Biotechnology Advances 28 (2010) 489499

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Biotechnology Advances
j o u r n a l h o m e p a g e : w w w. e l s e v i e r. c o m / l o c a t e / b i o t e c h a d v

Research review paper

Nitric oxide in fruit ripening: Trends and opportunities

G. Manjunatha, V. Lokesh, Bhagyalakshmi Neelwarne
Plant Cell Biotechnology Department, Central Food Technological Research Institute, Mysore-570 020, India 1

a r t i c l e

i n f o

Article history:
Received 17 October 2009
Received in revised form 22 February 2010
Accepted 12 March 2010
Available online 20 March 2010
Keywords:
Nitric oxide
Fruit ripening
Ethylene
Signals

a b s t r a c t
Monitoring ethylene is crucial in regulating post-harvest life of fruits. The concept of nitric oxide (NO)
involvement in antagonizing ethylene is new. NO mediated physiologies casted through regulation of plant
hormones are widely reported during developmental and stress chemistry having no direct link with
ripening. Research in NO biology and understanding its interplay with other signal molecules in ripening
fruits suggest ways of achieving greater synergies with NO applications. Experiments focused at convincingly
demonstrating the involvement of NO in altering ripening-related ethylene prole of fruits, would help
develop new processes for shelf life extension. This issue being the central theme of this review, the putative
mechanisms of NO intricacies with other primary and secondary signals are hypothesized. The advantage of
eliciting NO endogenously may open up various biotechnological opportunities for its precise delivery into
the target tissues.
2010 Elsevier Inc. All rights reserved.

Contents
1.
2.
3.
4.
5.

Introduction . . . . . . . . . . . . . . . . . . . . . . .
Rationale behind NO usage for ripening control . . . . . .
Nitric oxideethylene cross talks . . . . . . . . . . . . .
NO versus ROS . . . . . . . . . . . . . . . . . . . . . .
Interplay of NO with other signals . . . . . . . . . . . .
5.1.
Salicylic acid (SA) . . . . . . . . . . . . . . . . .
5.2.
Jasmonic acid (JA) . . . . . . . . . . . . . . . . .
5.3.
Growth regulators . . . . . . . . . . . . . . . . .
6.
NO and polyamines (PAs) . . . . . . . . . . . . . . . .
7.
Application of NO and monitoring endogenous NO threshold
8.
Generation of endogenous NO by elicitations . . . . . . .
9.
Conclusions and future prospects . . . . . . . . . . . . .
Acknowledgements . . . . . . . . . . . . . . . . . . . . . .
References . . . . . . . . . . . . . . . . . . . . . . . . . .

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1. Introduction
Current increase in the awareness of the nutritional importance
and nutraceutical effects of edible fruits has called for more intense
research on their post-harvest biotechnology than ever before,
especially emphasizing technologies of relevance to their transport
and shelf life. Immediately after harvest, fruits undergo programmed
senescence accompanied by various physiological and biochemical

Corresponding author. Tel.: +91 821 2516501; fax: +91 821 2571223.
E-mail address: blakshmi_1999@yahoo.com (B. Neelwarne).
1
A constituent laboratory of the Council of Scientic and Industrial Research,
New Delhi.
0734-9750/$ see front matter 2010 Elsevier Inc. All rights reserved.
doi:10.1016/j.biotechadv.2010.03.001

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489
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changes occurring under highly coordinated genetic events as a

response to environmental cues (see Box 1 and Fig. 1). Among the
conserved developmental signals inuencing fruit ripening, the
participation of NO signal appears to be most interesting. Although
experiments focused on unveiling NO involvement in ripening have
provided a few insights into the biochemical events, comprehensive
genetic regulations are yet-to-be resolved. NO is known to be involved
in the growth and development of the plants starting from
germination and de-etiolation (Beligni and Lamattina, 2000), root
development (Correa-Aragunde et al., 2004), senescence (Leshem
et al., 1998), owering (He et al., 2004), reproduction (Prado et al.,
2004), fruit ripening (Leshem, 2000), biotic stresses of particular
relevance to disease resistance (Delledone et al., 1998; Manjunatha et
al., 2008), abiotic stress involved in drought resistance (Desikan et al.,

490

G. Manjunatha et al. / Biotechnology Advances 28 (2010) 489499

Box 1
Structural and biochemical changes in ripening fruits
Fruit ripening is a distinct aspect of plant development that signifies a large component of the food supply having direct relevance to human
health and nutrition. Fruits have been classified as climacteric and non-climacteric on the basis of patterns of respiration and of ethylene
production during ripening. In climacteric fruits, the massive burst of ethylene occurs at the respiratory peak period culminating in most
appropriate ripening (Giovannoni, 2004). In addition, ripening process involves a series of biochemical, physiological and structural
changes, albeit climacteric and non-climacteric fruits differ with each other (Prabha and Bhagyalakshmi, 1998; Bhagyalakshmi et al., 2002;
Antonio et al., 2009). Climacteric fruits ripen faster and are characterized with rapid senescence, deterioration of sensorial attributes with
reduced nutritive quality predisposing edible tissues vulnerable to microbes. Such losses of horticultural commodities are enormous,
particularly in tropical areas. Hence, extending the shelf life of fruits is one of the major concerns for delivering high quality commodities to
consumers. Generally, fruit quality is attributed by various parameters such as the cell wall/membrane integrity, cell homeostasis,
metabolism of reactive oxygen species causing oxidative burst, and peroxidation of membrane lipids. A classical strategy for extending the
post-harvest shelf life of fruits is the refrigerated storage, although its application for some of the climacteric fruits like banana and peach is
limited due to their sensitivity to chill injury. An alternative is to block the ethylene synthesis and/or prevent the ethylene action in
climacteric fruits for achieving ripening and senescence control.
2002), wounding (Huang et al., 2004), programmed cell death to
ferritin regulation and balancing cellular redox status (Beligni et al.,
2002; Murgia et al., 2004). Implications of NO in the modulation of
plant hormones such as cytokinin (Wilhelmova et al., 2006), auxin
(Otvos et al., 2005) and ethylene, the most important ripening
hormone (Leshem and Haramaty, 1996) have been the subject of
intense research. NO strongly induced a gene response that coded for
alternative oxidase, the latter in turn counteracted the cytochrome-Cdependent respiration as well as ethylene biosynthesis (Ederli et al.,
2006). There have also been observations on the enhancement of
secondary metabolites, that NO donors induced betalaine accumulation in Amaranthus seedlings (Scherer and Holk, 2000), enhanced
nutritionally important biological iron (Graziano et al., 2002) and
supported higher saponin accumulation in Panax ginseng cell suspensions (Hu et al., 2003). Chemical probes capable of monitoring
endogenous NO in vegetative and generative plant organs revealed
that temporal progress of maturation and senescence appeared to go

hand in hand with a signicant decrease of NO emission. The

extension of post-harvest life of fresh commodities primarily demand
reversal of senescence and suppression of ethylene formation, and
such properties are linked to endogenous NO formation; one can
derive a direct relationship between NO, ethylene and suppression of
fruit ripening (Fig. 2). Therefore, the use of this marvelous molecule
for monitoring post-harvest life of fruits and vegetables appears to
open up newer biotechnological avenues at commercial scales.
2. Rationale behind NO usage for ripening control
Methods currently practiced for the preservation and packaging of
whole fruits and vegetables to prolong the shelf life have inherent
limitations. The deterioration of fruit quality is caused by a range of
physiological processes associated with ripening, the major among
them are rapid respiration and senescence, allowing invasion of
opportunistic microbes. Although 1-MCP (1-methylcyclopropene), a

Fig. 1. Biochemistry of ethylene formation and functions in plants: S-adenosyl methionine (SAM) derived from Yang cycle produces 1-aminocyclopropane 1-carboxylic acid (ACC)
through ACC synthase (ACCS) and forms ethylene through ACC oxidase (ACCO) triggering a series of functional attributes.

G. Manjunatha et al. / Biotechnology Advances 28 (2010) 489499

491

Fig. 2. The mechanisms through which NO possibly antagonize ethylene causing ripening inhibition. (Dark lines with mark indicate the established phenomenon with indirect
link, red lines ended with mark indicate ethylene inhibition by NO and dotted lines indicate the unidentied mechanisms). Panel A shows pathways directly regulating the
negation of ethylene-induced effects by NO through transcriptional regulation of 1-Aminocycopropane 1-carboxylic acid oxidase (ACO) genes ACO1, ACOH2 and ACO4; Snitrosylation causing post-translational modication of methionine adenosyltransferase (MAT); inhibition of hydrogenation of ethane (C2H6) and stoichiometric reduction of ACC
into 1-malonyl aminocyclopropane-1-carboxylic acid (MACC); NO and ACC oxidase form a binary ACC oxidaseNO complex which is further chelated by ACC to produce a stable
ternary ACCACC oxidaseNO complex. Panel B shows indirect effects of NO causing ethylene regulation in tight co-ordination of other signal molecules; salicylic acid (SA) and NO
interact with each other synergistically through protein phosporylation and alteration of calcium inux during external stimuli, where SA acts on ACO to reduce ethylene;
polyamines (PA) also cross talk with NO through an unidentied mechanism (?) and reduce ethylene by inhibiting 1-aminocycopropane 1-carboxylic acid synthase (ACS) and by
competitive action on common precursor SAM; NO and cytokinin might also interact with each other through an unidentied (?) mechanism implicated in possible regulation of
ethylene.

