COMPARATIVE ANATOMICAL STUDIES OF THE STOMATA

OF CODIAEUM VARIEGATUM, ACALYPHA WILKESIANA AND

CROTON ZAMBESICUS SELECTED AS ORNAMENTALS

BY
EKILEONYEONWU, PROMISE UCHECHUKWU
2018/246777

A PROJECT SUBMITTED FOR THE AWARD OF BACHELOR

OF SCIENCE (B. SC.) DEGREE TO THE DEPARTMENT OF
PLANT SCIENCE AND BIOTECHNOLOGY

UNIVERSITY OF NIGERIA, NSUKKA

SUPERVISOR
DR. AYOGU, V. O.

OCTOBER, 2023

i
 TITLE PAGE

COMPARATIVE ANATOMICAL STUDIES OF THE STOMATA

OF CODIAEUM VARIEGATUM, ACALYPHA WILKESIANA AND
CROTON ZAMBESICUS SELECTED AS ORNAMENTALS

i
 CERTIFICATION
This is to certify that Ekileonyeonwu, Promise Uchechukwu an undergraduate student in the

Department of Plant Science and Biotechnology, with registration number 2018/246777 has

satisfactorily completed the research work and requirements for this project. This project is

therefore approved by the Department of Plant Science and Biotechnology for the award of

Bachelor of Science (B.Sc.) degree in the University of Nigeria, Nsukka. This project is original

and has not been submitted in part or full for any other diploma or degree of this or any other

university.

APPROVED BY:

……………………………………… …………………………………….

DR. AYOGU, V. O. PROF. ONYEKE, C. C.

(Supervisor) (Head of Department)

Date__________________________ Date_________________________

______________________________________

EXTERNAL EXAMINER

Date______________________

ii
 DEDICATION
This project work is dedicated to God Almighty whose strength and provision saw me through
this work. I also dedicate this project work to the loving memory of my dear mother late Mrs.
Angela Ekileonyeonwu, whose prayers and words of encouragement kept me going throughout
the period of this project work.

iii
 ACKNOWLEDGEMENT
My sincere gratitude goes to my supervisor, Dr. Ayogu, V. O. for guiding me through this
project work with patience, making himself available for supervision and corrections. His
insights and expertise played an indispensable role in shaping the direction of this project.

I also wish to thank my lecturers and colleagues at the Department of Plant Science and
Biotechnology because the knowledge acquired from their teachings and interactions at some
point played a notable role during the course of my project work.

My heartfelt appreciation goes to my family and friends especially Mr. Stephen Ekileonyeonwu
and Ngwu Darlington for their financial and emotional support which kept me going when this
work proved difficult at some point. Your believe in me and your support has been my greatest
motivation.

Lastly, I want to appreciate the anatomy laboratory attendant (Mrs Ann) who cooperated with me
and gave me guidelines when needed.

This project work would not have been completed without the involvement of the individuals I
mentioned above. I sincerely appreciate their contributions.

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 TABLE OF CONTENTS
Title page - - - - - - - - - - - i

Certification - - - - - - - - - - - ii

Dedication - - - - - - - - - - - iii

Acknowledgements - - - - - - - - - - iv

Table of Contents - - - - - - - - - - - v

List of Tables - - - - - - - - - - - viii

List of Plates - - - - - - - - - - - - ix

Abstract - - - - - - - - - - - - x

CHAPTER ONE: INTRODUCTION

1.1 Background of the study- - - - - - - - - - 1

1.2 Aim of the study - - - - - - - - - - 2

1.3 Objectives of the study - - - - - - - - - 3

1.4 Significance of the study - - - - - - - - - 3

1.5 Scope of study - - - - - - - - - - 3

1.6 Limitations - - - - - - - - - - 4

1.7 Practical applications and implications - - - - - - - 5

CHAPTER TWO: LITERATURE REVIEW

2.1 Overview of Codiaeum variegatum - - - - - - - - 6

2.1.1 Botanical description of Codiaeum variegatum - - - - - - 7

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2.1.2 Botanical classification of Codiaeum variegatum - - - - - 9

2.2 Overview of Acalypha wilkesiana - - - - - - - 9

2.2.1 Botanical classification of Acalypha wilkesiana - - - - - 10

2.3 Overview of Croton zambesicus - - - - - - - - 11

2.3.1 Botanical classification of Croton zambesicus - - - - - - 11

2.4 Stomata morphology - - - - - - - - - 12

2.4.1 Stomata morphology of Codiaeum variegatum - - - - - 13

2.4.2 Stomata morphology of Acalypha wilkesiana - - - - - - 14

2.4.3 Stomata morphology of Croton zambesicus - - - - - - 14

2.5 Correlation between anatomical traits of ornamental plants and carbon sequestration - 15

2.6 Vessel size and pollutant translocation - - - - - - - 16

2.7 Balance between pollutant uptake and water conservation - - - - 17

CHAPTER THREE: MATERIALS AND METHODS

3.1 Plant collection - - - - - - - - - - - 19

3.2 Materials - - - - - - - - - - - 19

3.2.1 Laboratory equipment - - - - - - - - - 19

3.2.2 Microscopy settings - - - - - - - - - 20

3.3 Sample preservation - - - - - - - - - 20

3.4 Sample preparation - - - - - - - - - 21

3.4.1 Preparation of epidermal leaf strip - - - - - - - 21

3.4.2 Sectioning - - - - - - - - - - 22

3.5 Photographing - - - - - - - - - - 23

3.6 Stomatal size measurement - - - - - - - - 23

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3.7 Vessel size measurement - - - - - - - - - 24

CHAPTER FOUR: RESULTS

4.1 Anatomical analysis - - - - - - - - - 26

4.1.1 Anatomical analysis of stomata properties across different ornamental species - 26

4.1 2 Anatomical analysis of vessel properties across different ornamental species - - 28

4.2 Correlation coefficient of anatomical properties - - - - - - 31

4.2.1 Correlation coefficient of the relationship between stomata properties - - 31

4.2.2 Correlation coefficient of the relationship between stomata size and vessel pore size - - 32

4.2.3 Correlation coefficient of the relationship between vessel properties - - - 33

CHAPTER FIVE: DISCUSSION AND CONCLUSION

5.1 Discussion - - - - - - - - - - - 34

5.2 Conclusion - - - - - - - - - - - 36

REFERENCES- - - - - - - - - - - 38

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 LIST OF TABLES
Table 1: Comparison of stomata sizes and stomata density across different ornamental plants - -26

Table 2: Comparison of vessel size and vessel number across different ornamental plants - - 28

Table 3: Pearson correlation coefficient of the relationship between stomata properties - - 31

Table 4: Pearson correlation coefficient of the relationship between stomata and vessel properties 32

Table 5: Pearson correlation coefficient of the relationship between vessel properties - - 33

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 LIST OF PLATES
Plate 1: Codiaeum variegatum shrub - - - - - - - 8

Plate 2: Acalypha wilkesiana shrub - - - - - - - 10

Plate 3: Croton zambesicus tree - - - - - - - - - 12

Plate 4: T.S of leaf epidermis of Codiaeum variegatum showing the stomata - - 27

Plate 5: T.S of leaf epidermis of Acalypha wilkesiana showing the stomata - - 27

Plate 6: T.S of leaf epidermis of Croton zambesicus showing the stomata - - 27

Plate 7: T.S of stem section of Codiaeum variegatum showing the vessels - - 29

Plate 8: T.S of stem section of Acalypha wilkesiana showing the vessels - - 29

Plate 9: T.S of stem section of Croton zambesicus showing the vessels - - - 30

ix
 ABSTRACT
The increase in human activities majorly burning of fossil fuels especially in urban areas has led
to increased levels of carbon in the atmosphere. Ornamental plants play a role in removing
excess carbon from the atmosphere while performing their aesthetic function. Among the
extensive array of ornamental species, Codiaeum variegatum (garden croton), Acalypha
wilkesiana (copperleaf), and Croton zambesicus (African croton) stand out as exemplars of
botanical beauty. Their vibrant and intricately designed leaves have earned them a prominent
place in horticulture, landscaping. and indoor decor worldwide. The research work examined the
comparative anatomical studies of the stomata of Codiaeum variegatum, Acalypha wilkesiang
and Croton zambesicus selected as ornamentals. The plant specimens utilized in this study were
collected from the University of Nigeria, Nsukka metropolis, including the Botanical Garden of
the Department of Plant Science and Biotechnology UNN. Stomatal and vessel size
measurements were determined using standard methods by Harvanti and Tetrinica involving use
of stage and ocular micrometers. The anatomical stomatal properties showed that stomata size
(0.18 µm) was highest in Acalypha wilkesiang while stomata density (34.10 µm) was highest in
Codiaeum variegatum. The anatomical properties of the vessel showed that vessel size (6.86 µm)
was highest in Acalypha wilkesiang while vessel number (153) was highest in Croton
zambesicus. Correlation coefficient on the relationship between stomata and vessel properties
showed there was a strong negative correlation between stomata density and vessel size and it is
statistically significant. This finding implies that stomata density and vessel size which is
predominantly significant could be considered the most important anatomical parameter when
selecting ornamental plants for carbon sequestration. On this note, Acalypha wilkesiang with the
highest stomata size and vessel size values will be better suited as an ornamental plant for carbon
sequestration when compared to Codiaeum variegatum and Croton zambesicus at constant
temperature, humidity and ion concentration.

