RESIDUAL EFFECTS OF EUCALYPTUS

CAMULDULENSIS LEAF LITTER ON ZEA MAYS AND

PENNISITUM TYPHODIUM
A thesis submitted in partial fulfillment of requirement for the degree
of
BS Botany
By
ABDUL HAKEEM JOKHIO

2K10/BOT/1

FAHEEM BUGHIO

2K10/BOT/94

NAVEED ALI JHATIAL

2K10/BOT/57

AZIZULLAH HAJANO

2K10/BOT/91

MUHAMMAD AHSAN KHAN

2K10/BOT/103

Supervisor
Saeed Akhter Abro
DECEMBER 2013

INSTITUTE OF PLANT SCIENCES, UNIVERSITY OF SINDH

i

IN The NAME OF

The supremely merciful

The most kind
Whos help We solicit

ii

RESIDUAL EFFECTS OF EUCALYPTUS

CAMULDULENSIS LEAF LITTER ON ZEA MAYS
AND PENNISITUM TYPHODIUM
A thesis submitted for the fulfillment of requirements for the
degree of BS Botany

By
ABDUL HAKEEM JOKHIO

2K10/BOT/1

FAHEEM BUGHIO

2K10/BOT/94

NAVEED ALI JHATIAL

2K10/BOT/57

AZIZULLAH HAJANO

2K10/BOT/91

MUHAMMAD AHSAN KHAN

2K10/BOT/103

Supervisor
Saeed Akhter Abro

INSTITUTE OF PLANT SCIENCES, UNIVERSITY OF SINDH, JAMSHORO

iii

CERTIFICATE
This is to certify that the research work presented in this thesis entitled RESIDUAL
EFFECTS OF EUCALYPTUS CAMULDULENSIS LEAF LITTER ON ZEA
MAYS AND PENNISITUM TYPHODIUM has been carried out by ABDUL
HAKEEM JOKHIO (2K10/BOT/1), FAHEEM
BUGHIO (2K10/BOT/94),
NAVEED ALI JHATIAL (2K/10/BOT/57) AZIZULLAH
HAJANO
(2K10/BOT/91), MUHAMMAD AHSAN PATHAN (2K10/BOT/103) under my
supervision is of original nature. In my opinion it is satisfactory in scope and quality as
thesis for the degree of BS in Botany.

Thesis Supervisor:
Saeed Akhter Abro
Assistant Professor

................................

Institute of plant sciences

University of Sindh,
Jamshoro, Sindh,
Pakistan.

Thesis Review Committee

1. ---------------------------------------------------------------------------

2. ------------------------------------------------------------------------------------------

--------------------------------------

----------------------------------------------

Prof. Dr. Sher Mohammad Mangrio

Director
Institute of plant sciences
University of Sindh, Jamshoro,

iv

Dedicated to my parents
For their endless love, support and encouragement

Table of Contents
Contents

Page #

Acknowledgment.................................................................................................

VIII

Abstract...............................................................................................................

Abbreviations and symbols..............................................................................

XII

Chapter# 01 INTRODUCTION & LITERATURE REVIEW

1.1.

Introduction...............................................................

01

1.2

Allelopathy..............................................................

02

1.3.

Allelochemicals

04

1.4

Eucalyptus as Allelopathic plant..

04

1.5

Effects of allelopathy............................

05

1.6

Examples of Allelopathy..

06

1.7

Residual effects of Allelochemicals.

07

1.8

Objectives

08

Chapter#2. MATERIAL AND METHOD

2.1

Introduction..

09

2.2.

The Materials....

09

2.2.1.

The Bioassays Plants ...

09

2.2.2.

Soil and leaf litter formulations

09

2.2.3.

Eucalyptus leaf litter and its preparation..

09

The Methods.

10

2.3.1.

Experimental Design and treatment......

10

2.3.2.

Sowing of Seeds ...

10

2.3.3.

Data Collection and Analysis

11

Parameters used

11

Plant parameters

11

Germination .

11

Relative germination ratio

11

Percentage mortality rate

11

2.3.

vi

Fresh Weight ..

12

Dry Weight ..

12

Shoot length .

12

Relative biomass ......................

12

Chlorophyll content

12

Soil Parameters

13

pH

13

Electric
conductivity of soil

13

TDS...

13

Salinity .

14

Na+ and K+

14

Organic matter..

14

Soil water repellency ...

14

Chapter#3: Results and discussion

Zea mays..

16

3.1.1

Germination

16

3.1.2.

Mortality Rate...

16

3.1.3.

Relative germination ratio

16

3.1.4.

Relative elongation ratio of shoot.

16

3.1.5.

Shoot length..

17

3.1.6.

Fresh weight

17

3.1.7.

Dry Weight ..

17

3.1.8.

Relative biomass ratio..

17

3.1.9.

Chlorophyll content.

17

Soil properties...

20

3.2.1.

pH of soil .

20

3.2.2.

Electric conductivity of soil

20

3.2.3.

TDS..

22

3.2.4.

Salinity

22

3.1.

3.2.

vii

3.2.5.

Soil organic matter...

22

3.2.6.

Na+ and K+

22

3.2.7.

Water repellency...

23

Pennisitum typhoidum

23

3.3.
3.3.1.

Germination

23

3.3.2.

Mortality Rate..

23

3.3.3.

Relative germination ratio

23

3.3.4.

Relative elongation ratio of shoot.

24

3.3.5.

Shoot length.

24

3.3.6.

Fresh weight ...

24

3.3.7.

Dry Weight ..

24

3.3.8.

Relative biomass ratio..

25

3.3.9.

Chlorophyll content .

25

Soil parameters

26

3.4.1.

pH of soil .

26

3.4.2.

Electro conductivity of soil .

26

3.4.3.

TDS...

27

3.4.4.

Salinity .

27

3.4.5.

Soil organic matter...

27

3.4.6.

Na+ and K+

28

3.4.7.

Water repellency...

28

Conclusion........................................................

31

References

32

3.4.

3.5

viii

ACKNOWLEDGEMENT

First of all we would like to thanks of Almighty Allah who gave us brain covered
by hard skull by which we contemplate about his nature. This project was also minute
work that was completed by our narrow mind. Hazrat Muhammad (PBUH) whose
imminent teachings enlighten our mind to complete this hard work.

