J. Agr. Sci. Tech. (2022) Vol.

24(1): 213-226

Effect of Nanopriming with Zinc Oxide and Silver

Nanoparticles on Storage of Chickpea Seeds and
Management of Wilt Disease

G. Das1 and P. Dutta2*

ABSTRACT

Chickpea is an important pulse crop of India, but its productivity is quite low due to
several biotic and abiotic stresses. Low seed vigor is one of the issues that occur due to
changes in different biochemical properties during improper storage condition. To
overcome the biochemical activity during storage, seed priming is a promising strategy. In
the present study, two nanopriming agents viz., Zinc Oxide Nanoparticles (ZnONPs) and
silver Nanoparticles (AgNPs) were evaluated for biochemical activity (Peroxidase activity,
alpha amylase activity, total soluble sugar, total protein) of chickpea at 2, 4, 6 and 9
months storage period after priming for 1, 2, and 6 hours. Result showed increased
activity of Peroxidase (POX) with increase of storage time, but the rate of increase was
comparatively low when seeds were nanoprimed with ZnONPs. Similarly, alpha amylase
activity and protein content were recorded highest and Total Soluble Sugar (TSS) was
found lowest in ZnONPs primed seed. Out of the different priming times, 6 hrs was found
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to be the best at 9 months of storage with positive effect on biochemical parameters.

Among the biotic stresses, disease caused by Fusarium oxysporum f.sp. ciceri has been
considered as a destructive one, which causes yield loss up to 10% every year. To
overcome the biotic stress and enhance the storage life, chickpea seeds were primed with
ZnONPs and AgNPS at 100 ppm alone and in combination with each other. We found
positive effect on seed germination (%), plant growth, and yield attributing parameter
and negative effect on per disease incidence of F. oxysporum f.sp. ciceri.

Keywords: Biochemical parameters, Cicer arientinum L., F. oxysporum, Storage life.

INTRODUCTION variously known as Gram, Bengal gram,

Garbanzo bean as well as Egyptian pea.
Seeds are basic and key input for India produces about 150 million tons of
agriculture. Of all crops, pulses have a food grains per year, but losses have
specific importance for the vegetarian remained static at 10% (Ali et al., 2015).
population of our country as it act as major More than 60% of the net area is sown in
source of protein. Per capita availability of under rainfed system. It is quite appropriate
pulses per day is only 47 g, as against the to develop technologies for rainfed
minimum requirement of 104 g (WHO) agriculture. Major problems in chickpea
(Vishwas et al., 2017). Among the pulses, cultivation in rainfed system are poor seed
chickpea, Cicer arientinum L. (Family: germination, seed dormancy, less plant
Fabaceae, Sub-family: Faboidae) occupies vigor and disease and pest attack. Besides,
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a predominant position and is considered as improper storage is also another reason for
a king of pulses. Its different types are lower production and an average of 6% out
_____________________________________________________________________________
1
Department of Plant Pathology, Assam Agricultural University, Jorhat-785013, Assam, India.
2
School of Crop Protection, College of Post Graduate Studies in Agricultural Sciences, Central Agricultural
University, Umiam-793103, Meghalaya, India.
*Corresponding author; e-mail: pranabdutta74@gmail.com
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 ______________________________________________________________________ Das and Dutta

