VARIETAL RESPONSE AND BIOCHEMICAL ANALYSIS OF

DIFFERENT POTATO CULTIVARS AGAINST ROOT KNOT

NEMATODE
(MELOIDOGYNE
INCOGNITA)
AND
ITS
NUTRITIONAL MANAGEMENT

VARIETAL RESPONSE AND BIOCHEMICAL ANALYSIS OF DIFFERENT POTATO

CULTIVARS AGAINST ROOT KNOT NEMATODE (MELOIDOGYNE INCOGNITA) AND ITS
NUTRITIONAL MANAGEMENT
By
AMJAD SHAHZAD GONDAL
Thesis submitted in partial fulfillment of requirements for the degree of
MASTER OF SCIENCE (HONOURS) In PLANT PATHOLOGY DEPARTMENT OF PLANT
PATHOLOGY FACULTY OF AGRICULTURE, UNIVERSITY OF AGRICULTURE FAISALABAD,
PAKISTAN
2012
Dedicated to
My Beloved
Parents Brothers & Sisters
ACKNOWLEDGMENTS
In the name of alm
ACKNOWLEDGMENTS
In the name of almighty Allah, the merciful, the beneficent All praises (belong) to
Allah
alone, the Cherisher and Sustainer of the world. He is the First, he is the Last, He is thehidden, and He knows
about everything. He brings the night into the day and brings the dayinto the night, and He knows the thoughts of
hearts (Al-Quran). I have the pearls of my eyes to admire countless blessings of Allah Almighty becausethe words
are bound, knowledge is limited and time of life is too short to express His
dignity. It is the one of His infinite benedictions that He bestowed upon me with the potential and ability to complete
the present research program and to make a meek contribution to thedeep oceans of knowledge already existing. I
deem it my utmost pleasure to avail an opportunity to express my heartiest gratitudeand deep sense of obligation to
a very hardworking and personalized men and womenespecially my honorable supervisor,
Dr. Nazir Javed
(Professor) for his kind behavior,generous knowledge, moral support, constructive criticism and enlightened
supervisionduring the whole study period. His available words will always serve as a beacon of light throughout my
life. I express my deep sense of gratitude to my supervisory committee,
Dr. Sajid Aleem Khan
(Assistant Professor)
,
Department of Plant Pathology, UAF and
Dr. Waqas Wakil

(Associate Professor)
Department of Entomology, UAF. I also express my deep sense of gratitude to
Dr. Kashif Riaz
(Assistant Professor) Department of Plant Pathology, UAF. for positive criticism and enlightened supervisionduring
my research and thesis write-up. I am grateful to my brothers
Amir Shahzad Gondal
&
Muhammad Sohail
for their support during my studies. I am also cordially obliged to
Ishtiaq Ahmad Gondal
, for being so helpful to methroughout my research work. I am very thankful to all my
sincere friends and class fellows specially,
Abdul Rauf
,
Moaz Bin Riaz
,
Muhammad Shafiq
,
Abdur Rashid Khan
,
Ahmad Nawaz
,
Tahir Farooq
,
Sajjad Hyder
and
Saqlain Rasool. Amjad Shahzad Gondal

v
3.4.3.1 Leaf Potassium (K) percentage for FD-19-2 ................................................................ 57
3.4.3.2 Leaf Potassium (K) percentage for FD-8-1.................................................................. 57
3.4.3.3
Leaf Potassium (K) percentage for SH-692
................................................................. 57
3.4.3.4
Leaf Potassium (K) percentage for FD-69-1
................................................................ 58
3.4.3.5
Leaf Potassium (K) percentage for SH-5
...................................................................... 58
3.4.3.6
Leaf Potassium (K) percentage for FD-74-67
.............................................................. 59
3.4.3.7
Leaf Potassium (K) percentage for SH-332
................................................................. 59
3.4.3.8

Leaf Potassium (K) percentage for SH-339

................................................................. 59
3.4.3.9
Leaf Potassium (K) percentage for FD-49-62
.............................................................. 60
3.4.3.10
Leaf Potassium (K) percentage for FD-1-3
.................................................................. 60
4.4.1.1
Effect of different treatments on number of leaves per plant
.................................. 71
4.4.1.2
Effect of different treatments on average tuber weight (g)
...................................... 72
4.4.1.3
Effect of different treatments on average root weight (g)
......................................... 73
4.4.1.4
Effect of different treatments on average shoot weight (g)
...................................... 73
4.4.2.1
Effect of different treatments on root galls production
............................................. 75
4.4.2.2
Effect of different treatments on egg mass production
............................................. 76
4.4.2.3
Effect of different treatments on nematode population per 100 ml of soil
........... 77
4.4.2.4
Effect of different treatments on nematode population per root system
.............. 77
4.4.2.5
Effect of different treatments on nematode rate of reproduction.
......................... 78
vi
List of Tables
2.4.1
Effect of
Meloidogyne incognita
infection on plant growth parameners

