Icacina
trichantha Oliver is an underutilized weedy shrub with
future potentials as food, a quintessential medicinal tropical shrub and
industrial raw materials for bioplastic production. The research was set to
investigate the morphological, anatomical, and proximate properties of I. trichantha, belonging to the family
Icacinaceae. The plant samples were collected in the University of Port
Harcourt Campus. Measurement of morphological parts were ascertained with use
of the meter rule. The samples were fixed in FAA, dehydrated in ethanol
solutions (50%, 70%, 90% and absolute) and sectioned while those for
epidermal study were peeled using 50% Nitric acid. These were stained in 2%
aqueous solution of Safranin O, counter stained in Alcian blue for about 2-5
minutes and mounted on glycerine. Micro-photographs were taken using mounted
Sunny phone camera on Mono Ocular microscope. The proximate analysis were
conducted following AOAC method. Results revealed plant could grow up to
180±20cm in height with foliar organs broadly elliptical to abruptly acute at
the apex and rounded at the base with few branch growth. Epidermal studies
showcased paracytic and diacytic stomata which is hypostomatic with presence
of glandular and uniseriate trichomes. The anatomical sections revealed a row
of epidermal cells, hypodermis made of collenchyma and general cortex
predominated by parenchyma, with a ring of open vascular system. Proximate
composition were thus: Moisture content: 55.8%; Carbohydrate: 27.55%;
Proteins: 10.94%; Cyanide content: 8.00%; Ash: 4.42%; Lipid: 1.30% and Fiber
content: 0.01%. These findings would aid to further delimit the species.
The
tropical forest tree family, Icacinaceae, was first recognized by Miers (1864)
and then revised on the basis of DNA sequencing as given by Karehed (2001). The
genus Icacina comprised six accepted
species and eight synonyms (Anon, 2015), while the five main species recognized
in Africa are: Icacina claessensii De
Wild., I. guessfeldtii Asch. Ex Engl.
I. mannii Oliv. I. oliviformis (Poir.) J. Raynal and I. trichantha Oliv.), of these I.
oliviformis (Poir) J. Raynal is probably the best known as a food plant of
West Africa (Fay, 1987; Anon, 2008). I.
trichanthacan grow up to 2m in
height and could be processed in tograyish-white or creamy-yellowish flour (Umoh and Iwe, 2014) used as
source of emergency moisture and as famine food during long period of drought. In
Nigeria however, it is locally referred to as Gbegbe in Yorubas and the Igbos
called it Ibugo (Burkill, 1985). I.
trichantha has broadly elliptic simple leaves in alternate phyallotaxy, the
underground tuber is usually as large as yam (Timothy and Idu, 2011; Akobundu
and Agyakwa, 1998). The bitter
substances such as Hydrogen cyanide, oxalates, tannins, alkaloids and phytates
were found present in the flour including carbohydrate (Starch),proteins, lipids and other mineral elements
such as potassium, sodium and calcium (Udofia and Asuquoekpo, 2014). Hence in
Nigeria, I. trichantha is used as a
common household medicine for emergency and first-aid treatment for food
poisoning (Mbatchou and Dawda, 2012). False yam, Icacina trichantha tubers analyzed for proximate composition
revealed presence of carbohydrate 91.93% and proteins 5.25% (Sunday et al., 2016). In traditional and rural
setting in Nigeria, the foliar organs and underground tubers have folkloric
uses in the treatment of malaria, constipation and food poisoning (Che et al., 2016; Asuzu and Abubakar, 1995).
The nutritional contents of the seeds include: 13% moisture, 72% carbohydrate,
8 – 10% proteins, 0.1% fat (Fay, 1991). The leaves are thinly pilosed with simple,
fascicled hairs beneath while flowers are densely crowded and subsessile with
calyx nearly as long as the petals which are usually villous outside; the
fruits are tomentose on the surface, ellipsoid to globose in shape measuring up
to 2.5 cm in length (Hutchinson and Dalziel, 1958; Akobundu and Agyakwa, 1998).
The leaf epidermis properties of I.
trichantha include paracytic and diacytic stomatal types, irregular
epidermal cell shape together with angular or curved anticlinal wall patterns (Kadiri
et al., 2020).