x
 CHAPTER ONE

INTRODUCTION

1.1 BACKGROUND TO THE STUDY

Ornamental plants have long captivated the human imagination with their aesthetic appeal and

diverse foliage patterns. Among the extensive array of ornamental species, Codiaeum

variegatum (garden croton), Acalypha wilkesiana (copperleaf), and Croton zambesicus (African

croton) stand out as examples of botanical beauty. Their vibrant and intricately designed leaves

have earned them a prominent place in horticulture, landscaping, and indoor decor worldwide.

While these ornamental plants are often admired for their visual splendor, beneath their striking

appearances lie intriguing adaptations that enable them to flourish in diverse environmental

niches. One fundamental aspect of plant adaptation is the regulation of gas exchange and water

loss through stomata, the microscopic structures found on the epidermis of leaves, stems, and

other aerial plant parts. Stomata serve as gatekeepers for the exchange of gases, facilitating the

uptake of carbon dioxide (CO2) necessary for photosynthesis while regulating the release of

oxygen (O2) and water vapor (H2O) through transpiration (Raven et al., 2005). Stomata exhibit

remarkable diversity in terms of their density, size, distribution, and associated epidermal

features, all of which are influenced by an array of environmental factors, including light

intensity, humidity, temperature, and soil moisture (Franks and Beerling, 2009). Understanding

the stomatal adaptations of ornamental plants such as Codiaeum variegatum, Acalypha

wilkesiana, and Croton zambesicus can shed light on their ability to make our environment

healthier through the process of carbon sequestration and provide valuable knowledge for

horticultural practices. Codiaeum variegatum, commonly known as the garden croton, is

renowned for its vibrant and variegated leaves, featuring a palette of colors that can rival a

1
painter's palette. As it adapts to its diverse growing conditions, the stomatal characteristics of

Codiaeum variegatum play a crucial role in its ability to maximize photosynthesis while

conserving water. Acalypha wilkesiana also referred to as copperleaf or Jacob's coat, is another

captivating ornamental species known for its strikingly colored leaves. As with Codiaeum

variegatum, the stomata of Acalypha wilkesiana are integral to its ecological adaptability,

allowing it to strike a balance between photosynthesis and water conservation. Croton

zambesicus, the African croton, represents a more understated yet equally fascinating member of

the ornamental plant world. The stomata of Croton zambesicus contribute to its ability to thrive

under varying light conditions and manage its water resources effectively. This investigation

holds the promise of enriching horticultural practices, enabling gardeners, landscapers, and

botanists to make informed decisions about the cultivation and maintenance of these captivating

ornamental species. As we embark on this journey to delve into the stomatal adaptations of

Codiaeum variegatum, Acalypha wilkesiana, and Croton zambesicus, we seek to unveil the

secrets hidden within their leaves and gain a deeper appreciation for the intricate value of these

plants that have not only graced our world with their beauty but can also protect us from

consequences that comes with inhaling impure air. By conducting a comparative analysis, it

becomes feasible to discern differences in stomatal dimensions, size and density. These

characteristics contain valuable insights into a plant's efficiency and its potential effectiveness in

carbon sequestration to improve environmental quality (Brodribb et al., 2007).

1.2 AIM OF THE STUDY

Comparative anatomical studies of the stomata of Codiaeum variegatum, Acalypha wilkesiana

and Croton zambesicus to assess their suitability for environmental purification through carbon

sequestration.

2
1.3 OBJECTIVE OF THE STUDY

 Characterizing the stomatal size and distribution in the leaves of each species.

 Measuring vessel size in the vascular tissues of the studied plants.

 Investigating potential correlations between stomatal size, vessel size, and carbon

sequestration efficiency.

 Comparing the stomatal and vessel characteristics among the three species.

 Analyzing how these anatomical traits could influence pollutant uptake and transport.

 Discussing the feasibility and limitations of using Codiaeum variegatum, Acalypha

wilkesiana and Croton zambesicus for environmental purification based on their stomatal and

vessel features.

1.4 SIGNIFICANCE OF THE STUDY

This study carries importance in both environmental and horticultural realms. Investigating the

stomatal features of these ornamental plants adds to our comprehension of their potential as

agents for carbon sequestration. If their anatomical characteristics facilitate effective absorption

and transportation of pollutants, these plants could offer a dual benefit by not only beautifying

landscapes but also aiding in environmental cleansing. Such an approach is consistent with the

principles of sustainable urban development and the promotion of green infrastructure.

1.5 SCOPE OF STUDY AND LIMITATIONS

This study aims to conduct a comparative anatomical analysis of stomatal characteristics in

Codiaeum variegatum, Acalypha wilkesiana, and Croton zambesicus plants. Specifically, the

3
investigation will involve quantifying and comparing stomatal size and density on both leaf

surfaces and stems of these selected ornamental plant species. Additionally, it will explore vessel

size and distribution patterns to identify any noteworthy variations across different regions of the

stem. Throughout the research, the focus will be on contrasting and explaining the differences

and similarities in stomatal traits observed among these three chosen ornamental plant species,

with the objective of gaining insights into how these anatomical features of the plants will act as

organs of carbon sequestration.

1.6 LIMITATIONS

This research is subject to various limitations that define its scope and boundaries. Firstly, the

study is limited to geographic regions where Codiaeum variegatum, Acalypha wilkesiana, and

Croton zambesicus are commonly cultivated. Consequently, environmental variations across

different geographical areas may impact the observed stomatal characteristics. While the

research thoroughly examines stomatal density, size, and distribution, it does not encompass

other aspects of stomatal anatomy, such as the detailed morphological features of guard cells or

potential variations in subsidiary cell characteristics. Additionally, the study exclusively

concentrates on mature leaves of the selected ornamental plants, acknowledging the possibility

of distinct stomatal traits during various growth stages or under diverse environmental

conditions. Although genetic diversity within plant populations could influence stomatal

attributes, this research does not conduct comprehensive genetic analyses. Furthermore, it does

not engage in direct functional studies to investigate how stomata respond to specific

environmental stimuli, as this exceeds the primary focus of anatomical investigation. Lastly,

while the research recognizes the potential impact of environmental factors on stomatal traits, it

4
does not aim to comprehensively address all conceivable variables due to the intricate nature of

their interactions.

1.7 PRACTICAL APPLICATIONS AND IMPLICATIONS

Despite these limitations, this research holds substantial significance. The knowledge acquired

through the comparative analysis of stomatal characteristics in Codiaeum variegatum, Acalypha

wilkesiana, and Croton zambesicus plants can find practical applications. These findings have

the potential to inform decisions in ornamental horticulture, contribute to the thoughtful design

of landscapes, and provide valuable insights into the potential utilization of these ornamental

plants in carbon sequestration initiatives. Grasping the stomatal adaptations in these species lays

the groundwork for making informed choices in plant selection and management strategies. In

essence, this study serves as a bridge between the aesthetic and ecological facets of urban

environments, offering a foundation for enhancing the ornamental and functional roles of these

plants.