We would like to express our sincere gratitude and deep respect to our
supervisor, Sir Saeed Akhter Abro, Institute Of Plant Sciences, University of Sindh,
Jamshoro, whose expertise, understanding, and endurance, added considerably to our
graduate experience. We are highly grateful for his sincere guidance, keen interest,
valuable suggestions and moral support during the study period, and also for critically
reading various chapters of this thesis.

After that we would especially pay thanks to our parents and family members, sympathy
and courage they given us during the difficulties we faced for the completion of this
thesis, as well as for the completion of higher studies.

We would like to thanks lecturer Sir. Farooq Ali Bughio Institute of Plant science,
University of Sindh, Jamshoro, for allowing us to undertake the BS Final work, and
provide us the moral support, chemicals on laboratory on timely and encouragement. We
are also thankful to Muhammad Luqman Panhwar and Deedar Hussain Kaloi who
practically participated for fulfillment of project work.

There are many helping hands; we wish to express our deepest regards to all who
have helped us in this study.
viii

Additionally we offer our regards and blessings to all of those who supported us in
any respect during the completion of this research work.

Last but not the least we want to tender our sincere gratitude, respect, reverence to
our beloved Parents, brothers and sisters without their constant encouragement, support
and pray we would not have been successful in our aim.

ix

Abstract

The allelopathic effects of eucalyptus are widely reported and have effects on the
germination and growth of neighboring plants. To investigate the effects of leaf litter
residues, Zea mays and Pennisetum typhoideum were selected. The experiment was
conducted in green house, institute of plant sciences, university of Sindh, jamshoro. The
effect of two year old leaf litter mixed with soil in 10, 20, and 30% were observed. The
germination %, mortality rate, RGR, RERs, shoot length (cm), fresh and dry weight (g),
RBR, chlorophyll a, b, carotenoids (mg/g f.wt.) was calculated. In soil properties, pH, EC
(s/cm), TDS, salinity (ppt), OM (%), Na (ppt), K (ppt), soil water repellency was
analyzed. The results suggest that leaf litter slightly decrease the germination %, RGR of
maize and has stimulatory effect on germination %, RGR of Pennisetum typhoideum.
The eucalyptus leaf litter has stimulatory effect on RERs, shoot length, fresh weight, dry
weight, relative biomass ratio and chlorophyll a, b and amount of carotenoids of both
plants. The soil pH, K was almost same values in treatments, but second soil have more
values than initial soil. The soil ECe (s/cm), TDS, Salinity (ppt), OM (%), and Na (ppt)
was found in higher concentration in eucalyptus treatments and as concentration of leaf
litter increased, concentration of these values also increased in Zea mays soil. In
Pennisetum typhoideum soil pH was found 6.7 to 7.1 in initial soil and second soil pH
was almost same in treatments. The soil ECe (s/cm), TDS, salinity (ppt), OM (%), Na
(ppt), K (ppt), was found in high concentration in eucalyptus treatments than control. The
soil EC (ppt), TDS, salinity and K (ppt), was found in high concentration in initial soil
and OM (%), Na (ppt), was found in high concentration in second soil. In both plant soils,

treatment applied with eucalyptus leaf litter was found strongly to severely water
repellent as compared to control, which soil was non water repellent.

xi

Abbreviations and Symbols

Description

Abbreviation
pH

Potential or power of Hydrogen

ECe

Electro conductivity of extract

TDC

Total dissolved carbon

OM

Organic Matter

OC

Organic carbon

Na

Natrium / Sodium

Potassium

Treatment

kg

Kilogram

mg

Milligram

ml

Milliliter

Gn

Number of seeds germinated after 14 days

GN

The total number of seeds sown

GRt

Germination ratio of plants under treatment

GRc

Germination ratio of plants under control

Centigrade

RBR

Relative biomass ratio.

MR

Mortality rate

RGR

Relative germination Ratio

RERs

Relative elongation ratio of shoot

MBt

Mean biomass of plant under treatment.

MBc

Mean biomass of plant under control.

SL

Shoot length

FW

Fresh weight

xii

DW

Dry weight

WDPT

Water drop penetration time

OOC

Oxidizable organic carbon

TOC

Total organic carbon

PPT

Part per thousand

ppm

Part per million

WR

Water repellency

cm

Centimeter

Micro second

USA

United state of America

<

Greater than

>

Less than

Percentage

xiii

Chapter # 01
INTRODUCTION

1.1. INTRODUCTION
The Eucalyptus camaldulensis is a tree of the genus Eucalyptus,commonly known
as River Red Gum, It is one of around 800 in the genus. Eucalyptus is a long and
perennial tree belong to Myrtaceae family, it is a plantation species in many parts of the
world.

Height ranges between 45 to 65 meters and has essential oil at least 1%

(volume/weight) (Javanshir 1972), Found in tropical region, River Red Gum is subject to
regular flooding in its natural habitat. River Red Gum prefers soils with clay content. The
trees not only rely on rainfall but also on regular flooding, since flooding recharges the
sub-soil with water. River Red Gum seeds germinate readily after floods and require
regular spring floods throughout their life to survive.
It is native of Australia. Eucalyptus spp. in natural habitats Grow under a wide
range climatic and edaphic condition (Dawar et al . 2007). In Pakistan, a small nursery
of Eucalyptus globules in 1903 was raised by the forest department of Punjab (Siddique
and Hussain, 1980).This species has a high possible of allelochemicals and also essential
oils (Iqbal et al 2003). Researcher of India and Pakistan pointed out inhibitory effects of
this species on the germination rate and growth of seedling on many crops (khan et al .,
2008) Eucalypts can depart the soil unfavorable to growth of other plants eg, by directly
inhibiting under storey species or crop plants being grown in vicinity of a eucalypt stand.
Crop will not do well, at least for number of years where a eucalypt stand is harvested
and replaced by an agricultural crop (Fikreyesus et al ., 2011).

