World Research Journal of Agricultural Sciences

WRJAS

Vol. 2(2), pp. 022-027, November, 2015. www.premierpublishers.org. ISSN: 2326-7266x

Research Article

Identification and pathogenicity of fusarium and

phomopsis foliar diseases of Jatropha curcas L. in
North-west States of Nigeria
*1

A.M. Nasiru 2O.O. Banwo 1A. D. Isah and 2A. B. Zarafi

*1

Department of Forestry and Environment Usmanu Danfodiyo University, Sokoto, Nigeria.

Department of Crop Protection, Ahmadu Bello University, Zaria, Nigeria.

Research on foliage disease of Jatropha curcas was conducted in Sokoto, Kebbi and Zamfara
States of Nigeria to determine the occurrence, incidence and severity of the diseases. Fusarium
and Phomopsis species were the fungal pathogens found to be responsible for the disease on J.
curcas in the study area. A spore count of the isolates was made and used as inocula in the
pathogenicity trial in glasshouse of the department to prove Kochs postulate. Results from the
farmers field revealed that, highest incidence (81.00%) and severity (53.33%) of Phomopsis leaf
blight was recorded in Tsaki of Sokoto State, while Janbaki in Kebbi State had the highest
incidence (75.33%) and severity (60.00%) of Fusarium leaf blight. The surveys conducted
showed that, J. curcas planted in lowland areas tend to be more prone to the fungal leaf blight
particularly those close to water source. In the pathogenicity trial, results indicated that, there
was no significant difference in the methods of inoculation and number of days after inoculation
with respect to incidence and severity of leaf blight. It is recommended that fungicides that can
be used for the management of fungal leaf blight of J. curcas should be identified.
Keywords: Foliar diseases, jatropha curcas, fungal, blight, fusarium.

INTRODUCTION
Jatropha is a genus of approximately 175 succulent
plants, shrubs and trees (some are deciduous, like
Jatropha curcas) from the family Euphorbiaceae. The
name is derived from the Greek words jatros, meaning
physician, and trophe meaning nutrition, hence the
common name physic nut (Anon, 2009). There are
several different species of Jatropha which are commonly
grown in gardens or as ornamental plant. Many of these
Jatropha species produce very colourful flowers and are
huge butterfly and bee attractants. Examples include
Jatropha integerrina, J. cardiophylla, J. catharlica, J.
cinerea, J. cuneata, J. podagrica etc. Jatropha curcas is
a species of flowery plant and native to the American
tropics, most likely Mexico and Central America (Janick
and Robert, 2008).
Of the over 175 species in the genus Jatropha, Jatropha

curcas is the most popular, because the seeds are used

for biofuel and as alternative energy source (Joshua,
2010). About one-third of the energy in the fruit of
Jatropha can be extracted as oil that has a similar energy
value of diesel fuel and can be used directly in diesel
engines added to diesel fuel as an extender or transesterified to a bio-diesel fuel (Anon., 1998). Jatropha is
not browsed, for its leaves and stems are toxic to
animals, but after treatment, the seeds or seed cake
could be used as an animal feed. Being rich in nitrogen,
the seed cake is an excellent source of plant nutrients.
*Corresponding author: Department of Forestry and
Environment Usmanu Danfodiyo University, Sokoto,
Nigeria.
Email:
amnafad@yahoo.com
Tel:
+234(0)8034566086

Nasiru et al.

022

Various parts of the plant are of medicinal value; its bark

contains tannin, the flowers attract bees and thus the
plant has honey production potential (Mc Graw,
2003).Disease assessment also called phytopathometry
is the most important aspect in any crop loss programme
because it is the process that generates data to quantify
disease progress, three parameters used for the
assessment or measurement of plant diseases are;
disease incidence, disease severity and crop yield loss
(Marley, 2013). Disease incidence is the simplest method
of assessment which basically involves the counting of
the number of diseased plants and expressed as a
percentage, while disease severity is assessed by
estimating the proportion of total photosynthetic area that
is diseased. In other words, disease severity is a
quantitative trait which measures the amount of disease
on a plant in terms of intensity of symptoms or damage.
Among the most important challenges facing man and his
environment today is global warming which is attributed
to excessive exhausts from combustions in vehicles,
companies, industries, households etc. Among the
effects of global warming are drought, flood, erratic
rainfall, uncondusive atmosphere to man and other living
organisms due to high rate of radiation etc. The full
potential of Jatropha curcas is far from being realized
particularly in Nigeria because of several reasons as
pests and diseases, technical, economic, cultural and
institutional etc which need further discussion and
examination. The growing and management of Jatropha
curcas, either on private, public or community land is
poorly documented and there is little field experience that
is being shared especially in Nigeria where it is still grown
wild.
The aims of this study are to isolate and identify the foliar
diseases of Jatropha curcas, determine incidence and
severity of fungal foliar diseases on Jatropha curcas in
farmers field and pathogenicity of the isolated organisms