of total 10% loss takes place during storage environmental stress (Vanangamudi et al.,
of food grains. In storage condition, seeds 2008). Various priming methods like
lose their vigor due to different biochemical osmopriming, hydropriming, solid matrix
changes (Maiti and Moreno- Limon, 2001). priming, biopriming, nanopriming etc. are
Among the different diseases, wilt caused potentially used in seed technology.
by Fusarium oxysporum f. sp. ciceri has “Nanopriming” is a new and fascinating
been considered as a destructive one, which method that can be used for increasing
causes yield loss up to 10% every year seedling vigor index, improvement of
(Kumar et al., 2014). To overcome this germination percentage, and seedling
disease, seed priming is the most promising growth. Nowadays, engineered
strategy for management of modern crop nanomaterials are the most important index
and production. of the nanotechnology area, have entered
Seed priming is an effective tool for all aspects of human life and their various
increasing seed germination and plant applications are quickly expanded due to
growth that will event-ually increase their new characteristics compared to the
productivity under different environmental corresponding bulk materials (Roozbeh et
conditions and stresses (Khalaki et al., al., 2017). Seed nanopriming is mostly
2020). Efficient seed germination promotes done on the effect of germination process in
successful establishment and deep root the field of research. Silver (Ag), Zinc
system of plants. Seed priming methods, Oxide (ZnO), gold (Au) Nanoparticles
particularly nanopriming is more effective (NPs) etc. are some of the different
mainly because of its small size and unique nanoparticles used as a nanopriming agents.
physicochemical properties. Since plant We hypothesized that the nanoparticle as
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species are physiologically different, they priming agent will have positive effect on
differ in their uptake of nanoparticles in enhancement of storage life of chick pea
nanopriming, and, hence, in their rate and with more plant stand against Fusarium
manner of growth. Most previous studies wilt disease.
have separately investigated the effect of The aim of the present study was to
nanomaterial on seed germination, growth, investigate the effect of nanopriming agents
and development of one plant at a time (Ag and ZnO nanoparticles) on the
(Khalaki et al., 2016). However, very few biochemical parameters of chickpea seeds
studies have reported nanopriming effects at different storage periods and to study the
using different particles on seed performance of the nanoprimed seeds in pot
germination and seedling growth of forage conditions on per cent disease incidence of
and medicinal plants together. The need to Fusarium wilt and plant growth and yield
produce high seed quality efforts have been attributing parameters of chickpea.
directed towards the seed priming, which in
tern shorten germination time and helps to
MATERIALS AND METHODS
overcome the seed dormancy. Seed priming
advances seed germination process without
emergence of radical and enough to This research was carried out in the
activate the biochemical events. The Nanotechnology Laboratory of Department
product of these changes persists following of Plant Pathology, Assam Agricultural
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desiccation and are available on re- University (AAU), Jorhat, during 2017-2018
imbibition of water during seed planting, and 2018-2019. In the present study,
enabling completion of germination chickpea seeds (var. JG-16) were used to
rapidly, leading to a uniform crop stand and prime with two nanoparticles viz., Zinc
synchronized flowering and fruiting. Oxide (ZnO) and silver (Ag) Nanoparticles
Additionally, these physiological (NPs).
treatments induce tolerance to certain
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 Nanopriming Enhances Seed Storage and Plant Stand _____________________________

Source of the Nanoparticles Preparation of Pot Mixture for Pot

Experiment
The nanoparticles were collected from the
Nano Lab of Department of Plant Pathology, For pot experiment, pot mixture was
Assam Agricultural University, Jorhat, prepared by mixing soil (sandy loam), sand,
Assam. AgNPs and ZnONPs were and compost at 1:1:1 ratio and sterilized in
synthesized mediating Trichoderm autoclave (Make: Equitron) at 1210C at 15 lb
asperellum (Kaman and Dutta, 2019) and pressures for 15 minutes consecutively for
wet chemical process (Kaushik and Dutta, three days. Nutrients like N, P, K were
2017), respectively. Synthesized added in the form of Urea, SSP and MOP
nanoparticles were characterized by UV-VIS three days prior to the seed sowing.
spectroscopy, electron micrography (SEM
and TEM), FTIR, Zetasizer and DLS. For
Standardization of Priming Technique
AgNPs characteristic surface plasmon
absorption band was observed at 420 nm
with Zeta potential of -1.34 mV, size of For standardization of nanopriming agents,
27.64 nm and roughly spherical in seeds were treated with ZnONPs and AgNPs
morphology (Kaman and Dutta, 2018). On alone and in combination with each other for
the other hand, the used ZnONPs had 1, 2, and 6 hours and shade dried till
average size of 33.4 nm with PDI of 0.697, moisture reached 9%. The treatment
with negative zeta potential value of -20.7 combination used were: T1= Control, T2=
mV, spherical in shape with zinc and oxygen Recommended pesticide treatment
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as elemental composition (Kaushik and (Malathion 5% dust per 2.5 g kg-

