Optimization of Foliar Chitosan Doses as a Biofertilizer for Enhanced
Pepper Cultivation
Boran İkız
*a
, H. Yıldız Daşgan
b
, Cumali Yeniay and Muhammet Taş
Çukurova University, Department of Horticulture, 01330 Balcalı, Adana, Turkey
Keywords: Capsicum annuum L., Glucosamine, Sustanaible Agriculture, Biostimulant, Yield Optimization, Fruit
Quality.
Abstract: Chitosan is a water-soluble aminopolysaccharide obtained by the deacetylation of chitin, which is the second
most abundant biopolymer after cellulose. Chitin is primarily found in the exoskeletons of shellfish, and
chitosan is produced by converting the acetyl groups in the chemical structure of this natural polymer into
amine groups. The study aimed to evaluate the effects of different chitosan doses on plant growth, yield and
fruit properties of pepper. The experiment included a control group with no chitosan application, as well as
treatments with 150 ppm, 300 ppm, and 450 ppm of chitosan. The applications were made foliar once a week
The 150 ppm chtosan treatment ncreased yeld by 67.4%, and the 300 ppm treatment by 44.9%, compared
to the control. Chitosan treatments also significantly and positively influenced plant growth parameters, such
as plant height, plant width, and leaf number. Additionally, the chitosan applications increased, vitamin C,
total phenols and total flavonoids, EC (electrical conductivity) in the fruit juice.
This study’s objectives are
to ascertain the proper chitosan dosage for application and to track the beneficial benefits of employing
chitosan as a biostimulant.
1 INTRODUCTION
Chtosan s a naturally occurrng bopolymer wth a
wde range of possble uses n agrculture. In
partcular, chtosan s employed as a rhzosphere
modulator, plant growth regulator, and bopestcde.
When used as a bopestcde, chtosan effectvely
combats bactera, fungus, and vruses (Badawy et al.,
2011). It strengthens the structure of the sol, makes t
more capable of retanng water, and encourages
mcrobal actvty n the rhzosphere. It prevents harm
from heavy metals, whch enhances sol health
(Naeem ve et al., 2018). Because t encourages cell
dvson and plant growth, chtosan plays a sgnfcant
role n boostng frut and vegetable output yelds
(Khan et al., 2009). It s well known that the food and
agrcultural ndustres also employ chtosan as a flm
coatng. Bodegradable and antbacteral, chtosan-
based flms are also knd to the envronment.
Accordng to Rhm et al. (2020), ths s an addtonal
alternatve for food preservaton and plant protecton.
a
https://orcd.org/0000-0003-3012-4533
b
https://orcd.org/0000-0002-0403-1627
*
Correspondence Author
It can also have an effect on agrculture’s sustanablty
by beng economcal and envronmentally bengn
(Malerba et al., 2021). Pepper (Capscum annuum L.)
s an economcally mportant crop grown for ts
nutrtonal value, boactve compounds, antoxdant
propertes, and natural colors (Santosh, 2013).
Pepper
s wdely consumed worldwde both as a fresh
vegetable and as a processed product. Varetes of
peppers nclude sweet peppers (e.g., bell peppers) and
hot peppers (e.g., chl peppers). Wth ts hgh content
of vtamn C, vtamn A, and antoxdants, pepper s
consdered a hghly nutrtous vegetable and s
therefore regarded as an mportant source for a healthy
det (Sdhu et al., 2019).
Accordng to 2022 FAO data,
pepper s one of the most mportant vegetables,
cultvated on 2,020,816 hectares wth a producton of
36,972,494 tons (FAO, 2018).
Pepper s also used as a model plant for
botechnologcal and bostmulant applcatons that
enhance resstance to varous botc (pathogens,
nsects) and abotc (drought, salnty) stress factors
˙
Ikız, B., Da¸sgan, H. Y., Yeniay, C. and Ta¸s, M.
Optimization of Foliar Chitosan Doses as a Biofertilizer for Enhanced Pepper Cultivation.