MATERIALS AND METHODS

Study Area
The study was conducted in some parts of Sokoto, Kebbi
and Zamfara identified to have high population of
Jatropha curcas growing in the wild. The area lies in
0 1
north-west part of Nigeria between latitudes of 10 2 N
o
1
o 1
o 1
and 13 9 N and longitudes 3 7 E and 7 2 E (UNO,
2004). The areas exhibit tropical dry climate where
rainfall is 550-650mm recorded for only three to four
months (June September) while the rest of the year is
dry and hot (Anon., 2010). According to Nigerian
Metrological Service Report (2009), mean maximum
o
o
temperature ranges from 25 C to 43 C and mean
o
o
minimum temperature is between 19 C and 24 C. The
area falls within the Sudan savanna characterized by
short grasses and scattered thorny short trees (Anon.,
2011).

Sampling Technique
A purposive and multi-stage sampling was employed to
identify the Local governments areas and villages in
North-West States of Nigeria with high population of
Jatropha curcas from the list of assertions obtained from
the Institute of Agricultural Research (IAR) ABU, Zaria.
Tsaki, Kajiji and Barkeji in Kware, Shagari and Tambuwal
Local Government Areas respectively were selected from
Sokoto State, while in Zamfara State, Wanke, Nasarawar
wanke in Gusau Local Government Area and Yartukunya
in Bungudu Local Government Area were selected. In
Kebbi State were Basaura (Janbaki), Jega Birni (Gadar
Ruwan Kanwa) in Jega Local Government Area and
Gangije in Aliero Local Government Area were selected.
Disease Sample Collection
Occurrence of foliage diseases of Jatropha curcas was
determined in the selected areas, and disease incidence
and severity were determined. Ten diseased leaf
samples of Jatropha curcas were randomly collected and
labelled from three different sites in each of the locations
in line with Timothy et al. (1999) procedure and
preserved for isolation in the Pathology Laboratory,
Department of Crop Protection, ABU, Zaria.
Assessment of Incidence and Severity of Diseases of
Jatropha curcas
Disease incidence is the number of plant units infected,
expressed as a percentage of the total number of units
assessed as follows;
Disease incidence I =
Number of infected plant units
100.
Total number

healthy and infected of units assessed

Disease severity scores were obtained using 1-5 scale

adapted from Zelalem, et al. (2012), Ghosh, et al. (2010),
and Tarr, (1981) where;
Disease severity %
Sum of all leaves disease rating
=
Total number of leaves examined x maximum rating
100
1.
No symptoms on leaves
2.
1-25% number of leaves diseased
3.
26-50% number of leaves diseased
4.
51-75% number of leaves diseased
5.
76% or more number of leaves diseased
Disease severity %
Sum of all leaves disease rating
=
Total number of leaves examined x maximum rating
100
Isolation and Identification of Pathogens
The infected leaves were washed with distilled water, cut
into pieces and sterilized for five minutes using 0.5%

World Res. J. Agric. Sci.

sodium hypochlorite and rinsed thrice with sterile distilled
water. The pieces were then placed in 90 mm (diameter)
petridishes containing Potato Dextrose Ager with
streptomycin (PDAs) and labelled. The plates were
o
incubated at 28 C for 7 10 days. Fungal mycelia of the
isolated organisms were sub-cultured on fresh PDAs to
get pure culture. The cultural characteristics were noted
and detailed microscopic characteristics (morphological)
were observed. The fungi were identified using
Identification Manual of Barnett and Hunter (2006). The
isolated fungi were preserved in Marcoteny bottles
containing PDAs in a slanting position before the time of
inoculation.
Pathogenicity Trial in the Screen house