.................. 30
2.4.2
Effect of
Meloidogyne incognita
infection on nematodereproductionparameners
..................................................................................................... 35
4.4.1
Comparison of different treatments on the basis of plant growth parameters
....... 74
4.4.2
Comparison of different treatments on the basis of nematode reproductionparameters
.............................................................................................................................. 79

vii
List of Appendices
VARIETAL RESPONSE OF DIFFERENT POTATO CULTIVARS AGAINST ROOTKNOT NEMATODE (
MELOIDOGYNE INCOGNITA
) INFECTION.2.4.1.1
Analysis of variance for number of leaves
.................................................................. 104
2.4.1.2
Analysis of variance for tuber weight
.......................................................................... 104
2.4.1.3
Analysis of variance for root weight
............................................................................ 105
2.4.1.4
Analysis of variance for shoot weight
.......................................................................... 105
2.4.2.1
Analysis of variance for number of galls
.................................................................... 105
2.4.2.2
Analysis of variance for egg masses
............................................................................. 106
2.4.2.3
Analysis of variance for nematode population per root system
............................ 106
2.4.2.4
Analysis of variance for nematode rate of reproduction
........................................ 106
EFFECT OF MICRO-POWER, HUMIC ACID AND PLANT PROTECTORCONTAINING BENZOIC ACID AGAINST
ROOT KNOT NEMATODE(
MELOIDOGYNE INCOGNITA

) IN POTATO.
4.4.1.1
Analysis of variance for number of leaves
.................................................................. 107
4.4.1.2
Analysis of variance for tuber weight
.......................................................................... 107
4.4.1.3
Analysis of variance for root weight
............................................................................ 108
4.4.1.4
Analysis of variance for shoot weight
.......................................................................... 108
4.4.2.1
Analysis of variance for number of galls
.................................................................... 108
4.4.2.2
Analysis of variance for egg masses
............................................................................. 109
4.4.2.3
Analysis of variance for nematode population per 100 ml of soil
......................... 109
4.4.2.4
Analysis of variance for nematode population per root system
............................ 109
4.4.2.5
Analysis of variance for nematode rate of reproduction
........................................ 110

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VARIETAL RESPONSE
AND BIOCHEMICAL
ANALYSIS OF
DIFFERENT POTATO
CULTIVARS AGAINST
ROOT KNOT

NEMATODE
(MELOIDOGYNE
INCOGNITA) AND ITS
NUTRITIONAL
MANAGEMENT
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VARIETAL RESPONSE AND BIOCHEMICAL ANALYSIS OF DIFFERENT POTATO CULTIVARS
AGAINST ROOT KNOT NEMATODE (MELOIDOGYNE INCOGNITA) AND ITS NUTRITIONAL
MANAGEMENT
By...
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10
1.2.3.

Morphology
Meloidogyne

spp. are
sexually
dimorphic
(Wale
et al

., 2008).
Morphologicall
ydistinguishab
le males with
twisted bodies
are wormlike
having 1.2 to

1.5 mm length
and a
diameter of 30
to 36 m
(Singh, 2009).
Skeleton and
stylet and

labial region of
root knot

nematode are
tough with
short
hemispherical
tail having

strong
spicules while
bursa is
notpresent.
Females are
spherical
oval/globose/p

ear shaped
with about 0.4
to 1.3 mm
length
and0.27 to
0.75 mm
diameter

having a
cylindering
neck. Female
body has subterminal
vulvanear
anus;

annulated,
thin, whitish
Cuticle; short,
moderately
sclerotized
stylet and

labialregion
(Hunt
et al

., 2005;
Agrios, 2005).
An excretory
pore that is

positioned
near stylet
basebut in
some cases
anterior to
median bulb
valve plates is

found. In a
gelatinous
matrix,
eggsare
deposited
outside the
body on the

surface of
host plant
(Hunt
et al

., 2005; Luc
et al

.,2005;
Ashoub and
Amara, 2010).
1.2.4.
Host Range

The host
range of rootknot
nematodes is
so extensive
that it is
difficult to

findcommon
landscape and
garden plants
that are not
hosts. More
than 3000
plant

speciesincludi
ng vegetables,
bedding
plants, shrubs
and trees of
economic
importance

have
beenfound
susceptible to
root knot
nematode (
Meloidogyne

spp.) infection
(Agrios, 1997;
Abad
et al

., 2003).
1.2.5.