DOI: 10.5220/0014261000004738
Paper published under CC license (CC BY-NC-ND 4.0)
In Proceedings of the 4th International Conference on Research of Agricultural and Food Technologies (I-CRAFT 2024), pages 253-259
ISBN: 978-989-758-773-3; ISSN: 3051-7710
Proceedings Copyright © 2025 by SCITEPRESS – Science and Technology Publications, Lda.
253

(Trverd et al., 2020). Pepper s a plant that performs
best under warm clmate condtons. Factors such as
water stress and hgh temperatures have sgnfcant
mpacts on the productvty of pepper cultvaton. To
enhance the plant’s reslence to these envronmental
stresses, varous bostmulants and water management
strateges are employed.
There are over 200 common names used 2nda re
Capscum annuum speces. Among the most common
are chl pepper, red pepper (sweet varetes), bell
pepper, red pepper, jalapeños, and chltepn (hot
varetes), as well as Chrstmas peppers (ornamental)
(Latham et al., 2009; Zhang et al., 2002). In the past,
some knda forms of ths speces were referred to as
C. Frutescens, but the characterstcs used to
dstngush these 2nda re2e observed n many
populatons of C. Annuum, and there s no consstently
recognzable C. Frutescens speces (Zhang et al.,
2002). It can be dffcult to dfferentate Capscum
annuum from cultvated C. Chnense (the hottest
peppers) and C. Frutescens (tabasco peppers), as ther
morphologcal trats may overlap. These three speces
share the same ancestral gene pool 2nda re sometmes
referred to as the “annuum-chnense-frutescens
complex (Aracel et al., 2009).
Caps৻cum annuumL. Contans capsacnods
(capsacn, dhydrocapsacn), carotenods (luten,
zeaxanthn, capsorubn, β-carotene), flavonods
(quercetn, kaempferol, catechn, epcatechn, rutn,
luteoln), and sterodal saponns (capscosde E, F, G,
and capscdn). The man components of C. Annuum
essental ol, dentfed usng GC/MS, are capsamde
and acetc acd. C. Annuum has been reported to
possess varous bologcal actvtes, ncludng
antoxdant, antbacteral, antvral, antprolferatve,
ant-adpogenc, antmutagenc, enzyme nhbtory,
ant-nflammatory, hepatoprotectve, antdabetc,
renoprotectve, hypocholesterolemc, anttumor, ant-
obesty, analgesc, appette suppressant, and ant-
reflux propertes (Yuca, 2002).
2 MATERIALS AND METHODS
The research took place durng the summer growng
season of 2024 at the Unversty of Cukurova, Adana,
Turkye (36°59′N, 35°18′E, 20 m above sea level).
The temperature values n the regon fluctuated
throughout the days, wth daytme temperatures
rangng between 35-40°C. The row spacng was set to
50 cm, and the spacng wthn the rows was 90 cm.
Chtosan (Adaga-NanoWet) was appled to the plants
at doses of 150 ppm, 300 ppm, and 450 ppm per week
to determne the optmal dose after 15 days frst
plantng and per one week.
The pepper seeds used n
ths study were of the Semerkant F1 varety, developed
by the Nunhemscompany. After beng grown nto
seedlngs, they were transplanted nto the sol.).
Harvestng of the plants began 45 days after plantng
and was carred out fve tmes.
Plant heght, plant wdth, and plant canopy
measurements were taken usng a meter, whle frut
length, frut dameter, and frut wdth were measured
usng calpers.
Color measurements were performed
usng the FRU Precse color reader WR-18 color space
system. pH and EC measured by portable WTW
phmeter.
Chlorophyll content was measured usng a
handheld SPAD meter from the Mnolta brand.
Vtamn C content was carred out usng the adapted
approach outlned n Elgalan et al. (2017)’s study.
A randomzed block experment was conducted
usng a four-replcate desgn, wth 15 plants each
replcate. In the context of frut analyss,
measurements were performed on 10 fruts per repeat.