023

increase humidity. After 24 hours, the polythene bags

were removed for 20 minutes to aerate the seedlings and
were re-wet before replacement for another 24 hours and
finally removed. Seedlings were watered in the morning
and evening. Numbers of days as it affects blight
symptoms appearance on the leaves of Jatropha
seedlings with respect to the isolates were determined
through counting the number of infected leaves at seven
days. Effect of the methods of inoculation as it affects the
symptoms appearance was also evaluated.
Leaves showing blight symptoms were collected and
taken to laboratory for re-isolation and subculture to
obtain pure culture of the organism and then compared
with original preserved culture to confirm Kochs
postulates.

Certified seeds of Jatropha curcas were obtained from

Institute for Agricultural Research (IAR), ABU Zaria. The
seeds were soaked in sodium hypochlorite for five
minutes then washed with sterile water; again they were
washed with 20 ml of alcohol and rinsed with sterile water
to ensure safety against dust and other pathogens that
may be present in the surface. Thirty-nine clay pots with
diameter and depth of 25 cm and 24 cm respectively
were washed, filled with heat sterilized soil and watered.
Two seeds were sown in each pot and watered for 28
days under aseptic condition to prevent contamination.
The seedlings were later thinned to one.

Statistical Analysis

Inocula Preparation and Inoculation of Seedlings

Incidence and severity of Phomopsis and Fusarium leaf

blight on leaves of Jatropha curcas in the study area is
presented in Table 1. The results depicts that, among the
locations, Janbaki in Kebbi state recorded the highest
incidence of Phomopsis blight (81.67%), this was
statistically similar to those obtained from Tsaki in Sokoto
State and Gadar Ruwan-Kanwa in Kebbi State which
recorded 81.00% and 71.67% respectively, while lowest
incidence was obtained in Wanke (33.00%)
and
Nasarawar wanke (34.00%) in Zamfara State. Severity of
Phomopsis blight on leaves of Jatropha curcas in Table 1
revealed that, highest value was obtained in Gadar
Ruwan Kanwa, Janbaki and Tsaki which recorded
53.33% each. This was followed by the value obtained
from Kajiji (40.00%) that was significantly higher (P <
0.05) than those obtained from Gangije (33.33%) and
Barkeji (26.67%); which were statistically similar. Wanke
and Nasarawa Wanke had the least severity of
Phomopsis leaf blight (20.00%) each.

The preserved pure cultures of the isolated pathogens

were grown on PDA in the laboratory until they
sporulated. 10 ml of sterilized distilled water was added
to each petridish and grown mycelia mat from the culture
was harvested using a sterile scalpel. The mycelia were
blended in an electric blender for five minutes, 200 ml of
sterile distilled water was added in 500 ml conical flask
and filtered using a double layer muslin cloth. Spores
count was made using haemocytometer and compound
microscope using the equation;
4
Spores/ml = (n) x 10
where n = the average cell count per square of the four
corner squares counted.
Jatropha seedlings were inoculated outside the glass
house to avoid contamination using three methods viz;
smear, spray and soil inoculations.
Smearing was
conducted through direct application of harvested mat on
leaves using finger tips fitted with gloves. The soil
inoculation was made by pouring 20 ml of inocula
suspension at the base of stem (root collar) of each plant.
Inoculation through spray was carried out using hand
atomizer to spray on the leaves. Seedlings for control
were treated with sterile distilled water only. One seedling
per pot and three pots for each inoculation method for the
four isolated pathogens were labelled and arranged in
completely randomized design. After the inoculation,
seedlings were covered with wet polythene bags to

The data collected were subjected to analysis of variance

(ANOVA) procedure using SAS (2012) software.
Significant difference in the treatment means were
separated using Duncan Multiple Range Test (DMRT).

RESULTS
Incidence and Severity of Fungal Blight on Leaves of
Jatropha curcas

Pathogenicity of the Isolated Organisms

The values of spores count obtained from each of the
fungal isolates used for inoculation were as follows;
4
- Fusarium species = 1180 x 10 spores/ml
4
- Phomopsis species = 34.75 x 10 spores/ml
Results on leaf blight symptoms appearance on the
leaves of Jatropha curcas seedlings at 7 days after
inoculation is presented in Table 2. The results revealed

Nasiru et al.

