Dagon University Commemoration of 25th Anniversary Silver Jubilee Research Journal 2019, Vol.9, No.

2 321

Study on Morphology, Anatomy, Preliminary Phytochemical Test,

Nutritional Values and Antimicrobial Activities of leaves of
Ludwigia octovalvis (Jacq.) Raven

Dr Lai Win Aung1, Daw Khin Ei Chaw2

Abstract
Ludwigia octovalvis (Jacq.) Raven is a perennial herb and it belongs to the family Onagraceae in
order Myrtales. It is locally known as Taw-Lay-Nyin and collected from North Dagon Township,
Yangon Region. The morphological characters of vegetative and reproductive parts of the plants
wereidentified and classified by the use of available literatures. Then, the leaves samples were
dried powdered and kept in airtight container to diagnose characters of powdered drug. In
microscopical study, the stomata (anomocytic type)was found abundantly on the lower surface of
the leaves. Phytochemical analysis of powdered leaves indicated that the presence of alkaloids,
saponins, reducing sugar, carbohydrates, glycosides, flavonoids, tannins, phenolic compounds,
terpenoids and steroids. The nutritional values of powdered leaves such asprotein, fat, fiber,
carbohydrate and energy value was conducted at Department of Research and Innovation Analysis
Department. By using paper disc diffusion method,antimicrobial activities of various crude
extracts were carried out with seven test organisms. The ethanolic and methanolic extracts of
exhibited the highest against on different pathogenic microorganisms. Therefore, it was observed
that Ludwigia octovalvis (Jacq.) Raven is endowed with antimicrobial activities, nutritional value
and bioactive constituents provided to be used in traditional medicine.
Keywords: Morphology, Anatomy, Phytochemical, Nutritional value, Antimicrobial activity

Introduction
Plants are well known as a major source of modern medicines. Medicinal plants are an
essential part of the traditional health care systems. There are more than 8000 plants species in
South Asia with well-known medicinal uses.Ludwigia octovalvis (Jacq.) Raven is erect, much
branched, perennial andundershrub. Leaves are alternate and nerve numerous. Flowers are
yellow, tetramerous, axillary and solitary. Fruits are sub-quadrangular, 8 ribbed, clove-like
appearance. Seeds are numerous, minute and ovoid (Kirtikar and Basu, 1975).Ludwigia
octovalvis (Jacq.) Raven belongs to the family of Onagraceae in order Myrtales and it has
about 20 genera and 650 species (Lawrence, 1964).
In microscopical characters, both upper and lower surface of epidermal cells are thin-
walled and wavy. Stomata(anomocytic type) is found abundantlyin lower surface. Transverse
section of midrib and petiole, vascular bundleswere found creasent in shape and collateral type.
Raphide presents in midrib and petiole (Trease and Evans 2002, Folorunso, 2015)
This plant is used as medicinal purposes such aschickenpox, lesions, diarrhea, dysentry,
anticancer, diuretic, glands in the neck, diabetes, skin diseases, cough, fever, fungal infection
of the toes and headache (Kirtikar and Basu, 1975 and Backer, 1965).
Phytochemical screening of Ludwigia octovalvis (Jacq.) Raven yielded to alkaloids,
flavonoids, tannin, saponin, phenol and steroids (Sikha and Rath, 2014).According to (Kadum

1
Dr. Assistant Lecturer, Department of Botany, Dagon University
2
Lecturer, Department of Botany, Taung-goke Degree College
322 Dagon University Commemoration of 25th Anniversary Silver Jubilee Research Journal 2019, Vol.9, No.2

et al. 2012) methanolic extracts of leaves showed activity against on Escherichia coli,
Pseudomonas aeruginosa and Staphylococcus aureus. Medicinal plants have very
highpotential antimicrobial drugs for treating various human diseases. Although a number of
plants have been screened, the search for antimicrobial substances from plants has continued
for a better and safer drug and combatting bacterial and fungal infections are still in
need.(Buwa and Staden, 2006).