gaseous compound known to efciently retard ripening by blocking

ethylene receptors and benets post-harvest quality of fruits, there
are a few disadvantages such as increased CO2 injury in apple (DeEll et
al., 2003; Zanella, 2003; Watkins and Nock, 2004), irradiation damage
aggravated in apple (Fan and Maththeis, 2001), increased chilling
injury in orange (Porat et al., 1999) and banana (Jiang et al., 2004),
increased disease susceptibility in avocado, custard apple, mango and
papaya (Hofman et al., 2001) and higher 1-MCP concentration
enhanced decay in strawberry (Ku et al., 1999). There have also
been a few other pathological storage distresses that are reviewed by
Watkins (2006). Extensive research has resulted in the development
of optimized modied atmosphere packaging (MAP), particularly the
CO2 and O2 levels inside the packaging that often maximize postharvest life of the horticulture produce. Nevertheless, there are
difculties in maintaining the specic MAP conditions during
transport through supply chains, and such difculties are further
aggravated by uctuating temperature. Inadequate control of MAP
may also lead to increased respiration, catalyzing undesirable
physiological events resulting in the development of off-avours,
browning and textural loss. Differences in the level of maturity of each
harvested produce demand variations in packaging designs to ensure
safe MAP.
The information of NO effect on fruit quality is sporadic and limited
to temperate fruits of non-climacteric nature. A few experiments
focused on the effects of NO on fruit quality have provided novel
information on its role in effectively delaying fruit softening, and
hence appear to play a signicant role in the transit and storage of
fruit commodities. Such effects were linked with the inhibition of
pectin depolymerisation and other polysaccharides with reduction
of chilling injury. NO treatment reduced the degree of disintegration

of the cell membrane with lesser electrolyte leakage which resulted in

better retention of the cellular components such as pigments,
titratable acidity, soluble solids and free antioxidant compounds,
particularly the ascorbic acid, which the fruits are most valued for
(Zhang et al., 2007a,b; Francisco et al., 2008; Flores et al., 2008). These
NO effects were paramount in climacteric fruits rather than in nonclimacteric ones (Zhu et al., 2006; Zhu et al., 2008). Table 1
summarizes the applications of NO in controlling ripening of various
fruit commodities. Post-harvest NO fumigation postponed ripening by
34 days and reduced chilling injury during cold storage up to
6 weeks in Japanese plums (Prunus salicina Lindell) (Singh et al.,
2009). Similarly in apple, 70% higher extension in post-harvest life
was evinced in treatment with NO donor DETANO (Pristijono et al.,
2008). Strawberry and kiwi fruits fumigated with NO resulted in a
marked extension of shelf life (Leshem et al., 1998). Even in
vegetables such as broccoli (Brassica oleracea), green bean (Phaseolus
vulgaris), and bok choy (Brassica chinensis) NO gas extended postharvest life signicantly (Soegiarto and Wills, 2004). Furthermore, NO
gas was found benecial for protecting fruits and vegetables from
microbial infections by Aspergillus niger, Monilinia fructicola, Penicillium italicum and Rhizopus nigricans probably by imparting direct
inhibitory effects on such post-harvest pathogens (Lazar et al., 2008;
Fan et al., 2008). That apart, it is very well established that NO is a
highly potent molecule for induction of resistance in crop plants
against systemic infections (Manjunatha et al., 2008), and such effects
can be extended for protecting the fruits from a broad range of
phytopathogens. This new direction emanated from research on NO
would allow more orderly designed formulations for the marketing of
high quality fruits endowed with longer storage life. However, the
problem associated with NO application is its high reactivity;

492

G. Manjunatha et al. / Biotechnology Advances 28 (2010) 489499

Table 1
Application of NO for shelf life extension.
Crop/fruit

Treatment

Extent of shelf life extension

(%)

Specic conditions

Reference

Broccoli
Cucumber
Chinese-broccoli
Kiwi
Mushroom
Strawberry
Strawberry fruit
Carnations

NO gas

Not specied

Leshem et al. (1998)

No signicance of storage temperature

20 C in humidied air
Treatment solution requires pH 6.5
Storage at 20 C

Wills et al. (2000)

Bowyer et al. (2003)

NO gas and DETANO

182
67
71
160
73
147
50
30
50
14
50
50

Treatment in solution with citric acid and

starch followed by storage at 10 C

Wills et al. (2007)

NO gas and DETANO

SNP
DETANO
NO gas

40
Up to 120 days
170
70

Storage at 2 C with 8590% RH

Treatment solution preferably acidic
No signicance of storage temperature and
treatment conditions

Zhang et al. (2007b)

Pristijono et al. (2008)
Wills et al. (2008)

DETANO solution

100

Green bean
Broccoli
Strawberry
Mushroom
Plum
Apple
Lettuce

NO gas
DETANO gas
DETANO in solution
NO gas

Soegiarto and Wills (2004)

DETANO diethylenetriamine-nitric oxide; SNP sodium nitroprusside.

therefore delivery of NO to biological items is a great challenge,

needing a strategic approach for developing formulations. Nevertheless, the possibility of elicitation of endogenous NO appears
advantageous, as discussed in Section 8 of this review.
3. Nitric oxideethylene cross talks
The earliest milestone report of 1940 indicated that NO caused
disturbances in ethylene formation by inhibiting hydrogenation of
ethylene from ethane (Burnham and Pease, 1940). This was later
established with data on ethylene being inversely regulated by NO in
growing plants (Leshem and Haramaty, 1996), as well as in fruits
during temporal progression of maturation and senescence as
demonstrated in avocado (Persea Americana), Citrus spp. Kiwi
(Actinidia chinensis), tomato (Solanum lycopersicum) and strawberries
(Leshem and Wills, 1998; Leshem and Pinchasov, 2000). NO
effectively prevented the autocatalytic ethylene biosynthesis in
climacteric peaches. The probable mechanism proposed here is the
binding of NO to ACC oxidase forming a binary ACC oxidaseNO
complex, which is then chelated by ACC to produce a stable ACCACC
oxidaseNO complex. This ternary complex, in turn, causes decrease
in the ethylene production. An irreversible transformation of ACC to
1-malonyl aminocyclopropane-1-carboxylic acid (MACC) has also
been noticed in NO-treated peach fruit. Yet another mode being that
the NO and/or peroxynitriles formed during interaction of NO with
H2O2 can also inhibit the activities of key enzymes of ethylene
pathway ACC synthase (ACS) and ACC oxidase (ACO), by oxidative
inactivation of co-factors resulting in an overall reduction in the turnover of ethylene (Zhu et al., 2006) (Fig. 2). Thus, the exogenous
application of NO markedly delayed maturation and senescence in
fruits where NO effect was stoichiometrically related to ethylene
suppression (Leshem et al., 1998). Similarly, the endogenously
generated NO also suppressed ethylene formation in situ impairing
progression of ripening in apple fruit. In addition, with the linear
generation of NO the ethylene generation decreased exponentially
indicating that a few molecules of NO are sufcient enough to
accomplish suppression of large amount of ethylene (Rudell and
Mattheis, 2006). In addition, the simultaneous treatment of peach
with NO and ethylene generating compounds decreased the ethylene
level in vivo, suggesting that NO competitively inhibits the actions of
both external as well as endogenous ethylene (Meng-chen et al.,
2007). Nevertheless, certain tissues displayed contrasting effects. For
instance, NO treatment enhanced ethylene during breaking of embryo

dormancy in apple (Gniazdowska et al., 2007). In tobacco plants NO

up-regulated the mRNA accumulation of ethylene biosynthetic gene
ACS2 in a concentration dependent manner resulting in higher
ethylene (Ederli et al., 2006). In tomato, the expressions of ACS
homologues such as ACS, LeACS2, LeACS4, LeACS6, LeACSH6A and
LeACSH6B were unaffected by NO whereas the expressions of ACO
genes such as LeACO1, LeACOH2 and LeACO4 were decreased/delayed
by prior treatment with NO during ripening (Eum et al., 2009). These
results are indicative that the ethylene pathway regulation by NO is
chiey through the suppression of ACO rather than ACS. In
Arabidopsis, NO markedly reduced ethylene by down-regulating
methionine adenosyl transferase1 (MAT1) activity through posttranslational S-nitrosylation regulation, accounting for the reduction
of the pool size of the chief ethylene precursor, the S-adenosyl
methionine (SAM), thus affecting the overall turn-over of ethylene
biosynthesis (Lindermayr et al., 2006). However, in banana, NO
altered ethylene production through alterations in the expression
levels of homologues of ACO and ACS genes (Cheng et al., 2009) (Fig. 2
and Table 1).
4. NO versus ROS
The oxidative burst in plants is mainly caused by reactive oxygen
species (ROS) that are established to have many cellular implications
in the cell cycle, programmed cell death, hormone signaling, biotic
and abiotic responses, including fruit ripening. Their excessive
quantities result in stress due to altered redox homeostasis, aberrant
cell signaling and massive disturbances of orchestrated cellular
functions (Bailey-Serres and Mittler, 2006). ROS in above threshold
levels can damage cellular composition such as protein and lipids,
which leads to the loss of membrane integrity and functionality
having special signicance in fruit ripening as senescence triggering
factor. Hence the maintenance of ROS at cellular level in balanced and
optimum level is necessary for maintaining the quality of fruits.
Therefore, maintenance of cellular homeostasis of ROS is very vital for
retarding their deleterious effects, and there are ample evidences
indicating NO's critical role in suppressing ROS molecules (Cheng et
al., 2002; Guo and Crawford, 2005), although their mutual effects are
paradoxical depending on respective concentrations (Delledonne et
al., 2001). In systems where ROS toxicity is incurred, NO can act as a
reaction cascade breaker and prevent damage (Farias-Eisner et al.,
1996; Durzan and Pedroso, 2002) (Fig. 3). This is perhaps one of the
modes by which browning of harvested fruits and senescence of

