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Agro-morphological Characterization and Fatty Acid Composition Analysis of

Selected Sunflower Accessions

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Philippine Journal of Science
150 (5): 1255-1264, October 2021
ISSN 0031 - 7683
Date Received: 01 Mar 2021

Agro-morphological Characterization and Fatty Acid

Composition Analysis of Selected Sunflower Accessions

Christine Joy S. Manalili1,2, Erick Allain C. Flores1,

Paula Blanca V. Gaban1,3, and John Dave C. Aquino1,2*

1University
Research Center, Research and Extension,
Central Luzon State University Science City of Muñoz, Nueva Ecija 3120 Philippines
2Department of Crop Science, College of Agriculture, Central Luzon State University
Science City of Muñoz, Nueva Ecija 3120 Philippines
3Science Education Institute, Department of Science and Technology
Bicutan, Taguig City 1630 Philippines

Sunflower is one of the most important oil crops, with the essential fatty acids of its oil considered
to be important in the human diet. Various sunflower accessions necessitate morphological and
oil research. This study evaluates the agro-morphological characteristics of sunflower using the
guidelines developed by the International Union for the Protection of New Varieties of Plants
(UPOV). Moreover, it also assessed the distinctness, uniformity, and stability among accessions
through the agglomerative clustering method, with the morphological diversity estimated using
the Shannon-Weiner diversity index that ranges from low to moderate. A significant relationship
was observed between pollen diameter and sunflower oil. Twenty (20) accessions were extracted
to determine the oil content and fatty acid composition using gas chromatography–mass
spectrometry (GC-MS). Out of 20 sunflower germplasms, 18 accessions had oil extracted. CL-
SF20 obtained the highest amount of extracted oil. GC-MS analysis revealed that CL-SF14
has 71.5% oleic acid, 6.86% palmitic acid, and the lowest polyunsaturated/saturated fatty acid
(P/S) index of 0.4228.

Keywords: fatty acid composition, morphological traits, oleic acid, sunflower oil

INTRODUCTION oil sources (Gunstone 2002). In the Philippines, sunflower

Oilseed crops are broadly developed around the world
and are considered one of the foremost imperative crops
due to their financial esteem. They are developed for
different purposes and accumulated more consideration
due to the expanding request for solid vegetable oils,
animal bolsters, pharmaceuticals, biofuels, and other
mechanical employments (Rahman and de Jimenez 2016).
Soybean, palm, rapeseed/canola, and sunflower seed are
considered the world’s top four most important vegetable
*Corresponding Author: jdaveaquino@clsu.edu.ph
was introduced during the 1970s (dela Cruz 1986), and
Central Luzon State University (CLSU) is one of the
leading institutions that work on sunflower researches
(Aganon et al. 1996).
Sunflower (Helianthus annuus), with 2n = 34, is an annual
plant that belongs in the family Asteraceae (Luqueño
et al. 2014). The flower head (inflorescence) consists
of disk florets tightly packed in the receptacle and ray
florets around the head’s edge. Disk florets are perfect
flowers that produce seeds (achenes) (Seiler and Jan