1
Dutta, 2017) +Carbendazim with 0.1% concentration),
T3= Nanopriming with ZnONPs at 100 ppm
(for 1, 2 and 6 hours), T4= Nanopriming
Source of the Pathogen and Preparation with AgNPs at 100 ppm (for 1, 2 and 6
of Inoculum for Pot Experiment hours), and T5= Nanopriming with ZnONPs
at 100 ppm+AgNPs at 100 ppm (for 1, 2 and
The pathogen Fusarium oxysporum f. sp 6 hours). Treated seeds were packed in
ciceri was collected from the culture of HDPE semipermeable bag and stored for 2,
Department of Plant Pathology, AAU, 4, 6 and 9 months. The experiment was
Jorhat, Assam and maintained the pure arranged in Completely Randomized Design
culture by periodic transfer to fresh PDA (CRD) with 6 replications.
slants and stored in refrigerator at 4oC. Mass
culture of F. oxysporum was done in Effect of Nanopriming on Biochemical
sterilized Maize meal Sand Media (MSM).
Parameters and Storage Life
Two mycelial disc (5 mm diam.) of F.
oxysporum from fresh PDA medium was
transferred aseptically in Erlenmeyer flasks Estimation of Total Soluble Sugar (TSS)
(500 mL) containing MSM and was The total soluble sugar content was
incubated in BOD incubator (Make: REICO) estimated by using the method of Yemm and
for seven days at 25±10C. On 8th day, MSM Willis (1954). For the estimation of TSS of
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with fungal inoculum containing fungal storage chickpea seeds, 20 mg was

mycelium, micro and macrop conidia etc. homogenized in 5 mL of 80 per cent hot
was inoculated to pot mixture at 0.2% (w/w) ethanol and centrifuged at 4,000 rpm for 20
seven days prior to the seed sowing so that minutes. The supernatant was collected and
the pathogen settles well in the pot mixture. the residue was again centrifuged with 5 mL
of 80 per cent hot ethanol at 4,000 rpm for
20 minutes. Both supernatants were mixed
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 ______________________________________________________________________ Das and Dutta

together. The known amount of ethanol centrifuged with 5 mL of phosphate buffer at

extract was evaporated to dryness in a test 4,000 rpm for 20 minutes. Both supernatants
tube on a water bath and cooled at room were mixed together. In 0.2 mL of
temperature. One mL of distilled water was phosphate extract, 5 mL of Carbonate- Cu++
added in a test tube and mixed thoroughly. solution was added and mixed well by
To each test tube, 4 mL of anthrone reagent vortexing. After 10 minutes, 0.5 mL of
(200 mg of anthrone reagents was dissolved Folin- Ciocalteu reagent was added and
in 100 mL of concentrated sulfuric acid) was mixed well vortexing and absorbance was
added and mixed thoroughly and gently measured at 660 nm against blank reagent.
heated on water bath at 100oC for 10 The amount of total protein was calculated
minutes, cooled rapidly under running tap by using standard curve prepared from
water, and absorbance was measured at 630 bovine serum albumin stock solution and
nm against reagent blank. The amount of expressed as mg/g or 100 g sample.
total soluble sugar was calculated using
standard curve prepared from graded Estimation of Alpha-Amylase Activity
concentration of glucose. Calculation of the
(Bernfield, 1955)
concentration of TSS in the test tube were
determined from the standard curve and
expressed as per cent or mg g-1 dry weight. The estimation of alpha-amylase activity
was measured by the method of Bernfield
(1955) and expressed in milligram of
Estimation of Total Protein (TP) maltose equivalent. The reagents used for
doing alpha-amylase were: (a) 0.1M Sodium
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The total protein content was estimated by Acetate buffer (pH- 4.7), (b) Acetic acid
using the method of Lowry et al. (1951). 0.2M (1.5 mL glacial acetic acid was added
The reagents used for estimation of total in 100 mL distilled water), (c) 1% Starch
protein were: (a) Phosphate buffer (0.1M, solution (1 g starch was dissolved in 100 mL
pH 7.6), (b) Solution A: 2% Na2CO3 in 0.1N of 0.1M Sodium acetate buffer), (d)
NaOH (20 g of sodium carbonate and 4 g Dinitrosalicylic (DNS) acid reagent (1 g
sodium hydroxide was dissolved in distilled DNS, 0.2 g crystalline phenol and 0.05g
water of volume 1 liter), (c) Solution B: 1% sodium sulfite was dissolved in 100 mL of
CuSO4.5H2O solution (1 g of copper sulfate 1% NaOH),(e) 40% Potassium Sodium
was dissolved in 100 of distilled water) and Tartrate (PST), (e) Standard maltose
2% potassium sodium tartrate was dissolved solution (50 mg maltose was dissolved in 50
in 100 mL of distilled water. Both 1% mL of distilled water).
CuSO4.5H2O solution and 2% potassium For estimation of alpha-amylase activity,
sodium tartrate were mixed in equal volume the storage chickpea seeds were
before use, (d) Solution C: Carbonate-Cu++ homogenized in 5 mL of ice cold 10 mM
solution (this solution was prepared by Calcium chloride solutions over night at 40C
mixing 50 mL of Solution A and 1 mL of or 3 hours at room temperature and
Solution B), (e) Solution D: Folin- Ciocalteu centrifuged the extract at 6,000 rpm at 40C
Reagent (equal amount of water was added for 10 minutes, and the supernatant was
to the reagent before used), (f) Standard made up to 10 mL with 10 mM calcium
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protein solution: 25 mg of bovine serum chloride. The supernatant was used as