G. Manjunatha et al. / Biotechnology Advances 28 (2010) 489499

Fig. 3. Rare interactions of NO with ROS. NOS, an enzyme involved in biosynthesis of NO

predominantly uses arginine as substrate. ROS-generation is generally catalyzed by
NADPH oxidase. Under conditions of arginine depletion the same enzyme is responsible
for the formation of H2O2; in rare situation (shaded box) it is established that
Diphenylene iodonium, a specic inhibitor of ROS-generating NADPH oxidase, is also
responsible for the inhibition of NOS activity thereby inhibiting the H2O2-generation.

ornamentals are prevented. The other mode established is during

stress metabolism induced by abiotic (Orozco-Cardenas and Ryan,
2002) and biotic stresses (Maolepsza and Rozyalska, 2005) where ROS
effects are reversed by NO by directly suppressing either the activities
of ROS enzymes or relevant signaling cascade in a tightly coordinated
manner (Clark et al., 2000). For instance, AHb1, is a nonsymbiotic
hemoglobin that accomplishes the detoxication of NO to nitrate in
Arabiodopsis, and the rate of reaction here is so low that it does not
interfere when a burst of NO occurs during hypersensitive response
against pathogen (Perazzolli et al., 2004). Therefore it is very essential
to precisely monitor both NO and ROS thresholds compatible with the
metabolism of cellular compartment, calling for maintaining a specic
level of NO for modulating fruit ripening. The major enzymes involved
in ROS removal are catalase (CAT), peroxidases (POD) and superoxide
dismutase (SOD), which are signicantly up-regulated by NO in
peaches (Flores et al., 2008) and kiwi fruit (Zhu et al., 2008). Whereas
in plum, NO failed to suppress activities of polyphenol oxidase (PPO),
POD and phenylalanine ammonia lyase (PAL), although it caused a
delay in total phenol formation (Zhang et al., 2007b). Longan, litchi
and jujube fruits treated with NO showed inhibition of the activities of
PPO, POD and PAL and maintained a high total phenol homeostasis
throughout storage (Duan et al., 2007; Kramchote et al., 2007; Zhu et
al., 2009). The ROS effects were signicantly reversed by NO via
genetic up-regulation of SOD and CAT, and suppression of lipoxygenase (LOX) in kiwi fruit resulting in maintenance of vitamins C and E
(Zhu et al., 2008). Even in case of crops like potato, NO strongly
countered ROS mediated cytotoxic processes and prevented loss of
chlorophyll caused by methyl viologen herbicide diquat (a ROS
generator) (Beligni and Lamittina, 1999). Avoiding the formation of
one of the most deleterious reactive oxygen species, the hydroxyl
radical (OH), (Wink et al., 1995) has been linked to the scavenging of
iron by NO, thus causing prevention of Fenton reaction. In this
reaction NO easily forms ironnitrosyl complex with haem iron and
affects the functioning of plant POD involved in the cell wall
lignication (Ferrer and Ros Barcelo, 1999). Further, free radical
scavenging ability of NO may act preferentially in quenching of oquinone radicals resulting in the interruption of normal browning
reactions at the cut surfaces of fruits (Pristijono et al., 2008).
Conversely, the protection offered by NO to organic acids such as
ascorbic acid may also complement the prevention of browning.
Above all, one cannot ignore the inhibition of PAL by NO, the rst key
enzyme involved in the biosynthesis of phenolic compounds that

493

cause browning in fruits, which is triggered by various stress

conditions (Nguyen et al., 2003) (Box 2).
The biochemistry of reactive NO and ROS is closely inter-twined.
NOS, involved in NO biosynthesis using arginine as substrate, under
conditions of arginine depletion, synthesizes H2O2 (Stamler et al.,
1992). On the other hand, diphenylene iodonium, an inhibitor
considered specic for the ROS-generating NADPH oxidase and
H2O2-generating activity of peroxidases, also inhibits NOS activity
(Bolwell, 1999; Zhang and Zhao, 2008) (Fig. 3). Lipoxygenase (LOX) is
a key enzyme in lipid peroxidation and contributes to the development of oxidative stress occurring in fruit ripening and senescence.
Inhibition of LOX activity was proposed due to binding sites of NO to
Fe3+ from the LOX active site which is then reduced to Fe2+
inactivating the catalytic activity of the enzyme (Zhu et al., 2006).
LOX being a catalyst for the formation of jasmonic acid (JA) and JA
being a growth regulator/signal molecule physiologically implied
with ethylene (Zhu et al., 2006), its involvement in ripening is
anticipated. The inhibition of LOX by NO has been reported in kiwi
fruit during storage (Zhu et al., 2008). NO can also react with proteins
by nitrosation. Once the thiol group in protein is nitrosated, it is
possible for the exchange of NO between thiol groups by transnitrosation (Bruckdorfer, 2005). LOX and ACC oxidase could be
nitrosated by associating NO, and the nitrosation process could
continue within the proteins in peaches during storage. NO might
have an effect on enzymes by these processes during the extended
phase of storage (Zhu et al., 2006). Since ROS and NO exert reciprocal
control on each other in several ways, the possibility of regulating ROS
levels by modulating the activities of antioxidant enzymes nudges
ripening control by NO. Alternatively, the non-enzymatic small
molecules which ably reduce free radicals by self oxidation and
their stable oxidized forms and other molecules such as ascorbic acid,
carotenoids, phenolic compounds, and transition metals like iron,
copper and zinc, can also inuence endogenous NO during the
oxidation.
The other mechanism associated with oxidative burst in higher
plants is the lignication process which strengthens the cell wall by
cross-links with other proteins and polysaccharides, and offer
protection during abiotic and biotic stresses. For enhanced lignications the role of NO has been studied during abiotic stresses,
indicating possible similar implications for enhanced shelf life of
horticultural commodities (Ferrer and Ros Barcelo, 1999). These
studies are indicative of the fact that interaction of NO with ROS is still
far from being clearly elucidated.
5. Interplay of NO with other signals
The growing evidence suggests that NO as a signal molecule
interacts with various stress responsive signal networks, mainly SA, JA
and ethylene. That apart, NO also interacts with other secondary
signal molecules such as cADP ribose, cGMP and Ca2+, which are
intimately involved in H2O2 metabolism and phenyl propanoid
pathways (Durner et al., 1998; Wendehenne et al., 2001; Pagnussat
et al., 2003; Pagnussat et al., 2004; Lamotte et al., 2004; Romeropuertas et al., 2004). These interactions either directly or indirectly
modulate the transcription of specic genes (Parani et al., 2004)
involved in ripening and senescence. It has been shown that cGMP
triggers genes in encoding chalcone synthase and ferredoxin NADP+oxidoreductase and initiates anthocyanin biosynthesis in soybean
(Bowler et al., 1994) (Fig. 4).
5.1. Salicylic acid (SA)
SA is a signal molecule affecting a number of physiological
processes in plants, including thermogenesis, ethylene synthesis,
and fruit ripening (Rhoads and McIntosh, 1992). Treatment of tobacco
and A. thaliana leaves with donors of NO resulted in the induction of a

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G. Manjunatha et al. / Biotechnology Advances 28 (2010) 489499

Box 2
Nitric oxide generation in plants
NO is formed as a part of nitrogen cycle. The nitrificationdenitrification cycles liberate NO as a by-product (Goretski and Hollocher, 1988).
In addition to inorganic form, NO could be generated by non-enzymatic mechanisms, e.g. via chemical reduction of NO2 by light mediated
reactions of carotenoids (Cooney et al., 1994). In living systems, NO is a highly reactive molecule that rapidly diffuses and permeates the
cell membranes, the formation from being nitrite or nitrosothiols functioning as NO precusors helps in supplying NO for various biological
functions. In recent years, research on NO has been the subject of intensive investigation in animal subjects and humans, and to a lesser
extent in plant systems, probably due to its extensive versatility and complex interplay in vivo in a number of signaling pathways that
respond to various physiological demands. Hemoglobins are well known regulators of NO homeostasis, functioning through either
detoxification (Joshi et al., 2002) or transnitrosylation reactions (Gow et al., 1999) in humans. In bacteria, flavohemoglobins consume NO
enzymatically (Gardner et al., 1998; Hausladen et al., 2001). Yeast flavohemoglobin possesses the same NO reductase/denitrosylase
activity as in bacteria (Liu et al., 1999). NO was first identified as a unique diffusible molecular messenger in animals, from which emerges
the knowledge in plants accomplishing similar signal transductions. As in mammalian systems, endogenous NO generation in plants occur
due to the catalytic action of two main enzymes, namely NO synthase (NOS) and nitrate reductase (NR). In general NOS family catalyzes
the NADPH-dependent formation of NO (Furchgott, 1995; Li and Poulos, 2005). But in plants, there is no clear homologue of animal NOS
in the genome (The Arabidopsis Genome Initiative, 2000 www.genomenewsnetwork.org). Thus the plant enzyme displaying NOS-like
activity is structurally different from classical mammalian NOS. However, mammalian NOS inhibitors have been successfully used to
suppress NO production in plants under biotic and abiotic stresses (Zhang et al., 2007a,b; Zhao et al., 2007; Zottini et al., 2007),
indicating the mimicking nature of plant NOS with mammalian NOS.
In an attempt to further characterize NO generation in plants, a novel enzyme AtNOS1 of A. thaliana was identified having no similarity
with mammalian NOSs family. The protein was shown to produce NO from L-arginine in mitochondria (Guo et al., 2003; Guo and
Crawford, 2005), and this pathway has further been confirmed after elicitation with lipopolysaccharides (LPS) and virulent pathogens
(Zeidler et al., 2004). However, catalytic activity of AtNOS1 for generation of NO similar to mammalian systems was questioned and
further it has been renamed as A. thaliana nitric oxide associated-1 (AtNOA1) (Crawford, 2006; Zemojtel et al., 2006). Through
pharmacological approach using specific inhibitors, the existence of NOS in different plants was demonstrated, although the genetic
evidences for its regulation are far from being clear.
Using nitrite as substrate, plants generate NO by enzymatic as well as non-enzymatic routes. The former route is mainly catalyzed by
nitrate reductase (NR) a key enzyme of nitrogen assimilation (Yamasaki and Sakihama, 2000). NR is also the main enzyme for generation
in certain NO metabolic pathways, often intercepting with NOS. That apart, a root-specific plasma membrane bound nitriteNO reductase
(NINOR) was shown to catalyze the reduction of apoplastic nitrite into NO (Stohr et al., 2001). Other sources, including a yet-to-be
identified enzyme catalyzing a mitochondrial electron transport-dependent reduction of nitrite to NO (Planchet et al., 2005) and an
apoplastic non-enzymatic conversion of nitrite to NO, occurring at acidic pH in the presence of reductants such as ascorbic acid, have been
noted (Bethke et al., 2004). Recently, it was demonstrated that NO generation was associated with IRT1 which encodes a major plasma
membrane transporter for iron and this is responsible for the NO generation in Arabidopsis during cadmium toxicity (Besson-Bard et al.,
2009). NO was also generated by xanthine oxidoreductase (Harrison, 2002), horseradish peroxidase and other hemoproteins (Boucher et
al., 1992).