024

Table 1. Incidence and severity of Phomopsis and Fusarium Leaf blights

Locations
Kajiji
Tsaki
Barkeji
Wanke
N/ Wanke
Yartukunya
Janbaki
Gangije
Gadar Ruwan Kanwa
SE
Significance

Phomopsis
Incidence (%)
67.672.52ab
81.008.19a
43.3315.28cd
33.002.65d
34.001.00d
36.674.16cd
81.672.52a
52.3319.66bc
71.6710.12a
22.03
*

Severity (%)
40.000.00ab
53.3311.55a
26.6711.55bc
20.000.00c
20.000.00c
22.000.00c
53.3311.55a
33.3311.55a
53.3311.55a
57.75
*

Fusarium
Incidence (%)
74.334.04ab
62.6715.14ab
70.6710.59ab
66.6713.32ab
70.6731.09ab
62.678.14ab
75.3311.59a
71.676.03ab
45.0018.03b
39.32
*

Severity (%)
60.000.00a
46.6711.55ab
53.3311.55ab
46.6711.55ab
53.3330.55ab
46.6711.55ab
60.0020.00a
40.000.00ab
26.6711.55b
36.1
*

Means followed by the same letter(s) do not differ significantly according to Duncan Multiple Range Test (DMRT) at 5% level of
significance.

Table 2. Leaf blight symptoms appearance on

the leaves of Jatropha curcas seedlings in
glasshouse at 7 Days After Inoculation

Pathogens
Fusarium species
Phomopsis species
Control
SE
Significance

Occurrence (%)
0.671.03a
0.390.78ab
0.000.00b
1.05
*

Figure 1. Phomopsis leaf blight on seedling of Jatropha curcas planted in glasshouse at

IAR, Zaria

that, Phomopsis species (Figure 1) require fewer days

(0.39) to appear on the leaves after inoculation. This
could be attributed to the continuous wetting of incubated
seedlings through watering which favours the germination
and penetration of conidia. Fusarium (Figure 2) require
longer time (0.67) days to manifest their symptoms on the
leaves of seedlings. This could be due to the fact
Fusarium is within the soil inhabiting fungi (Agrios, 2004)
in which it starts inflicting from the roots through the stem
to the leaves, while on the other hand, pathogenicity trial
was conducted when the temperature was low at IAR,
Zaria and seedlings were under regular watering. Control
seedlings did not produce any disease symptoms during
the period under observation.

Effect of methods of inoculation on the appearance of

fungal pathogens is shown in Table 3. It was revealed in
the result that, there was no significant difference among
the methods (soil inoculation, spray and smear) used in
the pathogenicty trial. Similarly, results of general blight
symptoms appearance on leaves of seedlings of
Jatropha curcas as influenced by number of days after
inoculation irrespective of the pathogen type is presented
in Table 4. It shows that, there was no significant effect of
number of days after inoculation and the fungal pathogen
appearance on the leaves of Jatropha seedlings. Though
not significant, 17 Days After Inoculation (DAI) recorded
the highest value of fungal appearance (0.73) which was
closely followed by 11 DAI. The least fungal pathogen

World Res. J. Agric. Sci.

025

Figure 2. Fusarium leaf blight on seedling of Jatropha curcas planted in glasshouse at IAR,
Zaria.

Table 3. Effects of inoculation methods on fungal blight appearance on the leaves of Jatropha curcas
seedlings at IAR, Zaria

Inoculation Methods
Soil inoculation
Spray
Smear
SE
Significance

Occurrence (%)
0.470.90
0.370.62
0.330.71
1.29
Ns

Table 4. Results on the general symptoms appearance of fungal pathogens on the leaves of Jatropha curcas seedlings
as influenced by number of Days After Inoculation (DAI)

Days After Inoculation (DAI)

7
9
11
13
15
17
SE
Significance

symptom on the leaves of seedlings was recorded at 9

and 13 DAI with 0.20 each.

