ESTIMATION OF CHLOROPHYLL CONTENT OF

SELECTED PLANTS SPP FROM POLLUTED AND

NON POLLUTED AREA

BY

BISMA LIAQAT
18-F-UH-ZAGGPC-2975
LUBNA BIBI
18-F-UH-ZAGGPC-3007
AQSA HAMEED
18-F-UH-ZAGGPC-292976

BS BOTANY

DEPARTMENT OF BOTANY
THE UNIVERSITY OF HARIPUR,
KHYBER PAKHTUNKHWA,
PAKISTAN
AUGUST, 2022

i
ESTIMATION OF CHLOROPHYLL CONTENT OF
SELECTED PLANTS SPP FROM POLLUTED AND
NON POLLUTED AREA

BY

BISMA LIAQAT

LUBNA BIBI

AQSA HAMEED

A thesis submitted in partial fulfillment of the requirement for the degree of

BS BOTANY

DEPARTMENT OF BOTANY
THE UNIVERSITY OF HARIPUR,
KHYBER PAKHTUNKHWA,
PAKISTAN
AUGUST, 2022

ii
 ESTIMATION OF CHLOROPHYLL CONTENT OF SELECTED
PLANTS SPP FROM POLLUTED AND NON POLLUTED AREA

BY
BISMA LIAQAT, LUBNA BIBI, AQSA HAMEED

A thesis submitted to University of Haripur

in partial fulfillment of the requirement for the degree of
BACHOLR OF SCIENCES IN BOTANY

Approved By:

_________________ Principal
Miss Ghazala Saheed

_________________ Supervisor
Miss Sidra Khalid

_________________ HOD
Botany Department
Miss Shahana Saheed

DEPARTMENT OF BOTANY
THE UNIVERSITY OF HARIPUR,
KHYBER PAKHTUNKHWA,
PAKISTAN
AUGUST, 2022

iii
 ESTIMATION OF CHLOROPHYLL CONTENT OF SELECTED
PLANTS SPP FROM POLLUTED AND NON POLLUTED AREA

BY

BISMA LIAQAT, LUBNA BIBI, AQSA HAMEED

A thesis submitted to University of Haripur

in partial fulfillment of the requirement for the degree of
BACHOLR OF SCIENCES IN BOTANY

Approved By:

_________________ External Examiner

Prof. Dr. ABCD
Designation and address

_________________ Internal Examiner (Supervisor)

Prof. Dr. ABCD
Designation and address

DEPARTMENT OF BOTANY
UNIVERSITY OF HARIPUR, KHYBER
PAKHTUNKHWA, PAKISTAN
AUGUST, 2022

iv
 DEDICATION

To my loving and sympathetic parents, my friendly brothers and sisters whose sincere

suggestions, actions and prayers have guided me through life’s difficult stages.”

v
 ACKNOWLEDGEMENTS:

All praise to Allah Almighty, The most beneficent, the most merciful, who gave me strength
and enabled me to undertake and execute this research task. Countless salutations upon the Holy
Prophet Muhammad (SAWW), source of knowledge for enlightening with the essence of faith
in Allah and guiding the mankind, the true path of life.

We would like to extend my deepest appreciation to those people, who helped me in one way
or other in planning and executing this research work and writing up this thesis manuscript. I am
thankful to HOD Miss Shahana, Miss Sidra Khalid lecturer of botany for extending the research
facilities of the department to complete this work. I feel highly privileged in taking opportunity
to express my deep sense of gratitude to my supervisor for his scholastic guidance and valuable
suggestions throughout the study and presentation of this manuscript. I am thankful to him for
his inspiration, reassurance and counseling from time to time. He is not only a supervisor to me,
he is a good mentor ever I seen in my life.

We are very grateful to HOD ma’am Shahana, Sidra Khalid lecturer of Botany for his
skillful guidance and support throughout the study.

We are thankful to our research colleagues specially Farhana Hafeez for their moral support
and wonderful company throughout my experimental work.

Solemn gratitude to our Fathers, Mothers, Brothers and Sisters who deserve special
mention for their inseparable support and prayers. They have always been my source of strength
and love. It wouldn’t have been this bearable if I didn’t have these in my life. Thank you for your
unconditional support with my studies. Thank you for giving me a chance to prove and improve
myself through all walks of my life. Finally, I would like to thank everybody who was important
to the successful completion of thesis, as well as expressing my apology that I could not mention
personally one by one.

