Studies on Growth Disorders of Pistachio Caused by Root-knot Nematode
(Meloidogyne javanica ) in Mahvelat Iran
A. Fazeli Salmani, A. Tehranifar, G. Davarynejad, E. Mahdikhani and A. Mokaram
Department of Horticultural Science, Ferdowsi University of Mashhad, Mashhad, Iran
Department of Plant Protection, Ferdowsi University of Mashhad, Mashhad, Iran
Keywords: pistachio, Meloidogyne javanica, Razavi Khorasan province
Abstract:
Pistachio is one of the most important of horticultural products in Iran.
Numerous of diseases are affected to pistachio in the world. Gummosis, verticillium wilt
and root-knot nematodes are the most important diseases in the world. Root-knot
nematodes, Meloidogyne species, are endoparasitic nematodes that reduced products
and cause structural, physiological and biochemical disorders in plants. Severity of
damage is depended on species and intensity population of nematode. In this research,
pistachio gardens of Mahvelat region (Razavi Khorasan province – Iran) with two
cultivars, Ohadi and Badami, were studied. Meloidogyne javanica was found in the area
with high populations. The experiments were setup with 50 replications in 2×5 factorial
experiment in an imbalanced Completely Randomized Design. Number of galls and egg
mass on the roots were measured. Amount of leaf chlorophyll, leaf area, leaf necrosis,
tree height, east-west spread, north-south spread, growth of annual branch, diameter of
annual branch and fruit yield were determined. Results showed that with increasing of
gall index/egg mass, growth characteristics of plant were significantly reduced
according to Tukey test. There was no difference among the treatments for leaf necrosis
and diameter of annual branch. Amount of chlorophyll and fruit yield in Badami
cultivar were decreased significantly comparing with Ohadi cultivar.
ITRODUCTION
Pistachio is a member of family Anacardiaceae and genus pistacia. About 11 species are
recognized in this genus that P. vera is the most important commercial cultivar (11).
Production level (more than 350,000 hectares), high internal consumption and considerable
situation in worldwide market have made pistachio one of the most important agricultural
products in Iran. Also Iran is the biggest producer and first exporter of pistachio in the world
(9). Nematodes are plant parasites especially root-knot nematode (Meloidogyne spp.) that
widely distibuted and cause to reduction of yield (19). It is estimated that loss of yield
approximately $100 billion in the world (13). Meloidogyne spp. are the most important
nematode from aspect of lesion, they are one of fifth factor of plant diseases in the world
1
(3,21). Approximately 30 diseases are reported for pistachio in the world. Soil originated
diseases especially Gomoz, Vertisiliomic wiliting and root-knot nematode of Meloidogyne
spp. are the most important pistachio diseases in the world (17). Root-knot nematodes were
recognized in pistachio gardens of Rafsanjan in 1345 for the first time. Farivar mahin did
some researches from aspect of biology, chemical treatment and susceptibility of different
kinds rootstock pistachio to root-knot nematode during 1986-1987 and result showed that
species of M. incongita, M. arenaria and M. javanica present in Kerman(7). Sajoudi et al.
(2002) with collection of pistachio roots contaminated with root-knot nematode in Neyriz
Fars recognized M.halpa race 1, M. arenaria race 2and M. javanica(20). Askarian et al.
realized that dominant species in Kerman was M. javanica and it was 88% of nematode
population (2).
Due to root-knot nematode can cause disorder in absorption and transportation of water and
nutrients by plant, they can cause stress in plant even in low contamination. Inoculation of
root-knot nematode in eggplant without humidity stress resulted in colorosis and reduction of
pigment synthesis (19). Amount of plant destruction influenced by species and population of
nematode. Khorasan provinces have got third place in terms of pistachio production in Iran.
Developing position of pistachio production in Razavi Khorasan province makes some
researches about restriction factors like root-knot nematode necessary.
The objective of this research was to study of growth disorders in pistachio trees
contaminated with root-knot nematode in Mahvelat region (Razavi Khorasan province –
Iran).
MATERIAL AND METHODS
Sampling did in orchards of Mahvelat of Razavi Khorasan, Iran in 2009. Orchards with two
cultivars of Ohadi and Badami were selected. Root samples at shadow zone of contaminated
trees from subsidiary roots with soil were collected. All criteria about collected sample were
written on the bag. Samples were transferred to laboratory and stored at 4˚c.
Recognition of root-knot nematode species: At first, root samples were washed and female
nematode was extracted and after preparation of cuticle reticulum of hind body, species was
realized.