substantial increase in endogenous SA (Durner et al., 1998; Huang et

al., 2004). Alternatively, the effect of SA in inducing NO in Arabidopsis
was found to be associated with the dosage affecting a variety of
patho-physiological responses where the involvement of calcium and
casein kinase2 (CK2) suggests that SA induces NO partly through the
activity of a NOS-like enzyme (Zottini et al., 2007). In transgenic
tobacco NO effects were abolished by the expression of nahG, which
encodes an SA degrading enzyme salisylate hydroxylase (Song and
Goodman, 2001) indicating that SA and NO work synergistically in
certain plants. The mechanism of disrupted expression of ethylene
biosynthesis by SA in tobacco plants during incompatible interaction
with pathogens was found to be controlled by salicylate hydroxylase
gene expression through transcriptional control (Mur et al., 2008).
Several models suggest that redox signaling through NO and ROS is
enhanced by SA in a self-amplifying process (Klessig et al., 2000),
which may be one of the mechanisms of SA-mediated ripening control
in kiwi fruit and suppression of ethylene in banana (Srivastava and
Dwivedi, 2000; Yu et al., 2003). NO and SA synergistically caused
inhibition of JA with a concomitant decrease in ethylene and lipid
peroxidation after treatment with pyraclostrobin, a strobulirin, which
further supports antagonistic effects of NO on ethylene (Venancio et
al., 2003). Another study demonstrated NO and SA synergistic actions
affecting the ethylene metabolism, which then inuences the kinetics
of defense responses in tobacco disease resistance supporting NO's
intricacy in the alteration of ethylene formation (Mur et al., 2008)
(Fig. 4).

5.2. Jasmonic acid (JA)

JA, being a well-accepted signal molecule, triggers a cascade of
reactions, of which a few have been linked with NO. NO and JA were
found involved in defense responses in plants, such as the inhibition
of wounding-induced H2O2 production and proteinase inhibitor
synthesis in tomato leaves (Orozco-Cardenas and Ryan 2002), and
methyljasmonate-induced accumulation of H2O2 and malondialdehyde (MDA) in rice leaves (Hung and Kao, 2004). NO mediated the
fungal elicitor-induced hypericin production in Hypericum perforatum
cell suspension cultures through a JA-signal pathway (Xu et al., 2005)
and a strong NO burst was induced by JA as well by wounding in
Arabidopsis plants (Huang et al., 2004). Although synergistic relation
of NO and JA in wounding is established, contradictory inferences
have been derived in some studies where NO down regulated the LOX
(Zhu et al., 2008), a biosynthetic enzyme of JA having a negative
interference with the ripening syndrome in peach fruit (Ziosi et al.,
2008). While these studies have shed some light on the involvement
of various signal molecules, further elucidation is required at genetic
level for obtaining a comprehensive picture to extrapolate the results
and apply the same for fruit ripening (Fig. 4).
5.3. Growth regulators
In addition, another set of signaling molecules is endogenous
growth regulators such as abscisic acid (ABA) and indole-3-acetic acid

G. Manjunatha et al. / Biotechnology Advances 28 (2010) 489499

495

Fig. 4. A possible link between nitric oxide (NO) and other networks implicated in regulating ethylene in fruit ripening through identied (straight lines) and unidentied (dotted
lines) mechanisms. NO formed in the cell organelles by different routes interacts with primary and secondary signal molecules during plant growth and development and stress
stimuli. NO regulates H2O2 generated from NADPH oxidase during abiotic stresses, acts as antioxidant where NO quench pro-oxidants, NO is also involved in cell death through the
execution of systemic acquired resistance (SAR) against post-harvest pathogens, NO is also known to regulate MAP kinase for the cell wall cross linking and gene expression. NO
interactions with other secondary signal molecules like cGMP and cADRP and cascades of Ca++. Calmudulin, the calcium signal protein activates the hormones/systemic signals like
salicylic acid (SA), cytokinin and jasmonic acid (JA). Polyamines and NO regulation are less characterized, albeit together involved in the ethylene regulation.

(IAA) which are known to modulate climacteric fruit ripening (Pathak

and Sanwal 1999; Jiang et al., 2000) were also modulated by NO (Guo
et al., 2003; Pagnussat, et al., 2004). The mechanism underlying this
antagonism has not been elucidated, albeit the pharmacological
evidence in plant cell culture system prevailed indicating that NO is
involved in such responses (Carimi et al., 2003). The role of NO in
cytokinins signal transduction has also been documented without any
comprehensive inference. Such NO-cytokinin mediated synergistic
properties are reected in a study where NOS inhibitors inhibited
cytokinins-induced betalaine accumulation in Amaranthus seedlings
(Scherer and Holk, 2000) and that cytokinins-induced NO synthesis in
tobacco, parsley and Arabidopsis cell cultures (Tun et al., 2001). In case
of cytokinins-induced programmed cell death in plants, NO mediated
such events leading to cell death (Carimi et al., 2003) (Fig. 4).
Gibberellic acid (GA) is known to play a signicant role in the control
of ripening of several fruits, however, NO's role in interfacing GA
effects has never been recorded.
6. NO and polyamines (PAs)
PAs are basic molecules positively charged at physiological pH and
hence bind strongly with negatively charged nucleic acids (Feuerstein
et al., 1991), acidic phospholipids and many types of proteins (Carley
et al., 1983). Such ionic interactions are important in regulating the
structure and functions of biological macromolecules, as well as their
synthesis in vivo. It is well known that supplemental PAs provide antisenescence effects in many plant species (Smith, 1985). Such effects
have relevance in the context of fruit ripening (Malik and Singh, 2005)
since ripening is also a senescence phenomenon. In the case of tomato
fruit, PAs inhibited the transcript accumulation of the wound
inducible ACC synthase (Li et al., 1992). PA treatments in pomegranate enhanced the shelf life as well as quality attributes (Mirdehghan et
al., 2007), establishing the relationship between ethylene and PAs,
and the latter's suppressive effects on fruit ripening and senescence
(Katoh et al., 1987). The mechanism of PAs effects here is linked
chiey to the reduction of ethylene (as both share the common
precursor SAM) by post-transcriptional modication rather than the

reported feedback inhibition of enzymes of ethylene biosynthesis

(Pandey et al., 2000). Involvement of NO in this context is partially
understood when PA treatment induced the NO burst (Tun et al.,
2006). Putrescine (Put) application prior to storage suppressed
ethylene biosynthesis and retarded fruit softening during low
temperature storage in plum, albeit NO's involvement in this case
was not followed (Khan et al., 2007). Similarly, spermidine (Spd) and
spermine (Spm) were shown to reduce ethylene synthesis by
inhibition of ACC synthase (Bouchereau et al., 1999). Transgenic
tomato engineered to accumulate higher levels of Spd and Spm in
fruits, even at late ripening phases, was associated with activation of a
wide array of genes regulating transcription, translation, signal
transduction, chaperone activity, stress proteins, amino acid biosynthesis, ethylene biosynthesis and action, isoprenoid pathway and
avonoid biosynthesis (Mattoo and Handa, 2008). In addition, NO
may act downstream of PAs during stress phenomena of plants
(Arasimowicz-Jelonek et al., 2009). That apart, various reports also
suggest that PAs act as substitute/secondary messengers and activate
other signaling molecules, such as plant hormones and NO, which
further activate a vast genetic network involved in growth regulation
and senescence. In our study, treatment with Put signicantly
suppressed softening of banana with concomitant increase in
endogenously formed NO as well as Put, where the mechanism
involved is yet to be established (our unpublished data).
7. Application of NO and monitoring endogenous NO threshold
The diverse bio-regulatory functions of NO in metabolic responses
of plants are induced experimentally by the administration of
exogenous NO, using NO donor compounds. The NO molecules
released by its donors are able to either mimic an endogenous NOrelated response or substitute for an endogenous NO deciency. The
pathways leading to the formation of NO from a donor compound
differ greatly, while some require specic co-factors, others require
catalytic support of an enzyme, whereas, sodium nitroprusside (SNP)
requires photo-oxidation, thus posing a challenge for judicious
selection of NO donors and their application conditions. In addition