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Philippine Journal of Science
Vol. 150 No. 5, October 2021
2002), which leads to oil production (Palmer 1998).
There is an expanding trend of sunflower oil demand due
to its dietary properties (Renganayaki et al. 2008). The
nutritional qualities of vegetable oils depend mainly on
their fatty acid composition. Sunflower seed oil quality
is linked with the concentration of oleic and linoleic acid
(Zambelli et al. 2015). High oleic acid is well-preferred
due to its benefits to human health, decreasing the risk of
coronary heart disease and other cardiac diseases (Salas
et al. 2015).
Additionally, oils with high oleic acid content are resistant
to heat oxidation and have a longer shelf life, low
cholesterol effect, and lesser linolenic acid content (Harun
2019). The information of the fatty acid composition
makes oil production potential for intended purposes. For
that reason, it is imperative to analyze the oil content and
composition of CLSU sunflower germplasm for future
breeding of sunflower with stable and healthy oil.
Figure 1. Plant Materials. A) CL-SF1, B) CL-SF2, C) CL-SF3, D) CL-SF11, E) CL-SF13, F) CL-SF14, G) CL-SF16, H) CL-SF17, I) CL-
SF18, J) CL-SF19, K) CL-SF20, L) CL-SF21, M) CL-SF23, N) CL-S24, O) CL-SF26, P) CL-SF27, Q) CL-SF30, R) CL-SF31,
S) CL-SF32, and T) CL-SF33.
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Manalili et al.: Agro-Morphological Characterization
MATERIALS AND METHODS
Collection of Plant Materials
Sunflower (Helianthus annuus L.) accessions used in this
study were obtained from all different sources across the
globe. Figure 1 shows the CLSU sunflower collection.
Sunflower Crop Management
All the agronomic practices recommended for the
region using the CLSU sunflower production manuals
were followed to reach the optimum quality of the crop
(CLSU 2004). Sunflower crops were cultivated during the
production season of December–May. The experimental
field was plowed and harrowed using a rotoridger and
was irrigated afterward. Final harrowing was done after
three days to ensure complete land preparation. Plots were
prepared with a size of 0.5 m x 1 m and a distance of 0.5
m between plots. Seeds were pre-germinated in Petri
plates for 2 d under a 37 °C temperature, then planted in
Philippine Journal of Science
Vol. 150 No. 5, October 2021
seedling trays. Soil medium was prepared with a mixture
of 1:1:1 ratio of garden soil, carbonized rice hull, and
organic soil. Soil mixing was done twice a day (morning
and afternoon) for 14 d to prevent soil-borne pathogens.
Ten (10) seedlings per variety were planted per plot with
a distance of 0.5 m between plants. About 0.2 g of urea
(46-0-0) was applied basally after transplanting. At 7 d
after transplanting, spot application of 2.0 g urea was
done and repeated weekly during the vegetative stage.
The experimental area was irrigated weekly, and manual
watering was done whenever necessary. Insect pests and
disease incidence were monitored daily, while manual
hand weeding was done weekly.
Harvesting, Drying, and Threshing
Harvesting was done when the sunflowers’ receptacle head
turned brownish-yellow or when the flower reached its
post-harvest maturity. It was done by cutting the sunflower
heads using sharp pruning shear. The heads were sundried
for 2–3 d before threshing. Threshing was done by rubbing
the heads with an improvised tool. Seeds were cleaned
and subjected to oil extraction.
Morphological Characterization
Sunflower accessions were seed increased through
selfing to ensure the uniformity of the planted accessions
prior to characterization. Sunflower plant descriptors
developed by UPOV (2018) were used in morphological
characterization with some modifications.
Oil Extraction
Seed extraction was carried out using a mini-oil extractor
(Labotech Trading) with a temperature requirement of 190
°C during extraction. Fifty grams (50 g) of seeds were
extracted. The extracted oil was filtered using a funnel and
filter paper until the oil was settled down. The extracted
oil was gathered and measured using the pipette.
GC-MS Analysis of Sunflower Oil Fatty Acids
Eighteen (18) sunflower oil samples were subjected to GC-
MS analysis. Prior to the analysis, the sunflower oil samples
were esterified following the official AOAC method (2012).
Materials and Reagents
Reagents for esterification such as boron trifluoride
(BF 3) in 14% methanol, 0.5 M methanolic sodium
hydroxide (NaOH), heptane, sodium chloride (NaCl), and
anhydrous sodium sulfate (Na2SO4) were of analytical
grade or prepared from analytical grade reagents. The
100-mg standard fatty acid methyl ester (FAME) for
the identification and quantification of fatty acids in the
sunflower oil was purchased from Sigma-Aldrich.
Manalili et al.: Agro-Morphological Characterization
Esterification of Fatty Acids
Three-hundred fifty (350) mg of oil is added to a flask, and
the methanolic NaOH was added. A reflux condenser was
attached to the flask and was gently heated at 60 °C until
the fat globules disappeared. The BF3 solution was added,
and the mixture was again boiled for 2 min. Five (5) mL
of heptane was added to the mixture to extract the FAMEs
and was boiled for 1 min. The solution was then removed
from heat and 15 mL of the saturated NaCl solution was
added, followed by vigorous shaking. Additional NaCl
solution was added to float the heptane solution containing