2
Evaluation of growth factors in plants: tree height, east-west spread, north-south spread,
growth of annual branch, diameter of annual branch and fruit yield were recorded. Amount of
leaf chlorophyll, total leaf area and leaf necrosis were measured by SPAD and leaf area
meter, respectively.
Calculation of gall index and egg mass: A 5g sample of subsidiary roots was selected from
each tree (replicate) and number of egg mass based on index of 0-4 was evaluated. Root gall
intensity was evaluated as o: without gall, 1: 1-25%, 2: 26-50%, 3: 51-75%, 4: 76-100%
galls. The experiment was conducted as a 2×5 factorial in an imbalanced Completely
Randomized Design (CRD). Analysis of variance was conducted using GLM procedure of
SAS 9.1(2004). Tukey’s test was used to compare means.
RESULTS
Microscopic observations showed that M. javanica was species of root-knot nematode in this
study. M. javanica is one of four important species of nematode that is universally spread. It
is distributed among different horticultural crops throughout Iran (16).
Leaf chlorophyll: These results indicated that there was significant difference (p<0.01)
between two cultivars. Amount of chlorophyll in Ohadi was more than Badami (Table 2).
Also, it was observed that there was significant difference (p<0.01) between contaminated
trees and control. The lowest chlorophyll was observed in indexes 3 and 4(Table 3).
Tree height, north- south and east-west spread: ANOVA showed that effect of gall/egg mass
on tree height was significant (p<0.01). The least height was observed in indexes of 3, but
there was not significant difference between indexes 3 and 4 (Table 3). There was no
significant difference between cultivars and interaction effect between gall/egg mass index
and cultivar (Table 1). Measurements of north-south spread indicated that there was
significant difference (p<0.05) among gall indexes and the least spread was for index 4
(Table 3). But there was no significant difference between two cultivars and interaction.
There was significant difference (p<0.01) about east-west spread for gall effect and effect of
cultivar (p<0.05), but their interaction was not significant (Table 1).
3
Leaf necrosis and diameter of annual branch: ANOVA showed that main effects and their
interaction (gall/egg mass × cultivar) were not significant for leaf necrosis and diameter of
annual branch (Table 1).
Leaf area: These results indicated that effect of gall/egg mass on leaf area was significant
(p<0.01) and indexes of 3 and 4 had the least leaf area. There was not significant difference
between these two indexes (Table 3).
Growth of annual branch: results showed that effects of gall or egg mass and their interaction
with cultivar were significant (p<0.05) (Fig1).
Yield: results showed that effect of gall/egg mass (p<0.01), cultivar and its interaction with
gall/egg mass were significantly different (p<0.05) (fig 2).
Table 1 Mean square ANOVA growth factors in two cultivars with five gall/egg mass index
S.O.V
DF
tree
height
(cm)
leaf
chlorop
hyll
diameter
of
annual
branch
(mm)
growth
of
annual
branch
(cm)
leaf area
leaf
necrosis
northsouth
spread
(cm)
eastwest
spread
(cm)
Yield(kg)
Cultivar
1
426/1 ns
389/3**
1/76 ns
./44 ns
6154/6 ns
323/81 ns
3255/3 ns
6866/7*
23/61*
Gall/egg
mass
index
cultivar×
gall/egg
mass
Error
4
2920/9**
360/9**
./64 ns
148/6**
55730/5**
3168/9 ns
4192/9*
4208/2**
168/65**
4
597/5 ns
46/8 ns
./38 ns
29/2*
10763/5 ns
2061/7 ns
305/9 ns
216/5 ns
13/30*
40
537/8
38/1
./51
10/88
4969/22
1472/7
1265/9
1071/7
4/92
Table 2 Mean comparison between cultivars for growth factors
Cultivar
tree
leaf
diameter growth leaf
height(cm) chlorophyll of
of
area
annual
annual
branch
branch
(mm)
(cm)
4
leaf
northeast-west
Yield(kg)
necrosis south
spread(cm)
spread(cm)
Ohadi
211/9 a
43/57 a
4/86 a
a
b
a
10/44 a 264/54 48/82 a
226/31 a
206/03 b
5/18 a
243/18 a
230/53 a
3/75 b
a
Badami
205/8
37/74
5/26
10/64
a
241/34 54/4 a
a
indexs
Table 3 Mean comparison of gall/egg mass index levels for growth factors
tree
leaf
diameter growth leaf
leaf
northeast-west
Yield(kg)
height(cm) chlorophyll of
of
area
necrosis south
spread(cm)
annual
annual
spread(cm)
branch
branch
(mm)
(cm)
0
229/29 a
48/3a
5/32a
16/87a
365/45a 29/56a
262/02a
246/91a
10/08a
1
226/67 ab
45/38ab
5/34a
12/09b
269/60b 39/46a
243/66ab
223/27ab
7/93b
2
204/87bc