BISMA LIAQAT, LUBNA BIBI, AQSA HAMEED,

vi
 ABSTRACT
Pollution effects the plants growth include: mottled foliage, “burning” at leaf tips or

margins, twig dieback, stunted growth, premature leaf drop, reduced chlorophyll content and

yield or quality. Typically air pollution is caused by anthropogenic activities such as releases

from factories, cars, planes, or aerosol cans. In the present study, comparative studies have been

done, to find the effect of air pollutants generated from the exhaust of industries and automobiles

on the chlorophyll content of leaves. Significant changes in the level of chlorophyll “a”,

chlorophyll “b” and total chlorophyll “a+b” were observed in the leaves of six plant species

namely Eribotrya japonica, Morus serrate, Ficus benghalensis, Prunus armeniaca, Psidium

guajava and Eucalyptus Camaldulensis sp. collected from polluted sites (Tarbela Road Ghazi) as

compared to control site (village Jammu).In this study the non-polluted area leaves showed

higher chlorophyll content in comparison to polluted area leaves. This study clearly showed that

the vehicular activities induced air pollution problem and affects the level of chlorophyll content

in plants exposed to road side pollution.

Key words: Eribotrya japonica, Morus serrate, Ficus benghalensis, Prunus armeniaca,

Psidium guajava , Eucalyptus Camaldulensis.

vii
 TABLE OF CONTENTS

DEDICATION ......................................................................................................................... V
ACKNOWLEDGEMENTS:.................................................................................................. VI
ABSTRACT .......................................................................................................................... VII
TABLE OF CONTENTS .................................................................................................... VIII
INTRODUCTION .................................................................................................................... 1
LITERATURE REVIEW ........................................................................................................ 6
MATERIAL AND METHODS ............................................................................................. 13
SURVEY OF SAMPLING SITES ...................................................................................................... 13
COLLECTION OF SAMPLES .......................................................................................................... 13
TRAFFIC DENSITY ...................................................................................................................... 14
PHOTOSYNTHETIC PIGMENTS ..................................................................................................... 14
RESULTS ................................................................................................................................ 16
DISCUSSION.......................................................................................................................... 18
CONCLUSION: ............................................................................................................................ 22
RECOMMENDATION: .................................................................................................................. 23
REFERENCES ....................................................................................................................... 24

viii
 Chapter#1

INTRODUCTION
Environmental pollution due to rapid urbanization and industrialization has become a

major concern for living organisms almost all over the world. Anthropogenic activities emit a

number of pollutants in huge quantity into the environment (Hadayat, 2018). In urban

environment, tree play an important role in improving air quality by taking up gases and

particles.(Giri, Shrivastava, Deshmukh, & Dubey, 2013).Plants are the primary source of

organic matter and energy that support soil microbial activity and maintain many important

soil ecosystem functions.(Vezzani et al., 2018) It is important for urban planners when

selecting plants for heavily trafficked roads keeping in view their role in environmental

quality maintenance and improvement and their ability to tolerate stress.(Khalid, Masood,

Noman, Aqeel, & Qasim, 2019). Leaves of the plants in particular provide a large surface area

for the deposition of atmospheric pollutants and subsequently act as a sink.(Bharti, Trivedi, &

Kumar, 2018).

Environmental pollution has been a matter of concern for many years. The Mellon

Institute of Pittsburgh, PA, USA, sponsored the first broad scientific study of smoke

abatement, which resulted in legislation designed to decrease the effects of smoke. It is now

well known that environmental contamination impacts on health; the World Health

Organization estimates that every year, 2.4 million people die from causes associated with air

pollution.(SIERRA‐VARGAS & Teran, 2012) . Air pollution can be distinct as the human

interference into the atmosphere this may be of chemicals, particulate matter or biological

materials that cause harm or discomfort to human health, or other living organism or damage

the environment.(Leghari, Zaidi, Ahmed, & Nazim, 2011).

1
 The effect of air pollution on plants can be seen in a variety of ways from visible

markings on the foliage to reduced growth and yield, or even to premature death of the plant.

The level of injury depends on the concentration of the particular pollutant and other factors,

such as the length of exposure, the plant species and its stage of development and the

preconditioning of the plant, which make it either susceptible or resistant to injury. Some

plants are more tolerant to air pollution than others. The London plane tree, for example,

sheds its bark and so stops air pollution reaching the inner trunk of the tree. However, the

short, stiff hairs shed by the young leaves and the dispersing seeds of the London plane are an

irritant and can exacerbate breathing difficulties for people with asthma.(Tiwary & Williams,

2018)

Chlorophyll pigments play an important role in the plant productivity system.

chlorophyll is the chief pigment present in the chloroplast of leaves which adjusts the plant

productivity. It is the main pigment that gives the plants their characteristic green color. Plant

productivity and photosynthetic rate depend on the quantity of chlorophyll per unit area of the

chloroplast. Percentage of chlorophyll content in the leaf tissues is influenced by the

obtainability of nutrients and the environmental stress such as crude oil pollution of soil and

drought.(Baruah, Saikia, Baruah, & Deka, 2014) .