1.2. Allelopathy:
Allelopathic studies were undertaken earlier (Schreinerand Reed, 1907, 1908) in the
history of allelopathy one of the defining experiments was carried out by (Massey
1925).Allelopathy is defined as any process involving secondry metabolite produced by
plants, algae bacteria and fungi that influence the growth and development of agriculture
and biological system Anonymous;(1996). A term Allelopathy from two Greek words of
Allello and pathy meaning mutual harm (Molisch (1937). A term allelochemical
derived from allelochemics was firstly used by chou and Waller and has become popular
in agriculture science, Chou and Waller (1983). Any direct or indirect effects (stimulation
or inhibitory) by one plant including microorganisms, on another through production of
chemicals compounds that escape into environment, Rice , (1984) Allelopathy is
fascinating and confusing subjects that concern with the interaction

of plants as

influenced by the chemical substance that they release into environment (Bais et al .,
2003,Wills 2004 and Machado 2007) Allelopathy is the fabrication and release of
chemicals that damage or otherwise decrease the fitness of other plants (Hierro and
Callaway, 2003) , it is Capable of suppressing the germination and growth of various test
species. Allelopathic effects depended upon the parts assayed, test species and
physiological process involved .The rapid advances in chemistry, plant physiology,
biochemistry and ecology the role of allelopathy has increasingly become in investigated
Anon ( 2000), Malik (2002).
1.3. Allelochemicals
Essential oil neither stays long in soil nor penetrates underground water and never
causes environmental poisoning (Isman et al

2000) Allelochemicals are the small

2

molecular weight compounds excreted from plants during the process of secondary
metabolism, Rice ,(1984) Allelochemicals are found to be released to environment
inappreciable quantities via root exudates, leaf leachates, roots and other degrading
plantresidues in the soil ,Chemically allelochemical compounds have been opened chain
molecular structures .Allelochemicals can be classified into terpaenes, glucocides,
coumanines, aldehyds and phenolic compounds (Alam and Islam 2002). These are
secondary metabolites that have role in plant plant, plant-soil, plant disease, plant-insect
and plant predator interaction that may be beneficial or detrimental to plant, Tang et al
.(1986),Yaduraju and Ahuja,(1996). 16 components in essential oil of E. camaldulensis
L., out of which five compounds (

6.0% -pinene,1.8%

3-carene ,7.94% -

phellandrene, 53.22% 1-8 cineole and 21.59% p-cymene) were indentified, Iqbal et al
(2003) . E. camaldulensis yield 1.37% oil and have 0.90 specific gravity ; Zafar Iqbal
(2003) Essential oil can be used to control weeds in organic farming systems Tworkoski
(2002).Concentration of chlorogenic acid in some plants may be increased because of the
shortage of water Li et al ., (2001). Some terpenes , such as -pinene, -pinene, cineole,
camphor and can, increase their volatile under dry conditions Shao-Lin et al . 2004.The
relationships between phenolics and soil microorganisms in spruce forests and found that
phenolics compounds can stimulate fungi and cellulose hydrolyzers in the winter, but
inhabit cellulose hydrolyzers in summer Souto et al

. 2000. Several laboratory

experiments indicate that mixture solutions of allelochemicals have greater effect than the
same concentrations of the compounds used separately Blum et al., 1999, Einhellig
1995, Chaves & Escudo 1997.

1.4. Eucalyptus as Allelopathic plant

Effect of Eucalyptus globulus essential oil on Triticum aestivum, Zea mays,
Raphanus sativus, Cassia occidentalis, Amaranthus viridis and Echinochloa crus-galli
have been proved as their radicle length, plumule length, germination percentage and
germination rate decreased under the effect of essential oil concentration (M. Rassaeifar
et al 2013) Maize seeds in fields surrounded by Eucalyptus trees significantly decreased
germination Blaise, et al., (1997). Aqueous extract of air dried leaf litter of E. citirodora
had inhibitory effect on the seed germination, in wheat, mustard and gram reported by
Singh et al.,

(1992). Plant height and root length of American cotton increased

significantly as the diatance from eucalyptus trees increased, Eucalyptus trees adversely
affected the plant population of cotton Reddy et al

(2006). Eucalyptus reduces the

growth of neighboring crops through the release of allelochemicals May and Ash, (1990).
aqueous extract from bark and leaf, and volatiles from leaves of Eucalytus citriodora
showed allelopathic effect on the growth of nine species, including the weeds Bidens
pilosa, Digitarie pertenuis, Eragrostics cilianesis, Setaria geniculata, and crops such as
corn, rice, cucumber, bean and Stylosanthes guianensi Cao and Luo (1996). In laboratory
bioassay germination, seedling length, chlorophyll content and respiratory ability of weed
plants was drastically affected, Batish et al, (2005). Eucalyptus produces prolific litter in
the form of dropping leaves and naturally flecked off bark. If allelochemicals are released
from Eucalyptus leaf litter its accumulation in the soil could result in poor development
of native flora Bughio et al , (2013). The concentration of chlorogenic acid in some
plants may be increased because of the shortage of water some terpencs , such as -

pinene, - pinene,,cincole, camphor and can increase their volatiles under dry conditions,
Dias and Dias (2000).
1.5. Effects of allelopathy
Allelochemicals significantly reduce the growth of other plants and the yields of
crop plants Rice (1984); Korner and Nicklish (2002), Leu et al., (2002) Inderjit and Duke
(2003), allelochemicals have significant effects on cell division, cell differentiation, ion
and water uptake, water status, phytohormone metabolism,respiration, photosynthesis,
enzyme function, signal transduction as well as geneexpression Singh and Thapar (2003);
Inderjit and Duke (2003); Belz and Hurle (2004),Science 2003). Any changes in Chl
content are expected to bring about change in photosynthesis. Allelopathy plays an
important role in biodiversity, it may regulate the density and production of plant
community under the canopy of a dominant species and limits the population of its
associated species (Chou., 1999), mostly seeds of plant species are not

killed by

allelopathic compound; Chou (1999). Using advanced biotechnology techniques

,scientists in the foreseeing future my be able to isolate allelopathic genes from one plant
and transfer the gene to another plant .genes such as trypsin inhibitor gene and ethylene
regulatory gene have been successfully isolated from one plant to another plant, (Yeh et
al .1997;Yang1998).Allelochemicals

activity varies with temperature, photoperiod,

water and soils during natural process, growing efforts are being made to examine the
role of allelopathy in controlling the spreads of weeds, pest insects and diseases. PENG
Shao-Lin et al . 2004. Allelochemicals are released and added to the soil over a time
period and also continually removed and/or immobilised from the soil solution by plant
uptake, adsorption to soil particles, and degradation by microorganisms Cheng (1995).
5