41/46b
4/81a
9/54bc
278/19b 45/72a
214/98b
225/21ab
3/49c
3
187/76c
33/48c
5a
7/48c
170/13c 75/35a
240/45ab
207/30bc
./75d
4
195/68c
34/68c
4/83a
6/70c
181/34c 67/33a
212/62b
188/70c
./07d
Table 4 Mean number of gall and egg mass in 5g root of two cultivars of pistachio
C1:Ohadi
g0: without
g1: 1-25%gall
g2: 26-50%
g3: 51-75%
g4: 76-100%
gall
Mean number of gall in
0
51
88/57
157/83
210/75
5g root
Mean number of egg
0
29
61/42
109/33
164
mass in 5g root
C2: Badami
g0: without
g1: 1-25%gall g2: 26-50%
g3: 51-75%
g4: 76-100%
gall
Mean number of gall in 0
28/85
88/33
181/33
222/8
5g root
Mean number of egg
0
10/85
61/83
136/3
167/6
mass in 5g root
5
growth of annual branch(cm)
20
a
18
16
ab
abc
14
12
bc
10
bc
bc
bc
8
bc
c
6
c
4
2
0
c1g0
c2g0
c1g1
c2g1
c1g2
c2g2
c1g3
c2g3
c1g4
c2g4
interactive effects of cultivar×gall/egg mass
Fig. 1 Interactive effects of cultivar×gall/egg mass on growth of annual branch
(c1: Ohadi, c2: Badami)
12
a
a
a
yeild(kg)
10
8
6
4
b
bc
2
bc
bc
bc
c
bc
0
c1g0 c2g0 c1g1 c2g1 c1g2 c2g2 c1g3 c2g3 c1g4 c2g4
interactive effects of cultivar×gall/egg mass
Fig. 2 Interactive effects of cultivar×gall/egg mass on yield
(c1: Ohadi, c2: Badami)
Discussion:
For optimum growth, a plant requires certain growth conditions both above and below the
soil surface, so that its physiological processes can produce new biomass. One of the
difficulties in examining the physiological processes is the complex nature of their
interactions and the influence of these interacting factors on host physiology and growth. The
complexity of these physiology processes is further influenced by nematodes and other
disease- causing agents. These effects vary with the species of host and parasite, population
size, host type and the age and stage of infection (15). The effect of Meloidogne on plant
growth and yield is varied and complex. The damage caused by this nematode on susceptible
plants probably involves several mechanisms. Nematodes remove plant nutrients, alter
nutrient flow patterns in plant tissue, and retard root growth, all of which may contribute to
6
suppressed plant yield (18). Root-knot nematode can cause reduction of plant growth. This
nematode is internal and non- invasive that can cause structural changes and physiological
and biochemical disorders in host plant and resulted in reduction of plant growth. Root
contamination cause to disorder in vascular system and prevent from absorption and
transportation of water and nutrients. Disorder in transportation system cause reduction of
foliar growth and leaf necrosis. Second age larve cause disorder in vascular system after
entrance to root and result is imbalance between root and stem and reduction of water and
nutrients absorption and photosynthesis (4,10). This reduction photosynthesis might be as a
result of two agents, at first, root lesion cause to reduction of water transportation. After that
leaf pores will closed and co2 entrance to leaf will reduced. Second agent is that reduction of
hormone synthesis and its transportation from root to other parts of the plant might be effect
upon photosynthesis (5). Wastage of net photosynthesis in tomato leaves, beans and grape is
affected by root-knot nematode (Melakeberhha et al. 1984). France et al. (1994) indicated
that contaminated bean with M. incongita significantly reduces chlorophyll, nitrogenous
compounds and carbohydrates, also in contaminated bean, absorption of water and nitrogen
reduced (8).
In our experiment in agreement with these results amount of chlorophyll
reduced in plants contaminated with M. javanica. Contamination of pistachio trees with rootknot nematode cause disorder in plant physiology and produce symptoms similar to nutrient
deficiency e.g. growth reduction, leaf necrosis or dwarfism (2). Also in this investigation,
height, growth of annual branch, diameter of annual branch, east-west spread, north-south
spread in contaminated trees reduced. Leaf necrosis was more in high polluted plants. In
agreement with our result, It has been reported that root-knot nematode of Meloidogyn Spp.
decreased fruit production in several commercial species of prunus like peach, almond, plum
and apricot (6).
Literature Cited
Agu, C.M. 2008. Effect of organic manure types on root – gall nematode disease and African
yam bean yeield. plant siences research 1(1):17-19.