albumin was dissolved in 100 mL of water. enzyme source. In a test tube, 1 mL of
For estimation of total protein, one gram supernatant was taken and added with 1 mL
of chickpea seeds were homogenized in 5 starch solution, 2 mL DNS solution and 1
mL of phosphate buffer and centrifuged at mL PST solution and mixed it properly by
4,000 rpm for 20 minutes. The supernatant using vortex mixture and heat in a boiling
was collected and the residue was again water bath for 5 minutes and dark orange red

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 Nanopriming Enhances Seed Storage and Plant Stand _____________________________

color was developed and absorbance was Based on the above biochemicals
measured at 560 nm against blank reagent. parameters, best priming time and highest
The alpha amylase activity was calculated storage months were selected and compared.
by using standard curve prepared form
maltose solution. Effect of Nanopriming against Wilt of
Chickpea and Plant Growth and Yield
Estimation of Peroxidase Activity Parameters

The peroxidase activity was estimated by The best treatment combinations found in
using the methods of Putter (1974). The the above experiments were selected for the
reagents used for doing peroxide activity pot experiment conducted in net house of
were: (a) Phosphate buffer 0.1M (pH 7.0), Department of Plant Pathology, Assam
(b) Guaiacol solution 20 mM (0.24 mL Agricultural University (AAU), Jorhat,
guaiacol was dissolved in 100 mL of Assam.
water), (c) Hydrogen peroxide solution The treatment combination used were: T1=
(0.14 mL of 30% H2O2 was diluted in 100 Uninoculated control (No seed
mL of water). treatment+No pathogen), T2= Inoculated
For estimation of peroxidase activity, one control (F. oxysporum f.sp. cicero at 0.02%),
gram of storage chickpea seeds were T3= Recommended pesticide treatment
homogenized in 3 mL of phosphate buffer (Malathion 5% dust per 2.5g kg-
1
by grinding in a pre-cooled mortar and +Carbendazim with 0.1%)+F. oxysporum f.
pestle and centrifuged the extract at 18,000 sp. ciceri, T4= Nanopriming with ZnONPs
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rpm at 50C for 15 minutes. The supernatant at 100 ppm (for 6 hours)+F. oxysporum f.sp.
was used as an enzyme source by making ciceri, T5 = Nanopriming with AgNPs at
10 mL with phosphate buffer. In a test tube, 100 ppm (for 6 hours)+F. oxysporum f. sp.
0.1 mL of enzyme extract was taken and ciceri and T6= Nanopriming with ZnONPs
added with 3 mL of phosphate buffer, 0.05 at 100 ppm+AgNPs at 100 ppm (for 6
mL guaiacol solution and 0.03 mL hours)+F. oxysporum f. sp. ciceri.
hydrogen peroxide solution. The mixture Observation on seed germination percentage
was well shaken and placed in a (%), shoot length and root length (cm),
spectrophotometer by putting the mixture in number of branches per plant, fresh and dry
a cuvette. The time required for the mixture weight of plant at final harvest (g), seedling
optical density to be increased by 0.1Δt at vigor index, number of pods per plant, seeds
436 nm was recorded and used in per pod, seed yield per plant (g plant-1),
calculations. 1,000 seed weight (g) and disease incidence
(%) etc. were recorded as per following
procedure.
Calculation of Peroxidise Activity
Physiological Parameters of Priming
Storage Chickpea Seeds
Since the extinction coefficient of guaiacol
dehydrogenation product at 436 nm under
the condition specified is 6.39 per Seed germination percentage and
moisture content
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micromole, the enzyme activity per liter of

extract was calculated as, Enzyme activity
(U L-1)= (3.18×0.1×1000/6.39×1×Δt×0.1)= Seed germination was studied following
500/Δt. the between paper methods (ISTA, 1985).
Δt=time change in minute Fifty seeds were placed equidistantly
between two seeds of germination paper
soaked in water, rolled and tagged and