5
6
 CHAPTER TWO

LITERATURE REVIEW

2.1 OVERVIEW OF CODIAEUM VARIEGATUM

Croton, Codiaeum variegatum, is a common houseplant grown for its striking foliage. It is one of

six species of broadleaf evergreen perennials, shrubs, and small trees in this genus in the

euphorbia family (Euphorbiaceae) which are native to tropical Asia and the western Pacific

region (not to be confused with Croton, another genus of more than 700 species in the same

family, in which it was formerly incorrectly classified as Croton variegatum). This evergreen

shrub or small tree varies widely in leaf color and pattern and numerous cultivars have been

developed. It is a tender perennial, hardy only in zones 11-12. In subtropical and tropical

climates, they are frequently used as landscape shrubs for dramatic hedges, bold focal points in

gardens, or potted specimens around buildings. In tropical climates, crotons make attractive

hedges and potted patio specimens, valued for their striking foliage. They only survive outdoors

where temperatures do not normally drop below 10 to 13 °C (50 to 55 °F) in winter; colder

temperatures can cause leaf loss. In colder climates, the plants are grown in greenhouses or as

house plants. The cultivated garden crotons are usually smaller than the wild plant, rarely over

1.8 m (5.9 ft) tall, and come in a wide diversity of leaf shapes and colours. They are sometimes

grouped under the name Codiaeum variegatum. The several hundred cultivars are selected and

bred for their foliage. Depending on the cultivar, the leaves may be ovate to linear, entire to

deeply lobed or crinkled, and variegated with green, white, purple, orange, yellow, red, or pink.

The colour patterns may follow the veins or the margins, or be in blotches on the leaf. The large,

thick, leathery leaves are 2 to 12 inches long and quite variable. The shiny, alternate leaves may

be linear to oval, have a smooth or lobed margin (sometimes deeply cut to the midrib), and some

7
are wavy or twisted into a spiral. The foliage colour ranges from green variegated with white,

pink, orange, red, yellow, or purple in various combinations that may change as the leaves age.

Markings may follow along the major veins or may be blotches on any part of the leaf blade in

regular or random patterns. When grown as a houseplant it rarely flowers. When growing in the

ground in frost-free climates in spring it produces insignificant flowers in long axillary racemes.

The white male flowers with five small petals and 20-30 stamens that give a starburst-like effect

are produced on separate inflorescences than the yellowish, petal-less female flowers. Pollinated

female flowers are followed by fruits which are a small tripartite capsule about a third of an inch

in diameter containing three small seeds. Croton is typically offered as an indoor plant but can

also be used outdoors as a seasonal accent plant in containers or plantings of annuals or mixed

ornamentals. Croton does best in fertile, well-drained, moist soil. They need bright, indirect light

when grown indoors. Outdoors they thrive in partial shade, and in cool climates can tolerate full

sun if kept moist (and are acclimated first when moved from inside). Higher light produces more

vibrant color in the leaves and a more compact plant. The colorful leaves may revert to shades of

green in insufficient light, while too much direct sun makes the leaves gray and dull looking.

These plants have moderate water needs and should be watered only when the top half-inch to an

inch of soil dries out. Reduce watering in the winter. Plants will drop leaves if they are too wet or

too dry for extended periods. As a tropical plant, it does best with moderate to high humidity and

warm temperatures. The evergreen leaf is simple and spiral with lobed and undulated leaf

margin. Leaf shape is ovate with blade length between 4 to 8 inches. Fruit shape is round, brown-

coloured and dry (Gilman, 1999).

2.1.1 Botanical Description

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It is a tropical, evergreen, monoecious shrub growing to 3 m (9.8 ft) tall and has large, thick,

leathery, shiny evergreen leaves, alternately arranged, 5–30 cm (2.0–11.8 in) long and 0.5–8 cm

(0.20–3.15 in) broad. The leaf blades can, for example, be ruler-lanceolate, oblong,

elliptic, lanceolate, ovate inverted, ovate spatulate, or violin-shaped and coloured green, yellow,

or purple in various patterns, depending on the variety. The petiole has a length of 0.2 to 2.5 cm.

The inflorescences are long racemes, 8–30 cm (3.1–11.8 in) long, with male and female flowers

on separate inflorescences; the male flowers are white with five small petals and 20–30 stamens,

pollens are oval approximately 52x32 microns in size. The female flowers are yellowish, with no

petals. The flowering period is usually in early autumn. The fruit is a capsule 9 mm (0.35 in) in

diameter, containing three seeds that are 6 mm (0.24 in) in diameter. When cut, stems bleed a

milky sap like many of the Euphorbiaceae (Bingtao and Gilbert, 2008).

9
Plate 1: Codiaeum variegatum shrub

2.1.2 BOTANICAL CLASSIFICATION OF CODIAEUM VARIEGATUM

Kingdom: Plantae

Phylum: Tracheophytes

Subphylum: Angiosperms

Clade: Eudicots

Clade: Rosids

Order: Malpighiales

Family: Euphorbiaceae

Genus: Codiaeum

Species: C. variegatum

Source: https://en.wikipedia.org/wiki/File:Colpfl05.jpg

2.2 OVERVIEW OF ACALYPHA WILKESIANA

Acalypha wilkesiana is indigenous to Fiji and the surrounding South Pacific islands. It is a

tropical shrub or subshrub that grows to 10-15' tall and as wide in frost-free environments. In St.

Louis, it is commonly cultivated as an annual, growing 2-4' tall in a single growing season. This

shrub is grown for its striking foliage, which has lustrous, coarsely-toothed, oval to elliptic

bronze-green leaves (5-8 inches long) mottled with red and purple or red and bronze. Green

flowers, occasionally tinted with reddish-pink, occur in slender spikes up to 8 inches long. It is a

fast-growing evergreen shrub which provides a splash of colour in the landscape with bronze red

10
to muted red, the leaves appear as heart shaped with combination of colour like green, purple,

yellow, orange, pink or white depending on cultivation. However, few studies have mentioned

the phytochemical constituents and elemental studies of A. wilkesiana. Akinde (1986) reported

the presence of sesquiterpenes, monoterpenes, triterpenoids and polyphenols.

2.2.1 BOTANICAL CLASSIFICATION ACALYPHA WILKESIANA

Kingdom: Plantae
Phylum: Tracheophytes
Subphylum: Angiosperms
Clade: Eudicots
Clade: Rosids
Order: Malpighiales
Family: Euphorbiaceae
Genus: Acalypha
Species: A. wilkesiana
Source: https://en.wikipedia.org/wiki/Acalypha_wilkesiana

11
Plate 2: Acalypha wilkesiana shrub

2.3 OVERVIEW OF CROTON ZAMBESICUS

Croton zambesicus is a shrub or small tree up to 16 m high, of fringing forest and savanna. It is

predominant in Gambia and Nigeria, and widely distributed elsewhere in tropical Africa. The

tree has a scaly bark and silvery leaves, rusty-scaly below, and has an attractive appearance. It is

often planted in towns and villages. In times past it was planted as a fetish tree. It has a

reputation of conferring protection, and is often planted near the entrance of houses to ward off

evil influence. It is particularly protective against witches. Boyom et al. (2002) studied the

composition of essential oils from the leaves, stem and roots of Croton zambesicus and found the

three types of oils to be similar in composition, with those from the leaves and stem rich in

monoterpenes, while that of the root bark contains sesquiterpenes. The root and stem bark oils

were found to be rich in oxygen-containing compounds, with spathulenol and linalool as major

components. Studies have reported on the antimicrobial properties of the leaf and stem

(Papazoglou et al., 2017).

2.3.1 BOTANICAL CLASSIFICATION

Kingdom: Plantae
Phylum: Tracheophytes
Subphylum: Angiosperms
Clade: Eudiocots
Clade: Rosids
Order: Malpighiales
Family: Euphorbiaceae
Subfamily: Crotonoideae
Genus: Croton

12
Species: Croton zambesicus

Sources: https://en.wikipedia.org/wiki/Croton_(plant)

Fig 5: Croton zambesicus

2.4 STOMATA MORPHOLOGY

Stomata, minute openings on the surfaces of plant leaves, stems, and other aerial parts, play a

pivotal role in plant physiology and adaptation. The morphological characteristics of stomata are

essential for understanding their functions and ecological significance. Stomata consist of two

specialized guard cells surrounding a pore, and their morphology is subject to variations

influenced by genetic, environmental, and developmental factors. These structural variations can

profoundly impact a plant's ability to adapt to its surroundings. Stomatal density, the number of

13
stomata per unit area, is a crucial morphological trait. Research by Hetherington and Woodward

(2003) suggests that stomatal density influences a plant's capacity to exchange gases and regulate

water loss. High stomatal density may enhance photosynthetic efficiency but could increase

water loss, while low density conserves water but may limit carbon dioxide uptake. Stomatal

size, another critical aspect, varies between plant species and can be influenced by factors such

as light intensity (Lake et al., 2001). Larger stomata may enhance gas exchange efficiency in

low-light conditions, but smaller stomata can reduce water loss, particularly in arid

environments. The distribution pattern of stomata across leaf surfaces is also significant.