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G. Manjunatha et al. / Biotechnology Advances 28 (2010) 489499

to treatment conditions, the storage atmosphere after the NO

application would also inuence the shelf life and quality of the
produce (Table 2). In a treatment involving a mixture of citric acid and
starch saturated with gaseous NO, maintenance of storage temperature at 10 C was highly effective for strawberry and mushroom (Wills
et al., 2007). Similarly, treatment of apple slices with DETANO in acidic
condition was found useful for extending the storage life by 170%
(Pristijono et al., 2008). Broccoli, green bean, bok choy and carnations
treated under slightly acidic pH and storage at 20 C were more
benecial than treatment done at normal pH and storage at room
temperature (Soegiarto and wills, 2004; Bowyer et al., 2003)
(Table 2).
Several studies have established that NO absorption is faster than
its oxidation in the air and that the rate of NO absorption by each
horticulture produce is variable depending on the surface area of the
produce. For example, the absorption rate of NO per min per 100 g of
different horticulture produce is as fallows; lettuce-91.7; bok choy71.7; carnation-42.3; lily-29.2; strawberry-14.7; apple-8.7 banana5.3; tomato-5.1 and lime-0.7 (Soegiarto et al., 2003). Various other
unknown implications of biochemical nature within each commodity
cannot be ignored since the rate of NO absorption is almost
immediate. This also indicates the need for experimental data for
the use of not only the NO donors and co-factors, but also on the
effective use of NO scavengers and related enzyme inhibitors. Thus for
post-harvest treatments, NO concentration appears to play a key role
and application of NO gas per se and its donors to treat horticultural
materials appears economically feasible. The effect of cytoprotective
or cytotoxic action of NO on plant metabolism depends to a large
extent on the local concentration of the molecule and is affected by
the rate of synthesis, displacement and efciency of removal of this
reactive nitrogen species (Romero-Puertas et al., 2004). Therefore, NO
homeostasis holds an important key in biological functions of plants
in general, and in fruits in the present context. High concentrations
(4080 pphm) inhibited tomato growth, while low concentrations
(up to 20 pphm) enhanced it (Hufton et al., 1996), as with lettuce and
pea plants (Leshem and Haramaty, 1996). Therefore, delicate balance
between ROS and NO production in horticultural produce is crucial for
orchestrating large networks of genes, so that NO generating
compounds need to be administered with prior data of its consequences in each fruit commodity. However, NO delivery to organs
such as ripening fruit in the desired concentrations indicate a need for
highly judicious selection of compounds and treatments as well as
storage conditions for the development of effective formulations.

8. Generation of endogenous NO by elicitations

It is widely accepted that several metabolites in plants can be
induced in vivo/in planta through appropriate elicitation. NO can also
be similarly generated by a wide array of elicitors of abiotic and biotic
origin. Oligogalacturonic acid stimulated the NO generation in the
growth medium of Panax ginseng through activation of NOS (Hu et al.,
2003). Phyto-oligosaccharides stimulated the release of NO from
murine effector cells (Hirazumi and Furusawa, 1999) suggesting
potential interplay between NO and the cell wall oligosaccharins. The
commercially available chitosan as well as proteinaceous elicitor
derived from the cell wall of endophytic Penicillium sp. induced the
accumulation of NO in pearl millet plants and this elevated NO has
been implicated in one of the factors responsible for imparting disease
resistance (Manjunatha and Shetty, 2006; Manjunatha et al., 2009).
Elicitor, prepared from the cell walls of Penicillium citrinum induced
multiple responses in Catharanthus roseus suspension cells (Xu and
Dong, 2005a), one of which being the rapid generation of NO, causing
enhancement of catharanthine production (Xu and Dong, 2005b).
Fungal elicitor prepared from the cell walls of Aspergillus niger
induced NO generation in Hypericum perforatum cells (Xu et al., 2005).
Treatment of tobacco culture with Fungal elicitor from Phytophthora
cryptogea, strongly increased intracellular NO (Foissner et al., 2000).
Exogenously supplied MJ, a widely used elicitor, induced the rapid
production of NO in Taxus cell cultures and NO elevation has been
correlated with higher metabolite production of taxol, an anticancer
agent (Wang and Wu, 2005). Absciscic acid-induced H2O2 production
mediating NO generation, which, in turn, activated mitogen activated
protein kinase (MAPK) resulting in the transcriptional up-regulation
of the activities of antioxidant enzymes (Zhang et al., 2007a). Further,
along with a fungal elicitor cryptogein, NO has been shown to regulate
the ethylene responses in tobacco cells (Lamotte et al., 2004). These
studies provide an insight for the possible development of formulations targeted at in vivo NO generation in various fruits.
9. Conclusions and future prospects
The chemical biology of NO in plants is rapidly advancing,
unraveling various hidden facets hitherto unknown. The enormous
current developments with mammalian systems and the existence of
analogy in plants reveal other promising prospects for future studies.
We are at the beginning of the road in this NO-induced web of signal
cross-talks, having several questions to be answered through

Table 2
NO associated post-harvest biochemical changes.
Commodity

Treatment type

Biochemical effects

References

Strawberry
Broccoli
Cucumber
Kiwi
Mushroom
Bartlett Pears
Peaches

NO gas

Inhibition of ethylene production

Leshem et al. (1998)

NO gas
NO gas

Strawberry

SNP

Longan
Plum fruit

SNP
SNP donor

Apple slices
Banana slices

DETANO
SNP

Jujube fruit

NO fumigation

Inhibition of ACC oxidase, thus redirection of ethylene to

MACC production and inhibition of lipoxygenase
Inhibition of ethylene production, respiration rate, activity of
ACC synthase and reduction of ACC content
Modulation of the phenolic metabolism for reduced browning
Inhibition of pectin solubilisation and depolymerisation, reduced
phenolic content, increase in titratable acidity, lower DPPH
radical scavenging activity, chilling injury protection.
Inhibition browning of fruits
Inhibition of ACO activity and transcription of gene MA-ACO1, PG, PME,
and endo--1,4-glucanase and maintained higher contents of ASP and starch
Inhibition of PPO and PAL activities, maintained a low total anthocyanin
content and a high total phenol content, and delayed the increase of
soluble solids and decrease of vitamin C.

DPPH: ,-diphenyl--picryl hydrazyl; ASP: acid-soluble pectin.

Sozzi et al. (2003)

Zhu et al. (2006)
Zhu and Zhou (2007).
Duan et al. (2007)
Zhang et al. (2007b)

Pristijono et al. (2008)

Cheng et al. (2009)
Zhu et al. (2009)

G. Manjunatha et al. / Biotechnology Advances 28 (2010) 489499

judicious experiments with utmost care. Nonetheless, available data

have established NO's role in senescence-delay, acting primarily but
not solely, by limiting ethylene emission resulting in delayed ripening.
Most of the recent studies related to NO and ripening are intriguing, as
fruit ripening has broader inuence on aspects of climacteric
metabolism and senescence thereafter. Therefore, the mechanisms
of NO involvement between both climacteric and non-climacteric
have to be interpreted appropriately for exploiting any commercial
benets. The ultimate results obtained with NO treatment are similar
to that of 1-MCP although 1-MCP the works by blocking the ethylene
perception and NO works by suppressing ethylene generation per se in
various horticultural produce. It is heartening that ethylene suppression by endogenous NO regulation is possible through elicitor usage.
Also, the application of nano technology for NO delivery with
precision may open up enormous opportunities. Undoubtedly, the
genetic links among pathways of NO generation, signaling networks,
possible linking cross-talk pathways such as defense and ROS
homeostasis need to be established through transcriptome analysis
for efcient ethylene control in fruits.
Acknowledgements
First and second authors acknowledge the Department of
Biotechnology and Council of Scientic and Industrial Research,
India, respectively for fellowship. Encouragement by Dr. G. A.
Ravishankar, Head, Plant Cell Biotechnology Department, Central
Food Technological Research Institute (CFTRI) and Dr. V. Prakash,
Director, CFTRI are gratefully acknowledged.
References
Antonio MJ, Nigel GE, Mi-Young C, James GJ, Jocelyn RKC. Biology and genetic
engineering of fruit maturation for enhanced quality and shelf-life. Curr Opin
Biotechnol 2009;20:197203.
Arasimowicz-Jelonek M, Floryszak-Wieczorek J, Kubis J. Interaction between polyamine
and nitric oxide signaling in adaptive responses to drought in cucumber. J Plant
Growth Regul 2009;28:17786.
Bailey-Serres J, Mittler R. The roles of reactive oxygen species in plant cells. Plant
Physiol 2006;141:3116.
Beligni MV, Lamattina L. Nitric oxide stimulates seed germination and de-etiolation and
inhibits hypocotyl elongation: three light-inducible responses in plants. Planta
2000;210:21521.
Beligni MV, Lamattina L. Nitric oxide counteracts cytotoxic processes mediated by
reactive oxygen species in plant tissues. Planta 1999;208:33744.
Beligni MV, Fath A, Bethke PC, Lamattina L, Jones RL. Nitric oxide acts as an antioxidant
and delays programmed cell death in barley aleurone layers. Plant Physiol
2002;129:164250.
Besson-Bard A, Antoine G, Pierre R, Pascaline A, Duc, Frederic GC, et al. Nitric oxide
contributes to cadmium toxicity in Arabidopsis by promoting cadmium accumulation in roots and by up-regulating genes related to iron uptake. Plant Physiol
2009;149:130215.
Bethke PC, Badger MR, Jones RL. Apoplastic synthesis of nitric oxide by plant tissues.
Plant Cell 2004;16:33241.
Bhagyalakshmi N, Prabha TN, Yashodha HM, Prasanna V, Jagadeesh BH, Tharanathan
RN. Biotechnological approach for textural regulation in banana and mango fruit
during ripening. Acta Hort 2002;575:71724.
Bolwell GP. Role of reactive oxygen species and nitric oxide in plant defense responses.
Curr Opin Plant Biol 1999;2:28794.
Boucher JL, Genet A, Vadon S, Delaforge M, Mansuy D. Formation of nitrogen oxides and
citrulline upon oxidation of N-hydroxy-L-arginine by hemeproteins. Biochem
Biophys Res Commun 1992;184:115864.
Bouchereau A, Aziz A, Larher F, Martin-Tanguy J. Polyamines and environmental
challenges:recent developments. Plant Sci 1999;140:10325.
Bowler C, Neuhaus G, Yamagata H, Chua N. Cyclic GMP and calcium mediate
Phytochrome transduction. Cell 1994;77:7381.
Bowyer MC, Wills RBH, Badiyan D, Ku VVV. Extending the postharvest life of carnations
with nitric oxide comparison of fumigation and in vivo delivery. Postharvest Biol
Technol 2003;30:2816.
Bruckdorfer R. The basics about nitric oxide. Mol Aspects Med 2005;26:3-31.
Burnham HD, Pease RN. Inhibition of the hydrogenation of ethylene by nitric oxide.
J Am Chem Soc 1940;62:453.
Carimi FM, Zottini E, Formentin M, Terzi, Lo Schiavo F. Cytokinins, new apoptotic
inducers in plants. Planta 2003;216:41321.
Carley E, Wolosiuk RA, Hertig CM. Regulation of the activation of chloroplast fructose1,6-bis phosphatase (E.C 3.1.11). Inhibition by spermidine and spermine. Biochem
Biophys Res Commun 1983;115:70710.

