Physiol Biochem
2010;48:60211.
Sexton R, Roberts JA. Cell biology of abscission. Annu Rev Plant Physiol 1982;33:13362.
Shan X, Wang J, Chua L, Jiang D, Peng W, Xie D. The role of Arabidopsis rubisco activase
in jasmonate-induced leaf senescence. Plant Physiol 2011;155:275164.
Sharabi-Schwager M, Lers A, Samach A, Guy CL, Porat R. Overexpression of the CBF2
transcriptional activator in Arabidopsis delays leaf senescence and extends plant
longevity. J Exp Bot 2010;61:26173.
Shiu SH, Bleecker AB. Expansion of the receptor-like kinase/Pelle gene family and
receptor-like proteins in Arabidopsis. Plant Physiol 2003;132:53043.
Simeonova E, Sikira S, Charzynska M, Mostowska A. Aspects of programmed cell death
during leaf senescence of mono- and dicotyledonous plants. Protoplasma
2000;214:93-101.
Smart CM. Gene expression during leaf senescence. New Phytol 1994;126:41948.
Smykowski A, Zimmermann P, Zentgraf U. G-Box binding factor1 reduces CATALASE2
expression and regulates the onset of leaf senescence in Arabidopsis. Plant Physiol
2010;153:132131.
Sobieszczuk-Nowicka E, Di Sandro A, Del Duca S, Serani- Fracassini D, Legocka J.
Plastid-membrane-associated polyamines and thylakoid transglutaminases during
etioplast- to chloroplast transformation stimulated by kinetin. Physiol Plant
2007;130:590600.
Sobieszczuk-Nowicka E, Krzesowska M, Legocka J. Transglutaminases and their sub-
strates in kinetin-stimulated etioplast-to-chloroplast transformation in cucumber
cotyledons. Protoplasma 2008;233:18794.
Sobieszczuk-Nowicka E, Wieczorek P, Legocka J. Kinetin affects the level of chloroplast
polyamines and transglutaminase activity during senescence of barley leaves. Acta
Biochim Pol 2009;56:2559.
Sood S, Nagar PK. Post-harvest alterations in polyamines and ethylene in two diverse
rose species. Acta Physiol Plant 2008;30:2438.
Swartzberg D, Hanael R, Granot D. Relationship between hexokinase and cytokinin in
the regulation of leaf senescence and seed germination. Plant Biol 2011;13:
43944.
Swidzinski JA, Leaver CJ, Sweetlove LJ. A proteomic analysis of plant programmed cell
death. Phytochemistry 2004;65:182938.
Tang DZ, Innes RW. Overexpression of a kinase decient form of the EDR1 gene en-
hances powdery mildew resistance and ethylene-induced senescence in
Arabidopsis. Plant J 2002;32:97583.
Tenea GN, Bota AP, Raposo FC, Maquet A. Reference genes for gene expression studies
in wheat ag leaves grown under different farming conditions. BMC Res Notes
2011;4:373.
Thomas H, Ougham HJ, Wagstaff C, Stead AD. Dening senescence and death. J Exp Bot
2003;54:112732.
Thomas H, Stoddart J. Leaf senescence. Annu Rev Plant Physiol 1980;31:83-111.
Thompson JE, Froese CD, Madey E, Smith MD, Hong YW. Lipid metabolism during plant
senescence. Prog Lipid Res 1998;37:11941.
Thomson WW, Plat-Aloia KA. Ultrastructure and Senescence in Plants. In: Thomson
WW, Nothnagei EA, Huffaker RC, editors. Plant senescence: its biochemistry and
physiology. Rockville: Am. Soc. Plant Physiol.; 1987. p. 2030.
Tiburcio AF, Altabella T, Masgrau C. Polyamines. In: Bisseling T, Schell J, editors. New
Developments in Plant Hormone Research. New York: Springer-Verlag; 2002.
Torrigiani P, Bressanin D, Ruiz KB, Tadiello A, Trainotti L, Bonghi C, et al. Spermidine ap-
plication to young developing peach fruits leads to a slowing down of ripening by
impairing ripening-related ethylene and auxin metabolism and signalling. Physiol
Plant 2012;146:8698.
2007;6:43745.
Trivellini A, Ferrante A, Vernieri P, Serra G. Effects of abscisic acid on ethylene biosyn-
thesis and perception in Hibiscus rosa-sinensis L. ower development. J Exp Bot
2011;62:543752.
Tuteja N. Abscisic acid and abiotic stress signaling. Plant Signal Behav 2007;2:1358.
Uberti-Manassero NG, Lucero LE, Viola IL, Vegetti AC, Gonzalez DH. The class I protein
AtTCP15 modulates plant development through a pathway that overlaps with the
one affected by CIN-like TCP proteins. J Exp Bot 2012;63:80923.