Research by Franks et al. (2009) indicates that varying stomatal distribution patterns can reflect a

plant's adaptation to specific ecological niches. Stomatal morphology is a dynamic and essential

aspect of plant anatomy that influences a plant's ability to balance gas exchange and water

conservation. Understanding these morphological traits is pivotal for comprehending plant

adaptation to diverse environments and their responses to changing environmental conditions.

2.4.1 STOMATA MORPHOLOGY OF CODIAEUM VARIEGATUM

Codiaeum variegatum, commonly known as garden croton, exhibits distinctive stomatal

morphology that reflects its adaptation to various environmental conditions. Stomata in

Codiaeum variegatum is primarily located on the lower leaf surface, a characteristic often

observed in plants from tropical regions to minimize water loss through transpiration. The

stomata of Codiaeum variegatum typically consist of two kidney-shaped guard cells surrounding

a central pore. Recent research on stomatal morphology, although not specific to Codiaeum

variegatum, highlights the critical role of these structures in regulating gas exchange, particularly

carbon dioxide uptake for photosynthesis and the control of transpiration rates (Franks and

Beerling, 2009). Stomatal size, density, and distribution are vital aspects of Codiaeum

14
variegatum's stomatal morphology, and these traits may vary in response to environmental

factors, including light intensity and humidity (Mudhoo et al., 2017).

2.4.2 STOMATA MORPHOLOGY OF ACALYPHA WILKESIANA

Acalypha wilkesiana, commonly referred to as copperleaf or Jacob's coat, possesses distinct

stomatal morphology that reflects its adaptation to various environmental conditions. Stomata in

Acalypha wilkesiana are primarily found on the lower leaf surface, a characteristic commonly

observed in plants from tropical and subtropical regions to minimize water loss through

transpiration. The stomata of Acalypha wilkesiana typically consist of two specialized guard

cells encircling a central pore. Recent research on stomatal morphology, though not specific to

Acalypha wilkesiana, underscores the crucial role of these structures in regulating gas exchange,

particularly carbon dioxide uptake for photosynthesis and the control of transpiration rates

(Franks and Beerling, 2009). Stomatal size, density, and distribution are significant components

of Acalypha wilkesiana's stomatal morphology, and these traits can vary in response to

environmental factors, including light intensity, humidity, and temperature (Mott and Peak,

2010).

2.4.3 STOMATA MORPHOLOGY OF CROTON ZAMBESICUS

Croton zambesicus, also known as African croton, exhibits notable stomatal morphology that

reflects its adaptation to varying environmental conditions. Stomata in Croton zambesicus are

predominantly found on the lower leaf surface, a feature commonly observed in plants from

tropical and arid regions as an adaptive strategy to minimize water loss through transpiration.

The stomata of Croton zambesicus typically comprise two specialized guard cells surrounding a

15
central pore. Recent research emphasizes the essential role of stomata in regulating gas

exchange, particularly carbon dioxide uptake for photosynthesis and the control of transpiration

rates (Franks and Beerling, 2009). Stomatal size, density, and distribution are significant

components of Croton zambesicus's stomatal morphology. These traits can vary in response to

various environmental factors, including light intensity, humidity, and water availability. The

size of stomata in Croton zambesicus can vary depending on environmental factors. Stomata in

this plant are typically of moderate size, allowing for efficient gas exchange. Stomatal size can

be influenced by light intensity, where lower light levels may result in larger stomata to facilitate

increased gas absorption (Mott and Peak, 2010). The distribution of stomata on the leaf surface

of Croton zambesicus follows a specific pattern. These stomatal characteristics collectively

represent an adaptation strategy that allows Croton zambesicus to thrive in a variety of

environmental conditions, including arid and tropical regions. By adjusting stomatal size,

density, and distribution in response to environmental cues, the plant efficiently regulates gas

exchange and conserves water, contributing to its resilience in challenging habitats. Typically, a

higher density of stomata is found on the lower leaf surface, which is a common adaptation in

many plants from arid regions. This distribution minimizes water loss through transpiration by

reducing the exposure of stomata to direct sunlight (Franks and Beerling, 2009).

2.5 CORRELATION BETWEEN ANATOMICAL TRAITS OF ORNAMENTAL

PLANTS AND CARBON SEQUESTRATION

The correlation between anatomical traits of ornamental plants and carbon sequestration is a

subject of increasing interest due to its potential applications in environmental management and

landscape design. Carbon sequestration, a sustainable and environmentally friendly approach,

employs plants to capture and store atmospheric carbondioxide with the goal of reducing global

16
climate change. Ornamental plants, often chosen for their aesthetic appeal, can also possess

anatomical traits that make them effective in carbon sequestration. This discussion explores the

key anatomical traits of stomata of ornamental plants that influence their carbon sequestration

capabilities and their broader implications for environmental sustainability. Stomatal density, the

number of stomata per unit leaf area, and their distribution on plant surfaces significantly affect

pollutant uptake. Higher stomatal density can enhance pollutant absorption, while specific

distributions may maximize exposure to contaminants. Research by McGrath and Zhao (2003)

highlights that plants with higher stomatal density tend to take up carbondioxide more

effectively. Therefore, ornamental plants with an appropriate stomatal pattern may excel in

carbon sequestration, as they can facilitate pollutant transport from the soil or water into plant

tissues. Leaf characteristics, such as leaf size, density and dimensions, affect transpiration rates

and the potential for air purification. This feature enhances the capacity of ornamental plants to

sequester contaminants effectively, making them valuable in carbon sequestration efforts. By

selecting ornamental species with favorable anatomical traits, landscape designers and

environmentalists can create aesthetically pleasing environments while simultaneously

addressing pollution issues. The use of ornamental plants for carbon sequestration aligns with the

principles of sustainable urban development and green infrastructure, offering a harmonious

blend of beauty and environmental remediation.

2.6 VESSEL SIZE AND POLLUTANT TRANSLOCATION

Vessel size, a crucial anatomical trait in plants, plays a significant role in pollutant translocation

and transport within the plant. Vessels, which are part of the plant's vascular system, are

responsible for the long-distance transport of water, nutrients, and other solutes. The size of these

vessels varies among plant species and can influence the plant's ability to absorb and translocate

17
pollutants. Plants with larger vessel sizes generally have an advantage in pollutant uptake and

translocation. Larger vessels facilitate the movement of water and dissolved substances,

including pollutants, more efficiently throughout the plant. This trait can enhance the plant's

capacity to absorb contaminants from the soil and transport them to various plant parts, including

the leaves and roots. Moreover, vessel size can influence the translocation of pollutants within

the plant. Smaller vessels may restrict the movement of pollutants to specific plant regions, while

larger vessels can distribute contaminants more evenly throughout the plant (Baker et al., 1994).

This has implications for carbon sequestration, where the goal is to reduce the amount of

carbondioxide in the atmosphere. The relationship between vessel size and pollutant

translocation underscores the importance of selecting appropriate plant species for carbon

sequestration purposes. Species with larger vessel sizes may be more effective at translocating

and accumulating pollutants, making them valuable candidates for environmental cleanup efforts

(Glick, 2010). Vessel size is a critical anatomical trait that influences the plant's ability to absorb

and translocate pollutants. Understanding this relationship is essential for selecting suitable plant

species for carbon sequestration projects, as it can impact the efficiency and effectiveness of

pollutant removal from contaminated environments.

2.7 BALANCE BETWEEN POLLUTANT UPTAKE AND WATER CONSERVATION

The balance between pollutant uptake and water conservation in plants is a crucial consideration

in carbon sequestration and ecological adaptation. Plants must efficiently absorb contaminants

from their surroundings while conserving water to ensure their survival. Several anatomical and

physiological adaptations contribute to this delicate equilibrium. Stomatal regulation, controlled

by guard cells, helps manage water loss during gas exchange. Plants can adjust stomatal aperture

to optimize pollutant uptake while minimizing transpiration (Jones, 2013). Root architecture also

18
plays a role, as plants with extensive root systems can access pollutants effectively without

compromising water conservation. Mycorrhizal associations further aid in nutrient and pollutant

uptake while enhancing drought tolerance (Smith and Read, 2008). Balancing pollutant uptake

and water conservation is essential for plant health, especially in contaminated environments.

Selecting plant species with suitable traits, like water-efficient stomatal control and efficient root

structures, can enhance both pollutant removal and the plant's vitality (Salt et al., 1995).