the FAMEs. A small amount of anhydrous sodium Na2SO4
was then added to the solution to remove water. Samples
for analysis were obtained by carefully pipetting out the
heptane layer using a pipettor.
GC-MS Analysis
Analysis of FAMEs was done using a gas chromatograph
coupled with a mass spectrometer (Shimadzu GCMS-
QP2020) equipped with an Rtx-5MS column of 0.25 mm
internal diameter, 30 m length, and 0.25 μm thickness.
The injector temperature was set to 250.0 °C. The column
flow was set to 0.89 mL/min (with a total flow of 51.2
mL/min). Split injection mode was applied with a split
ratio of 50. The oven temperature was programmed as
follows: the initial temperature was initially set at 100 °C,
then increased at a rate of 15 °C/min to 220 °C and held
for 5 min, then increased at a rate of 20 °C/min for a final
temperature of 260 °C, to a total program time of 15 min.
The carrier gas used was helium. Electron ionization was
the ionization method used, and the temperatures of the
interface and the ion source were set at 280 and 200 °C,
respectively. The mass-to-charge ratio (m/z) range used
for the analysis was between 100–400.
A solution of the FAME standard was first analyzed in the
GC-MS via scan mode to determine the corresponding
retention times and specific m/z ratios of each FAME
component. A calibration curve for each FAME was made
using five different concentrations of the FAME standard by
plotting the peak area in the chromatogram (y-axis) of each
FAME against the corresponding concentration (x-axis).
Once the retention time and the m/z for each FAME were
determined, the esterified sunflower oil samples were
analyzed in the GC-MS via the selected ion monitoring
mode.
Statistical Analysis
To assess the distinctness, uniformity, and stability among
accessions the agglomerative clustering method was used
to demonstrate the distribution among accessions, and
correlation analysis was cast out to analyze the correlation
of sunflower oil and its physical characteristics using
STAR (Statistical Tool for Agricultural Research).
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Philippine Journal of Science
Vol. 150 No. 5, October 2021
RESULTS
Morphological Traits and Clustering Analysis of the
Sunflower Germplasm
Sunflower has a very diverse germplasm across the
globe. Morphological characterization on the vegetative
to post-harvest is a basis for assessing the distinctness,
uniformity, and stability among accessions. The
agglomerative clustering method cast five clusters
representing the distribution among the similarities and
differences among 20 different accessions. Cluster A
(CL-SF1, CL-SF11, CL-SF13 CL-SF18, and CL-SF32)
was similar on the following characteristics: leaf wing
(none or very weakly expressed), stem color (green),
the shape of seeds (broad ovoid), and hairy structure
(strongly expressed). Cluster B (CL-SF16) formed
independently in a cluster due to its ray floret color
(dark red) and leaf blistering (weak). Cluster C (CL-
SF2, CL-SF3, CL-SF14, CL-SF17, CL-SF20, CL-SF21,
CL-SF23, CL-SF24, CL-SF27, and CL-SF33) showed
similarity in the presence of pollen production. Cluster
D (CL-SF26) differed on seed color index (white) and
seed stripes on a margin, among other accessions. Lastly,
Cluster E (CL-SF 19, CL-SF30, and CL-SF31) had
observed similarities of the following traits: leaf color
(medium), leaf shape (broad triangular), ray floret length
(medium), anthocyanin production (presence), type of
branching (overall), seed stripes (absence), and seed
shape (narrow ovoid).
Figure 2. Dendrogram using agglomerative clustering method
demonstrating the distribution of 20 CLSU sunflower
accession based on morphological traits.
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Manalili et al.: Agro-Morphological Characterization
Biodiversity Index
The measure of morphological diversity and equitability
index was estimated using the Shannon-Weiner diversity
index (H), whereas H = –å Pi(lnPi), where Pi is the
Figure 3. Morphological diversity and equitability index.
proportion of each species in the sample (Ortiz-Burgos
2016). Equitability index was found on the leaf (blistering
and wing), stem color, head shape, ray floret (density and
shape), disk floret color, type of branching, the position
of the lateral head, head attitude, seed stripes (on margin,
between margin and color), hairy structure, and shape of
the seeds – ranging from 0.71–0.95. Moderate diversity was
found mostly on leaf descriptor, flower heads (uniformity
and sizes), ray floret (length and color), anthocyanin of
the stigma, presence of branching and pollen, seed color
index, seed stripes, and seed spots on pericarp – ranging
from 0.57–0.66.
Figure 4. Oil content (µL/g) extracted from 20 different sunflower
accessions.
Oil Content
Sunflower seeds from 20 different sunflower accessions
were extracted using a mini-oil extractor. Among the
sunflower accessions, 18 contain oil while the remaining
two accessions have no oil gathered. The amount of oil
extracted per 1 g of seeds ranged from 35.7–278.0 µL/g.
The highest oil content of 278.0 µL/g was collected from
CL-SF20 and followed by CL-SF2, which had 225.7 µL/g
in oil content. The least amount of oils was extracted from
CL-SF14 with 35.7 µL/g. On the other hand, no oil was
collected from two varieties such as CL-SF17 and CL-
SF24 (Figure 4).
Philippine Journal of Science Manalili et al.: Agro-Morphological Characterization
Vol. 150 No. 5, October 2021