Vehicular exhaust is one of the most serious issues and largest contributor of air

pollution these days. The rapidly growing number of vehicles coupled with rising

urbanization, industrialization and power consumption in Pakistan has increased air pollution

at an alarming rate. The major pollutants produced from the automobiles are various

hydrocarbons (HCs), Sulphur dioxide (SO2), carbon dioxide (CO2), carbon monoxide (CO),

oxides of nitrogen (NOx ), particulate matter (PM) and a number of toxic heavy metals.

2
Emission of these pollutants has been shown to have a variety of adverse effects on plants,

public health and natural environment.(Khalid, Noman, Sanaullah, Akram, & Aqeel, 2018).

Ficus benghalensis is a perennial woody species that can grow at least above breast

height or more and having a single woody trunk.(Basit, Khan, Sulaiman, & Shah, 2019). is a

fig species that is native to Asia, and widely distributed through the stretches of India.

Member of this family shows antibacterial activity against Bacillus cereus and Pseudomonas

aureus which pretend the resistance against the multi drugs.(Afzal, Ali, & Malik, 2020)

Eucalyptus Camaldulensis is an evergreen tree native to southern Victoria (Australia),

Tasmania and the Bass Strait Islands.(Deus, Silva, Marchante, Marchante, & Félix, 2018) E.

Camaldulensis belongs to the family of Myrtaceae; an evergreen broadleaf tree, with a

straight trunk, It is known as “the blue gum” or Tasmanian blue gum Eucalyptus

Camaldulensis is the main source of the Eucalyptus leaf oil used globally . Its leaves are

alternate, with yellowish petioles, precisely lanceolate, 10–30 cm long, 2.5–5 cm wide, shiny

dark green on both surfaces, and are a rich source of essential oil.(Shala & Gururani, 2021)

Guava (Psidium guajava) is a high-value tropical fruit suitable for niche markets, but

accession-level fruit quality data are lacking. Guava is native to tropical and warm subtropical

areas from Mexico to Peru and was expanded throughout the cool subtropical to tropical

regions of the world during the past 400 years .(Moon et al., 2018). Guava is known for its

hypoglycemic, antivirus, antibacterial, anti-inflammatory, antioxidant, and antitumor

properties. (Jiang, Lu, Qin, Jiang, & Wang, 2020)

Prunus genus is one amongst the 85 genera of Rosaceae family and is explored widely

by the scientists regarding oil characterization and medicinal importance of the fruits.

3
P.armeniaca.L is well known prunus specie famous due to its delicious, juicy fruit.

(P.armeniaca.L) Apricot a delicious table fruit, originated from China and later spread to

Europe . especially in Mediterranean countries such as Italy, France and Spain(Zhao et al.,

2022). the fruit used as the treatment of nervous system disorder, fight against anemia, relieve

constipation, prevent cancer whereas, the leaves in crude forms are the remedy for cough,

asthma, bronchitis and fever(Akhtar, Tariq, & Sultana).

Morus serreta belonging to the order Rosales, family Moraceae, and genus Morus is

distributed in a wide range of areas worldwide(Zeng et al., 2015) Mulberry (Morus) is

believed to have originated in the northern hemisphere, particularly in the Himalayan

foothills, and spread to the tropics of southern hemisphere. Leaves of wild mulberry species

such as M. laevigata, M. serrata, and M. tiliaefolia are considered too rough, leathery and

thick to be used for silkworm rearing(Vijayan et al., 2011) This plant also plays significant

ecological roles in the prevention and control of sand erosion, and in stony desertification and

saline-alkali land treatments (Zeng et al., 2015).

Eriobotrya japonica (Rosaceae) is native to southeastern China and evergreen large

shrub or small tree, with a rounded crown, short trunk and woolly new twigs. E.japonica

syrup is used in Chinese medicine for soothing the throat like a cough drop. The E.japonica is

having an anti-inflammatory, hypoglycemic, antioxidant, antitumor, antiviral, cytotoxic,

antimutagenic, chronic bronchitis, nephropathy, NF kappa B inhibitor, hypolipidemic activity.

Seeds and young leaves of the plant are slightly toxic, containing small amounts of

cyanogenic glycosides (including amygdalin) which release cyanide when digested, though

the low concentration and bitter taste normally prevents enough being eaten to cause

harm(Baljinder, Seena, Dharmendra, Vikas, & Bansal, 2010).