Furthermore, at the tme of harvest, 10 plants were
measured n each replcaton. Statstcal analyses were
performed wth the JMP software.
The am of ths
study s to observe the ameloratve effects of usng
chtosan as a bostmulant and to determne the
approprate chtosan dosage for applcaton.
3 RESULTS AND DISCUSSION
3.1 Plant Heght, Plant Dameter and
Stem Dameter
When examnng the plant heght n pepper plants, t
was observed that the use of chtosan as a bostmulant
ncreased plant heght. Plant heght ncreased n all
three doses compared to the control plants; however,
there was no statstcally sgnfcant dfference
between the treatments. In the control plants, the plant
heght was 48.08 cm, whle the plants treated wth 300
ppm chtosan as a bostmulant ncreased to 55.25 cm.
Smlarly, plants treated wth chtosan exhbted a
greater stem dameter compared to the control plants.
Although the dfference was not statstcally
sgnfcant, the plant dameter was found to be hghest
n the 300 ppm chtosan treatment, consstent wth
other parameters. Salachna and Zawadznska (2014)
demonstrated that chtosan, wth varyng molecular
weghts, can be used as a bostmulant n the
cultvaton of Freesa plants n pots. They observed
that, regardless of the compound’s molecular weght,
plants treated wth chtosan had more leaves and
shoots, flowered earler, and produced more flowers
and bulbs. Addtonally, Kumaraswamy et al. (2021)
I-CRAFT 2024 - 4th International Conference on Research of Agricultural and Food Technologies
254

reported that the applcaton of chtosan-slcon nano-
fertlzer on corn plants enhanced growth and yeld.
They observed that these bostmulants ncreased leaf
area as well as stem and root length. Perez et al. (2024)
conducted folar applcatons of chtosan,
brassnosterods, and thdazuron on strawberres,
demonstratng that chtosan treatment ncreased plant
heght, leaf count, leaf area, and frut frmness.
Table 1: Effects of chtosan doses on plant growth
measurements n pepper.
Apps.
Plant-
Dameter
(
cm
)
Plant
Heght
(
cm
)
Stem
dameter
(
mm
)
Control 45.54 48.08 b 12.83 b
150 ppm
Chtosan
53.46 55.00 a 17.42 a
300 ppm
Chtosan
55.08 55.25 a 19.70 a
450 ppm
Chtosan
52.00 54.50 a 17.76 a
P0.05 N.S 0.02 0.002
LSD N.S 6.14 2.80
There s no sgnfcant dfference between means wth the
same letter n the same column, LSD the least sgnfcant
dfference, NS: Non sgnfcant, Apps: Applcatons.
3.2 Yeld
The 150 ppm chtosan treatment ncreased yeld by
67.4%, the 300 ppm treatment by 44.8%, and the 450
ppm treatment by 17.7%, compared to the control
(Fgure 1). Abdel-Mawgoud et al. (2010) appled
chtosan to strawberry plants and observed an ncrease
n the number of fruts per plant as well as
mprovements n yeld parameters per plant. Plant
growth parameters, chlorophyll content, and
consequently photosynthetc capacty, as well as
Vtamn C content an mportant factor n pepper plants
showed better results n all chtosan treatments
compared to the control.
Fgure 1: Effects of chtosan doses on yeld n pepper.
However, t was observed that hgher chtosan
doses led to a reducton n yeld. Usng the lowest
dose of chtosan s also economcally advantageous.
When examnng other parameters, no sgnfcant
dfferences were observed among the dfferent
chtosan treatments.
3.3 Chlorophyll Content
In terms of chlorophyll content, the use of chtosan,
bo-fertlzer yelded hgher results compared to the
control plants. Snce an ncrease n chlorophyll
content s assocated wth photosynthess, t was
observed that chtosan enhances photosynthess,
whch n turn nfluences the ncrease n growth
parameters such as plant heght and stem dameter.