Ueda J, Kato J. Identication of a senescence-promoting substance from wormwood
(Artemisia absinthum L.). Plant Physiol 1980;66:2469.
Uzelac B, Janosevi D, Budimir S. In situ detection of programmed cell death in
Nicotiana tabacum leaves during senescence. J Microsc 2008;230:13.
Van der Graaff E, Schwacke R, Schneider A, Desimone M, Flugge UI, Kunze R. Transcrip-
tion analysis of Arabidopsis membrane transporters and hormone pathways during
developmental and induced leaf senescence. Plant Physiol 2006;141:77692.
van Doorn WG, Woltering EJ. Senescence and programmed cell death: substance or se-
mantics? J Exp Bot 2004;55:214753.
van Doorn WG, Yoshimoto K. Role of chloroplasts and other plastids in ageing and
death of plants and animals: a tale of Vishnu and Shiva. Ageing Res Rev 2010;9:
11730.
van Leyen K, Duvoisin RM, Engelhardt H, Wiedmann M. A function for lipoxygenase in
programmed organelle degradation. Nature 1998;395:3925.
Vanacker H, Sandalio LM, Jimnez A, Palma JM, Corpas FJ, Meseguer V, et al. Roles for
redox regulation in leaf senescence of pea plants grown on different sources of ni-
trogen nutrition. J Exp Bot 2006;57:173545.
Vardhini BV, Rao SS. Acceleration of ripening of tomato pericarp discs by
brassinosteroids. Phytochemistry 2002;61:8437.
Walbot V, Hoisington DA, Neuffer MG. Disease lesion mimic mutations. In: Kosuge T,
Meredith C, editors. Genetic Engineering of Plants. New York: Plenum Publishing
Company; 1983. p. 43142.
Wang W. Regulatory RNA-binding proteins in senescence. Ageing Res Rev 2012;11(4):
48590.
Wang X, Chory J. Brassinosteroids regulate dissociation of BKI1, a negative regulator of
BRI1 signaling, from the plasma membrane. Science 2006;313:111822.
Weaver LM, Gan S, Quirino B, Amasino RM. A comparison of the expression patterns of
several senescence-associated genes in response to stress and hormone treatment.
Plant Mol Biol 1998;37:45569.
Wei Q, Guo Y, Kuai B. Isolation and characterization of a chlorophyll degradation regu-
latory gene from tall fescue. Plant Cell Rep 2011;30:12017.
Wingler A, von Schaewen A, Leegood RC, Lea PJ, Quick WP. Regulation of leaf senescence
by cytokinin, sugars, and light. Effects on NADH-dependent hydroxypyruvate reduc-
tase. Plant Physiol 1998;116:32935.
Wingler A, Purdy S, MacLean JA, Pourtau N. The role of sugars in integrating environ-
mental signals during the regulation of leaf senescence. J Exp Bot 2006;57:
3919.
Woo HR, Chung KM, Park JH, Oh SA, Ahn T, Hong SH, et al. ORE9, an F-Box protein that
regulates leaf senescence in Arabidopsis. Plant Cell 2001;13:177990.
Woo HR, Goh CH, Park JH, Teyssendier de la Serve B, Kim JH, Park YI, et al. Extended leaf
longevity in the ore4-1 mutant of Arabidopsis with a reduced expression of a plas-
tid ribosomal protein gene. Plant J 2002;31:33140.
Woo HR, Kim JH, Kim J, Kim J, Lee U, Song I, et al. The RAV1 transcription factor posi-
tively regulates leaf senescence in Arabidopsis. J Exp Bot 2010;61:394757.
 M. Sarwat et al. / Biotechnology Advances 31 (2013) 11531171
Woo HR, Kim JH, Nam HG, Lim PY. The delayed leaf senescence mutants of Arabidopsis,
ore1, ore3, and ore9 are tolerant to oxidative stress. Plant Cell Physiol 2004;45:
92332.
Xiao S, Dai LY, Liu FQ, Wang ZL, Peng W, Xie DX. COS1: An Arabidopsis coronatine in-
sensitive1 suppressor essential for regulation of jasmonate-mediated plant senes-
cence and defense. Plant Cell 2004;16:113242.
Xiao S, Gao W, Chen Q, Chan S, Zheng S, Ma J, et al. Overexpression of Arabidopsis Acyl-CoA
binding protein ACBP3 promotes starvation-induced and age-dependent leaf senes-
cence. Plant Cell 2010;22:146382.
Xiao S, Chye ML. Overexpression of Arabidopsis ACBP3 enhances NPR1-dependent plant
resistance to Pseudomonas syringe pv tomato DC3000. Plant Physiol 2011;156:
206981.
Xu F, Meng T, Li P, Yu Y, Cui Y, Wang Y, et al. A soybean dual-specicity kinase, GmSARK,
and its Arabidopsis homolog, AtSARK, regulate leaf senescence through synergistic
actions of auxin and ethylene. Plant Physiol 2011;157:213153.