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 Activity (enzyme unit/mg of tissue)
______________________________________________________________________ Das and Dutta

Figure 1. Effect of time of nanopriming (1, 2, and 6 hourrs) on peroxidase activity of chickpea seeds at
different storage periods (2, 4, 6, and 9 months).

calculated. The Seedling Vigor Index (SVI)

incubated inside germinator at 300C. Final was calculated using the following formula:
count was taken on 8th day. Observation was SVI= Seedling length×Germination (%)
recorded on seed germination percentage.
RESULTS
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Speed of Germination
Results obtained on different biochemical
Seeds of different treatments were also parameters like peroxidase activity; alpha
evaluated in terms of Speed of Germination amylase activity, total soluble sugar, and
Index (SGI). Fifty seeds were placed total protein on different months of storage
equidistantly between two seeds of are presented as follows:
germination paper soaked in water, rolled
and tagged and incubated inside germinator
at 30 0C (ISTA, 1985). Number of seeds Effect of Nanopriming on Peroxidase
germinated daily was counted till Activity (POX) in Chickpea Seeds
completion of germination. From the
observation data, speed of germination index In this study, it was found that the POX
was calculated as per the following formula: enzyme activity increases with increase of
Speed of germination= X1/n1+(X2– storage periods. POX activity was highest in
X1)/n2+(X3–X2)/n3....(Xn–Xn-1)/nn the control (non-primed seeds), but the rate
Where, X1, X2, X3.....= Number if of increase was comparatively lower when
seedling germinated on n1, n2, n3........days. seeds were primed with ZnONPs at 100 ppm
Shoot Length, Root Length, Seedling in all periods of storage for 1, 2 and 6 hours.
Fresh Weight and Seedling Dry Weight and This was followed by seeds treated with
Seedling Vigor Index both ZnONPs at 100 ppm and AgNPs at 100
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To study the effect of priming seedling (Figure 1).

vigor index, normal seedling showing
normal root and shoot growth were
randomly selected. The root and shoot Effect of Nanopriming on Alpha
portion were separated, their length was Amylase Activity in Chickpea Seeds
measured, and mean seedling length was

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 Nanopriming Enhances Seed Storage and Plant Stand _____________________________
Activity (mg of maltose equivalent)

Figure 2. Effect of time of nanopriming (1, 2, and 6 hours) on alpha amylase activity of chickpea seeds
at different storage periods (2, 4, 6, and 9 months).

Alpha amylase is an indicator of seed seeds primed with both ZnONPs at 100 ppm
germination. The data recorded showed that and AgNPs at 100 ppm (Figure 4).
the alpha amylase activity decreases with
increase of storage period. The alpha
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Effect of Nanopriming on Physiological

amylase activity was recorded highest in
Parameters of Chickpea Seeds in
seeds treated with ZnONPs at 100 ppm in all
Suppression of Wilt Disease
the periods of storage for 1, 2 and 6 hours.
This was followed by seeds treated with
both ZnONPs at 100 ppm and AgNPs at 100 The study on the effect of nanopriming
ppm (Figure 2). The lowest concentration of showed that all the treatments had positive
alpha amylase activity was recorded in the effect on plant growth and yield parameters.
control, where seeds were non-primed. Seeds primed with nanoparticles for 6 hours
caused significant reduction in seedling
infection by the pathogen (Table 1). Highest
Effect of Nanopriming on Total Soluble seed germination percentage were recorded
Sugar and Total Protein in Chickpea for seeds nanoprimed with ZnONPs at 100
Seeds ppm (85.46%), followed by seeds treated
with both ZnONPs and AgNPs at 100 ppm.
Highest protein content was recorded Amongst the nanoprimed seeds, the
when ZnONPs was used as nanopriming minimum seed germination (65.00 %) was
agent in all periods of storage in all priming observed in seeds treated with AgNPs at 100
times (1, 2 and 6 hours.) (Figure 3). This ppm with lowest in the inoculated control
was followed by nanopriming with ZnONPs (50.76%). Seeds treated with the
at 100 ppm+AgNPs at 100 ppm. The lowest recommended chemical showed a
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protein content was recorded in the control germination percentage of 71.56% (Table 1).
with highest Total Soluble Sugar (TSS). TSS Lowest percent disease index (PDI) (6.16%)
content was at the lowest concentration in was recorded in seeds priming with ZnONPs
seeds primed with ZnONPs at 100 ppm in at 100 ppm. This was followed by seed
all the period of storage in all priming time primed with both the combination of
(1, 2 and 6 hours.). This was followed by ZnONPs at 100 ppm and AgNPs at 100 ppm

