Vol.4, No.11, 608-613 (2013) Agricultural Sciences
http://dx.doi.org/10.4236/as.2013.411081
Induced systemic resistance to Meloidogyne spp by
β,amino butyric acid in tomato
Shamaael S. Mutar, Farkad A. Fattah*
Department of Plant Protection, College of Agriculture, University of Baghdad, Baghdad, Iraq;
*Corresponding Author: farkad.fatah@gmail.com
Received 17 September 2013; revised 16 October 2013; accepted 27 October 2013
Copyright © 2013 Shamaael S. Mutar, Farkad A. Fattah. This is an open access article distributed under the Creative Commons At-
tribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is prop-
erly cited.
ABSTRACT
β,amino butyric acid (BABA) induced resistance
against Meloigogyne spp in tomato. Signifi-
cantly (p = 0.05) less, 41.11 second stage juve-
niles (J2) enter the roots of treated than, 116.66
J2 in untreated control plants. Root fresh and
dry weight (Rfw, Rdw) were 2.87 and 0.12 g in
treated compared with 4.78 and 0.30 g in nema-
tode infected control plants respectively, 30 d
after nematode inoculation. Sfw and Sdw were
8.62 and 0.60 g compared with 3.94 and 0.22 g in
control plants. Foliage spray at 40 and 20 mM of
BABA was more effective than 10 and 5 mM
treatments. The former two concentrations re-
corded the lowest, average gall index, 2.11 com-
pared with 3.33, 4.11, and 5 for the latter two
concentrations and nematode infected control
respectively. Results also indicated that treat-
ments with BABA prior to nematode inoculation
were superior in inducing resistance to Meloi-
dogyne spp over treatment at the time or after
nematode inoculation.
Keywords: Induced Resistance; β,Amino Butyric
Acid; Meloigogyne spp; Tomato
1. INTRODUCTION
Tomato, Solanum lycopersicum L., formerly known as
Lycopersicon esculentum Mill., is the second most im-
portant vegetable crop after potato with annual world
production of about 152.9 million ton in 2009 [1]. An-
nual tomato production in Iraq was estimated at 830.000
tons in 2008 [2]. Tomato plants are subjected to infection
by many important plant pathogens including the root
knot nematodes, Meloidogyne spp which are considered
as the most important nematode species worldwide and
in Iraq. Many effective control measures were used to
manage these pathogens such as soil solarization chemi-
cal and biological control. Induced systemic resistance to
plant pathogen provides an ideal control measures
against plant pathogens [3]. Chemical, physical and bio-
logical inducers to control the root knot nematodes were
used [4]. In recent years, various chemical, physical and
biological agents were used to induce resistance to many
plant pathogens. β-1,3 amino butyric acid (BABA), Ben-
zol [1-3] thaiadiazole-7-carbothionic acid S-methyl ester
and salicylic acid are among the most widely studied
chemical inducers [5-7]. These chemicals activate resis-
tance against different plant pathogens like fungi, bacte-
ria, viruses and nematodes [8]. BABA was reported to
induce resistance against members of solanacea, cucur-
bitacea, legumenacea, cruciferea, and rosacea [9]. Jakob
et al. [10], reported that BABA induced resistance to dif-
ferent plant pathogens such as fungi, bacteria, viruses,
and plant parasitic nematodes. Only few studies on in-
duced resistance by BABA against plant nematodes are
reported. BABA was reported to reduce damage due to
root knot nematode on tomato [11] and M. javanica and
Rotylenchulus reniformis on pineapple [12] and induce
resistance against M. javanica in cucumber [4].
This study was undertaken to assess the ability of β,
aminobutyric acid to induce acquired systemic resistance
against the root knot nematodes, Meloidogyne spp in to-
mato plants.
2. MATERIALS AND METHODS
Experiments were performed in a greenhouse (27 ± 5
C) and tomato, Solanum lycopersicum L. cv. “Supper Re-
gina” highly susceptible to Meloidogyne spp was used.
Plants were grown and maintained in1kg plastic pots
throughout th e experiments.
2.1. Nematode Inoculation
Soil from cucumber grown plastic house heavily in-
Copyright © 2013 SciRes. OPEN ACCESS
S. S. Mutar, F. A. Fattah / Agricultural Sciences 4 (2013) 608-613 609
fested with Meloidogyne spp (predominantly M.javanica)
were collected and stored at 4 C in polyethylene pages
until use to inoculate tomato seedlings.The nematode
infested soil was mixed with peat moss in 1:1 ratio and
used as nematode inoculums.