1.6. Examples of Allelopathy

Numerous crops have been investigated more or less thoroughly for allelopathic
activity towards weeds or other crops. A suppressive effect on weed, possibly mediated
by the release of allelochemicals has been reported for a wide range of temperate and
tropic crops. These include alfalfa (Medicago sativa), barley (Hordeum vulgare), clovers
(Trifolium spp., Melilotus spp.) oats (Avena sativa) pearl millet (Pennisetum glaucum),
rice (Oryza sativa) rye (Secale cereale), sorghums (Sorghum spp.), sunflower
(Helianthus annuus), sweet potato (Ipomoea batatas) and wheat (Triticum aestivum), ,
Weston 1996, Narwal 1996, Narwal et al . 1998, Miller 1996, , Dilday et al . 1994.
Reduced chlorophyll content in allelochemical-treated plants has been frequently
reported. ZHOU, Y.H. and YU, Einhellig and Rasmussen (1979) and Patterson (1981) all
found that treatment Of soybean plants with phenolic acids such as ferulic, p-coumaric,
and vanillic acids greatly decreased the biomass associated with reduced chlorophyll
content in leaves.Similar results are also found in species such as Parthenium
hysterophorus,Kanchanand Jayachandra, (1980), Cucumis sativus Pramanik et al .
(2001); Yu et al .unpublished. In contrast, growth inhibition by allelopathic agents in
grain sorghum seedling was not followed by decreased chlorophyll content in some cases
Einhellig, (1986) Theophrastus (372-285) reported the inhibitory effect of pigweed on
alfalfa Jelenic, (1987). The negative impact of such species on native biodiversity has
been well documented DAntonio and Mahall 1991, Ridenour and Callaway 2001).
Species X. strumarium secretes allelochemical compounds that influence both the
germination of the seedes and the growth of vegetative mass in culture plants and weeds
6

(Bozsa and Oliver 1993; Sondhia and Saxena 2003; Sinha and Samart 2004; Dvid et al.,
2005; Tanveer and collaborators. 2008). Many crop plants, including chick pea (Cicer
arietinum) and barley (Hordeum vulgare), inhibit the growth of weeds and crop plants
other than barley (Rice 1984). The two alkaloids, gramine (N, N-dimethyl-3-aminomethylindole) and hordenine (N, N dimethyltyramine) have been confirmed to play an
important role in the phytotoxic ability of barley (Lovett & Hoult 1995, Overland 1966).
1.7. Residual effects of Allelochemicals
Inhibitory effects on germination and establishments of crops caused by residues
of either crops or weeds have lead to investigation of the release of toxic compounds
from such residues. For example, the allelopathic interference of both living plant and of
plant residues of the highly aggressive weed Elytrigia repens, quackgrass, has been
strongly indicated (Weston & Putnam 1985). Residues from several crop species have
been examined for their potential to reduce weed germination (Creamer et al . 1996,
Moyer & Huang 1997). Most work concerning allelopathic effects of rye has been carried
out using residues. Rye residues have been employed as mulches or cover crops in notillage cropping systems to suppress certain weed species (Barnes & Putnam 1986). In
contrast, results obtained by Creamer et al . (1996) by leaching rye of its water soluble
allelopathic compounds and using it as an inert material, indicated that the physical
suppression of rye was responsible for the reduced emergence of two weedy species,
eastern black night shade (Solanum ptycanthum) and yellow foxtail (Setaria glauca). The
release of allelochemicals from living wheat plants has also been documented (Peth
1992a).

Literature survey suggest two directions one in which allelopathy are beneficial
as weed control on other hand it effects the germination ,division, growth and
photosynthesis rate of the other species but mostly adverse effects of allelopathy was
observed by surveying the literature. New leaf litter have allelopathy effects is observed
by conducting experiment by different scientists but to observe the effects after keeping
that leaf liter mixed soil for some year , Experiment was conducted to observe the
allelopathy effects of Eucalyptus camaldulensis leaf litter on Zea mays and Pennisetum
typhoids.
1.8. Objectives
The primary objective of this study was to assess the allelopathic effects of eucalyptus
leaf litter residue on the germination and growth of two cereals Zea mays and Pennisetum
typhodeum. Since the leaf litter of eucalyptus decomposes slowly in the soil environment
thus its residue remain in soil for longer periods. The study was designed to analyze the
effects of residue on plants and soil properties.

Chapter # 02
Material and methods
2.1. Introduction
In this present research leaf litter of Eucalyptus camaldulensis as allelopathy was
used to analyze the effects on the Zea mays and Pennisetum typhodeum (bioassay). This
present research provides vital information about plant parameters, e g germination rate,
mortality, shoot length, fresh weight, dry weight, water repellency and Soil properties, e
g. pH, EC, TDC, PPT, OM, Na and K. The experimental work was carried out in the
green house of Institute of Plant Sciences, University of Sindh, Jamshoro. The study
comprised of the following procedures and experimentation.
2.2. The Materials
2.2.1. The Bioassays Plants
In this experiment Zea mays and Pennisetum typhodeum were used as bioassay
plants. Seeds were purchased from the market, and were primed in water for eleven hours
then in the morning five seeds per pot was sown.
2.2.2. Soil and leaf litter formulations
Object of this research was to analyze that how long effects of Allelopathy of
Eucalyptus camaldulensis remain in the soil, for this purpose such soil was required in
which leaf litter must have mixed for some years, therefore formulation of soil and leaf
litter was used formed by previous worker, who Already worked three years ago on
Allelopathy, formulations is following.
2.2.3. Eucalyptus leaf litter and its preparation
9

The dropped dried leaves of Eucalyptus camaldulensis Dehnh., were collected

from Eucalyptus camaldulensis monoculture stands growing at Pai- Forest, District:
Shaheed

Benazirabad,

Sindh,

Pakistan.

dropped

dried

leaves

of

eucalyptus

cammaldulensis washed by distal water, dried one week of room temperature then
grounded and passed through 2.0 nm sieve, powder was used in soil as leaf litter w/w
(except control), eucalyptus leaf litter powder of leaves was mixed in these three
concentration i.e 10, 20 & 30% (w/w) in sandy silt soil Bughio et al., (2013). The soil
was analyzed for the physical and chemical parameters.