Oil Composition DISCUSSION

The sunflower oil analyses aimed to identify and quantify
the amount of long-chain unsaturated fatty acids, mainly
Morphological Traits and Clustering Analysis of the
oleic and linoleic acids, which were ideal fatty acids
Sunflower Germplasm
sought in vegetable oils. The total monounsaturated fatty
Dendrogram using the agglomerative clustering method
acids (MUFA), total polyunsaturated fatty acids (PUFA),
was presented in Figure 2 for the demonstration
total saturated fatty acids (SFA), and the P/S index were
of the distribution among traits. Five clusters were
also calculated from the obtained data.

Table 1. Fatty acid composition of sunflower oils (8- to 16-chain).

Fatty acid composition (%)
Variety Caprylic Capric Lauric Tridecylic Myristoleic Myristic Pentadecanoic Palmitoleic Palmitic
(C8:0) (C10:0) (C12:0) (C13:0) (C14:1) (C14:0) (C15:0) (C16:1) (C16:0)

CL-SF1 0.0054±0.0014 0.0042±0.0009 0.0083±0.0020 0.0057±0.0015 0.0042±0.0004 0.0710±0.0036 0.0217±0.0023 0.1600±0.0514 7.9667±1.4620

CL-SF2 0.0000±0.0000 0.0000±0.0000 0.0073±0.0005 0.0051±0.0006 0.0041±0.0006 0.0913±0.0213 0.0246±0.0036 0.1557±0.0283 9.5632±1.5219

CL-SF31 0.0060±0.0007 0.0043±0.0012 0.0061±0.0013 0.0043±0.0010 0.0045±0.0011 0.0966±0.0178 0.0278±0.0053 0.1631±0.0205 9.8656±2.2235

CL-SF13 0.0043±0.0004 0.0059±0.0010 0.0046±0.0003 0.0049±0.0012 0.0044±0.0013 0.0451±0.0093 0.0213±0.0017 0.1307±0.0189 7.3120±0.6086

CL-SF27 0.0048±0.0012 0.0252±0.0024 0.0076±0.0013 0.0052±0.0008 0.0062±0.0005 0.0903±0.0166 0.0303±0.0052 0.1510±0.0105 9.2671±1.4942

CL-SF18 0.0049±0.0007 0.0049±0.0002 0.0092±0.0021 0.0053±0.0008 0.0000±0.0000 0.0852±0.0085 0.0353±0.0059 0.1923±0.0309 8.8575±2.0381

CL-SF26 0.0046±0.0009 0.0098±0.0020 0.0051±0.0007 0.0049±0.0008 0.0058±0.0005 0.0922±0.0177 0.0349±0.0027 0.1272±0.0270 8.6008±0.3513

CL-SF33 0.0048±0.0009 0.0048±0.0010 0.0078±0.0016 0.0054±0.0005 0.0000±0.0000 0.1443±0.0363 0.0215±0.0046 0.2561±0.0527 11.8436±0.919

CL-SF30 0.0047±0.0003 0.0064±0.0006 0.0108±0.0024 0.0074±0.0023 0.0059±0.0011 0.0795±0.0215 0.0428±0.0071 0.0878±0.0183 8.1636±2.1159

CL-SF3 0.0050±0.0007 0.0061±0.0014 0.0064±0.0011 0.0059±0.0009 0.0064±0.0018 0.0758±0.0125 0.0411±0.0070 0.0923±0.0101 8.5192±2.0014

CL-SF21 0.0000±0.0000 0.0042±0.0009 0.0053±0.0003 0.0041±0.0004 0.0041±0.0011 0.0647±0.0123 0.0367±0.0067 0.1551±0.0174 8.0927±2.0450

CL-SF20 0.0000±0.0000 0.0000±0.0000 0.0076±0.0008 0.0057±0.0010 0.0033±0.0006 0.1008±0.0152 0.0229±0.0044 0.2143±0.0518 9.8635±1.1167

CL-SF19 0.0035±0.0004 0.0043±0.0003 0.0073±0.0017 0.0043±0.0006 0.0037±0.0009 0.0908±0.0086 0.0333±0.0062 0.2076±0.0581 9.6021±0.5489

CL-SF16 0.0060±0.0017 0.0051±0.0005 0.0061±0.0017 0.0090±0.0020 0.0053±0.0007 0.0755±0.0226 0.0396±0.0024 0.2054±0.0489 9.2493±1.4964

CL-SF14 0.0050±0.0010 0.0043±0.0009 0.0054±0.0012 0.0046±0.0008 0.0057±0.0017 0.0587±0.0135 0.0188±0.0024 0.1169±0.0321 6.8569±1.5911

CL-SF11 0.0056±0.0012 0.0048±0.0008 0.0063±0.0008 0.0081±0.0005 0.0058±0.0010 0.0751±0.0232 0.0316±0.0008 0.1724±0.0254 8.9188±2.5936

CL-SF32 0.0070±0.0007 0.0067±0.0012 0.0073±0.0005 0.0074±0.0014 0.0058±0.0006 0.0848±0.0156 0.0320±0.0074 0.1956±0.0350 9.1075±1.7354

CL-SF23 0.0066±0.0010 0.0057±0.0002 0.0071±0.0018 0.0063±0.0010 0.0062±0.0011 0.0810±0.0147 0.0340±0.0055 0.1888±0.0370 9.1383±1.7975

Table 2. Fatty acid composition of sunflower oils (17- to 22-chain).