497

Cheng FY, Hsu SY, Kao CH. Nitric oxide counteracts the senescence of detached rice
leaves induced by dehydration and polyethylene glycol but not by sorbitol. Plant
Growth Regul 2002;38:26572.
Cheng G, En Y, Wangjin L, Yongxia J, Yueming J, Xuewu D. Effect of nitric oxide on
ethylene synthesis and softening of banana fruit slice during ripening. J Agric Food
Chem 2009;57:5799804.
Clark J, Durner D, Navarre A, Klessig DF. Nitric oxide inhibition of tobacco catalase and
ascorbate peroxidase. Mol PlantMicrobe Int 2000;13:13804.
Cooney RV, Harwood PJ, Custer LJ, Franke AA. Light-mediated conversion of nitrogen dioxide to nitric oxide by carotenoids. Environ Health Perspect 1994;102:
4602.
Correa-Aragunde N, Graziano M, Lamattina L. Nitric oxide plays a central role in
determining lateral root development in tomato. Planta 2004;218:9005.
Crawford NM. Mechanisms for nitric oxide synthesis in plants. J Exp Bot 2006;57:
4718.
DeEll JR, Murr DP, Wiley L, Porteous MD. 1-Methylcyclopropene (1-MCP) increases CO2
injury in apples. ActaHortic 2003;600:27780.
Delledone M, Xia Y, Dixon RA, Lamb C. Nitric oxide functions as a signal in plant disease
resistance. Nature 1998;394:5858.
Delledonne M, Zeier J, Marocco A, Lamb C. Signal interactions between nitric oxide and
reactive oxygen intermediates in the plant hypersensitive disease resistance
response. Proc Natl Acad Sci USA 2001;98:134549.
Desikan R, Grifths R, Hancock J, Neill S. A new role for an old enzyme:nitrate
reductase-mediated nitric oxide generation is required for abscisic acid-induced
stomatal closure in Arabidopsis thaliana. Proc Natl Acad Sci USA 2002;99:163148.
Duan X, Su X, You Y, Qu H, Li Y, Jiang Y. Effect of nitric oxide on pericarp browning of
harvested longan fruit in relation to phenolic metabolism. Food Chem 2007;104:
5716.
Durner J, Wendehenne D, Klessig DF. Defense gene induction in tobacco by nitric oxide,
cyclic GMP, and cyclic ADP-ribose. Proc Natl Acad Sci USA 1998;95:1032833.
Durzan DJ, Pedroso MC. Nitric oxide and reactive nitrogen oxide species in plants.
Biotechnol Genet Eng Rev 2002;19:293337.
Ederli L, Morettini R, Borgogni A, Wasternack C, Miersch O, Reale L, et al. Interaction
between nitric oxide and ethylene in the induction of alternative oxidase in ozonetreated tobacco plants. Plant Physiol 2006;142:595608.
Eum HL, Kim HB, Choi SB, Lee SK. Regulation of ethylene biosynthesis by nitric oxide in
tomato (Solanum lycopersicum L.) fruit harvested at different ripening stages. Eur
Food Res Technol 2009;228:3318.
Fan XT, Maththeis JP. 1-Methylcyclopropene and storage temperature inuence
responses of Gala apple fruit to gamma irradiation. Postharvest Biol Technol
2001;23:14351.
Fan B, Shen L, Liu K, Zhao D, Yu M. Interaction between nitric oxide and hydrogen
peroxide in post harvest tomato resistance response to Rhizopus nigricans. J Sci
Food Agric 2008;88:123844.
Farias-Eisner R, Chaudhuri G, Aeberhard E, Fukuto JM. The chemistry and tumoricidal
activity of nitric oxide/hydrogen peroxide and the implications to cell resistance/
susceptibility. J Biol Chem 1996;271:614451.
Ferrer MA, Ros Barcelo A. Differential effects of nitric oxide on peroxidase and H2O2
production by the xylem of Zinnia elegans. Plant Cell Environ 1999;22:8917.
Feuerstein BG, Williams LD, Basu HS, Marton LJ. Implications and concepts of
polyamine-nucleic acid interactions. J Cell biochem 1991;46:3747.
Flores BF, Sanchez-Bel P, Monika V, Felix R. Maria, Isabel EM. Effects of a pretreatment
with nitric oxide on peach (Prunus persica L.) storage at room temperature. Eur
Food Res Technol 2008;227:1599611.
Foissner I, Wendehenne D, Langebartels C, Durner J. In vivo imaging of an elicitorinduced nitric oxide burst in tobacco. Plant J 2000;23:81724.
Francisco FB, Paloma S, Monika V, Felix R, Isabel EM. Effects of a pretreatment with
nitric oxide on peach (Prunus persica L.) storage at room temperature. Eur Food Res
Technol 2008;227:1599611.
Furchgott RF. Special topics:nitric oxide, arginase-negative mutants of Arabidopsis
exhibit increased NO signaling in root development. Annu rev Physiol 1995;57:
65982.
Gardner PR, Costantino G, Salzman AL. Constitutive and adaptive detoxication of nitric
oxide in Escherichia coli. J Biol Chem 1998;273:2652833.
Giovannoni J. Genetic regulation of fruit development and ripening. Plant Cell 2004;16:
17080.
Gniazdowska A, Urszula D, Tomasz B, Renata B. Breaking the apple embryo dormancy
by nitric oxide involves the stimulation of ethylene production. Planta 2007;225:
10517.
Goretski JT, Hollocher TC. Trapping of nitric-oxide produced during denitrication by
extracellular hemoglobin. J Biol Chem 1988;263:2316-232.
Gow AJ, Luchsinger BP, Pawloski JR, Singel DJ, Stamler JS. The oxyhaemoglobin reaction
of nitric oxide. Proc Natl Acad Sci USA 1999;96:902732.
Graziano M, Beligni MV, Lamattina L. Nitric oxide improves iron availability in plants.
Plant Physiol 2002;130:18529.
Guo FQ, Crawford NM. Arabidopsis nitric oxide synthase1 is targeted to mitochondria
and protects against oxidative damage and dark-induced senescence. Plant Cell
2005;17:343650.
Guo F, Okamoto M, Crawford NM. Identication of a plant nitric oxide synthase gene
involved in hormonal signaling. Science 2003;302:1003.
Harrison R. Structure and function of xanthine oxidoreductase: where are we now?
Free Radic Biol Med 2002;33:77497.
Hausladen A, Gow A, Stamler JS. Flavohemoglobin denitrosylase catalyzes the reaction of a
nitroxyl equivalent with molecular oxygen. Proc Natl Acad Sci USA 2001;98:1010812.
He Y, Tang RH, Hao Y, Stevens RD, Cook CW, Ahn SM, et al. Nitric oxide represses the
Arabidopsis oral transition. Science 2004;305:196871.