Materials and Methods

(i) Botanical study of leaves of Ludwigia octovalvis (Jacq.) Raven
The selected plants Ludwigiaoctovalvis(Jacq.) Raven was collected from North Dagon
Township, Yangon Region. The morphology study of the plant was undertaken with the help
of available literatures; Kirtikar and Basu, (1975), Lawrence (1964), Ceylon (1995) and Flora
of Hong Kong (2008).The collected parts of the plants were recorded with images.
(ii) Microscopical characters study of leaves of Ludwigia octovalvis (Jacq.) Raven
Microscopical characters of leaveswere examined by free hand sections according to
the methods given by Metcalfe and Chalk (1950), Easu (1965) and Trease and Evans (2002).
The fresh leaves were sliced into fine cutting by free hand cutting. The cut specimens were
cleared by using chloral-hydrate solution as a clearing agent then stained with suffranin. Then,
the stained specimens were examined under light microscopes.
(iii) Chemical study constituents of leaves of Ludwigia octovalvis (Jacq.) Raven
Phytochemical examination was carried out in Botany Department, at the University of
Dagon, according to the methods of Central Council for Research in Unani Medicine (1987),
Marini Bettoloet al., (1981). The leaves were dried at the room temperature for one week.
When samples were completely dried,they were pulverized by grinding machine to get
powdered and stored in airtight containers.
(iv) Nutritional values study of leaves of Ludwigia octovalvis (Jacq.) Raven
Nutritional values of energy, carbohydrate, fiber, protein and fat were investigated
according to the method of Association of Official Agricultural Chemists (A.O.A.C) Horwitz,
1997. The experiments were carried out at Department of Research and Innovation Analysis
Department at the Union of Myanmar under the Ministry of Education.
(v) Microorganism and Antimicrobial activity determination use for the tests
Preparation of nutrient agar and antimicrobial activities were carried out by using
Cruickshank (1968) and Atlas (1993) methods. Nutrient Agar Medium was prepared by Agar -
25 g, Peptone - 0.5 g, Yeast - 0.2 g, Distilled water - 100 L and pH - 7.0 ±.The antimicrobial
activity of the 50 ml of acetone, ethanol, ethylacetate, methanol, pet-ether and water extract
was determined by individual testing on Bacillus subtilis, Candida albican, Escherichia coli,
Pseudomonas aeruginosa, Saccharomyces cerevisiae, Staphylococcus aureus and Micrococcus
luteus.
After autoclaving nutrient agar 20 - 25 ml of the medium was poured into petri-dishes
and made plating by using 0.1 to 0.2 ml to test organisms. These plates were allowed to set for
2 - 3 hours. And then, 10 mm paper disc was absorbed about 0.5 - 0.8 ml of extract. 10 mm
paper disc was introduced into agar plate and incubated at room temperature for one day. The
clear zones appeared around andthe paper disc indicated the presence of antimicrobial activity.
Dagon University Commemoration of 25th Anniversary Silver Jubilee Research Journal 2019, Vol.9, No.2 323

The extent of antimicrobial activity shown by clear zone was measured with the help of
clipper.

Results
(i) Morphological characters of Ludwigia octovalvis (Jacq.) Raven
Ludwigia octovalvis (Jacq.) Raven is perennial herb. The leaves are alternate and
simple. Inflorescences are axillary and cymes. Flowers are yellow, calyx tube slender, corolla
broadly obovate. The eight stamens are almost equal filament white andanther yellow. There
are four Carpels , axile placentation as one ovule in each locule. Fruits are capsule, clove-like
appearance. Seeds are numerous brown. Flowering periodis from June to Octoberwhereas the
Fruit period is from June to February.

(ii) Anatomical characters of leaves of Ludwigia octovalvis (Jacq.) Raven

Stomata(anomocytic type) was present on both surfaces but abundantly found on the
lower surface. In transerve section of lamina, one layer of palisade parenchyma cells were
elongated in shape and contain numerous chloroplasts. Two to three layers of spongy
mesophyll cells were loosely arranged. In the surface view of midrib and petiole, epidermal
cells were thin walled, irregular and some cells were barrel shaped on both the surfaces.
Transverse section of midrib and petiole, the cortex layer consists of collenchymatous and
parenchymatous cells. The vascular bundles were arranged collateral type and crescent in
shape. Raphides were present in midrib and petiole. In the diagnostic characters of powdered
leaves were foundon the fragment of surface view, vessel, pitted vessel, scalariform vessel,
fiber and raphides.

(iii) Preliminary Phytochemical examination of leaves of Ludwigia octovalvis (Jacq.)

Raven
In preliminary phytochemical investigation, the presence of alkaloids, glycosides,
tannins, starch, steroids and terpenoids were observed in leaves. Tannins, glycosides,
terpenoids and phenolic compounds were mostly observed.

(iv) Nutritive values of powdered leaves of Ludwigia octovalvis (Jacq.) Raven

Nutritional evaluation on the powdered leaves of Ludwigia octovalvis (Jacq).Raven was
investigated in energy 111.92 Kcal / 100 g, carbohydrate 15.88%, fibre 10.08%, fat 1.16% and
protein 9.49% at Department of Research and Innovation Analysis Department. Energy was
greater than the other components.