2.3. The Methods

2.3.1. Experimental Design and treatment
The complete block design was used with four treatments and four replications of each
treatment of both bioassays were made.
Table 1.Experiment design

Treatments
T0
T1
T2
T3

Control

Concentrations: Soil mixed dry leaf litter (w/w)

Soil without leaf litter
10%
20%
30%

2.3.2. Sowing of Seeds

The pots filled with soil (2kg) and leaf litter, was broken up earth and watered, one week
before sowing seeds for all the treatments for good germination. In each pot 5 holes of 1
inche was made and 1 seed were sown per hole in each pot of Zea mays and Pennisetum
typhodium. All the pots were provided homogenous conditions there was only one
variable in the experiment that was leaf litter of E. camaldulensis
10

2.3.3. Data Collection and Analysis

Parameters used
In experiment following parameters were recorded:
Plant parameters
In plant parameters following data parameters were recorded;
Germination %
Relative germination ratio
Percentage mortality rate
Fresh weight
Dry weight
Plant length
Relative biomass (RBR)
Chlorophyll content
(a) Germination % ( Scott et al ., 1984)
% =

100

Where Gn is the number of seeds germinated after 14 days and GN is the total number of
seeds sown.
(b)

Relative germination ratio; (Rho & Kill, 1986)

100

Using germination % data relative germination was calculated, GRt is germination ratio
of plants under treatment and GRc is the germination ratio of plants under control.
(c) Percentage mortality rate
It was counted that how many plants died after the germination.

11

100

MR = % mortality rate.
MRn = no of seeds died after germination.
GN = total no of seeds germinated.

(d) Fresh Weight (g/plant)

From each treatment leaves of plants were cut randomly after 20 days and were brought
in lab for fresh weight (biomass) and were recorded by using electric balance in grams.
(e) Dry Weight (g/plant)
These pre weighted leaves were then oven-dried at 65oC for 30 minutes and then data for
dry Leaves (biomass) were recorded.
(f) Shoot length (cm)
Three times (duration of a week) length of the plants were measured by using foot scale
length of the every plant was measured after 30 days data was recorded.
(g) Relative biomass (RBR)
=

100

Where;
RBR = relative biomass ratio.
MBt = mean biomass of plant under treatment .
MBc = mean biomass of plant under control.
(h) Chlorophyll content (mg g-1 )
The 1g of fresh leaves of Zea mays and Pennisetum typhodium were crushed in
mortar and pestle using 10 ml of 100% acetone. The chlorophyll extract was then filtered
by using what man filter paper .The extract was collected in a conical flask. The extract
was analyzed for absorbance using CIBA Corning 254 colorimeter. The chlorophyll
content was measured by using following formulae:

12

Chlorophyll a= [12.7(OD) 470-2.69(OD) 540] x V/1000xW

Chlorophyll b= [22.9(OD) 540-4.68(OD) 470] x V/1000xW

Soil Parameters
In order to measure the soil changes produced by Eucalyptus treatments on soil
properties, the following soil parameters were recorded;
pH of soil
Electro conductivity of soil ( ECe )
TDS
Salinity (PPT)
Soil organic matter
Na+
K+
Water repellency

(a) pH
The pH of saturated soil paste was measured using digital pH meter (JENCO- Models
5000 Series 301120).

(b)Electric conductivity of soil (ECe )

Electro conductivity can almost be varied as the quality of available nutrients in
soil, it indicates the presence or absence of salt. To measure the EC 25g of the soil was
taken and added 40ml distil water then stirrer and allowed to stand four hours, then
filtered extract was used for EC meter (Thermo scientific, Orion 5 star (USA).

13

(c)TDS
Total Dissolved Solids (TDS) is the amount of mineral and salt impurities in the
water. For measurement of TDS same extract was used (Thermo scientific, Orion 5 star
(USA).
(d) Salinity (PPT)
It is measurement of salinity, ppt stand for parts per thousand. The salt concentration
is usually expressed in parts per thousand (permillle, ) or parts per million (ppm). The
United States Geological Survey classifies saline water in three salinity categories. Salt
concentration in slightly saline water is around 1,000 to 3,000 ppm (0.1-0.3%), in
moderately saline water 3,000 to 10,000 ppm (0.3-1%) and in highly saline water 10,000
to 35,000 ppm (1-3.5%). Seawater has a salinity of roughly 35,000 ppm, (From
Wikipedia, 20 December 2013) (Thermo scientific, Orion 5 star (USA).

(e) Na+ and K+

Sodium and Potassium are the important nutrients of the soil that affects the soil
properties; therefore Na+ and K+ were also analyzed. To analysis of Na and K 10g soil
was taken, then 100ml distil water was added and stirred for half an hour
(f) Organic matter
Measurement of organic matter from the soil 1g of soil was taken to every
treatment. Soil organic matter was calculated by the procedure, which involves reduction
of potassium dichromate (K2Cr2O7) by Organic carbon (OC) and subsequent
determination of the unreduced dichromate by oxidation reduction titration with ferrous
ammonium sulfate (Walkley, 1947).

Calculation:
14

Percentage organic matter in soil

10

1. =
2. % () =

0.3

Where;
Wt = weight of soil
0.3 = 3 10-3 100 where 3 is equivalent weight of Carbon.
3. = 1.334 % %
4. % = 1.724 %

(h) Soil water repellency

Soil passed through 2mm sieve, after drying samples WDPT test was conducted
by placing a water dropper on the soil surface and recording the time taken for water to
penetrate the soil, five drops of distill water were applied with hypodermic syringe to the
surface of soil samples, the penetration time for each drop was recorded and average
penetration time taken as representative of the WDPT for each sample.