498

G. Manjunatha et al. / Biotechnology Advances 28 (2010) 489499

Hirazumi A, Furusawa E. An immunomodulatory polysaccharide-rich substance from

the fruit juice of Morinda citriora (Noni) with antitumour activity. Phytother Res
1999;13:3808.
Hofman PJ, Jobin-Decor M, Meiburg GF, Macnish AJ, Joyce DC. Ripening and quality
responses of avocado, custard apple, mango and papaya fruit to 1-methylcyclopropene. Aust J Exp Agric 2001;41:56772.
Hu X, Neill SJ, Cai W, Tang Z. Nitric oxide mediates elicitor-induced saponin synthesis in
cell cultures of Panax ginseng. Fun Plant Biol 2003;30:9017.
Huang X, Stettmaier K, Michel C, Hutzler P, Mueller MJ, Durner J. Nitric oxide is induced
by wounding and inuences jasmonic acid signaling in Arabidopsis thaliana. Planta
2004;218:93846.
Hufton CA, Besford RT, Wellburn AR. Effects of NO (+NO2) pollution on growth, nitrate
reductase activities and associated protein contents in glasshouse lettuce grown
hydroponically in winter CO2 enrichment. New Phytol 1996;133:495501.
Hung KT, Kao CH. Nitric oxide acts as an antioxidant and delays methyl jasmonateinduced senescence of rice leaves. J Plant Physiol 2004;161:4352.
Jiang Y, Joyce DC, Macnish AJ. Effect of abscisic acid on banana fruit ripening in relation
to the role of ethylene. J Plant Growth Regul 2000;19:10611.
Jiang YM, Joyce DC, Jiang WB, Lu WJ. Effects of chilling temperatures on ethylene
binding by banana fruit. Plant Growth Regul 2004;43:10915.
Joshi MS, Ferguson TBJ, Han TH, Hyduke DR, Liao JC, Rassaf T, et al. Nitric oxide is
consumed, rather than conserved, by reaction with oxyhaemoglobin under
physiological conditions. Proc Natl Acad Sci USA 2002;99:103416.
Katoh Y, Hasegawa T, Suzuki T. Fujji. Effect of 1-aminocyclopropane 1-carboxylic acid
production on the changes in the polyamine levels in Hiproly barley callus after
auxin withdrawal. Agric Biol Chem 1987;51:245763.
Khan SA, Singh Z, Nadeem A. Pre-storage putrescine application suppresses ethylene
biosynthesis and retards fruit softening during low temperature storage in
Angelino plum. Postharvest Biol Technol 2007;46:3646.
Klessig DF, Durner J, Noad R, Navarre DA, Wendehenne D, Kumar D, et al. Nitric oxide and
salicylic acid signaling in plant defense, Proc Natl Acad Sci USA 2000;97:884955.
Kramchote S, Wongs-Aree V, Srilaong A, Uthairatanakij S, Kanlayanarat. Inuence of
nitric oxide on pericarp browning of litchi cv. chakrapad. ISHS Acta Horticulturae
804: Europe-Asia Symposium on Quality Management in Postharvest Systems
Eurasia 2007.
Ku VVV, Wills RBH, Ben-Yehoshua S. 1-methylcyclopropene can differentially affect the
postharvest life of strawberries exposed toethylene. Hort Sci 1999;34:11920.
Lamotte O, Gould K, Lecourieux D, Sequeira-Legrand A, Lebrun-Garcia A, Durner J, et al.
Analysis of nitric oxide signaling functions in tobacco cells challenged by the
elicitor cryptogein. Plant Physiol 2004;135:51629.
Lazar EE, Wills RBH, Ho BT, Harris AM, Spohr LJ. Antifungal effect of gaseous nitric oxide
on mycelium growth, sporulation and spore germination of the postharvest
horticulture pathogens, Aspergillus niger, Monilinia fructicola and Penicillium
italicum. Lett Appl Microb 2008;46:68892.
Leshem YY, Wills RBH, Ku VVV. Evidence for the function of the free radical gas nitric
oxide (NO) as an endogenous maturation and senescence regulating factor in
higher plants. Plant Physiol Biochem 1998;36:82533.
Leshem YY. Nitric oxide in plants: occurrence, function and use. Kluwer; 2000.
Dordrech, The Netherlands.
Leshem YY, Haramaty E. Characterisation and contrasting effects of the nitric oxide free
radical in vegetable stress and senescence of Pisum sativum Linn. foliage. J Plant
Physiol 1996;148:25863.
Leshem YY, Pinchasov Y. photoacoustic spectroscopic determination of relative
endogenous nitric oxide and ethylene content stoichiometry during the ripening
of strawberries Fragaria anannasa (Duch.) and avocados Persea Americana (Mill.). J
Exp Bot 2000;51:14713.
Leshem YY, Wills RBH. Harnessing senescence delaying gases nitric oxide and nitrous
oxide: a novel approach to postharvest control of fresh horticultural produce. Biol
Plant 1998;41:1-10.
Li H, Poulos TL. Structure-function studies on nitric oxide synthases. J Inorg Biochem
2005;99:293305.
Li N, Parsons BR, Liu D, Mattoo AK. Accumulation of wound inducible ACC synthase
transcripts in tomato fruit is inhibited by salicylic acid and polyamines. Plant Mol
Biol 1992;18:47787.
Lindermayr C, Saalbach G, Bahnweg G, Durner J. Differential inhibition of Arabidopsis methionine adenosyltransferases by protein S-nitrosylation. J Biol Chem 2006;281:428591.
Liu L, Zeng M, Stamler JS. Hemoglobin induction in mouse macrophages. Proc Natl Acad
Sci Usa 1999;96:66437.
Malik AU, Singh Z. Pre-storage application of polyamines improves shelf-life and fruit
quality of mango. J Hort Sci Biotechnol 2005;80:3639.
Manjunatha G, Shetty HS. Involvement of nitric oxide signaling in induction of systemic
resistance in pearl millet by endophytic cell wall protein of Penicillium sp. J Mycol
Plant Pathol 2006;36:392404.
Manjunatha G, Niranjan Raj S, Nandini S, Shetty HS. Nitric oxide donor seed priming
enhances defense responses and induces resistance against pearl millet downy
mildew disease. Pestic Biochem Physiol 2008;91:1-11.
Manjunatha G, Niranjan-Raj S, Geetha NP, Deepak S, Amruthesh KN, Shetty HS. Nitric
oxide is involved in chitosan-induced systemic resistance in pearl millet against
downy mildew disease. Pest Manag Sci 2009;65:73743.
Maolepsza, Rozyalska S. Nitric oxide and hydrogen peroxide in tomato resistance nitric
oxide modulates hydrogen peroxide level in hydroxyethylorutin-induced resistance to Botrytis cinerea in tomato Plant. Physiol Biochem 2005;43:62335.
Mattoo AK, Handa AK. Higher polyamines restore and enhance metabolic memory in
ripening fruit. Plant Sci 2008;174:38693.
Meng-chen L, Wei-hon S, Shu-hua Z, Jie Z. Effects of nitric oxide and exogenous ethylene
treatments on ethylene biosynthesis in Feicheng peach. Agric Sci China 2007;6:2905.