Fatty acid composition (%)

Variety Margaric Linoleic Oleic/elaidic Linolenic Stearic Gondoic Eicosanoic Erucic Behenic
(C17:0) (C18:2) (18:1) (C18:3) (C18:0) (C20:1) (C20:0) (C22:1) (C22:0)

CL-SF1 0.0640±0.0069 20.9009±3.7547 58.9832±11.5155 0.0297±0.0052 8.8900±1.4269 0.4067±0.0477 0.9688±0.1904 0.0052±0.0011 1.5044±0.3349

CL-SF2 0.0590±0.0080 19.8445±3.7241 58.3700±17.1912 0.0204±0.0018 9.2405±1.5077 0.3745±0.0365 1.0084±0.1737 0.0036±0.0004 1.2278±0.0543

CL-SF31 0.0832±0.0174 26.2594±3.9274 53.0223±8.0756 0.0319±0.0035 6.7000±1.2433 0.3805±0.0524 0.8270±0.2496 0.0043±0.0005 2.5131±0.2723

CL-SF13 0.0862±0.0175 17.3261±0.6764 64.0512±13.8724 0.0258±0.0040 6.9517±0.2955 0.4760±0.0754 0.7983±0.1621 0.0045±0.0008 2.7469±0.0854

CL-SF27 0.0828±0.0020 15.4910±2.8606 63.0980±11.7048 0.0244±0.0030 8.2526±1.3667 0.3677±0.0565 0.9863±0.1813 0.0000±0.0000 2.1093±0.4154

CL-SF18 0.0575±0.0096 20.8059±5.7090 60.2287±8.6955 0.0359±0.0055 6.3902±1.2341 0.3717±0.0829 0.8069±0.0868 0.0049±0.0009 2.1038±0.5264

CL-SF26 0.0674±0.0149 18.1851±2.4826 62.4264±14.7544 0.0210±0.0050 7.2730±1.8039 0.3969±0.0501 0.8629±0.1899 0.0047±0.0009 1.8774±0.4454

CL-SF33 0.0887±0.0141 29.3833±3.8839 51.6587±9.9162 0.0300±0.0041 3.8555±0.9217 0.4578±0.0958 0.4585±0.0907 0.0044±0.0012 1.7746±0.3549

CL-SF30 0.1048±0.0168 13.5873±3.1140 65.5988±11.5543 0.0323±0.0052 8.8321±1.6415 0.3649±0.1210 0.9934±0.1706 0.0056±0.0006 2.0718±0.1938

CL-SF3 0.0798±0.0109 21.1450±4.8233 57.4790±11.5048 0.0234±0.0039 9.0119±0.8773 0.4394±0.0751 1.0650±0.1503 0.0050±0.0011 1.9934±0.2920

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Vol. 150 No. 5, October 2021

Fatty acid composition (%)

Variety Margaric Linoleic Oleic/elaidic Linolenic Stearic Gondoic Eicosanoic Erucic Behenic
(C17:0) (C18:2) (18:1) (C18:3) (C18:0) (C20:1) (C20:0) (C22:1) (C22:0)

CL-SF21 0.0720±0.0135 20.0900±1.4504 61.0219±9.1565 0.0268±0.0034 7.8402±2.2135 0.3690±0.0822 0.9336±0.2511 0.0040±0.0008 1.2758±0.3451

CL-SF20 0.0664±0.0201 19.4353±3.2751 57.4734±17.6244 0.0185±0.0060 9.5329±1.7479 0.2772±0.0131 0.9304±0.1033 0.0041±0.0010 2.0439±0.3592

CL-SF19 0.0894±0.0237 18.7762±4.8404 60.0638±11.1374 0.0309±0.0078 7.9090±1.1094 0.3322±0.0652 0.9738±0.0804 0.0059±0.0010 1.8619±0.3292

CL-SF16 0.075±0.01650 19.3079±1.9983 59.9079±13.4374 0.0288±0.0055 6.6256±0.7470 0.3387±0.0508 0.7776±0.0591 0.0000±0.0000 3.3368±0.3993

CL-SF14 0.0809±0.0155 8.3284±1.2180 71.5058±6.8813 0.0134±0.0028 8.8742±1.4384 0.2966±0.0705 0.8761±0.1874 0.0045±0.0011 2.9439±1.0020