19
 CHAPTER THREE

MATERIALS AND METHODS

3.1 PLANT COLLECTION

The plant samples utilized in this study were collected from Nsukka metropolis, including the

Botanical Garden of the Department of Plant Science and Biotechnology UNN. The selection of

plant species, including Codiaeum variegatum, Acalypha wilkesiana, and Croton zambesicus

was based on their prevalence in Nsukka area of Enugu state and their suitability as ornamental

plants for carbon sequestration. The taxonomic identification of the collected plant specimens

was conducted and authenticated at the herbarium unit of Department of Plant Science and

Biotechnology, University of Nigeria, Nsukka. The sample collection process was conducted in

accordance with ethical guidelines and university policies. For each plant species, both mature

leaves and stem parts were carefully selected, ensuring that they were in a healthy and

representative condition.

3.1 MATERIALS

3.1.1 Laboratory Equipment

20
The laboratory work for this study was conducted at the Central Faculty Laboratory and at the

anatomy lab of the department of Plant Science and Biotechnology within the University of

Nigeria Nsukka (UNN). The Key laboratory equipment and materials used in our study included:

- Leica DM2500 light microscope equipped with high-resolution imaging capabilities for

stomatal analysis and vessel size measurements.

- Ocular micrometer

- Stage micrometer

- Microscope slides and coverslips for sample preparation.

- Fixative solution (400 ml Formaldehyde Acetic Acid) for preserving the plant samples.

- 360 ml of ethanol series for sample dehydration during the preparation process.

- Stains, include safranin, for staining plant tissues during the microscopic analysis.

- Colourless nail varnish

- Fine-tip forceps.

- Razor blade.

- Petri dishes.

- Water

3.1.2 Microscopy Settings

Microscopic analysis of stomatal characteristics and vessel sizes was conducted using a Leica

DM2500 light microscope. The microscope was configured to a magnification of X400 for

21
stomatal examinations and X100 for vessel size measurement with proper adjustments for

contrast and illumination. The microscope settings were maintained consistently throughout the

analysis to ensure the reliability of our measurements.

3.2 SAMPLE PRESERVATION

Leaves and stem segments from each of the three plant species (Codiaeum variegatum, Acalypha

wilkesiana, and Croton zambesicus) were carefully selected based on their health and uniformity.

Formaldehyde Acetic Acid (FAA) solution was chosen as the preserving agent. The FAA

solution used in this study consisted of a mixture of Formaldehyde, Acetic Acid, and ethanol in

specific proportions. Each selected leaf and stem segment was submerged in the FAA solution

immediately after collection and care was taken to ensure that the plant material was fully

immersed in the fixative to allow proper preservation.

3.3 SAMPLE PREPARATION

3.3.1 PREPARATION OF EPIDERMAL LEAF STRIP

Procedures

1. Leaf Selection

Mature leaves from healthy plant specimens were selected for the epidermal leaf strip

preparation. These leaves were chosen to ensure that the stomata were fully developed and

representative of the respective species.

2. Leaf Surface Cleaning

22
To remove any surface contaminants or dust particles, the selected leaves were gently washed

with distilled water and blotted dry using a clean paper towel. This step ensured that the nail

varnish adhered effectively to the leaf surface.

3. Application of Nail varnish

A thin layer of transparent nail polish was carefully applied to the adaxial (upper) surface of the

leaf using a clean, fine-tip brush. The nail polish was spread evenly. A small, discrete area was

chosen to ensure that the resulting epidermal strip contained an adequate number of stomata for

analysis.

4. Drying and Hardening

The nail varnish-coated leaf section was allowed to air-dry completely in a dust-free

environment for approximately 10-15 minutes. Once dried, the nail varnish hardened, forming a

transparent film over the leaf surface.

5.Peeling the Epidermal Strip

Using fine-tip forceps, a small incision was made at the edge of the nail varnish film. Care was

taken to gently lift the edge of the nail varnish film without damaging the underlying leaf tissue.

The film was then carefully peeled off the leaf surface, resulting in the collection of an epidermal

strip.

6. Mounting on Microscope Slides

23
The collected epidermal strip was mounted, nail varnish side down, onto a clean microscope

slide, stained using safranin and a coverslip was gently placed over the epidermal strip to secure

it in place.

3.3.2 SECTIONING

The plant stems were sectioned at the Anatomy laboratory of Plant Science and Biotechnology

Department. Transverse and longitudinal sections of the stems of the plants were made. All these

sections were made using the Reichart sledge microtone, the blade was well sharpened using

electric sharpener. The sections were collected in different petri dishes and 70% alcohol was

poured into the petri dishes to preserve the sections for further anatomical studies and prevent the

slides from decay. The petri dishes were then properly labelled according to the content inside.

3.4 PHOTOGRAPHING

After carefully mounting the plant samples on microscope slides, they were observed and the

images were captured. Using an attached digital camera, high-resolution images of the stomata

on leaf surfaces were captured. Attention was paid to lighting, focusing, and magnification

settings to ensure clear and informative visuals. All images were digitally recorded and labeled

for future reference, allowing for data verification and presentation in the scientific report.

3.5 STOMATAL SIZE MEASUREMENT.

The method of stomatal measurement refers to the research conducted by Haryanti and Tetrinica

(2009). Measuring stomata using stage and ocular micrometers were carried out. For each

section of observation, ten replications were made. Stomatal measurements were carried out in

24
two stages:

a. Micrometer Calibration:

Ocular micrometer was put on the ocular lens giving a picture of line score. The stage

micrometer was then put on the stage. The line coming out from the stage micrometer was

positioned in line together with the line of the ocular micrometer (Agustini, 1994). The

conversion factor at x400 magnification is:

100 of ocular = 4 of stage, 1 = x.

X = 4/100 = 0.04

b. Measuring Stomatal Length and Width:

 After the scale value of the ocular micrometer was known, the stage micrometer was

taken out and replaced with the microscope slides of stomata to be measured. The ocular

micrometer stayed in its original place.

 The magnification of the objective lens used for measurement was at x400 magnification,

this was the same as the lens used for calibration.

 The scale of the ocular lens was put on the position of the scale preserved slides so that

the length and the width of stomata can be measured.

 Length and width were measured using the scale multiplied by calibration value.

 The standard errors were calculated for each length and width of the plant.

 The stomata number for each field of view was also counted and recorded.

3.6 VESSEL SIZE MEASUREMENT

25
The compound light microscope equipped with both ocular and stage micrometers was set to the

desired magnification of x100, a critical parameter for this measurement process. For each

section of observation, ten replications were carried out. Vessel measurements were carried out

in two stages:

a. Micrometer Calibration:

Ocular micrometer was put on the ocular lens giving a picture of line score. The stage

micrometer was then put on the stage. The line coming out from the stage micrometer was

positioned in line together with the line of the ocular micrometer (Agustini, 1994).

The conversion factor at x100 magnification is:

100 of ocular =97 of stage, 1 = x.

X = 97/100 = 0.97

b. Measuring vessel length and width:

 The measurement process began with the placement of the prepared vessel sample on the

microscope stage, ensuring that the vessel of interest was within the microscope's field of

view.

 The microscope's focus controls were used to obtain a sharp and clear image of the

vessel.

 Measurements of vessel diameter or length were then conducted using the scale

multiplied by the calibration value.

26
  The ocular micrometer was employed by aligning its scale with the vessel in focus, and

the number of divisions spanning the vessel's dimensions was recorded.

 The number of vessels in each field of view was also counted and recorded

CHAPTER FOUR

RESULTS AND STATISTICAL ANALYSIS

4.1 ANATOMICAL ANALYSIS

4.1.1 Anatomical analysis of stomata properties across different ornamental species

The variations in stomata properties across different ornamental plants is shown in Figure 1. The

stomata size ranged from 0.005± 0.000 in Croton zambesicus to 0.180 ± 0.169 in Acalypha

wilkesiena for the selected ornamentals. Acalypha wilkesiana had the highest mean value for

27
stomata size while the lowest mean value was recorded in Croton zambesicus. There was no

significant difference observed in the stomata size of the three plants.

The mean value of stomata density ranged from 16.100 ± 0.100 observed in Acalypha wilkesiana

to 34.100 ± 0.100 observed in Codiaeum variegatum. There was a significant difference in

stomata density among the three plant species (Table 1).