Mirdehghan SH, Rahemi M, Castillo S, Martinez-Romero D, Maria S, Valero D. Prestorage application of polyamines by pressure or immersion improves shelf-life of
pomegranate stored at chilling temperature by increasing endogenous polyamine
levels. Postharvest Biol Technol 2007;44:2633.
Mur LAJ, Lucas JJ, Laarhoven, Frans JM, Harren, Michael AH, et al. Nitric oxide interacts
with salicylate to regulate biphasic ethylene production during the hypersensitive
response. Plant Physiol 2008;148:153746.
Murgia I, de Pinto MC, Delledonne M, Soave C, de Gara L. Comparative effects of various
nitric oxide donors on ferritin regulation, programmed cell death, and cell redox
state in plant cells. J Plant Physiol 2004;161:77783.
Nguyen TBT, Ketsa S, van Doorn WG. Relationship between browning and the activities
of polyphenol oxidase and phenylalanine ammonia lyase in banana peel during low
temperature storage. Postharvest Biol Technol 2003;30:18793.
Orozco-Cardenas ML, Ryan CA. Nitric oxide negatively modulates wound signaling in
tomato plants. Plant Physiol 2002;130:48793.
Otvos K, Pasternak TP, Miskolczi PM, Domoki M, Dorjgotov D, Szucs A, et al. Nitric oxide
is required for, and promotes auxin-mediated activation of, cell division and
embryogenic cell formation but does not inuence cell cycle progression in alfalfa
cell cultures. Plant J 2005;43:84960.
Pagnussat GC, Lanteri ML, Lamattina L. Nitric oxide and cyclic GMP are messengers in the
indole acetic acid-induced adventitious rooting process. Plant Physiol 2003;132:
12418.
Pagnussat GC, Lanteri ML, Lombardo MC, Lamattina L. Nitric oxide mediates the indole
acetic acid induction activation of a mitogen-activated protein kinase cascade
involved in adventitious root development. Plant Physiol 2004;135:27986.
Pandey S, Ranad SA, Nagar PK, Kumar N. Role of polyamines and ethylene as modulators
of plant senescence. J Biosci 2000;25:2919.
Parani M, Rudrabhatla S, Myers R, Weirich H, Smith B, Leaman DW, et al. Microarray
analysis of nitric oxide responsive transcripts in Arabidopsis. Plant Biotechnol J
2004;2:35966.
Pathak N, Sanwal GG. Regulation of the ripening of the banana (Musa acuminata) fruits
by chemicals. Indian J Agric Sci 1999;69:1720.
Perazzolli M, Dominici P, Romero-Puertas MC, Zago EZ, Sonoda M, Lamb C, et al.
Arabidopsis nonsymbiotic haemoglobin AHb1 modulates nitric oxide bioactivity.
Plant Cell 2004;16:278594.
Planchet E, Gupta KJ, Sonoda M, Kaiser WM. Nitric oxide emission from tobacco leaves
and cell suspensions: rate-limiting factors and evidence for the involvement of
mitochondrial electron transport. Plant J 2005;41:73243.
Porat R, Weiss B, Cohen L, Daus A, Goren R, Droby S. Effects of ethylene and 1methylcyclopropene on the postharvest qualities of Shamouti oranges. Postharvest Biol Technol 1999;15:15563.
Prabha TN, Bhagyalakshmi N. Carbohydrate metabolism in ripening banana fruit.
Phytochem 1998;48:9159.
Prado AM, Portereld DM, Feijo JA. Nitric oxide is involved in growth regulation and reorientation of pollen tubes. Development 2004;131:270714.
Pristijono P, Wills RBH, Golding JB. Use of the nitric oxide-donor compound,
diethylenetriamine-nitric oxide (DETANO), as an inhibitor of browning in apple
slices. J Hort Sci Biotech 2008;83:5558.
Rhoads DM, McIntosh L. Salicylic acid regulation of respiration in higher plants:
alternative oxidase expression. Plant Cell 1992;4:11319.
Romero-Puertas MC, Perazzolli M, Zago ED, Delledonne M. Nitric oxide signaling
functions in plant-pathogen interactions. Cell Microb 2004;6:795803.
Rudell RD, Mattheis PJ. Nitric oxide and nitrite treatments reduce ethylene evolution
from apple fruit disks. Horti Sci 2006;41:14625.
Scherer GFE, Holk A. NO donors mimic and inhibit cytokinins in betalaine accumulation
in Amaranthus caudatus. Plant Growth Regul 2000;32:34550.
Singh SP, Singh Z, Swinny EE. Post harvest nitric oxide fumigation delays fruit ripening
and alleviates chilling injury during cold storage of Japanese plums (Prunus salicina
Lindell). Postharvest Biol Technol 2009;53:1018.
Smith TA. Polyamines. Annu Rev Plant Physiol 1985;36:11743.
Soegiarto L, Wills RBH. Short term fumigation with nitric oxide gas in air to extend
the postharvest life of broccoli, green bean, and bok choy. Horti Technol 2004;14:
53840.
Soegiarto L, Wills RBH, Serberry JA, Leshem YY. Nitric oxide degradation in oxygen
atmosphere and rate of uptake by horticulture produce. Postharvest Biol Technol
2003;28:32731.
Song FR, Goodman M. Activity of nitric oxide is dependent on, but is partially required
for function of salicylic acid in the signaling pathway in tobacco systemic acquired
resistance. Mol Plant Microb Int 2001;14:145862.
Sozzi GO, Trinchero GF, Fraschina AA. Delayed ripening of Bartlet'pears treated with
nitric oxide. J Hort Sci Biotechnol 2003;78:899903.
Srivastava MK, Dwivedi UN. Delayed ripening of banana fruit by salicylic acid. Plant Sci
2000;158:8796.
Stamler JS, Singel DJ, Loscalzo J. Biochemistry of nitric oxide and its redox-activated
forms. Science 1992;258:1898902.
Stohr C, Strube F, Marx G, Ullrich WR, Rockel P. A plasma membrane-bound enzyme of
tobacco roots catalyses the formation of nitric oxide from nitrite. Planta 2001;212:
83541.
Tun NN, Holk A, Scherer GFE. Rapid increase of NO release in plant cell cultures induced
by cytokinin. FEBS Lett 2001;509:1746.
Tun NN, Santa-Catarina C, Vanildo S, Walter H, Eny Iochevet SF, Gunther FES.
Polyamines induce rapid biosynthesis of nitric oxide (NO) in Arabidopsis thaliana
seedlings. Plant Cell Physiol 2006;47:34654.
Venancio WS, Marco A, Rodrigues T, Begliomini E, de Souza NL. Physiological effects of
strobilurin fungicides on plants. Publ. UEPG Ci. Exatas Terra, Ci. Agr. Eng., Ponta
Grossa 2003;3:5968.

G. Manjunatha et al. / Biotechnology Advances 28 (2010) 489499

Wang JW, Wu JY. Nitric oxide is involved in methyl jasmonate induced defense
responses and secondary metabolism activities of Taxus cells. Plant Cell Physiol
2005;46:92330.
Watkins BC. The use of 1-methylcyclopropene (1-MCP) on fruits and vegetables.
Biotechnol Adv 2006;24:389409.
Watkins CB, Nock JF. SmartFresh(1-MCP) the good and bad as we head into the
2004 season. New York Fruit Quarterly 2004;12:38.
Wendehenne D, Pugin A, Klessig DF, Durner J. Nitric oxide: comparative synthesis and
signaling in animal and plant cells. Trends Plant Sci 2001;6:17783.
Wilhelmova H, Fuksova M, Srbova D, Mikov Z, Mytinova D, Prochazkova R, et al. The
effect of plant cytokinin hormones on the production of ethylene, nitric oxide, and
protein nitrotyrosine in ageing tobacco leaves. Biofators 2006;27:20311.
Wills RBH, Ku VVV, Leshem YY. Fumigation with nitric oxide to extend the postharvest
life of strawberries. Postharvest Biol Technol 2000;18:759.
Wills RBH, Soegiarto L, Bowyer. Use of solid mixture containing diethylnetriamine/
nitric oxide (DETANO) to liberate nitric oxide gas in the presence of horticultural
produce to extend postharvest life. Nitric Oxide 2007;17:449.
Wills RBH, Pristijono P, Golding JB. Browning on the surface of cut lettuce slices
inhibited by short term exposure to nitric oxide (NO). Food Chem 2008;107:
138792.
Wink DA, Cook JA, Pacelli R, Liebmann J, Krishne MC, Mitchell JB. Nitric oxide (NO)
protects against cellular damage by reactive oxy gen species. Toxicol Let 1995;82:
2216.
Xu M, Dong J. Nitric oxide stimulates indole alkaloid production in Catharanthus roseus
cell suspension cultures through a protein kinase dependent signal pathway.
Enzyme Microb Technol 2005a;37:4953.
Xu M, Dong J. Elicitor-induced nitric oxide burst is essential for triggering catharanthine
synthesis in Catharanthus roseus suspension cells. App Microb Biotech 2005b;67:
7829.
Xu MJ, Dong JF, Zhu MY. Nitric oxide mediates the fungal elicitor-induced hypericin
production of Hypericum perforatum cell suspension cultures through a jasmonicacid-dependent signal pathway. Plant Physiol 2005;139:9918.
Yamasaki H, Sakihama Y. Simultaneous production of nitric oxide and peroxynitrite by
plant nitrate reductase: in vitro evidence for the NR-dependent formation of active
nitrogen species. FEBS Let 2000;468:8992.
Yu z, Kun-Song C, Qing-Jun C, Shang-Long Z, Yi-Ping R. Effects of acetylsalicylic acid
(ASA) and ethylene treatments on ripening and softening of postharvest kiwi fruit.
Acta Bot Sin 2003;45:144752.

499

Zanella A. Control of apple supercial scald and ripening a comparison between 1methylcyclopropene and diphenylamine postharvest treatments, initial low
oxygen stress and ultra low oxygen storage. Postharvest Biol Technol 2003;27:
6978.
Zeidler D, Zahringer U, Gerber I, Dubery I, Hartung T, Bors W, et al. Innate immunity in
Arabidopsis thaliana: lipopolysaccharides activate nitric oxide synthase (NOS) and
induce defense genes. Proc Natl Acad Sci USA 2004;101:158116.
Zemojtel T, Frochlich A, Palmieri MC. Plant nitric oxide synthase: a never-ending story?
Trends Plant Sci 2006;11:5245.
Zhang L, Zhao L. Production of nitric oxide under ultraviolet-b irradiation is mediated
by hydrogen peroxide through activation of nitric oxide synthase. J Plant Biol
2008;51:395400.
Zhang A, Jiang M, Zhang J, Ding H, Xu S, Hu X, et al. Nitric oxide induced by hydrogen
peroxide mediates abscisic acid-induced activation of the mitogen-activated
protein kinase cascade involved in antioxidant defense in maize leaves. New
Phytol 2007a;175:3650.
Zhang DD, Cheng GP, Li J, Yi C, Yang E, Qu HX, et al. Effect of Nitric oxide on disorder
development and quality maintenance of plum stored at low temperature. ISHS
acta horticulture 804: EuropeAsia symposium on quality management in
postharvest systems Eurasia; 2007b.
Zhao MG, Tian QY, Zhang WH. Nitric oxide synthase-dependent nitric oxide production
is associated with salt tolerance in Arabidopsis. Plant Physiol 2007;144:20617.
Zhu S, Zhou E. Effect of nitric oxide on ethylene production in strawberry fruit during
storage. Food Chem 2007;100:151722.
Zhu S, Liu M, Zhou J. Inhibition by nitric oxide of ethylene biosynthesis and
lipoxygenase activity in peach fruit during storage. Postharvest Biol Technol
2006;42:418.
Zhu S, Lina S, Mengchen L, Jie Z. Effect of nitric oxide on reactive oxygen species and
antioxidant enzymes in kiwi fruit during storage. J Sci Food Agric 2008;88:
232431.
Zhu S, Sun L, Zhou J. Effects of nitric oxide fumigation on phenolic metabolism of
postharvest Chinese winter jujube (Zizyphus jujuba Mill. cv. Dongzao) in relation to
fruit quality. LWTFood Sci Technol 2009;42:100914.
Ziosi V, Bonghi C, Maria BA, Trainotti L, Biondi S, Sutthiwal S, et al. Jasmonate-induced
transcriptional changes suggest a negative interference with the ripening
syndrome in peach fruit. J Exp Bot 2008;59:56373.
Zottini M, Costa A, De Michele R, Ruzzene M, Carimi F, Lo Schiavo F. Salicylic acid
activates nitric oxide synthesis in Arabidopsis. J Exp Bot 2007;58:1397405.