219
 ______________________________________________________________________ Das and Dutta
Concentration (mg.g-1)

Figure 3. Effect of time of nanopriming (1, 2, and 6 hours) on total protein (mg g -1) of chickpea seeds at
different storage periods (2, 4, 6, and 9 months).
Concentration (mg.g-1)
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Figure 4. Effect of time of nanopriming (1, 2, and 6 hours) on total soluble sugar (mg g-1) of chickpea
seeds at different storage periods (2, 4, 6, and 9 months).

(6.16%). Seeds primed with recommended

chemical check showed PDI of 7.03%.
Study on effect of nanopriming of plant Effect on Seedling Vigor Index
growth parameter showed that when ZnONP
at 100 ppm was used as priming agents, it The highest seedling vigor index
showed the highest plant growth parameters (6085.60) was recorded when the seeds were
like root length (15.21 cm), shoot length primed with ZnONPs at 100 ppm (for 6
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(56.00 cm), number of branches (5), fresh hours)+F. oxysporum f. sp. ciceri (Table 1).
weight of roots (0.77 g) and shoots (31.62 g) This was followed by seeds treated with
and dry weight of roots (0.64 g) and shoot both ZnONPs at 100 ppm and AgNPs at 100
(7.19 g). This was followed by seeds treated ppm with seedling index of 4224.27. Lowest
with both the combination of ZnONPs at seedling vigor index (2535.71) was recorded
100 ppm and AgNPs at 100 ppm and seeds in inoculated control and seeds primed with
primed with the recommended chemicals.
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 Nanopriming Enhances Seed Storage and Plant Stand _____________________________

the recommended chemical check showed best which results in biochemical parameters
the seedling vigor index of 3446.17. of chickpea at 9 months of storage.
Peroxidase enzyme activity was the lowest
in seeds that were nanoprimed with ZnONPs
Effect of Nanopriming on Seed
Production of Chickpea Seeds at 100 ppm. This indicated that, due to
priming, firmness of seed increases resulting
the more storage life. On the other hand,
In nanoprimed seeds, the highest pods per alpha amylase activity was lower in
plant was recorded when seeds were primed ZnONPs treated seeds. This indicates that
with ZnONPs at 100 ppm (23.00) (Table 2). the low rate of germination percentage of
This was followed by seeds treated with chickpea seeds in ZnONPs primed seeds
both ZnONPs @ 100 ppm and AgNPs at 100 reduces compared to other treatments.
ppm. The lowest pods/plant (16 nos.) was Ramegowda (1992) reported that a decrease
recorded in inoculated control. Data in the activity of enzymes viz., alpha
presented in Table 2 shows the highest seeds amylase, and peroxidase coupled with
per pod in T4 and T6 where seeds were progressive ageing of rice seeds. He further
treated with ZnONPs at 100 ppm alone and authenticated that alpha amylase and
in combination with AgNPs at 100 ppm. peroxidase enzymes were more directly
Similarly, seed yield per plant (2.19 g) and involved in the maintenance of better
1000 seed weight (319.66 g) were recorded germination of differentially aged seeds.
highest in seeds treated with ZnONPs at 100 Zafar et al. (2016) reported that treatment
ppm. This was followed by seeds treated with ZnONPs increases the biochemical
with both ZnONPs at 100 ppm and AgNPs parameters in Brassica nigra seedlings.
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at 100 ppm. Lowest seed yield per plant Biochemical studies have been done to
(1.68 g) and 1,000 seed weight (117.16 g) amylase the effect of nanoparicles on
were recorded in inoculated control. Seeds biochemical changes of storage chickpea
primed with the recommended chemical seeds, which were treated for three different
check had a seed yield per plant and 1,000 hours (2, 4, and 6 hours) and stored for 9
seed weight of 1.79 and 219.83 g, months. Result on POX activity showed that
respectively. with increase of storage time, POX activity
increased. However, the rate of increase was
DISCUSSION comparatively low when seeds were nano-
primed with ZnONPs. In case of alpha
amylase activity, lowest concentration was
Seed priming is considered to be the recorded in the control, but seeds nano-
important approach in emergence of crops as primed with ZnONPs showed the highest
well as in storage condition (Pill et al., 2009; activity of alpha amylase. Highest protein
Rakshit et al., 2014). Maintenance of good content was recorded when ZnONPs was
quality seeds is the clear understanding of used as nano-priming agents. On the other
the biochemical events during storage. In the hand, the highest TSS was recorded in the
present study we found that, with the control with lowest in nano-primed with
increase in storage periods, the biochemical ZnONPs.
parameters like alpha amylase activity and During our experiment, we found
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total protein decreased and peroxidase significantly the lowest Percent Disease
activity and Total Soluble Sugar (TSS) Incidence (PDI) when seeds were primed for
increased. This may be due to the reduction 6 hrs with ZnONPs at 100 ppm followed by
of concentration of protein that leads to seeds primed with ZnONPs at 100
increase in TSS concentration as observed in ppm+AgNPs at 100 ppm. The former
the present study. Out of different treatment also showed significantly positive
nanopriming time, 6 hrs was found to be the effect on germination percentage, plant
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222
______________________________________________________________________ Das and Dutta
 Nanopriming Enhances Seed Storage and Plant Stand _____________________________