(v) Antimicrobial activity of different solvent extracts of leaves of Ludwigia octovalvis

(Jacq.) Raven
In this experiment, the best antimicrobial activity effecting leaves extracts were found
at ethanolic, methanolic and aqueous extracts against Bacillus subtilis, Candida albican,
Escherichia coli, Pseudomonas aeruginosa, Saccharomyces cerevisiae, Staphylococcus aureus
and Micrococcus luteus. Methanolic and ethanolic extract the highest and it indicated that
324 Dagon University Commemoration of 25th Anniversary Silver Jubilee Research Journal 2019, Vol.9, No.2

antimicrobial activity is against the Escherichia coli, Pseudomonas aeruginosa and

Staphylococcus aureus.

Habit Upper and Lower surface of leaves Inflorescence Flowers

L.S of flower Fruits Seeds T.S of ovary

Figure 1.Morphological characters of leaves of Ludwigiaoctovalvis(Jacq.) Raven

Upper epidermis Lower epidermis Lamina Upper and lower surface

of midrib

T.S of midrib Upper surface of Lower surface of T.S of petiole

petiole petiole
Figure 2. Anatomical characters of Ludwigiaoctovalvis(Jacq.) Raven

Vessel Vessel Pitted vessel Scalariform Fiber Raphide Surface view

Figure 3.Diagnostic characters of powdered of leaves of Ludwigiaoctovalvis(Jacq.) Raven

Dagon University Commemoration of 25th Anniversary Silver Jubilee Research Journal 2019, Vol.9, No.2 325

Figure 4. Nutritional values of powdered of leaves of Ludwigiaoctovalvis(Jacq.) Raven

1 1 1
5 5 5
6 2 6 2 6
4 4 4 2
3 3 3

Control Bacillus subtilis Control Candida albican Control Escherichia coli

1
1 1
5 5 5
6 2 6 6 2
4 4 2
4
3 3 3

Control Micrococcus luteus Control Pseudomonas Control Saccharomyces

aeruginosa cerevisiae

1
5 1. Acetone
6
4 2 2. Ethyl acetate
3 3. Ethanol
4. Methanol
5. Pet-ether
Control Staphylococcus
6. Aqueous
aureus

Figure 5. Antimicrobial activities of various solvents extracts from leaves of

Ludwigiaoctovalvis(Jacq.) Raven

Discussion and Conclusion

The current investigation involved the morphology study of vegetative and
reproductive parts as well as the microscopical examination of leaves. In this study, Ludwigia
octovalvis (Jacq.) Raven is perennial herb. The leaves are alternate and simple. Inflorescences
are axillary and cymes. Flowers are yellow, calyx tube slender, corolla broadly obovate. The
eight stamens are, almost equal filament white andanther yellow. There are four Carpelsaxile
326 Dagon University Commemoration of 25th Anniversary Silver Jubilee Research Journal 2019, Vol.9, No.2

placentation as one ovule in eachlocule. Fruits are capsule, clove-like appearance andseeds are
numerous brown. These characters were agreed with Lawrence (1964), Ceylon (1995), Flora of
Hong Kong (2008) and Kirtikar and Basu (1975).
In anatomical study, the stomata was found abundantly present on lower surface
(anomocytic type). In transverse section of lamina, palisade parenchyma was one layer and
spongy mesopyll cells were two to three layers. Surface view of midrib and petioles, irregular
and some cells were rectangular in shape. In transverse section of midrib and petiole, vascular
bundle was creasent in shape and collateral type. Raphideswere found in midrib and petiole.
These characters were agreed with Kirtikar and Basu (1974) and Folorunso (2015).
In the diagnostic characters of powdered leaves, on the fragment of surface view,
vessel, pitted vessel, scalariform vessel, fiber and raphides were found.
The preliminary phytochemical test was carried out on Ludwigia octovalvis (Jacq.)
Raven showed that secondary metabolites as glycosides, alkaloids, flavonoids, saponins,
tannins, reducing sugars, phenolic compounds, carbohydrates, steroids and terpenoids. These
phytochemical constituents of leaves of Ludwigia octovalvis (Jacq.) Raven was agreed with
those mentioned by Sikha and Rath (2014). Alkaloids, flavonoids and terpenoids are the main
secondary metabolites that exhibit many pharmacological and properties in living cells
(Rhodes, 1994). Phenolic compounds in plants are essential part of human diet and it has
antioxidant properties (Khan and Islam, 2011).
According to the result from the Department of Research and Innovation Analysis
under the government of the Republic of the Union of Myanmar, the nutritional values of
powdered leaves contained energy 111.92% Kcal/100g, carbohydrate 15.88%, fiber 10.08%,
protein 9.49% and fat 1.16%.Living organisms require available energy to stay a live; humans
get such energy from food. Daily food intake of a normal adult: 6-8 MJ.Many carbohydrates
and fats processing involved for energy production.Protein intake excess of that required to
build muscle. There six main classes of nutrients that the body needs are carbohydrates,
proteins, fats, vitamins, minerals and water.It is important to consume these six nutrients on a
daily basis to build and maintain healthy function of the body. (Smith and Crosbie,1998).
In this experiment, methonolic and ethanolic extracts showed the highest activity
against on Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus. The present
results were agreed with Kadum et al. (2012). He reported thatLudwigia octovalvis of leaves
extracts which exhibited significantly has higher animicrobial activity. Methanolic extracts of
leaves showed activity against on Escherichia coli, Pseudomonas aeruginosa, Staphylococcus
aureus.
In conclusion, the selected plant is resistant to some strains of clinical microorganisms
such as bacteria and fungi. Thus, Ludwigiaoctovalvis(Jacq.) Raven could be used for
preparation of effective antimicrobial source.Plant can be used as herbal medicine due to the
result of the enrichment of bioactive, antimicrobial activity and having nutritional value. The
above data would be helpful in further study of the research and potential new durg
development for the prevention and treatment of infection diseases.
Furthermore,the pharmacological activitiesof Ludwigia octovalvis (Jacq.) Raven should
be undertaken.
 Dagon University Commemoration of 25th Anniversary Silver Jubilee Research Journal 2019, Vol.9, No.2 327