Xu Y, Gianfagna T, Huang B. Proteomic changes associated with expression of a gene
(ipt) controlling cytokinin synthesis for improving heat tolerance in a perennial
grass species. J Exp Bot 2010;61:327389.
Xu Y, Huang B. Effects of foliar-applied ethylene inhibitor and synthetic cytokinin on
creeping bentgrass to enhance heat tolerance. Crop Sci 2009;49:187684.
Yang SD, Seo PJ, Yoon HK, Park CM. The Arabidopsis NAC transcription factor VNI2 in-
tegrates abscisic acid signals into leaf senescence via the COR/RD genes. Plant Cell
2011;23:215568.
Yang TF, Gonzalez-Carranza ZH, Maunders MJ, Roberts JA. Ethylene and the regulation of
senescence processes in transgenic Nicotiana sylvestris plants. Ann Bot 2008;101:
30110.
Yoon EK, Yang JH, Lim J, Kim SH, Kim SK, Lee WS. Auxin regulation of the
microRNA390-dependent transacting small interfering RNA pathway in Arabidopsis
lateral root development. Nucleic Acids Res 2010;38:138291.
Yoshida S. Molecular regulation of leaf senescence. Curr Opin Plant Biol 2003;6:7884.
Yoshimoto K, Jikumaru Y, Kamiya Y, Kusano M, Consonni C, Panstruga R, et al. Autoph-
agy negatively regulates cell death by controlling NPR1-dependent salicylic acid
signaling during senescence and the innate immune response in Arabidopsis.
Plant Cell 2009;21:291427.
Yu K, Wang Y, Wei J, Ma Q, Yu D, Li J. Improving rhizome yield and quality of Paris
polyphylla through gibberellic acid-induced retardation of senescence of aerial
parts. Plant Signal Behav 2009;4:4135.
1171
Yun-xia G, Li-jun Z, Feng-hai L, Zhi-bin C, Che W, Yun-cong Y, et al. Relationship be-
tween jasmonic acid accumulation and senescence in drought-stress. Afr J Agric
Res 2010;5:197883.
Yupsanis T, Kefalas PS, Eleftheriou P, Kotinis K. RNase and DNase activities in the alfalfa
and lentil grown in iso-osmotic solutions of NaCl and mannitol. J Plant Physiol
2001;158:9217.
Zapata JM, Guera A, Esteban-Carrasco A, Martin M, Sabater B. Chloroplasts regulate leaf
senescence: delayed senescence in transgenic ndhF-defective tobacco. Cell Death
Differ 2005;12:127784.
Zeevaart JAD, Creelman RA. Metabolism and physiology of abscisic acid. Annu Rev Plant
Physiol Plant Mol Biol 1988;39:43973.
Zentgraf U, Hemleben V. Molecular cell biology: are reactive oxygen species regulators
of leaf senescence? In: Luttge U, Beyschlag W, Murata J, editors. Progress in Botany,
69. Berlin: Springer; 2008. p. 11738.
Zentgraf U, Jobst J, Kolb D, Rentsch D. Senescence-related gene expression proles of
rosette leaves of Arabidopsis thaliana: leaf age versus plant age. Plant Biol
2004;6:17883.
Zhang K, Xia X, Zhang Y, Gan S. An ABA-regulated and Golgi-localized protein phospha-
tase controls water loss during leaf senescence in Arabidopsis. Plant J 2011;69:
66778.
Zhao H, Zhang ZB, Shao HB, Xu P, Foulkes MJ. Genetic correlation and path analysis of
transpiration efciency for wheat ag leaves. Environ Exp Bot 2008;64:12834.
Zhou JM, Trifa Y, Silva H, Pontier D, Lam E, Shah J, et al. NPR1 differentially interacts
with members of the TGA/OBF family of transcription factors that bind an element
of the PR-1 gene required for induction by salicylic acid. Mol Plant-Microbe Inter-
act 2000;13:191202.
Zhou C, Han L, Pislariu C, Nakashima J, Fu C, Jiang Q, et al. From model to crop: function-
al analysis of a Stay-Green gene in the model legume Medicago truncatula and ef-
fective use of the gene for alfalfa improvement. Plant Physiol 2011;157:148396.
Zhu T, Nevo E, Sun D, Peng J. Phylogenetic analyses unravel the evolutionary history of
nac proteins in plants. Evolution 2012;66:183348.
Zhu Z, Zhang Z, Qin G, Tian S. Effects of brassinosteroids on postharvest disease and se-
nescence of jujube fruit in storage. Postharvest Biol Technol 2010;56:505.
Zimmermann P, Zentgraf U. The correlation between oxidative stress and leaf senes-
cence during plant development. Cell Mol Biol Lett 2005;10:51534.