Citation
Oyeniyi, F. T., Bolaji, A. O., & Adeniran, O. I. (2026). SDS-PAGE characterization of green-stemmed and red-stemmed Basella alba L. International Journal of Research, 13(1), 394–405. https://doi.org/10.26643/ijr/2026/13
1Felix Timilehin OYENIYI, 1Abolade Oluremi BOLAJI and 2Oluwatobi Isaac ADENIRAN
1*Department
of Botany, Obafemi Awolowo University, Ile-Ife, Nigeria,
2Department of Biological Sciences, Adeleke
University, Ede Osun State.
*Corresponding author’s email: oyeniyiakanfe@gmail.com
ABSTRACT
The characterization of species by electrophoresis has played
significant roles in delimiting species over the years. In this study, the leaves and seeds obtained from matured green-stemmed and
red-stemmed Basella alba were
characterized using Sodium Dodecyl Sulphate (SDS-PAGE) Polyacrylamide Gel
Electrophoresis (PAGE) with
a view to investigate their taxonomic status. Data
obtained revealed high
level of similarity in the seed and leaf protein profile of
the two Basella morphotypes studied. The study revealed relatively high level of coefficient of similarity between the green stemmed and the red stemmed Basella
alba studied, with the seed protein
having 50% and the leaf protein having
75% similarity, indicating that a
strong degree of similarity exist between the green stemmed and the red stemmed Basella
alba studied irrespective of their stem
colors. Five common bands were found between the seed protein of the green stemmed and red stemmed
B. alba studied at 0.2cm – 0.4cm, 4.3 – 4.8 cm, 4.8 – 5.2, 5.5
cm – 5.8 cm, and 6.0 cm – 6.5 cm. The red stemmed B. alba seed
protein had 2 unique bands at 2.6 cm – 3.2 cm and 3.6 cm –
4.3 cm, while the green stemmed had 3 unique
bands at 2.8 cm –
3.2 cm, 5.2 cm – 5.3 cm, and 6.4 cm – 6.5 cm. Six common bands were found in the leaf protein of the two B.alba morphotypes studied at 2.3 – 2.6 cm,
4.5 cm – 4.8 cm, 5.0 – 5.4 cm, 6.0
cm – 6.3 cm, 6.4 cm – 6.5 cm, and 6.5 cm – 6.6 cm. The red-stemmed Basella alba leaf had one (1)
unique band at 0.2cm – 0.4cm, while the green-stemmed Basella alba leaf had one (1) unique band at 0.1cm. The study supported the taxonomic
classification of the green stemmed and red stemmed B.alba as
morphotypes or varieties belonging to same species.
KEY WORDS:
Basella alba, Electrophoretic characterization, Evolutionary relationship, Protein
bands
INTRODUCTION
The
green-stemmed and red-stemmed Basella alba are
vigorous, fast-growing, heat-tolerant, perennial tropical vines
(Chaurasiya et al., 2021; Tănase et al., 2020). They belong to
the family Basellaceae (Bolaji
et al., 2022a; Bose et al., 2023). Basella alba flowers are sessile, white
with pink tips, freely arranged in long-stalked spikes. The green-stemmed and
red-stemmed B. alba differ mainly in stem coloration and some floral and
fruit traits (Deshmukh & Gaikwad, 2020; Bolaji et al., 2022b; Bolaji et
al., 2023). Basella alba is widely cultivated as an annual leafy
vegetable and also grown as an ornamental vine (Tănase et al., 2020). It
is popular in many tropical regions, including southern and northern Nigeria,
where the leaves are cooked as vegetables and used in soups (Bose et al.,
2023; Bolaji et al., 2022c; Adegoke & Ojo, 2017). The leaves are
rich in vitamins, minerals, mucilage, and antioxidants, giving the plant
nutritional and functional food value (Kumar et al., 2021; Zhang et
al., 2024).