Acknowledgements
We would like to express our deep gratitude to Dr Myat Myat Moe, Professor, Head of Botany
Department, Dagon University, for allowing us to use the Departmental facilities and invaluable advices.
I also give my thanks to Dr Yee Yee Nwe, Lecture, Department of Botany, Taung-goke Degree College,
for her guidance and support in the test of antimicrobial activity.

References
Atlas, R. M. (1993). Handbook of MicrobiolocalMedia CRC Press, London.
Backer, C.A. and R.C. Bakhuizen-Van Den Brink, Jr. (1965). Flora of Java (Vol.II) N.V.P Noordhoff,
Groningen, The Netherland.
Central Council of Research in Unani Medicine, (1987). Physiochemical standards of unani formulation.
Ministry of Health and Family Welfare, New Delhi, India.
Cruickshank, R., J.P. Guguid and R.H.A., Swain (1968). Medical Microbiology (11th ed). The English Language
Book society and F. and S. Livingstone Ltd., London.
Dassanayake, (1995). Flora of Ceylon (Vol.IX), Published Amerid Publishing Co.Rvt.Ltd. New Delhi.
Easu, K.(1965). Plant anatomy ( 2nded). Toppan Company Ltd., Tokyo Japan.
Folorunso, (2015), Use of foliar and stem anatomy characters in the identification of Ludwigia (Linn) species
in Nigeria. Department of Botany, Faculty of Science, Obafemi Awolowo University, Ile- Ife, Nigeria.
Horwitz, W. (1997). Official methods of analysis of association of official analytical chemist (13th ed).
Washington DC: Benjiamin Franklin Station.
Hundley, H.G. and U Chit Ko Ko, (1961). List of Trees, Shrubs, Herbs and Principal Climbers etc. Yangon:
Supplt., Govt. Printin and Italy.
Kadum et al., (2012) Antioxidant and Antibacterial activity ofLudwigia octovalvis (Jacq.) Raven on some
Pathogenic Bacteia. School of Biological Sciences, Universiti Sains Malaysia, 11800 Miden, Penang,
Malaysia.
Kirtikar and Basu, (1975). Indian Medicinal Plant (Vol.III), Bishen Mahendra Pal Sigh, Dehradun, India.
Lawrence (1964). Taxonomy of vascular plants. The Macmillian Company,New York.
Melcalfe, C.R. and L. Chalk, (1950). Anatomy of the Dicotyledons.(Vol.I) Oxford University, London.
Qi-ming and WU Delin (SCBG), Hong Kong Herbarium (2008). Flora of Hong Kong (Vol III, Agriculture,
Fisheries and Conservation Department, Hong Kong.
Rhodes, M.J.C.(1994). Physiological roles for secondary metabolites in plants: some progress, many
outstanding problems, Plant Mol. Biology, 24,1-20.
Shils, et al., (2005). Modern Nutrition in Health and Disease. Lippincoff, Willians and Wilkins.
Sikha and Rath (2014). Phtochemical and Antioxidant activities ofLudwigia octovalvis (Jacq.) Raven
Department of Botany, Visva-Bharati University, West Bengal, India.
Smith, C.1998. The science of energy a cultural history, Energy Physics in Victorian Britain. The University of
Chicago Press.
Trease and Evans, (2002). Pharmacognosy. (15thed.), London, New York. Unani Formalations, New Delhi.