Table 1: Comparison of stomata sizes and stomata density across different ornamental
plant species

Stomata Stomata
Stomata length Stomata size
width density

Codiaeum
5.550 ± 0.217b 3.000 ± 0.247a 0.021 ± 0.002a 34.100 ± 0.100a
variegatum

Acalypha wilkesiana 8.100 ± 0.482a 1.350 ± 0.130b 0.180 ± 0.169a 16.100 ± 0.100c

Croton zambesicus 4.450 ± 0.138c 0.950 ± 0.050b 0.005 ± 0.000a 24.000 ± 0.149b

Means with different alphabet represents significant differences

28
 Guard cells

Stoma

Epidermal cell

Plate 4: T.S of leaf epidermis of Codiaeum variegatum showing the stomata

Stoma

Guard cell

Epidermal cell

Plate 5: T.S of Acalypha wilkesiana showing the stomata

Stoma

Plate 6: T.S of leaf epidermis of Croton zambesicus showing the stomata.

29
4.1.2 Anatomical analysis of vessel properties across different ornamental species

The variations in vessel properties across different ornamental plants is shown in Figure 2. The

result shows that the mean value for vessel size ranged from 4.608 ± 0.229 observed in

Codiaeum variegatum to 6.863 ± 0.162 observed in Acalypha wilkesiana. The highest mean

value which was recorded in Acalypha wilkesiana was significantly different from that of

Codiaeum variegatum and Croton zambesicus while the mean values for vessel size between

Codiaeum variegatum and Croton zambesicus were not significantly different from each other.

The mean value for vessel density ranged from 79.900 ± 0.100 to 153.000 ± 0.149. The highest

mean value for vessel density was observed in Croton zambesicus at 153.000 ± 0.149, followed

by Codiaeum variegatum at 109.100 ± 0.100. The lowest mean value for vessel density was

recorded in Acalypha wilkesiana at 79.900 ± 0.100. Although the vessel number of Codiaeum

variegatum and Croton zambesicus were higher when compared to Acalypha wilkesiana,

however, there was significant difference observed among the three plants (Table 2).

Table 2: Comparison of vessel size and vessel density across different ornamental plant
species

Vessel length Vessel width Vessel size Vessel number

Codiaeum variegatum 4.750 ± 0.291b 4.750 ± 0.227b 4.608 ± 0.229b 109.100 ± 0.100b

Acalypha wilkesiana 7.100 ± 0.163a 7.050 ± 0.263a 6.863 ± 0.162a 79.900 ± 0.100c

Croton zambesicus 5.200 ± 0.186b 5.330 ± 0.184b 5.107 ± 0.169b 153.000 ± 0.149a

Means with different alphabet represents significant differences

30
 Vessel

Plate 7: T.S of stem segments of Codiaeum variegatum showing the vessels

Vessel

Plate 8: T.S of stem sections of Acalypha wilkesiana showing the vessels

31
 Vessel

Plate 9: T.S of stem sections of Croton zambesicus showing the vessels

32
 4.2 CORRELATION COEFFICIENTS OF ANATOMICAL PROPERTIES

4.2.1 Correlation coefficient on the relationship between stomata properties

Correlation matrix was constructed to analyze the relationship between the stomata properties

(size and density) across different ornamental plants (Codiaeum variegatum, Acalypha

wilkesiana, and Croton zambesicus). From the table below, correlation coefficient between

stomata size and stomata density is -206, which indicates a weak negative correlation and the

correlation coefficient is not statistically significant (Table 3).

Table 3: Pearson correlation coefficient on the relationship between stomata properties

Stomata Stomata
Stomata Stomata Stomata Stomata
length x width x
length width size density
0.04 0.04

Stomata length 1

Stomata width .013 1

Stomata length x
.956** -.028 1
0.04
Stomata width x
-.012 .969** -.047 1
0.04

Stomata size .313 -.025 .298 -.008 1

Stomata density -.530** .707** -.588** .735** -.206 1

33
 4.2.2 Correlation coefficient on the relationship between stomata size and vessel pore size

Correlation matrix was constructed to analyze the relationship between the stomata and vessel

properties across different ornamental plants (Codiaeum variegatum, Acalypha wilkesiana, and

Croton zambesicus). The main properties of concern are the stomata size, stomata density, vessel

size and vessel number. Correlation coefficient on the relationship between stomata and vessel

properties showed that there was a weak positive correlation between vessel size and stomata

size and the correlation coefficient is not statistically significant. There was a strong negative

correlation between vessel size and stomata density and the correlation coefficient is statistically

significant at p < 0.01. The correlation coefficient between vessel number and stomata size is

-.221 which means there was a weak negative correlation and it is not statistically significant. A

weak positive correlation was observed between vessel number and stomata density with

coefficient value of .331, the value was not statistically significant (Table 4).

Table 4: Pearson correlation coefficient on the relationship between stomata and vessel
properties
Stomata
Stomata Stomata Stomata length Stomata Stomata
width x
length width x 0.04 size density
0.04

Vessel length .616** -.479** .690** -.492** .193 -.772**

Vessel width .639** -.438* .737** -.452* .195 -.772**

Vessel length x
.616** -.479** .690** -.492** .193 -.772**
0.97
Vessel width x
.639** -.438* .737** -.452* .195 -.772**
0.97

Vessel size .653** -.478** .742** -.491** .202 -.803**

Vessel number -.801** -.260 -.853** -.236 -.221 .331

34
4.2.3 Correlation coefficient on the relationship between vessel properties

Correlation matrix was constructed to analyze the relationship between the vessel properties

across different ornamental plants (Codiaeum variegatum, Acalypha wilkesiana, and Croton

zambesicus). The table below shows that the correlation coefficient between vessel size and

vessel number is -.567, which indicates that there is a weak negative correlation and the

correlation coefficient is statistically significant at p < 0.01 (Table 5).

Table 5: Pearson correlation coefficient on the relationship vessel properties

Vessel Vessel
Vessel Vessel Vessel Vessel
length x width x
length width size number
0.97 0.97

Vessel length 1

Vessel width .847** 1

Vessel length x
1.000** .847** 1
0.97

Vessel width x
.847** 1.000** .847** 1
0.97

Vessel size .962** .960** .962** .960** 1

Vessel number -.570** -.519** -.570** -.519** -.567** 1

CHAPTER FIVE
35
 DISCUSSION AND CONCLUSION

5.1 DISCUSSION

Global advancement in industrialization, urbanization, increase in human population as well as

economic growth is generally associated with increased demand for energy particularly from

fossil fuels which results in increased emission of toxic gases and other substances to the

environment (Ekpemerechi et al., 2017). Air quality is an important factor that plays a role in

public health as most people spend 80-90 % of their time doing activities indoors such as homes,

schools, offices, gymnasiums and means of transportation (Susanto et al., 2021). The United

States Environmental Protection Agency (USEPA) stated that exposure to indoor pollutants is

100 times more deleterious then exposure to outdoor pollutants due to the high concentration of

indoor pollutants (Pitarma et al., 2017). In accordance with outdoor air quality, bad indoor air

quality has the potential to cause health problem such as asthma, irritation, allergic reactions,

nausea, muscle aches, headaches etc., on a short-term basis, however continuous long-term

exposure may result to more severe health problems such as chronic pulmonary disease (COPD)

and lung cancer (Seguel et al., 2016). Generally, plants are considered as green liver of the earth

because they are the global repository for environmental chemicals. Ornamental plants on the

other hand, asides from their aesthetic value has the health benefit of absorption of air pollutants,

filtration of dust, biofiltration of volatile organic compounds (formaldehyde, benzene, xylene,

etc.), release of oxygen and increasing of humidity (Sayed, 2020). In this research, results of the

comparative anatomical studies of the stomata of Codiaeum variegatum, Acalypha wilkesiana

and Croton zambesicus showed variations in parameters measured. The findings showed that

Acalypha wilkesiana had the highest stomata size and vessel size values. Although Acalypha

wilkesiana had the highest value for the aforementioned parameters, when compared to reports

36
by other authors on ornamental plants, its anatomical values are very low, except for stomata

width in the study by Shaheen et al. (2016), which were slightly higher than those reported in

this study. Similar studies by Sayad et al. (2022) on the use of Golden pothos and Croton plants

sprayed with glutathione and bilirubin as indoor plants; Amujiri et al. (2022) on artisanal stone-

mining impacts on leaf microstructure and biochemical properties of some plants and

Susilowathi et al. (2022) on foliar stomata characteristics of tree species in a university green

open space all showed high anatomical values necessary for carbon sequestration. The stomata

are the main channels through which plants exchange water and gas with their surroundings and

are directly linked to plant physiological processes such as photosynthesis, respiration and

transpiration. Differing significantly in size and density, the maximum number of stomata per

leaf area is restricted by the size of the stomata, but there is a considerable variation in the

stomatal density and size between species leading to differences in rates of gas exchange

(Kardiman and Raebild, 2017). According to Drake et al. (2013 in kardiman), plants with large

and few stomata tend to have higher water-use efficiency, but also reduced rate of photosynthesis

when compared to plants that have many but smaller stomata. However, Susilowathi et al.