growth parameters, and chickpea seed was best, and among the different months of
production in pot condition as compared to storage, the highest was 9 months.
unprimed seeds by reducing seedling Therefore, all the studied biochemical
infection. The lowest PDI recorded in parameters were considered for the two best
ZnONPs primed and AgNPs primed seeds treatments (T3 and T5) and comparison was
may be due to the antifungal activity of the made with the control (T1). Results showed
nanoparticles against F. oxysporum as that seeds primed with ZnONPs at 100 ppm
reported earlier by Yehia et al. (2013), for 6 hrs showed highest biochemical
Karimiyan et al. (2015), and Ouda (2014). activities, which increased the storage life of
The reduction in disease indirectly helped in the chickpea seeds up to 9 months. This was
proper health management and ultimately followed by seeds nanoprimed with the
showed increased plant growth parameters combination of both ZnONPs at 100 ppm
and yield attributing characteristics. and AgNPs at 100 ppm (Figure 5).
Antifungal activity of ZnONPs against F. To conclude, nanopriming with ZnONPs
graminearum was also reported by Dimkpa for 6 hrs can help to store the chickpea seeds
et al. (2013). They reported that ZnONPs up to 9 months. We also found positive
was significantly more inhibitory to fungal effect of nanopriming with ZnONPs during
growth than micro-sized particles of ZnO, study on biochemical parameters like POX
although both types of particles released and alpha amylase activity, which helped in
similar levels of soluble Zn, indicating size- increasing the storage life of chick pea up to
dependent toxicity of the particles. AgNPs 9 months. Further, 9-months stored ZnONP
was reported to have antifungal activity nanoprimed seeds were also effective in
against a multiple fungal pathogen including suppressing the infection by F. oxysporum
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Fusarium spp. (Alananbeh et al., 2017). A and enhanced plant growth parameters and
study conducted to find out the effect of T. yield attributing parameters.
asperellum mediated AgNPs on physiology
of tea plant showed that AgNPs at 100 ppm ACKNOWLEDGEMENTS
can induce the plants in increasing
physiological parameters like chlorophyll
content, moisture content, relative water The authors are thankful to the Director of
content, TSS, total protein and biochemical Post Graduate Studies, and Professor and
parameters like lipid peroxidation, (MDA Head, Department of Plant Pathology,
content) and secondary metabolites like Assam Agricultural University, Jorhat,
phenol, alkaloid, and flavonoid content of Assam for their support and guidance during
treated plants without any harmful effect on the period of investigation. The help
the plant (Ahmed and Dutta, 2019). In an received from Dr. S. Dutta Deka, Principal
another study, we found that AgNPs (size Scientist, Department of Plant Breeding and
8.26 nm with polydispersity index of 0.857 Genetics, Assam Agricultural University,
and zeta potential value of -1.34 m) as soil Jorhat, Assam and Dr. D. Pathak, Principal
treatment has positive impact on soil pH, Scientist, Regional Agricultural Research
microbial count, organic carbon, and Station (RARS), Shillongani, Nagaon is also
microbial biomass carbon, except total duly acknowledged.
microbial count (Ahmed and Dutta, 2020);
[ DOR: 20.1001.1.16807073.2022.24.1.9.9 ]

and nuclear magnetic resonance REFERENCES

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223
 ______________________________________________________________________ Das and Dutta