Disease cycle
of Root knot
nematode

All nematodes
pass through
an embryonic
stage, four

juvenile
stages (J1-J4)
followedby an
adult stage.
Although root
knot
nematodes

are obligate
parasites, but
they have
abilityto stay
alive in the
form of eggs
in the soil for

several years.
Depending
upon the host
andenvironme
ntal
conditions,
female

nematode lay
about 2001500 eggs in
a gelatinous
matrixknown
as egg-mass
(Luc

et al

., 2005;
Ashoub and
Amara,
2010).The first
moult is
completed

inside the egg

and second
stage
Meloidogyne

juvenileparasi
tes, the
infective

stage, hatch
from the eggs
under
favorable soil
conditions and
movethrough
soil in search

of hosts.
Newlyhatched
juveniles have
a short freeliving stage in
thesoil, in the

rhizosphere of
the host
plants. Second
stage
juveniles do
not feed

during their
free
11living stage
and only use
lipids that are
stored in their
guts. They

invade in the
rootelongation
region and
migrate in the
root until they
became
sedentary. The

juveniles feed,
byinserting
their stylet, on
the giant cells
24 hours after
becoming
sedentary and

release
salivainto the
cell. Second
stage juvenile
then
undergone
morphological

changes and
becomesaccat
e. After three
moults, it
becomes adult
and a mature
globose

female with
400 to
1000m in
length is
formed after
four moults
(Mitkowski

and Abawi,
2003). In
mature
femalesthat
are spherical
oval shaped,
feeding is

resumed and
reproductive
system is
developed.
Thelife
duration of
mature female

may be
extended to
three months
depending
upon the
hostplant and
the

environmental
conditions.
Mature female
breaks the
root epidermis
of the
hostplant

through its
posterior
region and
eggs are
deposited
along with a
gelatinous

material
toform an egg
mass. Female
nematode
have the
ability to
continue egg

laying after
harvest
of aerial plant
parts and the
survival stage
between crops
is generally

within the
egg. Life
cycleof
Meloidogyne
incognita

is completed
in 27 days at

25 C, but the
duration of life
cycle maybe
shorter or
longer, below
or above this
temperature

(Agrios, 2005;
Hunt
et al

., 2005).
Figure 1.2.1:
Life/ disease
cycle of Root

knot
nematode (

Meloidogyne
spp.)Agrios
(2005)

12
1.2.6.

Symptomolog
y

Root-knot
nematode
infections
leads to the
decline in the

host plant,
and under
certainconditi
ons, may kill
the plant.
Plants
infected by

root knot
nematode
may show
stuntedgrowth
and chlorosis
and plant
usually wilt

easily. The
extent of
damage
caused by
rootknotnematode
infections

varies with
host, timing of
infection, soil
type,
environmental
conditionsand
cultural

conditions.Roo
t-knot
nematode
infection often
is easy to
identify
because of the

swellings
inroots which
look like
knots.
These knots
become large
and easy to

see on some
hosts suchas
potato, but
may be
smaller and
less obvious
on others

plant species
such as chili
pepper.Multipl
e infections on
one root result
in a swollen
and rough

appearance.
Rootknotnematode
s are
microscopic
organisms
that cannot be

seen through
necked eyes.
1.2.7.
Root knot
nematodes

and Plant
Damage

Root knot
nematodes
have a
needle-shaped
mouthpart

called as
stylet
Nematodepus
hes this stylet
into plant cells
and injects
enzymes that

break down
the cell
contents.
Itthen takes
out the liquid
contents with
the help of the

hollow stylet
to its digestive
tract(Crow,
2011). Mostly
all plantparasitic
nematodes

stay in the soil

as
endoparasites
orectoparasite
s and feed on
roots of the
host plan.

Foliar-feeding
nematodes
move
throughwater
films from soil
onto and into
stems or

leaves of the
host plants.
Some rootfeedingnemat
odes have the
ability to
attack other

underground
plant parts
such as bulbs
and
tubers(Agrios,
2005).Root
knot

nematodes
can attack on
a wide range
of crops
through
different ways
suchas

inducing their
host to
produce
nutrients for
nematode
survival, or
enlarging the

structuresin
which the
nematodes
live, or both.
Nematodes
can also
produce

metabolites
that
destroyhost
tissue
(Meadows
et al

., 1989). In
this way,
nematodes
remove
energy to
support
theiractivities.