Pharmacological studies of B. alba have demonstrated significant analgesic (antinociceptive),
anti-ulcer/gastroprotective, antioxidant, anti-proliferative, and
anti-angiogenic activities of B. alba extracts (Sheik et al., 2023;
Halayal et al., 2025). Comprehensive phytochemical and ethnomedicinal
reviews also confirm numerous traditional uses and pharmacological activities of B. alba, including antimicrobial, anti-inflammatory, wound-healing,
anti-ulcer, and fertility-related properties (Bose et al., 2021; Madhavi
et al., 2023).
Electrophoresis-based characterization of seeds and other plant parts have been utilized by researchers
in revealing interparental and interspecific diversity, phylogenetic
relationships, and trait associations, and gel-based proteomics remains a
valuable complement to newer methods (Pandey et al., 2025). There exist some inconsistencies in
the taxonomic classification of the green stemmed and the red stemmed B.
alba, with some researchers regarding them as being synonymous, some as
separate species, while others regard them as varieties (Warrier et al., 1996; Larkcom, 2007; Oladele and
Aborishade, 2009; Roy et al., 2010; Bolaji et al., 2022a). The objective of this study was therefore to
characterize the two Basella forms using SDS PAGE, so as to provide
useful information that could assist in understanding their taxonomic
relationship.
METHODOLOGY
SDS PAGE was carried out
on the crude proteins extracted from the
seeds and leaf of mature green-stemmed and red-stemmed Basella alba obtained from Ile-Ife
(7 30'42.3''N 4 32' 51.2''E), Osun State, Nigeria following the method of Oladejo et al. (2019). The
procedure was carried out in the Biotechnology
Laboratory, Department
of Animal Sciences, Faculty of Agriculture, Obafemi
Awolowo University,
Ile-Ife, Nigeria.
0.8 g of the plant samples (leaves and seeds
separately) were washed
with distilled water and macerated with sterile mortar and pestle in 0.8% phosphate buffered saline (PBS) containing 0.4 M NaCl
at pH 8.0. The extract was centrifuged at 5,000 rpm for 10 min and the
supernatant of each sample was collected. 15 µl of each of the extract was
subjected to electrophoresis in 12% polyacrylamide gel. Gels were stained with
0.3 % Coomassie brilliant blue. De-staining in methanol, acetic acid and
distilled water (1:3:5 v/v) was done overnight to reveal the protein bands for
scoring
Seven percent (7%) of
B-mercaptoethanol (Sigma) in sample buffer was used for the
preparation of each of
the plant samples under a fume hood. A
discontinuous system of employing 12% separation gel overlaid with a 4%
stacking gel was adopted. Sample buffer and B-mercaptoethanol was
added in addition to 30µl of high molecular and low molecular weight protein
markers following the method of Omitogun et al., 1999. The samples were
heated at 95 ⁰C for 5 min in a water bath. 25 µl of the heated sample was added to
the prepared SDS PAGE gels and sample buffer with 40% sucrose solution being
loaded in each well. The separation of protein was carried out with the use of
Bio-Rad Electrophoresis Power Supply Model 200/2.0 in the Bio-Rad Mini Protean
11 Cell at 150 Volts for 55 min. The gels were
stained with 0.3% Coomassie Brilliant blue for 18 hours. The gels were scanned
with HP 3320 scanner and the images stored for scoring.
Data were collected on the scanned gels by scoring the presence (1) or
absence (0) of protein bands directly from the computer screen. The protein
bands on each gel were compared with the known molecular weights (kDa) of the
standard. The similarity coefficient was then
calculated following the methods of Sneath and Sokal (1973)
as follows:
Coefficient of similarity
(CS) = 
Where CS
= coefficient of similarity between species 1 & 2
a =
number of bands common to 1 and 2
b =
number of bands in 1, not in 2
c =
number of bands in 2, not in 1
RESULTS
A close examination of the protein profile of the seed and leaf
protein of the Basella alba studied (Figures 1 and 2) revealed that the variation in the protein bands observed in this study were mostly qualitative,
having to do with the thickness of the bands and
staining intensity.