2.2. Preparation β,Amino Butyric Acid
(BABA)
Stalk solution of BABA, Mwt = 103 (Sigma Aldrich,
St Louis, Missouri, USA) was prepared by dissolving
412 mg in 100 ml of distilled water.
2.3. Foliage Treatments with BABA
Four concentrations of BABA, 5, 10, 20, and 40 mM
were used as foliage application. The experiment was
consisted of 13 treatments (one nematode inoculated
control sprayed with distilled water and 12 combination
of the 4 concentrations of BABA 3-d before, during and
7-d after nematode inoculation). Plants were carefully
sprayed till complete wetness using 1L hand sprayer.
Each treatment was replicated 4 times and arranged in
randomized complete blocks in the greenhouse. Thirty
days after inoculation, plants were assessed for growth
and nematode infection.
2.4. Root Treatments with BABA
In this experiment, 2 mL of 1, 2.5 and 5 mM of BABA
were used as soil drenching added around the stem of
single tomato plants growing in n ematode infested so il in
1 kg plastic pots. Ten treatments (one nematode inocu-
lated control watered with 2 ml of distilled water and 9
combinations of the 3 concentration of BABA applied
3-days before, during and 7-days after nematode inocula-
tion ). Thirty days after inoculation, plants were assessed
for growth and nematode infection. Each treatment was
replicated 4 times and arranged in a randomized com-
plete block in the greenhouse. Thirty days after inocula-
tion, plants were assessed for growth and nematode in-
fection.
2.5. Effect of BABA on Meloidogyne spp
Rate of Gall Index
The rate of infestation of the root knot nematodes is
determined using a 5 level scales [13]: 1 = no galls on
roots, 2 = galls on 1% - 25% of the root, 3 = galls on
26% - 50% of the root, 4 = galls on 51% - 75% of the
root, and 5, galls on 76% - 100% of the root.
2.6. Effect of BABA on Fresh and Dry
Weight of Shoot and Root Systems
Plants were carefully uprooted and roots were washed
under tap water to remove adhering soil. To determine
shoot and root dry and wet weight, shoots and roots were
separately weights and dried at 70 C for 48 h or until
weight fixed.
2.7. Effect of BABA on Nematode
Penetration
To determine the effect BABA treatments on nematode
penetration of tomato roots, roots were stained with acid
fuchsine [14], washed with water to remove excess stain
and 1 g of each root was individual examined under a
compound microscope to count nematodes inside the
roots.
3. STATISTICAL ANALYSIS
The data were subjected to analysis of variance and
means were separated by the least significant method at
(p = 0.05) using SAS, 200 4 [1 5].
4. RESULTS
4.1. Foliage Application
4.1.1. Effect of Foliage Treatment of BABA on
Meloidogyne spp Rate of Gall Index
BABA treatment induced resistant to Meloidogyne spp
in tomato plants (Ta bl e 1). The treatments significantly
(p = 0.05) reduced the rates of gall index of treated plants
compared with nematode infected control plants. While
40, 20, 10, and 5 nM of BABA treated plants recorded
low average gall indices, 2.11, 2.11, 3.33, 4.11 respec-
tively. BABA untreated plants recorded the highest av-
erage gall index, 5, 30 days after nematode inoculation.
The former two concentrations render tomato plant sig-
Table 1. Effect of spraying BABA on tomato, Solanum ly-
copersicum L. shoot on rate of root galls index of Meloidogyne
spp infected plants 30 days after nematode inoculation.
Rate of root gall index
Nematode inocu l ation
BABA Conc.
(mM)
7 days beforeDuring 3 days after
Mean
40 2.33 2.33 1.66 2.11
20 2.66 2 1.66 2.11
10 4 3 3 3.33
5 4 4 4.33 4.11
Control 5 5 5 5
Mean 3.60 3.26 3.13
LSD (P = 0.05) = Conc. = 0.35*,Trea. = 0.27*, Inter = 0.60*. Each number is
a mean of t hree r epli cates an d two plan t each. *Indicate significant deferens.
Nematode inoculums (nematode infested soil + peat moss, 1:1) were added
when plants were 4 - 5 true leaves. Gall index was according to 1 - 5 level
scale: 1 = no galls on the roots, 2 = galls on 1% - 25% of the root, 3 = galls
on 26% - 50% of the root, 4 = galls on 51% - 75% of the root, and 5 = galls
on 76% - 100% of roots. BABA = β,amino butyric acid.
Copyright © 2013 SciRes. OPEN ACCESS
S. S. Mutar, F. A. Fattah / Agricultural Sciences 4 (2013) 608-613
610
nificantly more resistant to the nematode than the two
latter concentrations (Table 1). The 40 and 20 mM
scored significantly (p = 0.05) the lowest nematode gall
index, 1.66, when BABA treatments exceeded nematode
inoculation by 3 days compared with other test treat-
ments.