Plant-parasitic
nematode
infection can
cause severe
damage that
leads to
yieldreduction

, poor market
value or even
total crop
failure (Osei
et al

., 2011).Root
knot

nematode
infection
cause
wounding that
can create
openings
throughwhich

other
pathogens
may enter
the plant
causing
physical
disruption of

the vascular
tissues.Plants
with damaged
roots are
unable to
accumulate
nutrients and

water
efficiently
making
13the plant
more
susceptible to
heat stress,

drought, and
nutrient
deficiencies
(Taylor,
2003).Damag
ed roots are
also

susceptible to
infection by
bacteria,
viruses and/or
fungi (Acular
et al

., 2011). Most
of the plant
parasitic
nematodes
can also serve
as a vector of
several

viruses(Calver
t, 2008).
Nematodes
infection leads
to the
diversion of
the nutrients

and
metabolicactiv
ities from
normal,
healthy
growth and
fruit

production
(Anwar,
1995).
1.2.8.
Losses
caused by

root knot
nematodes

Root knot
nematodes
are considered
as an
important

threat to
agricultural
productionall
over the world
including
Pakistan. They
are key

damaging
pests of
agricultural
crops,
thatattacks a
wide range of
crops (Luc

et al

., 2005).
Damage
caused by
root knot
nematode
isrecorded up

to 60-80
percent in
agricultural
plants
(Vasyukova
et al

., 2009). An
estimatedann
ual losses of
an amount of
$80 billion to
over $100
billion

annually has
been
reported(Agrio
s, 2005). In
heavily
infested fields,
root knot

nematode is
responsible to
cause
cropyield
losses from
90% to almost
total crop

failure (Agu,
2006).
Meloidogyne

spp. also
formssynergy
with
pathogenic

bacteria and
fungi which
cause
significant
yield losses
(Rivera
andAballay,

2008).Keeping
in view the
above
mentioned
facts, this
study aimed
at the

following
objectives:

Exploration of
source of
resistance
among

different local
cultivars of
potato
againstroot
knot
nematode (

Meloidogyne
incognita

).

Analysis for
biochemical
changes (NPK

contents) in
the leaves of
the
plantsinoculat
ed with root
knot
nematode (

Meloidogyne
incognita

).

Determination
of efficiency of
Humic acid,

plant
protector
containing
benzoic
acidand
micro-power
against root

knot
nematode (
Meloidogyne
incognita

).
14

CHAPTE
R

II
VARIETAL
RESPONSE
OF

DIFFERENT
POTATO
CULTIVARS
AGAINST
ROOT KNOT

NEMATODE
(
MELOIDOG
YNE
INCOGNITA
)

2.1.
ABSTRACT

Research wok
was designed
to identify
resistant

potato
germplasm
against root
knotnematode
(
Meloidogyne
incognita

) infection. A
field trial was
conducted in
the research
areaof
Department of
Plant

Pathology,
University of
Agriculture
Faisalabad.
Thirty six
(36)potato
verities/

cultivars
relocated five
times were
sown in four
years sick plot
containingroot

knot
nematode (
Meloidogyne i
ncognita

) in RCBD
layout. Root
knot

nematoderepr
oduction and
host damage
was accessed
by recording
nematode root
galls and egg

massindices,
root weight,
shoot weight, ,
number of
leaves, fruit
weight, rate of
reproduction

andfinal
population of
nematodes.
Experiment
revealed a
considerable
variation in

responseagain
st
Meloidogyne i
ncognita

infection
among the
genotype

tested but
none of the
singlecultivar
was immune.
The cultivar
FD-19-2 was
highly

susceptible
followed by
FD-8-1, SH692 and SH-5.
All other
cultivars had
less galling

index with low

fecundity rate
indicatingtheir
ability to
suppress the
adult female
reproduction.

The cultivar
FD-1-3 scored
leastnumber
of galls and
egg mass
indices
followed by

FD-49-62, SH339 and SH332.

Keywords:

root knot
nematode,

Meloidogyne
incognita,
screening,
potato.
2.2.

INTRODUCTIO
N

Potato (
Solanum
tuberosum

L.) is perennial
solanaceous
vegetable
crop also
knownas
nightshades.
It is grown in

140 countries
of the world
(Haase, 2008).
It endures
significantyiel
d losses due
to biotic and

abiotic
stresses
(Agrios, 2005).
It is attacked
by
numerousfung
al, bacterial,

viral and
nematode
diseases that
reduce the
both quality
and quantity
of theproduce.

Among
biological
agents,
nematodes
especially root
knot
nematodes

(RKN)
inflictheavy
yield
reduction
(Anwar
et al

., 2009). Root
knot
nematodes
are parasitic
on a
broadrange of
vegetable

crops. They
are considered
as the most
important
pathogens
found
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