CL-SF11 0.1054±0.0104 22.0247±4.4030 57.1104±2.5725 0.0264±0.0047 7.9733±1.5529 0.2978±0.0335 0.8456±0.1328 0.0000±0.0000 2.3880±0.3628

CL-SF32 0.0580±0.0021 21.2128±3.0987 58.8566±12.6064 0.0246±0.0058 6.3880±1.0537 0.3392±0.0546 0.8097±0.2277 0.0000±0.0000 2.8571±0.7922

CL-SF23 0.0808±0.0199 19.0027±5.2489 61.1049±11.6278 0.0253±0.0065 6.6693±1.0177 0.3281±0.0702 0.7279±0.2427 0.0048±0.0009 2.5823±0.7031

Table 3. Polyunsaturated/unsaturated fatty acid index of sunflower oils.

Total MUFA +
Variety Total MUFA (%) Total PUFA (%) Total SFA (%) P/S index
PUFA (%)
CL-SF1 59.5593 20.9306 19.5102 81.7903 1.0728
CL-SF2 58.9079 19.8649 21.2272 79.8660 0.9358
CL-SF31 53.5747 26.2913 20.1340 81.6394 1.3058
CL-SF13 64.6668 17.3519 17.9812 81.6709 0.9650
CL-SF27 63.6229 15.5154 20.8615 80.6346 0.7437
CL-SF18 60.7976 20.8418 18.3607 79.6375 1.1351
CL-SF26 62.9610 18.2061 18.8330 80.4899 0.9667
CL-SF33 52.3770 29.4133 18.2095 79.1905 1.6153
CL-SF30 66.0630 13.6196 20.3173 80.6608 0.6703
CL-SF3 58.0221 21.1684 20.8096 81.1671 1.0172
CL-SF21 61.5541 20.1168 18.3293 82.0187 1.0975
CL-SF20 57.9723 19.4538 22.5741 79.7940 0.8618
CL-SF19 60.6132 18.8071 20.5797 78.7728 0.9139
CL-SF16 60.4573 19.3367 20.2056 79.4203 0.9570
CL-SF14 71.9295 8.3418 19.7288 77.4261 0.4228
CL-SF11 57.5864 22.0511 20.3626 79.1383 1.0829
CL-SF32 59.3972 21.2374 19.3655 79.6826 1.0967
CL-SF23 61.6328 19.0280 19.3393 80.2713 0.9839

formed based on the similarities and differences to its
morphological characteristics among different accessions.
The environment is one of the contributing factors that
are responsible for the manifestation of leaf size and
shape. Increasing the rate of photosynthesis is one of the
main targets of sunflower breeding. Thus, leaves play an
imperative part in the process of photosynthesis (Hlandni
et al. 2017). It is additionally related to the plant species’
capability to obtain higher yields because it plays a vital
part in controlling water loss and affects crop growth
and development rates through photosynthesis (Kulkarni
et al. 2015). The marketing of the sunflower as the
ornamental cut flower influenced the size of the flower
head. Additionally, branching is an essential trait for the
ornamental use of sunflower. It can increase the florist’s
income due to having many flower heads even in just one
plant (Cjevic et al. 2017). Ray floret color was observed
during the full-bloom of the sunflower head. Different
results were obtained based on the inheritance of ray floret
color in sunflower, which can be accredited to the different
genetic backgrounds among individuals (Miller and Fick
1997; Hlandni et al. 2017). Accordingly, ray floret and
disk floret shape and color had no significant impact on
the crop yield except for attracting the pollinators and
involvement in the pollination process. The sunflower
head, which consists of disk florets, varies in color and

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Vol. 150 No. 5, October 2021