DISCUSSION
The high incidence and severity of Phomopsis leaf blight
obtained could be attributed to the fact that, Jatropha
curcas planted in those areas are in Fadama lands and
along streams; normally fungi are found in damp areas.
Also moisture is indispensable for the germination of
fungal spores and penetration of the host by the germ.
Agrios (2004) opined that moisture in such forms as
splashing rain and running water also plays an important
role in the distribution and spread of many pathogens like
bacteria, fungi and nematodes on the same plant and
their spread from one plant to another, he added that,
moisture increases the succulence of the host plants and
thus their susceptibility to certain pathogens, which
affects the extent and severity of disease. Shaner (1981)

Occurrence (%)
0.270.60
0.200.41
0.530.92
0.200.41
0.400.83
0.731.03
1.68
Ns

stated that, in fungal diseases, moisture affects spore

formation, longevity, and particularly the germination of
spore which requires a film of water covering the tissues.
Lowest incidence and severity of Phomopsis blight
obtained in Wanke, Nasarawa Wanke and Yartukunya in
Zamfara State could be attributed to the fact that,
Jatropha curcas in those areas were planted as fences in
upland farms assumed to have low soil moisture,
Shoeneweiss (1981) concluded that, the occurrence of
many fungal disease in a particular region is closely
correlated with the amount and distribution of rainfall
within the year, hence soil moisture can be said to be an
important factor for the growth and development of the
pathogenic fungi.
Fusarium leaf blight in Table 1 shows that, Janbaki
recorded significantly highest incidence (75.33%). This
was followed by Kajiji (74.33%), Gangije (71.67%),
Barkeji (70.67%), Nasarawa wanke (70.67%), Wanke
(66.67%), Tsaki and Yartukunya with 62.67% each which
were all statistically similar while lowest incidence was

Nasiru et al.

026

obtained in Ruwan-Kanwa with 45.00%. Fusarium blight

recorded the highest incidence in area away from water
source (stream), this shows that Fusarium species may
be among pathogenic fungi that do not require moisture
to induce infection as far as there is high relative humidity
in the atmosphere surrounding the plant. Agrios (2004)
stated that, some pathogenic fungi require free moisture
on the host or high relative humidity in the atmosphere for
spores release or for germination of their spores; he
added that most pathogens become independent of
outside moisture once they can obtain nutrient and water
from the host. The severity of Fusarium blight was
highest in Kajiji and Janbaki with 60.00% each. This was
followed by what was obtained from Barkeji, Nasarawa
Wanke, Tsaki, Wanke, YarTukunya and Gangije which all
recorded statistically similar values. The lowest severity
of leaf blight (26.67%) was obtained in Gadar Ruwan
Kanwa of Kebbi State. The severity of Fusarium blight
was statistically different within the locations possibly due
to aforementioned factors, but amongst the states the
difference was not significant probably because they
share the same climatic factors and vegetation (Sudan
Savannah). Fusarium species is within the soil inhabitant
fungi that are able to survive indefinitely as saprophytes
(Carly et al., 2001). Agrios (2004) stated that, the genus
Fusarium is a soil borne necrotrophic, plant pathogenic
fungi with many species that cause plant disease around
the world, the fungus can survive in the soil as mycelium
or as spores in the absence of its host.

CONCLUSION
Results of the study showed that, most of the leaves of
Jatropha curcas in the areas studied were infected with
fungal leaf blight caused by Fusarium and Phomopsis
species. Tsaki in Sokoto State recorded the highest
incidence and severity of Phomopsis leaf blights, while
Janbaki in Kebbi State had the highest incidence and
severity of Fusarium leaf blight. Jatropha curcas planted
in lowlands were found to be more prone to infection than
those planted in uplands, particularly those that are close
to streams. Based on the findings of the research,
following recommendations were made; Identify
fungicides that can be used for the management of leaf
blight disease of Jatropha curcas. Also In vitro and In vivo
evaluation of the chosen fungicides should be conducted
with the aim of identifying the most effective among them
and the appropriate dosage for application.

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Accepted 13 October, 2015.
Citation: Nasiru AM, Banwo OO, Isah AD, Zarafi AB
(2015). Identification and pathogenicity of fusarium and
phomopsis foliar diseases of Jatropha curcas l. in Northwest States of Nigeria. World Research Journal of
Agricultural Sciences 2(2): 022-027.

Copyright: 2015 Nasiru et al. This is an open-access

article distributed under the terms of the Creative
Commons Attribution License, which permits unrestricted
use, distribution, and reproduction in any medium,
provided the original author and source are cited.

027