Icacina
trichantha
is valued for its traditional uses, apart from the medicinal properties and as
a food energy source, it is still underutilized and could serve as a raw
material for bioplastic production in Nigeria, hence the objective focusses on:
“Studies on Anato-Morphological and Proximate Properties of Icacina trichantha Oliver.”
MATERIALS
AND METHODS
Morphological
Properties
The
meter ruler was used for measurement involving plant height from the
root-collar to the terminal bud. The leaf length from the leaf tip to the
petiole base. Leaf width across leaf lamina, from one margin to another at the
widest region. The root system, haustarium is embedded in host plant.
Epidermal
studies
The
foliar epidermal layers were peeled chemically using 50% Nitric acid. The
methods of Cutler (1978) was adopted.
Anatomical
Characteristics
The samples were fixed in Formaldehyde
Glacial Acetic acid 70% Alcohol in the ratio 1:1:18 following the methods of Johannsen (1940). Free hand section
was done following the method of Wahua
(2020). Thereafter, photo micrographs were taken from good slides.
PROXIMATE PROPERTIES
Proteins (Kjeldahl
method)
Stage
1: 0.1g of sample was weighed into a conical flask of 250ml capacity, 3g of
digestion catalyst was placed into the flask and 20ml conc. Sulphuric acid
added and heated to digest. Color change observed from black to sky-blue,
cooled to room temperature and then diluted to 100ml with distilled water.
Stage
2: 20ml diluted digest was measured into a distillation flask and held in place
on hot plate. The distillation flask was attached to a Liebig condenser
connected to a receiver containing 10ml of 2% boric acid indicator. 40ml NaOH
was injected into the digest, and heated to boiling and the distilled ammonia
gas via the condenser into the receiver beaker. The color of the boric acid
change from purple to green as ammonia distillate was introduced into the boric
acid.
Stage
3: The distillate was titrated with standard 0.1N HCl solution back to purple
from greenish. The volume of HCl added to effect this change was recorded as Titre
value.
Thus,
Where
1.4 = Nitrogen equivalent to the normality of the HCl used in the titration
0.1N
100
= the total volume of digest dilution
100
= percentage factor 0.1g of the sample
1000
= conversion from gram to milligram
20
= integral volume of digits analyzed or distilled
0.1g
= the weight of sample in gram digested
Carbohydrate
(Cleg Anthrone Method)
In
this regard, 0.1 of the sample was weighed into 25ml volumetric flask, 1ml
distilled water and 1.3ml of 62% perchloric acid was added and stirred for a
period of 20 minutes to homogenize completely. The flask was made up to 25ml mark
with distilled water. The solution was filtered with a glass filter paper and
allowed to sediment and then decanted. 1ml of the filtrate was collected and
transferred into a 10ml test tube which was diluted to volume with distilled
water. 1ml of the working solution was pipette into a test tube and made up to
volume with distilled water. 1ml of working solution was pipette into a test
tube and 5 ml anthrone reagent added. 1ml distilled water was added and 5ml
anthrone reagent mixed. Similarly, the whole mixture was read at 630nm
wavelength using the 1ml distilled water and 5ml anthrone reagent prepared as
blank. 0.1ml glucose was also prepared and was treated as the sample with
anthrone reagent. Absorbance of the standard glucose was read and the value of
carbohydrate as glucose was calculated as shown below:
Moisture (Air Oven
Method)
One
gramme (1g) of the sample was weighed in to a porcelain evaporating dish. This
was placed in an oven set at 1050C for 6 hours. The evaporating dish
was cooled in the desiccator to room temperature and reweighed. Thus, the
calculation of % moisture was as shown below:
Lipid (Soxhlet
Extraction Method)
Here,
2g of sample was inserted into a filter paper and was introduced into a soxhlet
extractor. The extractor was placed into a pre-weighed dried distillation
flask. Then the solvent (acetone) was added to the distillation flask through
the condenser end attached to the soxhlet extractor. The set-up was held in
place with a stand clamp and cooled water jet was allowed to flow into the
condenser and the heated solvent was refluxed as a result. The lipid in the
solvent chamber was extracted in the process of continuous refluxing. When the
liquid was observably extracted completely from the sample, the condenser and
the extractor were disconnected and the solvent was evaporated to concentrate
the lipid. The flask was then dried in the air oven to constant and reweighed
to obtain the weight of the lipid as thus calculated below:
Ash (Furnace Method)
One
gramme (1g) of dried sample was weighed in to a porcelain crucible which was
previously preheated and weighed. The crucible was inserted into a muffle
furnace set at 6300C for 3 hours and allowed to cool to room
temperature and reweighed. Thus % ash was calculated as shown below:
Crude Fibre
Crude
fibre observed as the insoluble, combustible organic residue which remained
after a sample was treated with light petroleum ether, diluted acid and alkali
(AOAC, 1990).