develops a male and female organ (Hlandni et al. 2017).
The inclined head position will increase the amount of
solar radiation intercepted by the flower head, positively
impacting seed setting and composition. However,
the most preferred by breeders is a head position at a
135–180 °C angle and firmly attached to the stem, as
this flower head lowers the frequency of birds feeding
and development of some diseases (Hlandni et al. 2017).
Sunflower seed is one of the high-value parts as it contains
oil and edible. The confectionary seed usually seeds
with white seed stripes (Hlandni et al. 2017). While oil-
type sources are those black with the absence of stripes,
they usually had significantly smaller seed size than the
confection-type (Nel 2001).
Biodiversity Index
An equitability index was found on 15 morphological
characteristics, which indicates that there is higher genetic
diversity in the aforementioned sunflower morphological
characteristics. Conversely, moderate diversity was found
mostly on leaf descriptor and 10 other reproductive and
post-harvest characteristics showed that lower genetic
diversity was found in these characteristics (Figure 3).
Leaf descriptor was found as the lowest diversity, seed
traits are moderated diverse, and the highest diversity
index was found for flower head characteristics (Hlandni
et al. 2017).
Correlation Analysis
The correlation between the individual oil content and
seeds morphological parameters (Table 4) showed that
there is a very weak linear association between oil content
and the morphological parameters such as 100-seeds
weight, seed length, seed width, seed thickness, equivalent
diameter, sphericity, the unit mass of the seeds, and
pollen diameter. Consequently, a weak linear association
between the oil content and percent seed set was observed.
However, pollen diameter shows a significant relationship
to the oil content. The seeds’ physical characteristics
have no significant relationship to the amount of fine
material that the seed can produce (Nel 2001). However,
fertile pollen is needed in the pollination process that is
eventually required in seed production for an increase
in percent seed setting (Bonciu 2013). Additionally,
the number of seeds is the most important indicator of
seed yield and oil content essential in production and oil
processing (Hlandni et al. 2017).
Oil Content
Sunflower germplasm was considered to be mostly oil-
type, having extracted sunflower oil from 18 among
20 accessions (Figure 4). Sunflower oil performs
to be superior among other oil crops containing the
highest percentage of long-chain MUFA and PUFA. It
is recognized as a high-grade vegetable oil in line for
high unsaturated linoleic/oleic fatty acid concentration,
absence of significant harmful compounds, and excellent

Table 4. Correlation matrix between oil content and seeds morphological traits.
Variables SS SW100 SL SW ST ED S UM OC
SS –0.45 –0.63 –0.38 –0.50 –0.69 –0.47 –0.04 0.32
SW100 0.73 0.97 0.92 0.87 0.95 0.45 0.08
SL 0.71 0.69 0.91 0.74 0.27 –0.02
SW 0.95 0.85 0.98 0.47 0.09
ST 0.87 0.98 0.37 0.09
ED 0.90 0.31 0.03
S 0.40 0.10
U.M. –0.01
PD 0.14*
0 – no linear association
0< p < 0.2 – very weak linear association
0.2 ≤ p ≤ 0.4 – weak linear association
0.4 ≤ p ≤ 0.6 – moderate linear association
0.6 ≤ p ≤ 0.8 – strong linear association
0.8 ≤ p ≤ 1 – very strong linear association
1 – perfect linear association.

Legend:
SS percent seed setting ED equivalent diameter
SW100 100 seeds weight(g) S sphericity
SL seed length (mm) UM unit mass (g)
SW seed width (mm) OC oil content (µl)
ST seed thickness (mm) PD pollen diameter (µm)