İnam et al. (2024) nvestgated the effects of chtosan
and znc oxde fertlzers on allevatng the severe
mpacts of drought on corn plants. They observed that
the applcaton of 1000 µg/L chtosan produced better
results than both the control and znc oxde
treatments.
Smlarly, n ther study on strawberres,
Perez et al. (2024) suggested that the combned
applcaton of chtosan and brassnosterods
sgnfcantly promoted crown dameter,
photosynthetc pgments, carotenods, and the fresh
and dry weghts of both roots and above-ground parts,
as well as maturaton.
In ther study, Tamer Khalfa et
al. (2024) nvestgated the nteractve effects of sol
mulchng materals (non-mulched, whte plastc, rce
straw, and sawdust) and chtosan folar spray
applcatons (control, 250 mg L⁻¹ regular chtosan,
125 mg L⁻¹ nano-chtosan, and 62.5 mg L⁻¹ nano-
chtosan) on the bochemcal sol propertes and
productvty of common beans grown n clay-salne
sol. Ther research emphaszed the adopton of eco-
frendly strateges to enhance the sustanablty of
agrcultural ecosystems. The hgher chlorophyll
content ndcates enhanced photosynthess, whch n
turn promotes ncreased plant growth and
development. Ths also explans the hgher yeld
observed n chtosan applcatons compared to the
control.
3.4 Frut Propertes
When examnng frut wdth, t s observed that the
use of chtosan shows an ncrease compared to the
control. The use of 150 ppm chtosan ncreased by
8.12% compared to the control, 300 ppm by 26.78%,
and 450 ppm by 11.22%. The use of chtosan has
generally ncreased the frut wdth. Frut sze has also
shown a sgnfcant ncrease n chtosan usage, just
lke frut wdth. The frut sze ncreased by 9.39% n
3801 c
6364 a
5506 ab
4473 b
0
2000
4000
6000
8000
Control 150 ppm
Chitosan
300 ppm
Chitosan
450 ppm
Chitosan
Yield(kg/m2)
Optimization of Foliar Chitosan Doses as a Biofertilizer for Enhanced Pepper Cultivation
255

the 150 ppm chtosan applcaton compared to the
control, 21.74% n the 300 ppm applcaton, and
7.65% n the 450 ppm applcaton.
No correlaton has
been establshed between chtosan applcaton and
dry matter producton percentage (Table 3)
. Wth the
use of chtosan as a bostmulant, not only qualty
parameters but also physcal measurements of the
frut show mprovement (Rahman et al., 2018).
Table 2: Effects of chtosan doses on chlorophyll content n
pepper.
Apps.
Chlorophyll
Content(SPAD)
Control 49.14 b
150
pp
m Chitosan 54.49 a
300
pp
m Chitosan 50.97 a
450
pp
m Chitosan 55.55 a
P
0.05
0.01
LSD 5.001
There s no sgnfcant dfference between means wth the
same letter n the same column, LSD the least sgnfcant
dfference
Table 3: Effects of chtosan doses on frut propertes n
pepper.
Apps. Frut
Wdth
(
mm
)
Frut
Heght
(
cm
)
Dry Matter
(%)
Control 25.5 b 11.50 b 5.31
150 ppm
Chtosan
27.57 b 12.58 ab 5.24
300 ppm
Chtosan
32.33 a 14.00 a 5.36
450 ppm
Chtosan
28.36 ab 12.38 ab 5.44
P
0.05
0.03 0.02 N.S
LSD 4.43 1.47 N.S
There s no sgnfcant dfference between means wth the
same letter n the same column, LSD the least sgnfcant
dfference
3.5 Frut Colour
The LAB color model ncludes L (lghtness), A (green
to red scale), and B (blue to yellow scale). Across all
treatments, the lghtness (L) values are smlar, wth a
slght ncrease observed wth chtosan applcatons.
The hghest lghtness occurs at 300 ppm (46.81),
whle the control group has the lowest value (45.19).
Although the dfferences n lghtness between the
control and chtosan-treated groups are 4ort, they
suggest a modest mprovement n brghtness wth
chtosan (Table 4).