4.1.2. Effect of BABA on Fresh and Dry Weight
of Root Systems
Results in Tabl e 2 sho wed that spraying tomato p lants
with BABA significantly (p = 0.05) affected root fresh
weight (Rfw) and root dry weight (Rdw) of Meloidogyne
spp infected tomato plants. The 40 mM recorded the
lowest average root weights, 5.74 and 0.34 g compared
with other test treatments 30 days after nematode inocu-
lation. Treatment with BABA 3 days before nematode
inoculation produced the lowest Rfw and Rdw, 3.96 and
0.29 g respectively 30 days after nematode inoculation.
The 40 mM concentration also recorded the lowest root
weights, 2.35 and 0.21 g compared with the highest root
weight, 12.98 and 0.80 g when the treatment was per-
formed 7 days after nematode inoc ul a t i on.
4.1.3. Effect of BABA on Fresh and Dry Weight
of Shoot Systems
Spraying of BABA on foliage of tomato plants signifi-
cantly (p = 0.05) increased root fresh and dry weight
(Sfw and Sdw) of nematode infected tomato plants 30
days after inoculation (Table 3). The average w eights f or
plants treated with 40 and 20 mM of BABA r ecorded the
highest, 21.40 and 2.10 g Sfw and Sdw compared with
the lowest average weights of 15.05 and 0.97 g in nema-
tode infected controls. Treatment with BABA 3 days be-
fore nematode inoculation caused significantly the high-
est weight, 20.99 and 2.24 g compared with 14.24, 0.75,
19.31 and 1.35 g when nematode was inoculated during
and 7 days after BABA treatment respectively. Treatment
with 40 and 20 mM of BABA 3 days prior to nematode
inoculation caused higher Sfw and Sdw, 25.32, 3.25 and
22.22, 3.90 g compared with, 12.86 and 0.51 g for nema-
tode infected control plants respectively.
4.2. Soil Application
4.2.1. Effectof Soil Drenching with BABA on
Meloidogyne spp Rate of Gall Index
Soil drenching with BABA significantly (p = 0.05)
reduced the root ga ll ind ex of to mato plan ts infected with
Meloidogyne spp (Tab le 4). Gall index was decreased as
the test BABA concentration was increased. The 5 mM
concentration produced significantly the lowest average
gall index, 2.33 compared with, 2.77, 3.55 and 5 for 2.5,
1 mM and control 30 days after nematode inoculation r e-
spectively. The lowest average gall index, 2.91 was re-
corded when BABA was applied 3 days before nematode
Table 2. Effect of spraying BABA on tomato, Solanum ly-
copersicum L. shoot on root weights of Meloidogyne spp in-
fected plants 30 days after nematod e inocu lation.
Root weight of infected plants (g)
Nematode inocu l ation
7 days afterDuring 3 days before
BABA
Conc.
(mM)
DryFreshDryFresh Dry Fresh
Mean
Dry Mean
Fresh
40 0.212.350.375.97 0.45 8.90 0.345.74
20 0.242.720.405.98 0.47 10.09 0.376.26
10 0.283.910.506.74 0.51 10.16 0.436.94
5 0.305.890.728.92 0.50 11.44 0.518.75
Control0.404.940.528.79 0.80 12.98 0.588.90
Mean 0.293.960.507.28 0.54 10.71
LSD (P = 0.05) Dry weight: Conc. = 0.065*, Treat. = 0.050*, Inter. = 0.11*.
LSD (P = 0.05) Fresh weight: Conc. = 1.78*, Traet. = 1.37*, Inter. = 3.08*.
Each number is a mean of three replicates and two plant each. *Indicate
significant deferens. Nematode inoculums (nematode infested soil + peat
moss, 1:1) were added when plants were 4 - 5 true leaves. BABA = β,amino
butyric acid.
Table 3. Effect of spraying BABA on tomato, Solanum ly-
copersicum L. shoot on shoot weights of Meloidogyne spp in-
fected plants 30 days after nematod e inocu lation.
Shoot weight of infected plants (g)
Nematode inocu l ation
3 days beforeDuring 7 dayd after
BABA
Conc.