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‫اثر نانوپرایمینگ با اکسیذ روی و ررات نانو نقره بر انبارداری دانه نخود و مذیریت‬
‫بیمای پژمردگی‬

‫ دوتا‬.‫ و پ‬،‫ داس‬.‫گ‬

‫چکیذه‬

‫ًخَد از حبَببت هْن ٌّذٍستبى است ٍلی بْرُدّی عولکرد آى بِ علت چٌذ تٌش زیستی ٍغیرزیستی‬
‫ ضعیف بَدى داًِ است کِ بِ خبطر تغییرات درخَاص‬،‫ یکی از هسبیل در ایي ارتببط‬.‫پبییي است‬
ُ‫ آهبد‬،‫ برای غلبِ برفعبلیتْبی بیَشیویبیی‬.‫بیَشیویبیی افتراقی درطی دٍراى اًببرداری ًبهٌبسب رخ هیذّذ‬
‫ دٍ عبهل ًبًَ پرایویٌگ بِ ًبم‬،‫ در پژٍّش حبضر‬.‫سبزی (پرایویٌگ) داًِ از راُکبرّبی اهیذ بخش است‬
‫( برای فعبلیت ّبی بیَشیویبیی ًخَد‬AgNPs) ُ‫( ٍ ررات ًبًَ ًقر‬ZnONPs)‫ررات ًبًَ اکسیذ رٍی‬
[ DOR: 20.1001.1.16807073.2022.24.1.9.9 ]

ُ‫ هب‬9 ٍ ،6 ،4 ،2 ‫ٍپرٍتئیي کل) در طی هذت‬،‫ کل قٌذ هحلَل‬،‫ آلفباهیالز‬،‫(شبهل فعبلیت پراکسیذاز‬

‫ ًتبیج حبکی از افسایش فعبلیت پراکسیذاز‬.‫ سبعت پرایویٌگ ارزیببی شذ‬6 ٍ ،2 ،1 ‫اًببرداری ٍ بعذ از‬
ZnONPs ‫) بب افسایش دٍرُ اًببرداری بَد ٍلی ًرخ افسایش در هَردی کِ داًِ ّب بب‬POX(
‫ فعبلیت آهیالز ٍ هحتَای پرٍتئیي در حذ بیشیٌِ ثبت شذ‬،‫ ّوچٌیي‬.‫ًبًَپرایویٌگ شذُ بَد ًسبتب کن بَد‬

225
 ‫‪______________________________________________________________________ Das and Dutta‬‬

‫ٍلی قٌذ هحلَل کل (‪ )TSS‬در داًِّبیی کِ بب ‪ ZnONPs‬پراین شذُ بَد در حذ کویٌِ بَد‪ .‬در هیبى‬
‫زهبًْبی هختلف پرایویٌگ‪ ،‬تیوبر ‪ 6‬سبعت پرایویٌگ برای ‪ 9‬هبُ اًببرداری بْتریي تیوبر بَد ٍ اثر ّبی‬
‫هثبتی رٍی پبراهترّبی بیَشیویبیی داشت‪ً .‬یس‪ ،‬در هیبى تٌش ّبی زیستی‪ ،‬بیوبری ایجبد شذُ بب‬
‫‪ Fusarium oxysporum f.sp. ciceri‬بِ عٌَاى یک بیوبری زیبًببر تلقی هی شَد زیرا ّر سبل‬
‫ببعث ‪ %11‬کبّش عولکرد هی شَد‪ .‬برای رفع تٌش زیستی ٍ ارتقبی شرایط اًببرداری‪ ،‬داًِ ّبی ًخَد بب‬
‫هبدُ ‪ AgNPs ٍ ZnONPs‬بِ تٌْبیی یب ّوراُ بب ّن بب ‪ 111‬قسوت در هلیَى (‪)ppm‬آهبدُ سبزی ٍ‬
‫پراین شذ‪ .‬در پی آى‪ ،‬اثرات هثبتی رٍی درصذ جَاًِ زًی داًِ‪ ،‬رشذ گیبُ‪ ٍ ،‬پبراهترّبی هربَط بِ‬
‫عولکرد ٍ اثرات هٌفی بر ابتال بِ بیوبری ‪ F.oxysporum f.sp. cicero‬هشبّذُ شذ‪.‬‬
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