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Vol. 150 No. 5, October 2021
nutritional properties. Although sunflower oil is more
expensive than other oil crops, the demand for sunflower
oil in the global marketplace is still rising due to its food
processors’ pursuit of trans-fat-free vegetable oil sources.
Sunflower oil is extensively used in cuisine for frying,
cooking, and dressing due to its nutritional values, pleasant
taste, and health benefits (FAO 2010). Oil concentration
in mature seeds differs on the specific cultivar due to
differences in protein body proportion, embryo, and oil
mass dynamics during achene growth, which underlie
differences in embryo oil concentration. All cell types
in the embryo – including the epidermal cells – formed
oil bodies, specifically in sunflower seeds (Mantese et
al. 2006).
Oil Composition
Shorter fatty acid chains were not apparent in all of the oil
samples (Table 1). Long-chain fatty acids were detected in
appreciable amounts starting from 16-carbon chains, and
the highest amounts were detected on 18-carbon chains
(linoleic, oleic/elaidic, and stearic). The predominant
SFA in the oil was palmitic (6.86% for CL-SF14 being
the lowest and 11.84% for CL-SF33 being the highest)
and stearic acids (3.86% for CL-SF33 being the lowest
and 9.53% for CL-SF20 being the highest). The highest
amount of linoleic acid was measured on CL-SF33
at 29.38%, while the rest of the accessions contained
8.33–26.26% linoleic acid. While the GC-MS column
used cannot distinguish oleic and elaidic acids from one
another, studies have already shown that sunflower oils
are rich in oleic acid. Therefore, it can be claimed with
certainty that the quantified C18:1 fatty acid were all oleic
acids. The highest oleic acid content was measured on CL-
SF14 with 71.5% oleic acid. The rest of the accessions had
oils containing 51–65% oleic acid (Table 2). High oleic
sunflower has greater MUFA content, which provides
better heat stability during frying compared to standard
sunflower oil. Moreover, its content in SFAs is also
reduced, whereas the vitamin E content does not change.
Linolenic acid, an omega-3 PUFA, was also detected in
small amounts ranging from 0.0134–0.0359%, but this
value is much lower than the 0.12–0.45% reported by
Chowdhury et al. (2007) and Zambiazi et al. (2007).
Kostik et al. (2013), however, did not detect any linolenic
acid at all in their sunflower oil samples. A diet rich in
linoleic and linolenic acids increases the levels of high-
density lipoproteins or “good” cholesterol (HDL) and
lowers the levels of low-density lipoproteins (LDL)
or “bad” cholesterol, while consumption of oleic acid
decreases only the LDL but does not affect the HDL levels
(Lawton et al. 2000). Fatty acid composition, nevertheless,
varies according to its genotypic properties (Chowdhury
et al. 2007; Akkaya 2018). There are three classes of
sunflower oil in the world market – namely, low oleic/
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Manalili et al.: Agro-Morphological Characterization
standard sunflower oil (10–29%), mid oleic (30–59%),
and high oleic (60–90%) (Lacombe and Bervillé 2001).
High oleic mutant (Pervenets) was first reported by
Soldatov (1976) with greater than 65% of oleic acid.
Behenic acid, a 22-carbon SFA, was also detected in
small amounts, with 1.23% in CL-SF2 being the lowest
value and 3.34% in CL-SF16 being the highest. This was
supported by the study of Kostik et al. (2013), which
claims that among their analyzed oil samples, only the
sunflower oil was detected for the presence of behenic
acid. Arachidic (20-carbon SFA), gondoic (20-carbon
MUFA), and erucic (22-carbon MUFA) acids were also
detected in the oil samples but in much smaller amounts.
The P/S index was calculated by dividing the PUFA by the
SFA. The highest calculated P/S index was 1.6153 for CL-
SF33, and the lowest is 0.4228 for CL-SF14 (Table 3). The
P/S index is an important indication of the nutritional value
of a particular oil, and oils and fats with indices greater
than 1 are considered to have nutritional values (Kostik
et al. 2013). Among the analyzed accessions, CL-SF33,
CL-SF31, CL-SF18, CL-SF21, CL-SF32, CL-SF11, CL-
SF1, and CL-SF3 have P/S indices that are greater than
1. Aside from nutritional values, however, higher values
of P/S mean lesser accumulation of lipids in the body
(Lawton et al. 2000). Moreover, the amounts of SFA in all
of the oil samples are all approximately one-fourth of the
total fatty acid content. High levels of SFA are favorable
for increasing the stability of the oil, but their excessive
consumption is related to an increase in cholesterol levels
and the tendency to become obese, as they are known to
increase the amount of LDL in the body (Przybylski and
McDonald 1995; Ristic and Ristic 2003; Dzisiak 2004).
It is recommended, therefore, to consume PUFA and
MUFA relative to SFA in the diet to appropriate the lipid
profile by inducing the oxidation of LDL and promoting
the synthesis of high-density lipoprotein, which in turn
lowers the triacylglycerol levels in the blood, which could
help in preventing heart diseases (Yu-Poth et al. 2000;
Kostik et al. 2013). The high levels of MUFA and PUFA
in sunflower oil (> 75%), therefore, make it suitable for
mass consumption (Kostik et al. 2013).
CONCLUSION
Sunflower is one of the most important oil crops as
the essential fatty acids of its oil is considered to be
important in the human diet. Some sunflower accessions
necessitate morphological and oil research. Among
the observed characteristics, five clustered was formed
using the agglomerative clustering method. The measure
of morphological diversity was estimated using the
Shannon-Weiner diversity index; low and moderate
Philippine Journal of Science
Vol. 150 No. 5, October 2021
diversity index was shown on the different morphological
traits. The correlation analysis of oil content reveals that
fertile pollen is necessary for seed and oil production.
The highest oil content was extracted from CL-SF20,
which can be recommended to sunflower growers for
oil production. GC-MS analysis revealed that CL-SF14
had 71.5% oleic acid and CL-SF33 had 29.38% linoleic
acid, both of which are the highest among the oil samples
analyzed. The accessions CL-SF33, CL-SF31, CL-SF18,
CL-SF21, CL-SF32, CL-SF11, CL-SF1, and CL-SF3 have
P/S indices that are greater than 1, which indicated a good
nutritional value. Moreover, further study is needed for
marker-assisted detection and gene sequencing analysis
in determining the type of mutation present. The mutation
may result in varying oil composition and assessment for
other oil genes responsible for oil-type sunflower seed to
evaluate different varieties that can be forwarded to oil
production and breeding purposes.
ACKNOWLEDGMENTS
This research study was supported by the CLSU Grants-
in-Aid Project. The authors acknowledge with thanks
the assistance of Ms. Xyrelle P. Juan and field support of
Mr. Albert H. Quero, Mr. Ramon R. dela Cruz, and Mr.
Jerry P. Salmasan.
STATEMENT ON CONFLICT OF
INTEREST
The authors declare no conflict of interest.
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