About 2g of sample was extracted with
petroleum ether (W1). Sample was boiled under reflux for 30 minutes
with 200ml of dilute HCl and filtered. The residue was thoroughly washed with
water until acid-free. The residue was transferred into a baker and boiled for
about 30 minutes with 200 ml of dilute NaOH solution, filtered and transferred
into ignition crucible. The residue was washed 3 times with 20 ml ethanol and 2
times with 10 ml ether. The residue was dried in an oven and cooled and weighed
(W2). The dried residue was transferred into a furnace and ignited,
cooled and weighed (W3). Thus %crude fibre was calculated as shown
below:
RESULTS
Morphological Properties
The
meter rule was used in measurements such as plant height, leaf length and width
etc. Plate 1.
Plate 1: Icacina trichantha Oliver. Leaves in opposite phyllotaxy,
larger at base while smaller towards apex. Scale bar represents 35 cm.
Plate 3: I. trichantha adaxial
epidermis revealing glandular trichomes (Gtr), unicellular trichome (Utr), and
irregularly shaped nucleated epidermal cell (Nep)
The
micro-morphological investigation revealed paracytic and diacytic stomata which
is hypostomatic. Glandular and
unicellular trichomes were observed present mostly in the adaxial foliar
epidermis.Plates 2 and 3.
Anatomical Study
The
vascular system is open and bicollateral. The hypodermis is made up of 4 to 5
rows of collenchymatous cells while the general cortex showcased 5 to 7 rows of
parenchymatous cells in the mid rib, the petiole revealed kidney shaped
vascular arc with numerous trichomes on epidermis. Nodal anatomy, showed
multilocunar pattern and the root anatomy central zone dominated by xylem in
alternating fashion with the phloem tissues. Plates 4, 5, 6 and 7.
Plate 4: Icacina
trichantha
Mid-rib Anatomy. Plate 5: I. trichantha Petiole Anatomy. Plate 6: I. trichantha Nodal Anatomy.
Plate 7: I. trichantha Root
Anatomy. Ep – Epidermis, Hy –Hypodermis, Co –Cortex, Ph –Phloem, Xy –Xylem, Pr
–Pericycle, Va –Vascular arc, Pt –Pith, Gc –General cortex, Pe –Petiole, Mvc
–Main vascular cylinder. Scale bar represents 2 mm
Proximate Composition
The
carbohydrate content was twice the protein while the hydro cyanide content was
almost about twice the ash content, and the fibre content was the lowest
followed by the lipid. Table 1.
Proximate contents
% Quantity
Ash
4.42
Moisture
55.80
Carbohydrate
27.55
Proteins
10.94
Lipid
1.30
Fibre
0.01
Cyanide in Mg/Kg
8.00
DICUSSION
The Icacina trichantha described conformed
to those of Timothy and Idu (2011), and Akobundu and Agyakwa (1998). The
epidermal study revealed paracytic and diacytic stomata which is hypostomatic, and
glandular and unicellular trichomes were observed present mostly in the adaxial
foliar epidermis in line with the work of Kadir et al. (2020). The proximate constituents conformed to those of
Sunday et al. (2016) and Fay (1991),
except that protein content was lower in concentration but approximately the
same for fay (1991) and carbohydrate was more than twice the concentration
detected;
CONCLUSION
Icacina
trichantha as
a potential food source and medicine and bioplastic properties is of relevance and
subjected to various lines of taxonomic evidence, and showed that I.
trichantha is bicollateral with open vascular system and multilacunar
node.
ACKNOWLEDGEMENT
We do
recognize and heartily appreciate effort made by Delekpe, V. in support to this
research investigation.
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Cite this Article: Wahua, C; Awogbayila,
OD (2024). Anato-Morphological and Proximate Properties of Icacina trichantha Oliver.Greener Journal of Biological Sciences, 14(1): 22-27.