4
1.1: Objectives:

1. Sample collection from polluted and non-polluted area.

2. Estimation of effect of pollution on the chlorophyll content of collected sample.

5
 Chapter#2

LITERATURE REVIEW
Hadayat reported the effect of air pollution on five wild plant species Calotropis

procera, Cenchrus ciliaris, Cynodon dactylon, Nerium oleander and Parthenium

hysterophorus generally growing near the two roads [a segment of Motorway (M-2) from

Pindi Bhattian to Kala Shah Kaku and a segment of Grand Trunk road (G.T. road) from

Lahore to Gujranwala] in Punjab, Pakistan. The plant leaves and soil samples were collected

from five different sites along each road. The control samples of leaves and soil were

collected at a distance of 50 meter away from roadside. He also examined certain

physiological parameters such as metal toxicity, gas exchange characteristic and chlorophyll

content i.e chlorophyll a, chlorophyll b, total chlorophyll and carotenoids.The higher contents

of all the metals were noted along G.T. road as compared to M-2. However, Kala Shah Kaku

site along M-2 and Muridke site along G.T. road appeared as the more polluted sites. The

contents of photosynthetic pigments were significantly lower whereas total antioxidant

activity were significantly higher in roadside plant species under metal stress..(Hadayat,

2018).

Khalid et al., 2018) reported the physiological, biochemical and defence responses of

Calotropis procera in response to vehicle exhaust pollution. They compared the chlorophyll

content i.e. chlorophyll a, chlorophyll b, total chlorophyll and carotenoid contents,

transpiration rate, stomatal conductance, photosynthetic rate, and total antioxidant activity of

different sample along two major road side i.e. Faisalabad to Sargodha road (FSR) and Pindi

Bhattian to Lillah motorway (M-2). M-2 chlorophyll a: 0.566 chlorophyll b: −0.556 Total

chlorophyll: −0.022 carotenoids 0.426 and FSR chlorophyll a: 0.965 chlorophyll b: 0.898
6
Total chlorophyll: 0.978 carotenoids 0.458. Their results showed they found their

considerable deposition in all the samples along roads. Inhibitory effects of roadside

pollutants were noted for photosynthetic pigments, stomatal conductance, transpiration rate,

photosynthetic rate. The stimulation of antioxidant enzymes activity revealed stress and

mitigation of reactive oxygen species. This study clearly signifies that C. procera has great

potential to endure the stress caused by roadside pollutants.(Khalid et al., 2018)

Khalid et al., 2019 Investigate the various physiological and biochemical responses of

two good biomonitor plant species i.e. Datura alba and Ricinus communis were studied along

two roads in the Punjab, Pakistan. Chlorophyll a, b, total chlorophylls, carotenoids, total

antioxidant activity, stomatal conductance, photosynthetic rate, transpiration rate, and water

use efficiency of D. alba and R. communis were examined at different sites along both roads.

Photosynthetic rate of both plant species to be affected. Reduced transpiration rate and

stomatal conductance and water use efficiency were also noted. We found a tremendous

increase in total antioxidant activity in both plant species .Hence, our results suggest that R.

communis is more resistant to urban roadside air pollution compared to D. alba.(Khalid et al.,

2019)

Pathak, Tomar, & Mahajan, 2015 reported the effects of air pollution on roadside

plants due to vehicular emission at Indore city .The study was conducted in Industrial area

pologround which is more polluted and less polluted area college campus.The two plant

species selected which are dominated on road side Bouganvillia spectabili and Delbergia

sisoo selected for study. The parameters like chlorophyll a, chlorophyll b, pH, Electrical

conductivity and optical properties of leaf wash from sample collected from less polluted and

7
more polluted area of road side. Depletion of chlorophyll, change in pH recorded from

polluted area. (Pathak, Tomar, & Mahajan, 2015)

Rachmadiarti, Purnomo, Azizah, & Fascavitri, 2019 studied that heavy metals cause

pollution in the environment, including plants. In Indonesia, Syzigium oleina and Wedelia

trilobata are grown on the main roads where vehicles frequently pass through; these plants are

exposed to heavy metals, such as lead. The levels of heavy metals in plant leaves were

measured by using an atomic absorption spectrophotometer, and chlorophyll content by using

a spectrophotometer. Data were analysed by two-way ANOVA. The results showed that the

location of plant species affected the content of Pb metal and chlorophyll in the leaves, but the

interaction of the location and the type of plant only affects the chlorophyll content of leaves

and not the Pb metal content.(Rachmadiarti, Purnomo, Azizah, & Fascavitri, 2019)

Houri et al., 2020 reported that vehicle pollution caused contamination of soil, air and

plants. They determine heavy metals contamination levels evaluated by a rare plant species