The A parameter, representng the green-red scale,
shows a gradual ncrease wth hgher chtosan
concentratons. The control group has an A value of -
5.18, whle the maxmum shft toward red occurs at
300 ppm (-6.92). Ths ndcates that the 300 ppm
chtosan applcaton slghtly enhances red
pgmentaton compared to other treatments and the
control (Table 4).
4ort he B parameter (blue-yellow scale), the
hghest value s recorded at 300 ppm (18.20 a), wth
both 300 ppm and 450 ppm treatments yeldng
statstcally sgnfcant ncreases n yellow
pgmentaton compared to the control (16.34 b). Ths
suggests that chtosan, partcularly at 300 ppm,
enhances the yellow coloraton of the frut,
potentally contrbutng to a more vsually appealng
appearance (Table 4).
The applcaton of 300 ppm chtosan s the most
effcacous n affectng frut coloraton, resultng n
substantal ncreases n the yellow (B) and red (A)
scales, thereby mprovng overall color
characterstcs. Ths concentraton has the greatest
lghtness, renderng the frut more lumnous.
Chtosan treatments, partcularly around 300 ppm,
sgnfcantly mprove the aesthetc qualty of the
fruts. Dulta et al. (2022) examned the mpact of ZnO
nanopartcle-nfused chtosan coatng on the post-
harvest qualty of eggplants. Improvements n both
the texture and color of the fruts were noted n
comparson to the control.
Table 4: Effects of chtosan doses on frut colour n pepper.
Apps. L(Frut) A(Frut) B(Frut)
Control 45.19 -5.18 16.34 b
150 ppm
Chtosan
46.61 -5.35 18.57 a
300 ppm
Chtosan
46.81 -6.92 18.20 a
450 ppm
Chtosan
46.16 -6.69 17.15 a
P
0.05
N.S N.S 0.01
LSD N.S N.S 1.98
There s no sgnfcant dfference between means wth the
same letter n the same column, LSD the least sgnfcant
dfference, NS: Non sgnfcant.
3.6 Qualty Paramaters
3.6.1 pH and EC n Frut Juce
Although there s a notceable ncrease n pH value,
no statstcally sgnfcant ncrease has been observed.
However, the EC value decreased as the chtosan dose
ncreased, specfcally showng a reducton at the 450
I-CRAFT 2024 - 4th International Conference on Research of Agricultural and Food Technologies
256

ppm applcaton (Table 5). In ther study publshed,
Daşgan et al. (2022) found the effects of bostmulant
applcaton on EC and pH wthn the same group.
Table 5: Effects of chtosan doses on pH and EC n pepper.
A
pp
s.
p
H EC
(
dSm
-1
)
Control 5.54 4.99 a
150 ppm
Chtosan
5.86 5.06 a
300 ppm
Chtosan
5.87 5.11 a
450 ppm
Chtosan
5.76 4.47 b
P
0.05
N.S. 0.01
LSD N.S. 0.51
(There s no sgnfcant dfference between means wth the
same letter n the same column, LSD the least sgnfcant
dfference, NS: Non sgnfcant).
3.6.2 Vtamn C
The control group exhbts the lowest Vtamn C
content at 69.6 (b), sgnfyng that n the absence of
chtosan treatment, Vtamn C levels are nferor to
those n treated groups. The 150 ppm chtosan
treatment demonstrates a consderable elevaton n
Vtamn C content to 76.1 (a), whch s statstcally
superor to the control, underscorng the benefcal
mpact of chtosan at ths dose. At 300 ppm, the
Vtamn C concentraton attans 73.0 (ab), surpassng
the control but not exhbtng a statstcally sgnfcant
dfference from the 150 ppm treatment. The 450 ppm
chtosan concentraton yelds the greatest Vtamn C
content at 78.2 (a), exhbtng a statstcally
sgnfcant dfference from the control, ndcatng that
elevated chtosan concentratons may further enhance
Vtamn C levels.