The SDS PAGE of the seed protein led to the detection of fifteen (15)
bands, two (2) of which were slow moving, three (3) were intermediate, and ten
(10) were fast moving (Table 1). The study revealed that for the seed
protein, the green stemmed B. alba had a total of eight (8) bands, while
the red stemmed had seven (7) bands (Table 1 ).
Figure 1: Electrophoregram of the seed
and leaf protein of the green-stemmed and red-stemmed Basella alba studied
M: Protein standard; A: Red-stemmed Basella alba seed; B: Red-stemmed Basella alba leaf; C: Green-stemmed
Basella alba seed; D: Green-stemmed
Basella alba leaf.
Figure 2: Schematic diagram of the
electrophoregram of the seed and leaf protein of the green-stemmed and
red-stemmed Basella alba studied.
A: Red-stemmed Basella alba seed; B: Red-stemmed Basella alba leaf; C: Green-stemmed Basella alba seed; D: Green-stemmed Basella alba leaf.
|
Table 1: Electrophoretic
profile of the seed and leaf proteins of the Basella alba morphotypes studied |
||||||
|
Protein type
|
Slow bands (0 – 1.9) cm |
Intermediate
bands
(2.0 – 3.9) cm |
Fast
bands (4.0 – 6.0) cm |
Total
number of bands |
Common bands |
Unique bands |
|
Green
stemmed seed |
1 |
1 |
6 |
8 |
5 |
3 |
|
Red
stemmed seed |
1 |
2 |
4 |
7 |
5 |
2 |
|
Green
stemmed leaf |
1 |
1 |
5 |
7 |
6 |
1 |
|
Red
stemmed leaf |
1 |
1 |
6 |
7 |
6 |
1 |
Five common bands
(62.5%) were found between the seed protein of
the green stemmed and red stemmed B. alba
studied at 0.2cm –
0.4cm, 4.3 – 4.8 cm, 4.8 – 5.2, 5.5 cm – 5.8 cm, and 6.0 cm – 6.5 cm (Figure
2). The red stemmed B. alba seed
protein had 2 unique bands at 2.6 cm – 3.2 cm and 3.6 cm –
4.3 cm, while the green stemmed had 3 unique
bands at 2.8 cm –
3.2 cm, 5.2 cm – 5.3 cm, and 6.4 cm – 6.5 cm (Figure
2).The SDS PAGE of the leaf protein also led to the detection of fifteen (15)
bands, two (2) of which were slow moving, three (2) were intermediate, and ten
(11) were fast moving (Table 1).
The analysis of the protein bands of
the leaf protein revealed that both the green stemmed and the red stemmed B.
alba had seven (7) bands. Six common bands
(85.7%) were found in the leaf protein of the two B.alba morphotypes studied at 2.3 – 2.6 cm, 4.5
cm – 4.8 cm, 5.0 – 5.4 cm,
6.0 cm – 6.3 cm, 6.4 cm – 6.5 cm, and 6.5 cm – 6.6 cm (Figure 2). The red-stemmed Basella alba leaf had one (1) unique
band at 0.2cm – 0.4cm,
while the green-stemmed Basella alba leaf
had one (1) unique band at 0.1cm (Figure 2). The Sokal and Sneath coefficient of
similarity revealed 50.0% similarity in the seed protein of the green
stemmed and red stemmed B. alba studied (Table 2), and 75% similarity in
the leaf protein contents (Table 3).