Askarian, H. 2005. Genetical variation among populations of different root-knote nematodes
using distinctional hosts test and moleculr methods in Kerman province. M.Sc. Thesis. plant
pathology. Isfahan University of Technology.
7
3- Baicheva, O., Salkova, D., and Palazova, G. 2002. Root-Knot nematodes (Meloidogyne,
Goeldi,1887) Species composition, pathogenicity, some problems for investigation.
Experimental pathology and parasitology 10: 21-24.
4- Bergeson, G.B., Van Gundy, S.D., and Thomason, I.J. 1970. Effect of Meloidogyne
javanica on rhizosphere microflora and Fusarium wilt of tomato. Phytopathology 60: 1245 9.
5- Brid, A .F. 1974. Plant response to root – knot nematode. Ann. Rev.phytopatol. 12:69-84.
6- Esmenjaud, D., Minor, J.C., Voisin, R., Pinochet, J., Simard, M.H. and Salesses, G. 1997.
Differential response to root-knot nematodes in prunus species and correlative genetic
implications. j.nematol,29 (3):370 -380.
7- Farivarmahin, H. 1986. Survey of root-knote nematodes (Meloidogyne spp.) in Kerman
province. Proceedings of 8th Iranian congress of plant pathology. p136.
8- France, R.A., and Abawi, G.S. 1994. Interaction between Meloidogyne incognita and
Fusarium oxysporum f. sp. phaseoli on selected been genotypes. J. Nematol. 26: 467-474.
9- Gholipour, Y.2005. Applied biology of pistachio. Vol. 1.Flowering and Fruit production.
Tehran Sadra Press. 112pp.
10- Haghighi, H., A. Taheri, S. Razavi, Z. Tanhamaafi, and M. Mamghni.2008. Greenhouse
survey of interactive effects among two breeds of root-knote nematode (Meloidogyne
incognita) and funges Verticillium dahlia cause to seedling olive(olea europaea) wilting in
Gorgan. J. Agri. Sci. Natur. Resou. Vol. 15. N. 4.
11- Hokmabadi, H. 1998. Effects of different sugars on guantitative and qualitative traits of
Kalleh Ghouchi pistachio. M.Sc. Thesis. Tehran University
12- Ibrahim, I.K.A., Rezk, M.A., and Khalil, H.A.A. 1982. Effects of Meloidogyne incognita
and Fusarium oxysporum f.sp. vasinfectum on plant growth and mineral content of cooton,
Gossypum barbadense. Nematologica 28: 298-302.
13- Keren-zur, M., Antonor, J., Berconvitz, A., Feldman, K., Husid, A. ,Keran, G.,
Morcov, N., Rebhun, M. 2000.Bacillus furmus formulations for the safe control of root –
knot nematode .47-52.In:BCPC conference on pestes and Diseass, Brighton,UK.
14- Melakeberhha.,N. W., Ebstejr. ,M . & Brooke, R. C. (1984 ).Improved techniques for
measuring the CO2 exchange rate of Meloidogyne infected bean plants. Nematologica, 30 :
213-221.
15- Melakeberhan, H. 1986. The influence of M. incongita the physiology and yield of
phaselous vulgaris. M.S. Thesis of physiology. Simon Fraser University.
16- Mehdikhani, E., Kheiri, A., Mohammadi, M., Eshtiaghi, H., and Okhovvat, M. 2003.
Three new records of Meloidogyne species for Iran. Iranian Journal plant Pathology 39 (3,4).
189-211.
17- Ogawa, J.M. and H. English.1991; Diseases of temperate zone tree fruit and nut crop.
University of California, Division of Agriculture and Natural Resources. 461 pp.
18- Sasser, J.N. and Carter, C.C. 1985. An advanced treatise on Meloidogyne . Vo.1. 422pp
19- Shaukat S.S., Moazzam A.K., Ahmad W. and Shahina F.2009.effect of M.javanica
and moisture strees on growth and physiological response of
brinjal.pak.j.nematol,27(2):281-296.
20- Sojudi, M., H. Eshtiaghi, and Sh. Barouti. 2002. Identification of species and race of
8
root-knot nematode in Neyriz region of Fars province. Proceedings of 15th Iranian congress
of plant pathology. p251.
21- Tanhamaafi, Z., and Mahdavian, S.I. 1997. Identification of spices and races of root-knot
nematode(Melodogyne spp) on kiwi and effect of M. incognita on kiwi seedling. Pests and
Plant Diseases of Iran 65:1-11.
22-Wallace, H. R. 1971. The influenence of the density of nematode populations on plants.
Nematologica 17:154-166.
9