15

Chapter # 03
Results and Discussion
3.1. Results Zea mays
3.1.1. Germination %:
The results presented in Table-1 shows Zea mays have been influenced but in
such a way that Treatment T0 (control) and T1 (10%) have same value of the germination
ratio and value of the germination ratio of Treatment T2 (20%) have been decreased and
T3 (30%) germination ratio have been increased by different leaf litter of E.
camaldulansis treatments. Overall the germination % is affected but there is little
difference in values as compare to the ratio of the leaf litter concentrations, it shows that
germination % have not been effected by treatment of leaf litter of E. camaldulansis 10%,
20%, and 30%, as compare to control, able-1.
3.1.2 Mortality Rate:
It is percentage seedling death after germination. In Table-1 MR ratio of T0 and
T1 is same and T2 and T3 has zero % MR, it shows that mortality rate (%) of Zea mays
have not been effected by treatment of leaf litter.
3.1.3 Relative germination ratio:
There in table-1 same sort of the values between percentage of leaf litter and
Relative germination ratios have been found, T0 and T1 have same value 100% and
Treatment T2(91.8%) have been decreased and RGR ratio of T3 (97.29%) have been
increased by leaf litter.
3.1.4 Relative elongation ratio of shoot:
The relative elongation ratio of shoot is increasing gradually T0 =100 and T1 =
140.6 but RERs value of T2 shows it have been decreased T2 = 130.7 and value of the T3
increased (189.0) (Table-1)
16

3.1.5 Shoot length:

The shoot length of the Zea mays was measured from base of the stem to tip of
the last leaf and their mean was calculated. Shoot length have been increased gradually
T0 = 22.75 and T1 = 32 but in T2 it have been decreased (T2 = 29.6) and value of the T3
have been increased 43.5 it shows that treatment of leaf litter have not been effected
(Table-1)
3.1.6 Fresh weight:
The fresh weight of the plants of Zea mays was measured in grams.The results are
presented in Tables-1. Fresh weight of treatment T1 have been decreased as compare to
other three treatments T0,T2, T3, and the value of T0,T2,T3

have bee increased

gradually. It shows that fresh weight have not been effected by the treatment of leaf litter
of E. camaldulansis
3.1.7 Dry Weight:
The dry weight of Zea mays was also measured in grams. Fresh weight of
treatment T0, T2, T3 has been increased gradually. T1 have been decreased as compare
to other three treatments T0, T2, T3.The results are presented in Tables-1
3.1.8 Relative biomass ratio:
Relative biomass ratio was calculated from the values of dry weight. The RBR
ratio of treatment T0 =100 and the value of the T1, T2, T3 have been gradually increased.
3.1.9 Chlorophyll content
The analysis of chlorophyll content samples were taken after the 30 days. shows
significant difference among values of treatments. The maximum value of total
chlorophyll content was observed in T0 control (32.07) and followed by T2 (27.04) and

17

the values of T1 (14.05), T3 (15.68). The minimum value of chlorophyll content was
observed in T1 and T3, (Table 3).

Table.1. Residual Effects of eucalyptus leaf litter on germination of Zea mays.

Treatments
Germination (%)
RGR
Mortality Rate
T0

92.5

100

5.0

T1

92.5

100

5.0

T2

85.0

91.8

0.0

T3

90.0

97.29

0.0

Mean

90.0

97.2

2.5

2.635231

SE

Table2. Residual Effects of eucalyptus leaf litter on growth of Zea mays.

Treatments
Shoot length
RERs
FW (g)
DW (g)

RBR

T0

(cm)
22.75

100

3.10

0.53

100.0

T1

32.00

140.6

2.60

0.44

83.90

T2

29.60

130.7

4.00

0.52

99.00

T3

43.50

189.0

5.18

0.69

130.0

Mean
31.90
140.0
0.901166
SE
RERs= Relative elongation ratio of shoot

3.70
0.54
0.239064 0.047817

103.0

FW= Fresh weight

DW= Dry weight
RBR= Relative Biomass Ratio

18

Table.3. Residual effects of Eucalyptus leaf litter on leaf Chlorophyll content in

Zea mays.
Chlorophyll A

Chlorophyll B

Carotenoids

(mg/g f.wt)

Total
(mg/g f.wt)

(mg/g f.wt)

T0

17.96

14.21

32.07

0.00000316

T1

7.99

6.06

14.05

0.00000041

T2

14.22

12.82

27.04

0.00000459

T3

7.79

7.89

15.68

0.00001931

Mean

12

10.2

22.2

0.00000068

Treatments

(mg/g f.wt)

19

3.2 Soil parameters

3.2.1 Soil pH
The pH of the soil was analyzed two times; firstly it was analyzed before the sowing of
seeds and after harvest. The analysis of initial soil pH per sample for individual
treatments T0, T1, T2, T3, shows no much differences among their values but with
difference of few points it have been increased and the value of the second soil sample
also shows no much differences among their values however there is difference between
the value of the initial soil and second soil by taking account their mean as I=6.3 and 7.4 .
(Table 2).
3.2.2 Electrical conductivity of soil ( ECe )
For electrical conductivity analysis soil sample was taken before the sowing seeds that
soil was named initial soil (I). The result was, treatment T0=229, T1=517, T2=576,
T3=620 for individual treatments values have been increased one by one. After duration
of one month soil was taken for second time analysis, there was not much difference in
Table 4. Residual Effects of eucalyptus leaf litter on Soil properties
Treatment

pH

EC (s/cm)

T0

5.9

7.4

T1

6.2

7.4

T2

6.5

T3
Mean

Salinity

TDS

WR

492

113

256

0.1

0.2

517

523

252

250

0.2

0.3

3:17

7.8

576

581

292

285

0.3

0.3

31:25

6.9

7.3

620

532

315

261

0.3

0.3

26

6.3

7.4

485

532

243

263

0.2

0.2

15:11

229

20

Table. 5. Residual Effects of eucalyptus leaf litter on Soil properties

Treatment

OM (%)

OOC (%)

TOC (%)

T0

2.6

2.2

0.75

1.05

1.00

1.4

T1

2.9

4.8

0.9

2.1

1.20

2.8

T2

3.8

5.5

1.05

2.43

1.4

3.2

T3

3.0

5.8

1.8

2.55

2.4

3.4

Mean

3.0

4.0

1.12

2.03

1.5

2.7

Ta Table.6. Residual Effects of eucalyptus leaf litter on Soil properties

Treatment

Na (ppm)

K (ppm)