Table
2: Coefficient of similarity of seed protein samples of Basella
alba morphotypes studied
|
|
Green stemmed seed |
Red stemmed seed |
|
Green stemmed
seed
|
_ |
50.0% |
|
Red stemmed
seed |
50.0% |
_ |
Table
3: Coefficient of similarity of leaf protein samples of Basella
alba morphotypes studied
|
Morphotypes |
Green stemmed leaf |
Red stemmed leaf |
|
Green stemmed leaf
|
_
|
75.0 %
|
|
Red stemmed
leaf |
75.0 % |
_ |
DISCUSSION
Identifying plant taxa using protein banding patterns allows
detection of genetic variation that may not be evident from gross morphological
characters such as habitat, stems, leaves and flowers, and has been widely
applied in plant systematics and diversity studies (Adhikari et al.,
2012; Bose et al., 2023). The results of the seed and leaf protein profile, as well as relatively
high coefficient of similarity (50% in the seed protein and 75% in the leaf
proteins) noted in this study is a strong indication that a strong
degree of similarity exist
between the green stemmed and the red stemmed Basella alba studied irrespective of their stem colors. The protein profile
revealed that most of the protein bands were common bands, with the seed
proteins comprising 62.5% common bands and the leaf proteins comprising 85.7%
common bands.
The presence of common protein
bands between the green stemmed and red stemmed Basella alba
suggests that the genes coding for these proteins are conserved and may
represent adaptive traits that have evolved and become fixed over time. Such
common bands support the classification of these morphotypes as the same
species and may reflect a shared evolutionary origin (Teniola et al.,
2021). Additionally,
these bands imply that the proteins they represent have identical amino acid
sequences. This suggest that the green stemmed and the red stemmed B. alba
are possibly not separate species, but morphotypes or varieties of same
species, hence supporting the taxonomic classification of the two as same
species. This is corroborated by previous reports by other researchers (Roy et
al., 2010; Bolaji et al., 2022a, Bolaji et al., 2023) who
noted that the green stemmed and the red stemmed Basella alba were quite
close genetically and are not separate species.
Notably, the presence of unique protein bands in the seed and leaf
proteins of the green stemmed and red stemmed B. alba points to intervarietal differences between
the two morphotypes.
Such variations in protein makeup have been widely reported as valuable markers
for differentiating and classifying members of a population into appropriate
taxonomic groups (Teniola et al., 2021). The unique bands at 2.6 cm – 3.2 cm and 3.6 cm –
4.3 cm in the seed protein of the red stemmed, and at 2.8 cm – 3.2 cm, 5.2 cm – 5.3 cm, and 6.4 cm – 6.5 cm for the
green stemmed seed protein, as well as the the unique band at 0.2 cm – 0.4 cm
for the red stemmed leaf protein and 0.1cm for the green stemmed leaf protein may be valuable for distinguishing the two morphotypes from each
another. Similar findings have been
reported in earlier studies (Oladejo et al., 2019; Shonde et al., 2023; Bolaji et
al., 2025),
supporting the use of unique bands as indicators of genetic differences. Protein electrophoresis has been widely reported as an effective
biochemical marker for assessing genetic relationships because differences in
band presence and absence correspond to variation in expressed gene products
(Abdelhaliem et al., 2023; Bolaji et al., 2025). Such variation can also be useful in crop improvement
programmes, as plant breeders can use these differences to select better traits
and exploit heterosis for improved crop development (Olanrewaju et al.,
2021; Olomitutu et
al., 2022, Bolaji et
al., 2023; Adeniran and Bolaji, 2024). The
predominance of fast moving bands in the seed and leaf protein of the
green stemmed and red stemmed B.alba studied implies that the two Basella
forms consist mainly of proteins with high molecular weight.
CONCLUSION
The study revealed relatively high level of coefficient of similarity between the green stemmed and the red stemmed Basella
alba studied, with the seed protein
having 50% and the leaf protein having
75% similarity, indicating that a
strong degree of similarity exist between the green stemmed and the red stemmed Basella
alba studied irrespective of their stem
colors. The study
supports the taxonomic classification of the green stemmed and red stemmed B.alba
as morphotypes or varieties belonging to same species.
Conflict of Interests
There are no conflict of interests before, during or after the work
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