Urginea maritima. The analysis of heavy metals is completed by a measurement of the

photosynthetic activity of the plant. As for photosynthetic pigments, the study showed an

adverse effect of heavy metals on ordinary photosynthetic activities of the plants. Plant

species loss Chlorophyll a and decline in activities the first year but then amplification, in

contrast of pheophytin which concentrations displayed a sharp rise especially in the most

closely site to the road. (Houri et al., 2020)

Amulya, Hemanth Kumar, & Jagannath, 2015 reported ``the air pollution effects on

micromorphological and biochemical parameters of Tabernaemontana divaricata

8
(Gentianales: Apocynaceae) and Hamelia patensi (Gentianales: Rubiaceae). They compare the

number of stomata, stomata clogged, stomata breadth number of subsidiary cells, trichome,

chlorophyll a, chlorophyll b, and total chlorophyll content from polluted and control sites.In

polluted area the number of stomata and clogged stomata were found to be higher than

control site. The stomatal breadth and pore length were found to be decreased in polluted area

in both the plants when compared to control. The number of subsidiary cells, trichome length

values was found to be less then control plants. The chlorophyll a, chlorophyll b, and total

chlorophyll content were found to be maximum in control samples when compare to polluted

samples. (Amulya, Hemanth Kumar, & Jagannath, 2015)

Khalid, etal reported that Pollution caused by vehicular exhaust emissions

detrimentally affect plants and other living beings. They investigate the effects of vehicular

exhaust pollutants on Parthenium hysterophorus at various sites along two major roads [Pindi

Bhattian to Lillah (M-2) and Faisalabad to Sargodha (FSR)] in the Punjab, Pakistan.

Chlorophyll contents, photosynthetic rate, transpiration rate, stomatal conductance, sub

stomatal CO2 concentration, and total antioxidant activity of P. hysterophorus were measured.

The results showed that significant reductions in chlorophyll a, chlorophyll b, total

chlorophyll and carotenoid contents of P. hysterophorus .the reduction in stomatal

conductance was also recorded which resulted in lowered photosynthetic and transpiration

rates. The limited photosynthesis resulted in increased levels of sub stomatal CO2

concentration. The elevated levels of total antioxidant activity were noted with the deleterious

effects of vehicular air pollutants. The significant correlations between various attributes of P.

hysterophorus with traffic density signifies the stress caused by vehicular emissions.(Khalid,

Hussain, Hameed, & Ahmad, 2017)

9
 Laghari, Zaidi, & Razaq appraised the effect of solid waste burning pollution on

carbohydrate, stomata and chlorophyll contents of seven different plant species. Leaf samples

of Artemisia maritima L., Fraxinus excelsior L., Amaranthus viridis L., Cynodon dactylon L.,

Chenopodium album L., Robinia pseudoacacia L., and Sophora mollis (Royle) Baker, were

collected from residential colony. Results exposed that the carbohydrate, chlorophyll ‘a and b’

and total chlorophyll contents in the leaves of selected plant species were found to be

significantly and source of pollution increased (500m & 1000m) these contents increased

consequently. The percentage of entirely and moderately clogged stomata was found higher

near the pollution source (1m distance). As respect to carbohydrate and chlorophyll contents,

the Artemisia maritima L., were found most sensitive to air pollution in all four directions at

1m distances as compared to the other species. While plant species, Cynodon dactylon L.

showed more resistant to air pollution effect as regard to carbohydrate contents and high

percentage of open stomata at 1m distances with respect to other species.(Laghari, Zaidi, &

Razaq, 2015).

Leghari, Zaidi, Ahmed, & Nazim, 2011 focuses on the determination of Air Pollution

Tolerance Indices (APTI) from twelve plant species growing along polluted site (the road side

of Quetta city) and non-polluted site (Botanical garden University of Baluchistan, Quetta).

They determined APTI by synthesizing the four different physiological and biochemical

parameters i.e. leaf relative water content (RWC), ascorbic acid content (AA), total leaf

chlorophyll (TCh) and pH of leaf extract..The maximum reduction (40.39%) of total

chlorophyll content was recorded in the leaves of Prunus armeniaca and minimum reduction

(4.48 %) was in the leaves of Morus alba of polluted site. There was maximum increment of

relative water content (38.87%) in the leaves of Fraxinus xanthoxyloids and minimum