(2022) affirms that plant species with larger stomata are more suitable to assign as carbon

sequesters as their large stomata would accommodate a higher rate of plant transpiration and a

resultant increased CO2 fixation. Variations in size and density of stomata may arise due to

differences in species, genetic factors and/or growth under different environmental conditions

(Khan et al., 2014 in susilowathi). Codiaeum variegatum and Croton zambesicus recorded the

highest values for stomata density and vessel number respectively. The stomata density is often

determined by the number of stomata on the leaf surface. The density of the stomata could

increase the evaporation in the leaves and accelerate CO 2 assimilation during the period of

37
stomata opening in the morning and evening. Croton zambesicus which had different vessel size

when compared to that of Codiaeum variegatum and Acalypha wilkesiana with similar vessel

size values has the potential for faster transpiration rate. Correlation coefficient on the

relationship between stomata size and stomata density showed that there was a weak negative

correlation between stomata size and density. Correlation coefficient on the relationship between

vessel properties showed that there was a weak negative correlation vessel size and vessel

number, and it is statistically significant at p < 0.01. Correlation coefficient on the relationship

between stomata and vessel properties showed that there was a strong negative correlation

between vessel size and stomata density, and the correlation coefficient is statistically significant

at p < 0.01. These finding implies that there is a significant relationship between the following

parameters: vessel size, vessel number and stomata density. Stomata size and vessel size which is

significant in all correlation coefficients could be considered the most important anatomical

parameter when selecting ornamental plants for carbon sequestration. On this note, Acalypha

wilkesiana with the highest stomata size and vessel size values will be better suited as an

ornamental plant for purification of air through carbon sequestration when compared to

Codiaeum variegatum and Croton zambesicus.

5.2 CONCLUSION

The comparative anatomical studies of the stomata of Codiaeum variegatum, Acalypha

wilkesiana and Croton zambesicus selected as ornamentals revealed that Acalypha wilkesiana

had the highest stomata size and vessel size values while Codiaeum variegatum and Croton

zambesicus recorded the highest values for stomata density and vessel number respectively.

Owning to the fact that plant species with larger stomata are more suitable to assign as carbon

sequesters as their large stomata would accommodate a higher rate of plant transpiration and a

38
resultant increased CO2 fixation, Acalypha wilkesiana with the highest values for the evaluated

parameters will be better suited for greening followed by Codiaeum variegatum and Croton

zambesicus.

39
 REFERENCES

Abdulmoniem M. A. (2011). Effects of auto-exhaust pollution on the micro-morphology and leaf

epidermal features of ornamental plants in Khartourn, Sudan. Journal of Applied Sciences
Research, 7(3): 270–273.

Adedeji O. and Jewoola O. A. (2008). Importance of leaf epidermal characters in the Asteraceae
family. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 36(2): 7–16.

Agrawal M. and Deepak S. S. (2003). Physiological and biochemical responses of two cultivars
of wheat to elevated levels of CO2 and SO2, singly and in combination. Environmental
Pollution, 121: 189–197.

Akinde B. E. (1986). Phytochemical and microbiological evaluation of the oils from the leaves
of Acalypha Wilkesiana. In: Safowora A. (ed). The State Medicinal Plant Reseaarch in
Nigeria. University of Ibadan Press, Nigeria. Pg. 362 – 363.

Akinyemi K., Oladapo, O. and Okawara, C. (2006), Screening of crude extracts of six medicinal
plants used in South-west Nigerian unorthodox medicine for anti-methicillin resistant
Staphylococcus aureus activity. BMC Complementary and alternative medicine, 1-7.

Amujiri, A. N., Nwafor, F. I., Ozokolie, C. B., Obayi, H. C., Asogwa, L. N. and Igwe, U. (2022).
Artisanal stone-mining impacts on leaf microstructure and biochemical parameters of some
plants at Eziani, Nsukka, Nigeria. Journal of Biological Research and Biotechnology, 20(3):
1740-1752.

Arun N., Peter G. A. and Udhaya A. (2003). Effective surface sterilization and callus induction
protocol for copper leaf (Acalypha Wilkesiana). International Journal of Applied
Biotechnology and Biochemistry, 3(1):37-49.

Baker A. J. M., McGrath S. P., Reeves R. D. and Smith J. A. C. (1997). Metal hyperaccumulator
plants: A review of the ecology and physiology of a biological resource for carbon
sequestration of metal-polluted soils. Carbon sequestration of Contaminated Soil and
Water, 85-107.

40
Baker A. J. M., Reeves R. D. and Hajar, A. S. M. (1994). Heavy metal accumulation and
tolerance in British populations of the metallophyte Thlaspi caerulescens and C. Presl
(Brassicaceae). New Phytologist, 127(1):61-68.

Baker E. A. and Hunt G. M. (1986). Erosion of waxes from leaf surfaces by stimulated acid rain.
New Phytologist, 102:161–173.

Bingtao L. and Gilbert M. B. (2008). Codiaeum. Flora of China, 11:268

Block S., Baccelli C., Tinant B., Meervelt L. V., Rozenberg R., Jiwan J. H., Llabres G., De
Pauw-Gillet M. and Quetin-Leclercg J. (2004). Diterpenes from the leaves of Croton
zambesicus. Phytochemistry, 65(8):1165-1171

Brodribb T. J., Field T. S. and Jordan G. J. (2007). Leaf Maximum Photosynthetic Rate and
Venation Are Linked by Hydraulics. Plant Physiology, 144(4):1890–1898

Chen, J. and Stamps, R. H. (2006). In: Dole, J. M. and Gibson, J. L. (Eds.) Cutting
Propagation: A Guide to Propagating and ProducingFloriculture Crops. Ball Publishing,
Batavia, IL, pp. 203–228.

Deng M., Chen J. J., Henny R. J. and Li Q. S. (2010). Genetic relationships of Codiaeum
variegatum cultivars analyzed by amplified fragment length polymorphism markers.
HortScience, 45(6):868-874.

Drake, P. L., Froend, R. H. and Franks, P. J. (2013). Smaller, faster stomata: Scaling of stomatal
size, rate of response and stomatal conductance. Journal of Experimental Botany, 64: 495-
505.

Echeverria, A., Anfodillo, T., Soriano, D., Rosell, J. A. and Olson, M. E. (2019). Constant
theoretical conductance via changes in vessel diameter and number with height growth in
Moringa oleifera. Journal of Experimental Botany, 70(20): 5765-5772.

Ekpemerechi, S. E., Ajao, A. A., Jimoh, M. A. and Saheed, A. (2017). Variation in leaf
anatomical characters in response to air pollution in some Euphorbiaceae species. West
African Journal of Applied Ecology, 25(1): 21-31.

41
Ekperemechi S. E., Ajao A. A., Jimoh M. A. and Saheed S. A. (2017). Variation in leaf
anatomical characters in response to air pollution in some Euphorbiaceae species. West
African Journal of Applied Ecology, 25(1):21–31.

Floral of Zimbabwe (2017). Croton zambesicus. Retrieved from: www.zimbabweflora.co.zw.

Accessed on 23th August, 2023.

Franks P. J. and Beerling D. J. (2009). Maximum leaf conductance driven by CO 2 effects on

stomatal size and density over geologic time. Proceedings of the National Academy of
Sciences, 106(25): 10343-10347

Franks P. J., Drake P. L., Beerling, D. J. and Chabot J. F. (2009). Stomatal acclimation to CO 2
within a C3/C4 intermediacy. Functional Plant Biology, 36(11):1037-1046.

Franks, P. J. and Beerling, D. J. (2009). CO2‐forced evolution of plant gas exchange capacity
and water‐use efficiency over the Phanerozoic. Geobiology, 7(2):227-236.

Gilman E. F. (1999). Codiaeum variegatum. University of Florida, Institute of Food and

Agricultural Science. 3Pp.

Giri S., Shrivastava D., Deshmukh K. and Dubey P. (2013). Effect of air pollution on chlorophyll
content of leaves. Current Agriculture Research Journal, 1(2): 93–98.

Glick B. R. (2010). Using soil bacteria to facilitate carbon sequestration. Biotechnology

Advances, 28(3):367-374.

Hetherington A. M. and Woodward F. I. (2003). The role of stomata in sensing and driving
environmental change. Nature, 424(6951):901-908.