The Vtamn C value showed an
ncrease of 9.34% at 150 ppm, 4.47% at 300 ppm, and
12.36% at 450 ppm compared to the control (Table 6).
The study demonstrates that chtosan treatments
markedly ncrease Vtamn C content. Whle 450 ppm
produces the maxmum concentratons, 150 ppm
provdes a sgnfcant enhancement wth only one-
thrd the quantty of chtosan relatve to 450 ppm. In
lght of the absence of a statstcally sgnfcant
dfference, 150 ppm may be regarded as the more
effectve opton. Plants subjected to chtosan exhbt
enhanced synthess of secondary metaboltes,
ncludng polyphenolcs, lgnn, flavonods, and
phytoalexns, alongsde Vtamn C, resultng n
mproved product qualty. Chtosan nanopartcles of
varyng szes have been employed to encapsulate
Vtamn C, prolong ts shelf lfe, and nvestgate ts
dstrbuton.
3.6.3 Total Phenols and Total Flavanods
Compounds
Total phenol and flavonoid contents in the fruit are
directly associated with quality, and it has been
observed that the use of chitosan in pepper fruit
increases these substances (Gholamipour et al.,
2009). In our study, when examining the effect of
chitosan use on the total phenolic content in pepper
plants, an increase of 15.69% was observed at a 150
ppm dose, 14.81% at a 300 ppm dose, and 12.39% at
a 450 ppm dose. Even though there is no statistical
difference, there is a decrease in the total phenol
content as the dose increases. When the total
flavonoid content was examined, compared to the
control, 150 ppm chitosan showed an increase of
22.21%, 300 ppm an increase of 11.31%, while the
application at the 450 ppm dose showed a decrease of
0.44%. A high dose of chitosan has caused a decrease
in the total flavonoid content (Table 7).
Table 6. Effects of chtosan doses on Vtamn C n pepper.
A
pp
s. Vt C
(
m
g
100
g
FW
−1)
Control 69.6 b
150 ppm Chtosan 76.1 a
300 ppm Chtosan 72.9 ab
450
pp
m Chtosan 78.2 a
P
0.05
0.01
LSD 4.41
There s no sgnfcant dfference between means wth the
same letter n the same column, LSD the least sgnfcant
dfference.
Table 7. Effects of chtosan doses on total phenols and
flavanods n pepper.
Apps. Total phenol
(mg GA/100 g
FW)
Total
flavonods
(mg
RU/100 g
FW)
Control 74.39 b 126.22 b
150
pp
m Chtosan 86.06 a 154.27 a
300
pp
m Chtosan 85.41 a 140.50 ab
450 ppm Chtosan 83.61 a 125.66 b
P
0.05
0.008 0.001
LSD 4.31 14.64
There s no sgnfcant dfference between means wth the
same letter n the same column, LSD the least sgnfcant
dfference.
Optimization of Foliar Chitosan Doses as a Biofertilizer for Enhanced Pepper Cultivation
257

4 CONCLUSIONS
Ths study conclusvely reveals that the applcaton of
chtosan greatly nfluences the growth, physcal
characterstcs, and yeld of pepper plants. The use of
150 ppm chtosan produced the maxmum yeld,
suggestng that lower concentratons are more
effcacous n mprovng yeld results n pepper
growng. Although 300 ppm chtosan was deal for
enhancng frut characterstcs ncludng breadth,
heght, and color pgmentaton, t dd not exceed 150
ppm n overall productvty. Moreover, elevated doses
such as 450 ppm shown no substantal advantages and
could even detrmentally affect yeld. The results
ndcate that chtosan s an excellent bostmulant for
pepper growth, wth 150 ppm dentfed as the
optmal concentraton for enhancng yeld and crop
qualty. Ths research underscores the promse of
chtosan as an economcal and envronmentally
sustanable bostmulant for mprovng pepper yeld.
Future nvestgatons should examne the mpact of
chtosan across dverse envronmental condtons and
varous pepper cultvars to enhance applcaton
methodologes.
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