T0

10

12

T1

10

11

T2

16

10

T3

17

13

Mean

11

13

5.7

3.5

I = Initial soil , S =Second soil

21

the values between treatments but EC of the second soil mean value have been increased
by leaf litter as compare to sample of initial soil. (Table 2).
3.2.3 TDS
The analysis of soil for total dissolved salts per soil sample for individual treatments
shows no significant differences among their values. As initial soil T0=113, T1= 252,
T2=292, T3=315.In second soil sample value of T2 have been increased, T1 value have
been decreased as compare to other treatments T0(256), T2 (285), T3(261) (Table 2).
3.2.4 Salinity (PPT)
The result of ppt also have been increased gradually from T0 to T3 treatment likewise, it
is give in table 2.
3.2.5. Soil organic matter
The analysis of soil for organic matter per soil sample for individual treatments of the
Zea mays shows gradually increased values from T0 (2.6) treatment to T3(3.0), T2 (3.8)
have high value as compare to other treatments. Values of second soil sample have been
increased one by one. However the mean value have been increased of second soil (4.0)
sample as compare to initial soil (3.0) value, (Table 2).
3.2.6 Na+ and K+
Soil was also analyzed for basic nutrients Na and K. The sample of the initial soil (I)
treatments for Na the values T0=10, T1=10, T2=9 have been increased but the value of
T3 is high (17) as compare to T0, T1, T2 treatments. In second sample values were
randomly as T0=12, T1=11, T2=16, T3=13. Mean value between initial soil and second
soil shows that mean value have been increased by leaf litter of second soil as compare to
initial soil.
22

The K analysis, the initial soil sample of the treatments T0, T2, T3, have equal values (6)
and value of the treatment T1was found low but the values of the second soil sample was
found randomly. By the mean, value of the initial soil (5.7) has been increased as
compare to second soil (3.5) sample.
3.2.7. Water repellency:
Low levels of water repellency have been observed in many soils (Hallett et al .,
2001). The biological origin of repellency suggests that it will have a high spatial and
temporal variability at very small scales, because of the submillimeter spatial ability of
organic matter, organisms and the microbial environment in soil (Nunan et al ., 2002).
WR ratio start from the lowest time 5 second of T0 control to 31 mints 25 seconds, WR
value has been increased gradually T0, T1, T2 and T3=26. Mean value was found 15:11
(Table 2)

3.3. Pennisitum typhodeum.

3.3.1. Germination %:
The results presented in Table-3 shows that germination rate have been increased from
treatment T0 to T3 of Pennisitum typhodeum .T0 and T1

have same rate of the

germination (95%) and T2 and T3 (100%). Rate of germination of T2 and T3 have been
increased as compare to T0 and T1.Mean value was found 97.5%
3.3.2. Mortality Rate:
Mortality rate of the Pennisitum typhodeum was not found. In all treatments there
was zero rate of the mortality, as shown in Table-4.
3.3.3. Relative germination ratio:

23

Relative germination ratio have been found, T0 and T1 have same value 100%
and Treatment T2 and T3 (105.2%) have been increased as compare to T0 and T1 102.6
was found their mean value (Table-4).
3.3.4. Relative elongation ratio of shoot:
The relative elongation ratio of shoot of the Pennisitum typhodeum have been
increased gradually T0 =100 and T1 = 128, T2=147, T3=163.Mean (134)(Table4)
3.3.5 Shoot length:
The shoot length of the Pennisitum typhodeum was also measured from base of
the stem to tip of the last leaf. Here the shoot length of the Pennisitum typhodeum have
been also increased gradually T0 = 28 and T1 = 36.4, T2=41.9, T3= 46.4 but the values
of the T1, T2, T3 have been increased as compare to control T0(Table 4)
3.3.6 Fresh weight:
The fresh weight of the Pennisitum typhodeum plants of was measured in
grams.The results are presented in Tables-3. Here the fresh weight of Pennisitum
typhodeum have been also increased gradually T0 = 0.20 and T1 = 0.6, T2=0.5, T3= 0.8
but the values of the T1, T2, T3 have been increased as compare to control T0 0.2 ,FW of
the control was found low (Table 4)
3.3.7 Dry Weight:
The dry weight of the Pennisitum typhodeum was measured in grams. dry
weight of Pennisitum typhodeum have been increased gradually T0 = 0.035 and T1 =
0.097, T2=0.092, T3= 0.011 but the value of the T3 was found high as compare to control
T0 0.035 ,DW of the control was found low 0.035 (Table 4)

24

3.3.8 Relative biomass ratio:

The RBR ratio of treatment was found T0 =100 and the value of the T1 = 278,
T2= 264, T3=321 have been gradually decreased. The high ratio 321 was found in T3 and
low ratio in control (Table 4)
3.3.9. Chlorophyll content (mg/g f. wt.)
The analysis of chlorophyll a, b and carotenoids shows that no significant
difference between the treatments was found (Table-5). It shows that eucalyptus leaf litter
stimulated the chlorophyll content of Pennisetum typhoideum.
Table 7. Residual Effects of eucalyptus leaf litter on germination of P. typhodium
Treatments
Germination (%)
RGR
Mortality Rate
T0

95

100

0.0

T1

95

100

0.0

T2

100

105.0

0.0

T3

100

105.0

0.0

Mean

97.5

102.6

0.0

1.86339

SE

Table.8. Residual Effects of eucalyptus leaf litter on growth of P. typhodium .

Treatments
Shoot length
RERs
FW (g)
DW
RBR
T0

(cm)
28.4

0.20

(g)
0.035

100

100.0

T1

36.4

128

0.60

0.097

278

T2

41.9

147

0.55

0.092

264

T3

46.4

163

0.8

0.112

321

Mean
38.2
134
1.348823
SE
RERs= Relative elongation ratio of shoot
FW= Fresh weight
DW= Dry weight
RBR= Relative Biomass Ratio

0.5

0.084

240

0.052434

0.0069

25

Table.9. Residual effects of Eucalyptus leaf litter on leaf Chlorophyll content in pearl
millet.
Chlorophyll A

Chlorophyll B

(mg/g f.wt)

T0

(mg/g f.wt)

Total
(mg/g f.wt)