Jones, H. G. (2013). Plants and Microclimate: A Quantitative Approach to Environmental Plant

Physiology (3rd ed.). Cambridge University Press. 428Pp.

Kardiman, R. and Raebild, A. (2017). Relationship between stomatal density, size and speed of
opening in Sumatran rainforest species. Tree Physiology, 38: 696-705.

Kathiresan P., Karar P. and Siva H (2011). Comparative micromorphological and phytochemical
studies on the roots of three Viburnum (Caprifoliaceae) species. Turkish Journal of Botany,
35: 663–670.

42
Khan, F., Yousaf, Z., Ahmed, H. S., Arif, A., Rehman, H. A., Younas, A., Rashid, M., Tariq, Z.
and Raiz, N. (2014). Stomatal patterning: An important taxonomic tool for systematical
studies of tree species of angiosperm. Annual Resource and Reviews in Biology, 4(24):
4034-4053.

Kumar S., Joseph L., George M., Lakhvir K. and Vasudha A. (2011). An overview - Croton
Zambesicus. International Journal of Pharmaceutical Research and Development, 3(4):
187-192.

Lake J. A., Woodward F. I. and Quick W. P. (2001). Long ‐distance CO 2 signaling in plants.
Journal of Experimental Botany, 52(356):895-906.

Liu Y. J. and Ding H. (2008). Variation in air pollution tolerance index of plants near a steel
factory: implication for landscape-plant species selection for industrial areas. WSEAS
Transactions on Environmental Development, 4: 24–32

Mansfield TA, Hetherington AM, Atkinson CJ (1990). Some current aspects of stomatal
physiology. Annual Review of Plant Biology, 41(1):55-75.

McGrath S. P. and Zhao F. J. (2003). Phytoextraction of metals and metalloids from

contaminated soils. Current Opinion in Biotechnology, 14(3):277-282.

Mollick A. S., Shimoji H., Denda T. and Yokota M. (2011). Croton Codiaeum variegatum L.
Blume cultivars characterized by leaf phenotypic parameters. Scientia Horticulturae,
132(1):71-79

Mott K. A. and Peak D. (2010). Stomatal responses to humidity in air and helox. Plant, Cell and
Environment, 33(2):260-270.

Mudhoo A., Sharma S. K., Jain, S. and Adegboye A. O. (2017). Phytoextraction strategies for
soils contaminated with heavy metals: Modifications and future perspectives. Journal of
Environmental Management, 186:263-276.

Ogundajo A. L., Ogunwande I. A., Gbadamosi H. G., Giwa R. and Flamini G. (2014). Chemical
Composition of the Leaf Essential Oils of Croton zambesicus Müll.-Arg. Grown in Lagos,
South-West Nigeria. European Journal of Medicinal Plants. 4(12): 1524-1533.

43
Ogundare C. S. and Saheed S. A. (2012). Foliar epidermal characters and petiole anatomy of four
species of Citrus L. (RUTACEAE) from south-western Nigeria. Bangladesh Journal Plant
Taxonomy, 19 (1):25–31.

Ogunwenmo, K. O., Idowu, O. A., Innocent, C., Esan, E. B. and Oyelana, O. A. (2007).
Cultivars of Codiaeum variegatum (L.) Blume (Euphorbiaceae) show variability in
phytochemical and cytological characteristics. African Journal of Biotechnology,
6(20):2400-2405

Okokon J. E. and Nwafor P. A. (2009). Antiplasmodial activity of root extract and fractions of
Croton zambesicus. Journal of Ethnopharmacology, 121(1):74-78

Okokon J. E., Nwafor P. A., Umoh E. E., Okokon P. J. and Udobang J. A. (2011). Antidiabetic
and hypolipidemic activities of ethanolic root extract of Croton zambesicus on alloxan
induced diabetic rats. Asian Journal of Pharmaceutical and Biological Research, 1(4):493-
499.

Olopete Q. R. and Ugbogu, O. A. (2019). Anatomical study of leaf and stem of Euphorbia
graminea (Euphorbiaceae). Journal Of Research In Forestry, Wildlife And Environment,
11(4):164-167

Oyelami, O. A. Onayemi, O. Oladimeji, A. and Onawunmi, O. (2003), Clinical Evaluation of

Acalypha ointment in the treatment of superficial fungal skin diseases. Phytotherapy
Research (Wiley Interscience), 17: 555-557.

Pandey S., Kumar N. and Kushwahar R. (2006). Morpho-anatomical and physiological leaf traits
of two alpine herbs, Podophyllum hexandrum and Rheum emodi in the western Himalaya
under different irradiances. Photosynthetic, 44: 11–16.

Papazoglou E. G. and Voulgaridou E. (2017). Trichomes and stomata as models for studying the
effects of heavy metal contamination on plant growth: a micromorphological survey.
Ecological Indicators, 74:178-187.

Pitarma, R., Marques, G., Ferreira, B. R. (2017). Monitoring indoor air quality for enhanced
occupational health. Journal of Medicine Systematics, 41: 23-30.

44
Raven P. H., Johnson G. B. and Losos J. B. (2005). Coerced group collaborative evolution as an
explanation for sexual reproduction’s prevalence. Natural Science, 2:30

Saheed S. A. and Illoh H. C. (2010). A taxonomic study of some species in Cassinae

(Leguminosae) using leaf epidermal characters. Notulae Botanicae Horti Agrobotanici Cluj-
Napoca, 38(1): 21–27.

Salt D. E., Smith R. D. and Raskin, I. (1995). Carbon sequestration. Annual Review of Plant
Biology, 46(1):443-448.

Sana S, Shiju M. and Krishnapriya R. S. (2012). Organogenesis and Somatic Embryogenesis in

Various Cultivars of Codiaeum variegatum (L.). Global Advanced Research Journal of
Biotechnology, 1(3):40-47.

Sayed, N. I. E. L. (2020). Spray Golden Pothos and Croton plants with glutathione and bilirubin
to purify the indoor air from pollutants. Journal of Plant Production, Mansoura
University¸11(5): 439-453.

Seguel, J. M., Merrill, R., Seguel, D. and Campagna, A. C. (2016). Indoor air quality. American
Journal of Lifestyle and Medicine, 11: 284-295.

Shaheen, A. M., Al-Toukhy, A. A. and Hajar, A. S. (2016). Effect of air pollution on leaf traits
of three species growing in the industrial zone of Jeddah, Saudi Arabia. JKAU: Metals in the
Environment and Arid Land Agricultural Science, 26(2): 33-40.

Shepherd T. and Griffiths D. W. (2006). The effects of stress on plant cuticular waxes. New
Phytologist, 171: 469–499.

Shimoji, H., Tokuda, G., Tanaka, Y., Moshiri, B. and Yamasaki, H. (2006). A simple method for
two-dimensional colour analyses of plant leaves. Journal of Plant Physiology, (53):139–147

Smith, S. E., & Read, D. J. (2008). Mycorrhizal Symbiosis (3rd ed.). Academic Press, London.
800Pp.

Stevovic S., Vesna, S. M. and Dusica C. (2010). Environmental impact on morphological and
anatomical structure of Tansy. African Journal of Biotechnology, 9(16): 2413–2421.

45
Susanto, A. D., Winardi, W., Hidayat, M. and Wirawan, A. (2021). The use of indoor plant as an
alternative strategy to improve indoor air quality in Indonesia. Reviews in Environmental
Health, 36(1): 95-99.

Susilowathi, A., Novriyanti, E., Rachmat, H. H., Rangkuti, A. B., Harahap, M. M., Ginting, I.
M., Kaban, N. S. and Iswanto, A. H. (2022). Foliar stomata characteristics of tree species in
a university green open space. Biodiversitas, 23(3): 1482-1489.

Tiwari S., Agrawal M. and Marshall F. M. (2006). Evaluation of ambient air pollution impact on
carrot plant at a sub-urban site using open top chambers. Environmental monitoring and
Assessment, 119:15–30.

Ugbabe G. E. and Dalyop J. D. (2022). Leaf microscopy, high performance liquid

chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) analyses of
Croton zambesicus Müll.-Arg. leaf (Family: Euphorbiaceae) in Nigeria. African Journal of
Biotechnology, 21(3):116-124

Usman L. A., Olawore N. O., Oladosu I., Hamid A. A. and Elaigwu S. E. (2009). Constituents of
leaf oil of Croton zambesicus Muell. Middle-East Journal of Science Resources, 4(4):242-
244.

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