10
increase (4.19%) was found in the leaves of Vitus vinifera of the polluted site. Ascorbic acid

contents showed maximum value (39.13 %) in the leaves of Ficus carica and minimum

amount (18.75 %) was recorded in the leaves of Vitis vinifera of the samples collected from

polluted site. The maximum pH (6.85 ± 1.00) was observed in the leaves of Vitis vinifera and

minimum value (5.44± 0.86) was observed in the leaves of Morus nigra of polluted site. The

results showed that significantly high level of pollution at the pollution sites, in all the

parameters studied mainly due to automobile exhaust.(Leghari, Zaidi, Ahmed, & Nazim,

2011)

Bharti, Trivedi, & Kumar, 2018 expresses that sensitive and tolerant plant species can

be identified by evaluating their air pollution tolerance index (APTI). They evaluating the pH,

ascorbic acid, total chlorophyll, and relative water content (RWC) of plant leaves. Their study

was designed to estimate the air pollution tolerance index (APTI) of 25 plant species growing

at Talkatora Industrial Area, Luck now Uttar Pradesh, India. The biochemical properties of

plant species ranged from; ascorbic acid: 0.6–19.6 mg/g, RWC 41.34%–98.62%, pH 4.5–8.2

and chlorophyll content 0.59–1.49 mg/g. Moringa oleifera leaves were found to have the

highest dust capturing potential (5.7 mg/cm2), whereas, the lowest was noticed in Acacia

nilotica (0.10 mg/cm2). Pearson correlation of biochemical parameters revealed that ascorbic

acid showed significant correlation (R2 =0.897) with APTI. The species having< 11 APTI

values may be used as a bio-indicator of air quality, while those having APTI ≥17 can be used

for green belt designing.(Bharti, Trivedi, & Kumar, 2018)

Muhammad, Shakeel, Khan, Hasnain, & Cheema, 2016 reported the Air Pollution

Tolerance Index (APTI) of commonly occurring tree of Mall road and Jail road of Lahore,

Pakistan. They were selected four species i.e. Alstonia scholaris (L.) R.Br., Callistemon

11
citrinus (Curtis) Skeels, Ficus religiosa L. and Morus alba L. The experiment was carried out

by taking four aspects viz. pH, ascorbic acid, total chlorophyll and leaf water content (%) of

leaf samples of these tree species.They collected plants from Mall road, Morus alba had the

highest value of chlorophyll content i.e. 0.99 mg/g and Alstonia scholaris had the lowest

chlorophyll value i.e. 0.31 mg/g. Ficus religiosa and Callistemon citrinus contains 0.67 mg/g

and 0.50 mg/g of chlorophyll content respectively. Among Jail road plants, A. scholaris

showed the highest chlorophyll content of about 0.44 mg/g followed by M. alba with

chlorophyll value of 0.19 mg/g, F. religiosa 0.18 mg/g and C. citrinus with the lowest value

i.e. 0.04 mg/g. Total ascorbic acid content was highest in C. citrinus and lowest in F. religiosa

among Mall road plants. It was found that the Ficus religiosa was the most tolerant having

APTI value 9.09, while Callistemon citrinus showed lowest value of 7.86 which indicated that

it was most sensitive to air pollution.(Muhammad, Shakeel, Khan, Hasnain, & Cheema, 2016)

12
 Chapter# 3

MATERIAL AND METHODS

Survey of sampling sites
Sections of (tarbela road) from nowshehra to tarbela ghazi in KPK, Pakistan were

selected for the phytomonitoring of metal pollution from vehicular sources. For data

collection survey was conducted during all the four seasons of the year i.e. winter, spring,

summer and autumn at regular intervals. The meteorological data of the study area has been

given hereunder (Table 3.1)

Table 3.1. Meteorological data

Winter Spring Summer Autumn

Average 15.4 20.8 32.5 27.9

temperature ( ˚C)

Average 20.2 27.9 90.3 62.2

precipitation (mm)

Collection of samples

i) Plants

The following wild plant species commonly growing along both the roads were chosen.

13
 Scientific name Common name

Eucalyptus Camaldulensis River Red Gum, Red Gum

Psidium guajava Guava

Prunus armeniaca Apricot

Ficus benghalensis Banyan tree

Morus serreta Himalayan mulberry

Eribotrya japonica Loquat

We collected these six plants that growing nearest to the roadside. Plant samples were

collected at a distance of 50 m away from the roadside. All the sampled leaves were packed in

labelled polythene zipper bags, kept in the cooling container and brought into the laboratory for

chemical analysis.

Traffic density
Traffic density (number of vehicles/day) was noted at all the selected sites adjacent to

road (Tarbela road) on specific days (weekends and midweeks) for 2 hours (morning, evening).