Carotenoids

42.64

41.53

84.17

0.00002421

T1

44.45

52.04

96.49

0.00003164

T2

26.29

21.28

47.57

0.00000544

T3

42.88

43.83

86.71

0.00002986

Mean

39.0

39.5

78.5

0.00000224

Treatments

3.4 Soil Parameters

3.4.1 pH of soil
The Ph of the soil was analyzed two time . The analysis of initial soil pH per sample for
individual treatments shows no much differences among their values but with difference
of few points it have been increased gradually T0=6.7, T1=6.8, T2=6.9, T3=7.1, T0
control 6.7 was found low value as compare to T3(7.1). In the second sample T0 was
found 7.7 and T3 7.8 high values, T1 (7.3), T2 (7.0).Mean value of the of the second soil
sample was found more (7.4) as compare to initial soil sample (6.8).
3.4.2 Electrical conductivity of soil ( ECe )
The analysis of electrical conductivity soil sample was taken initial and second The
T0=338, T1=448, T2=568, T3=750 values have been increased one by one. T0 have low
value and T3 high. Second time analysis, T0 was found 177.7 low as compare to T2

26

(428).Second soil, mean value have been decreased as compare to sample of initial soil.
(Table 5).
3.4.3 TDS
The values of total dissolved salts have been increased significantly. As initial soil
T0=166, T1= 220, T2=278, T3=367. Control was found low value of the TDS as
compare to other treatments .In second soil sample value of

T0 was found low

(166)value as compare to T3(367) been increased, (Table 5).

3.4.4 Salinity (PPT)

The result of ppt also have been increased gradually in initial soil sample T0 and T1 was
equal (0.2).Value of the T3 was found high . Mean value of initial soil was more (0.2) as
compare to second soil sample (table 5).
3.4.5 Soil organic matter
The organic matter is the resulting decomposed product from green manure, plant
residues and animal wastes accumulated in the surface soil layers and microbial debris in
various stages of the decay (Ndabakishye 1985; Leclerc 1995). Analysis of soil for
organic matter of initial soil per soil sample for individual treatments of the Pennisitum
typhodeum shows gradually increased values from T0 (1.6) treatment followed by T3
(4.0). T0 have lowest value as compare to treatments T1, T2, T3, whereas T3 have
highest value. Values of second soil sample have been also increased gradually T0=3.1,
T1=3.7, T2=4.0, T3=5.5. Control T0 has lowest value and T3 have highest value.
However the mean value have been increased of second soil (4.0) sample as compare to
initial soil (2.8) value, (Table 5).

27

3.4.6. Na+ and K+

Soil was analyzed for nutrients Na and K. The sample of the initial soil (I) treatments
values was found randomly for Na the values T0=4, T1=6, T2=4, T3 =6. In second
sample values have been increased gradually from control T0 to T3 (30% leaf litter), T0
have lowest value (7) and T3 highest value (12). Mean value between initial soil and
second soil value have been increased of second soil (11.7) as compare to initial soil (5).
Soil analysis for K, the initial soil sample of the treatment T1 have lowest value (9) and
T3 was found highest value (15), and the control T0 have (13) value. Second soil samples
T1and T3 have been increased (21) and the control was found lowest value. Mean, value
of the initial soil (12.2) and second soil (14.5).
3.4.7 Water repellency:
WR ratio have been increased gradually T0= 5 second, T1= 5 minutes and 50 second,
T2= 31 minutes and 25 and T3= 26 minutes and 13 seconds. Mean value was found
15:26 (Table 5)

28

Table.10. Residual Effects of eucalyptus leaf litter on Soil properties

Treatment

PH

EC (s/cm)

Salinity

TDS

WR

T0

6.7

7.7

338

177

166

87

0.2

0.1

T1

6.8

7.3

448

324

220

159

0.2

0.2

3:50

T2

6.9

7.0

568

428

278

210

0.3

0.2

31:25

T3

7.1

7.8

750

427

367

209

0.4

0.2

26:13

Mean

6.8

7.4

526

339

257

166

0.2

0.17

15:26

Table11. Residual Effects of eucalyptus leaf litter on Soil properties

Treatment

OM (%)

OOC (%)

TOC (%)

T0

1.6

3.1

1.2

1.35

1.6

1.8

T1

3.3

3.7

1.27

1.65

1.7

2.2

T2

2.3

4.0

1.72

1.77

2.3

2.3

T3

4.0

5.5

1.35

2.4

1.8

3.2

Mean

2.8

4.0

1.38

1.79

1.85

2.37

I = Initial soil , S =Second soil

29

Table.12. Residual Effects of eucalyptus leaf litter on Soil properties

Treatment

Na (ppm)

K (ppm)

T0

13

T1

11

21

T2

17

12

17

T3

12

15

21

Mean

11.7

12.2

14.5

30

Conclusion:
The results suggest that the eucalyptus leaf litter residues slightly decrease the
germination percentage of maize, the mortality rate was recorded in control and T1, in T2
and T3 no MR was found. Similarly RGR was decreased in eucalyptus treatments as
compared to the control. The eucalyptus leaf litter has stimulatory effect on RERs, shoot
length, fresh weight, dry weight, Relative biomass ratio and chlorophyll a, b and amount
of carotenoids of maize. The higher values of these parameters were found in eucalyptus
leaf litter than control. The soil pH, K was almost same values in treatments, but second
soil have more values than initial soil. Whereas EC, TDS, Salinity, OM, Na was found in
higher concentration in eucalyptus treatments as compared to the control, as
concentration of leaf litter increased, concentration of these values also increased. The
eucalyptus treatments soils were found strongly to severely water repellent as compared
to control soil, which was non repellent.
The eucalyptus leaf litter has stimulatory effect on germination percentage, RGR,
RERs, shoot length, fresh weight, dry weight, RBR, chlorophyll a, b and carotenoids of
Pennisetum typhoideum. These values were more in eucalyptus treatments than control
and were found increased as the leaf litter concentration increased. The soil pH was
found slightly in initial soil from 6.7 to 7.1 whereas in second soil pH was almost same in
treatments. The soil EC, TDS, salinity, OM, Na, K was found in high concentration in
eucalyptus leaf litter treatments than control. The soil EC, TDS, salinity and K was found
in high concentration in initial soil and OM, Na was found in high concentration in
second soil. The eucalyptus treatments soil was found strongly to severely water repellent
as compared to control, which soil was non water repellent.

31

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