Photosynthetic pigments
We can determine the contents of chlorophyll a, chlorophyll b and total chlorophyll. The

contents of pigments were measured in 1mg leaf fresh wt. Fresh plant leaves (0.5 g) were

crushed using pestle and mortar. The ground leaf samples were extracted with 10 mL of acetone

solution (80 %). Centrifugation of extract was carried out at 10,000 rpm for 5 min. at 4 ◦C. The

optical density (OD) of separated supernatant was measured at 663 and 645nm using

spectrophotometer. The contents of photosynthetic pigments were computed as follows:

14
Cchl a = 12.7A663 -2.69A645

Cchl b= 22.9A645- 4.68A663

15
 CHAPTER #04

RESULTS
Plants were collected from polluted (Tarbela road)and non polluted area(villages)

and check the chlorophyll content of these plants. Six plants were collected from polluted

and non polluted area and these six plants are Eucalyptus Camaldulensis, Psidium guajava,

Prunus armeniaca, Ficus benghalensis , Morus serreta , Eribotrya japonica and their

chlorophyll a measured at 645wavelength, chlorophyll b measured at 663wavelength and

total chlorophyll content was measured. The chlorophyll content of these six plants was

shown in graphs.

The minimum Chlorophyll content were measured in Prunus armeniaca from

polluted site chl a 2.3, chl b 1.6 and total chl 3.9 whereas maximum in non-polluted area Chl

a 6.3, chl b5.6 and total chl 12. The maximum chlorophyll content of Gavua (Psidium

guajava) from non-polluted area is chl a 4.3, chl b 3.6 and total chl 8 while minimum

chlorophyll content from polluted area is Chl a 1.6, chl b2.3 and total chl 4. The maximum

chlorophyll content of Morus serrata from non-polluted area chl a 8 chl b 8 and total chl 16

and minimum chlorophyll content from polluted area is Chl a 2.3,chl b 2 and total chl 4.3.

The maximum chlorophyll content of Eriobotrya japonica from non-polluted area is chl a

7.6, chl b 6.6and total chl 14 .3 whereas minimum in polluted area is Chl a 3,chl b 4 and

total chl 7.The maximum chlorophyll content of Ficus benghalensis from non-polluted area:

chl a 4.6,chl b 4.3and total chl 9 and minimum from polluted area: Chl a 2,chl b 1.4 and total

chl 3.4. The maximum chlorophyll content of Eucalyptus camaldualensis from non-polluted

area chl a 4.6, chl b 4.3and total chl 9 while minimum in polluted area: Chl a 1,chl b 1 and

total chl 2.
16
 8
7
6
5
4
3 chl a
2 chl b
1
0 total chl

Polluted Results Graph

18
16
14
12
10
8
6
4 chl a
2
0 chl b
total chl

Non-Polluted Results Graph

17
 Chapter#5

DISCUSSION
Pollution is a serious global environmental problem as it badly affects plant growth.

Gouda, Kerry et al. 2018)These gaseous pollutants, nitrogen oxides (NOx), carbon monoxide

(CO), carbon dioxide (CO2), sulfur dioxide (SO2) is combinations of trace elements cause

greater or synergistic effects to plants growth on the leaves affected their physiological

behavior. Automobiles are expected to have main effects on fruiting, flower development,

leaf senescence and leaf surface wax characteristics, seed germination, seedling growth,

physiological characteristics and plant development. (Iqbal, Shafiq et al. 2015)

Automobile-based pollutants deposit on leaves, blocking the stomata and ultimately

affecting transpiration. These depositions obstruct CO2 absorption and eventually decline

photosynthesis and affect the growth of plants and their productivity. Chlorophyll a, b, and

total chlorophyll contents have also been reported to be affected by air pollutants. Leaf

surfaces are also affected due to different trace elements settle down on the leaf surfaces and

because of gaseous discharges from road traffic. (Muthu, Gopal et al. 2021)

In the present study plants were collected from two different sites i.e. from polluted

and non-polluted sites and their chlorophyll content was measured and compared. The plants

were Eucalyptus Camaldulensis, Psidium guajava, prunus armeniaca, Ficus benghalensis,

Morus serreta, Eribotrya japonica.

The plants were collected from polluted area (tarbela road) are Eucalyptus

Camaldulensis , Psidium guajava, Prunus armeniaca ,Ficus benghalensis , Morus serreta ,

Eribotrya japonica while plants were collected from non-polluted area are Eucalyptus

18
Camaldulensis from Qazipur, , Psidium guagava and Prunus grameniaca from khalo ,Ficus

benghalensis from Jammu , Morus serreta and Eribotrya japonica from Hasanpur.

Giri, Shrivastava etal reported that Chl a, chl b and total chlorophyll content were

measured from these plants. Overall results showed that the chlorophyll content of the plants

collected from polluted area was lowered then the plants collected from non-polluted area

plants growing in industrial area showed decline in the chlorophyll content as compared to

the plants growing in fresh environment. (Giri, Shrivastava et al. 2013)

The maximum chlorophyll content from non-polluted sites was recorded in Morus

serreta, while the minimum chlorophyll content from polluted site was recorded in ficus

benghalensis.

The chlorophyll content of eucalyptus from polluted area (tarbela road) chl a1, chl b

1 and non-polluted area (qazipur) Chl a: 4.6, chl b: 4.3. similar findings have been reported

by previous scientist Iqbal etal measured the concentration of chlorophyll “a” “total

chlorophyll “in the leaves of Eucalyptus sp. at polluted site of Airport was recorded (0.258

mg/g) low as compared to control (0.271 mg/g).Low level of chlorophyll “a” was recorded in

leaf sample of Eucalyptus sp. (0.271 mg/g). High level of chlorophyll” (0.243 mg/g) was

found in leaf samples of Eucalyptus sp collected. The total chlorophyll “a+b” was recorded

in leaf sample of Eucalyptus sp. collected as compared to control 0.500 mg/g) site. (Iqbal,

Shafiq et al. 2015)

In present study the content of chlorophyll a, b, and total chlorophyll were

significantly lower in plant growing in all the sites along the road sides as compared to the

plants growing in non-polluted sites. (Rai and Panda 2014)

19
 In the present study plants were collected from two different sites i.e. from polluted

and non-polluted sites and their chlorophyll content was measured the chlorophyll content of

Ficus benghalensis from polluted area (tarbela road )chl a2,chl b 1.4 and Non polluted area

(jamu)Chl a4.6,chl b4.3. Similar findings have been reported by previous scientist measured

the concentration of Total chlorophyll = a+b Young leaf of Ficus benghalensis L. at polluted

site of college area was recorded (8.2709mg/g) low as compared to the Total

chlorophyll=a+b Adult leaf control (7.092mg/g. (Mulay and Kokate 2019)

In the present study plants were collected from two different sites i.e. from polluted

and non-polluted sites and their chlorophyll content was measured the chlorophyll content of

Gavua (Psidium guagava) from polluted area (tarbela road )chl a 1.6,chl b 2.3and Non

polluted area (khalo)Chla 4.3,chl b3.6. similar findings have been reported by previous

scientist measured the chlorophyll content in guava plants the chlorophyll content in young

leaves was Chl. a =6.27 mg pigment/m3 and Chl b =5.40 mg pigment/m3 while in adult

leaves it was Chl. a =3.91mg pigment/m3 and Chl. b=5.73 mg pigment/m3. Total Chl.

Content in guava young leaves: 12.54 while Total Chl. Content in guava adult leaves: 9.6.

20
1. Collection of plants 2. Cut leave with the help of
scissor

scis

3. Used pestle for extract 4. Extract Prepared for non-polluted

preparation leaves

5. Extract Prepared for polluted 6. Perform Experiment

leaves 21 for measuring chlorophyll
Conclusion:
Most air pollution is created by people, taking the form of emissions from factories, cars,

planes, or aerosol cans and this study has demonstrated a variety of responses showed by E.

Camaldulensis , P. guagava, P.armeniaca ,F .benghalensis , M. serreta , E. japonica to vehicular

exhaust pollutants at various sites along roadsides. These plants showed reduction in

photosynthetic pigments, photosynthetic rate, transpiration rate and stomatal conductance,

reduced chlorophyll content and yield or quality activities at various sites along roadsides.

Important changes in the level of chlorophyll “a”, chlorophyll “b” and total chlorophyll “a+b”

were establish in the leaves of six tree species (Eribotrya japonica, Morus serrate, Ficus

benghalensis, Prunus armeniaca , Psidium guagava and Eucalyptus Camaldulensis sp.)

collected from polluted sites (Tarbela Road Ghazi) as compared to control site (village

Jammu).These plants have grown in non-polluted sites and can perform roles i.e. photosynthetic

rate, transpiration rate, stomatal conductance, high chlorophyll content. The physiological

attributes measured in this study clearly indicate that they could be used as biomarker basis for

monitoring and prediction of early effects of air born vehicular exhaust pollution.

22
 Recommendation:

1- We should minimize the use of vehicles so that the plants are not damaged and our

environment is also cleaned.

2- We should use the vehicles as a minimum so that the chlorophyll of the plants does not

decrease and they continue to the process of photosynthesis.

3- We should use machinery in our vehicles that emits as little smoke as possible.

4- If we want green plants, we need to reduce the use of cars busses trucks etc.

23
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