(mM)
DryFreshDryFresh Dry Fresh
Mean
Dry Mean
Fresh
40 3.2525.321.1513.69 1.92 25.13 2.1021.38
20 2.9022.221.0017.92 1.26 24.06 1.7221.40
10 1.3921.230.5712.88 1.31 14.31 1.0916.14
5 2.2718.340.5313.88 1.24 18.58 1.3516.93
Control1.4017.830.5112.86 1.00 14.48 0.9715.05
Mean2.2420.990.7514.24 1.35 19.31
LSD (P = 0.05) Dry weight: Conc. = 0.2669*, Treat. = 0.2067*, Inter. =
0.4623*. LSD (P = 0.05) Fresh weight: Conc. = 3.20*, Treat. = 2 .4*, Inter. =
5.54*. Each number is a mean of three replicates and two plant each.
*Indicate significant deferens. Nematode inoculums (nematode infested soil
+ peat moss, 1:1) were added when plants were 4 - 5 true leaves . BABA= β
amino butyric acid.
inoculation compared to 3.25 and 4.08 when BABA was
applied during or 7 days after the nematode re spectively.
The lowest gall index, 1.66 was caused by application of
5 mM of BABA 3 days before nematode inoculation
compared with the highest gall index, 5 in nematode in-
fected control.
4.2.2. Effect of Soil Drenching with BABA on
Weight of Roots
Results in Ta bl e 5 showed that soil drenching with 5
Copyright © 2013 SciRes. OPEN ACCESS
S. S. Mutar, F. A. Fattah / Agricultural Sciences 4 (2013) 608-613 611
Table 4 . Effect of soil drenching of BABA on rate of root gall
index caused by Meloidogyne spp on tomato, Solanum lycoper-
sicum L. 30 days after nematode inoculation.
Rate of root gall index
Nematode inocu l ation
BABA Conc.
(mM)
3 days before During 7 days after
Mean
5 1.66 2.33 3 2.33
2.5 2 2.66 3.66 2.77
1 3 3 4.66 3.55
Control 5 5 5 5
Mean 2.91 3.25 4.08
LSD (P = 0.05) Conc. = 0.36*, Treat. = 0.31*, Inter. = 0.62* Each number is
a mean of th ree repli cates and two plan t each. *indicate significant deferens.
Nematode inoculums (nematode infested soil + peat moss, 1:1) were added
when plants were 4 - 5 true leaves. Gall index was according to 1 - 5 level
scale: 1 = no galls on the roots, 2 = galls on 1% - 25% of the root , 3 = galls
on 26% - 50% of the root, 4 = galls on 51% - 75% of the root, and 5 = galls
on 76% - 100% of roots. BABA = β,amino butyric acid.
Ta b l e 5 . Effect of soil drenching of BABA on root weights of
tomato, Solanum lycopersicum L. infected with Meloidogyne
spp 30 days after nematode inoculation.
Root weight (g)
Nematode inocu l ation
3 days before During 7 days after
BABA
Conc.
(mM)
Dry Fresh Dry FreshDry Fresh
Mean
Dry Mean
Fresh
5 0.27 3.25 0.34 3.720.39 9.86 0.355.61
2.5 0.16 2.50 0.39 8.980.51 9.44 0.356.97
1 0.32 3.37 0.49 11.39 0.52 9.43 0.438.06
Control 0.35 3.75 0.36 10.23 0.61 9.77 0.447.91
Mean 0.28 3.21 0.39 8.580.51 9.62
LSD (P = 0.05) Fresh weight: Conc. = 1.41*, Traet. = 1.22*, Inter. = 2.44*.
LSD (P = 0.05) Dry weight: Conc. = 0.0724*, Inter. = 0.1447*. Each number
is a mean of three replicates and two plant each.*Indicate significant
deferens. Nematode inoculums (nematode infested soil + peat moss, 1:1)
were added when plants were 4 - 5 true leaves. BABA = β,amino butyric
acid.
mM BABA significantly (p = 0.05) affected Rfw and
Rdw of Meloidogyne spp infected tomato plants. At this
concentration average Rfw and Rdw were 5.61 and 0.35
g compared with 7.91 and 0.44 g in control plants re-
spectively. However, 2.5 and 1 mM did not significantly
affect root weights. The 40 mM recorded the lowest av-
erage root weights, 5.74 and 0.34 g compared with other
test treatments 30 days after nematode inoculation. Treat-
ment with BABA 3 days before nematode inoculation
produced the lowest average Rfw and Rdw, 3.21 and
0.28 g compared with 8.58, 0.39, 9.62 and 0.51 g at the
time and 7 days after nematode inoculation respectively.
While the lowest root weights were recorded for the 5
and 2.5 mM, 3 days before nematode inoculation, 3.25,
0.27, 2.50, 0.16, the 1 mM concentration and control
recorded the highest root weights, 11.39, 0.49, 10.23,
0.36 g respectively when the treatment was performed at
the timeof nematode inoculation.
4.2.3. Effect of Soil Drenching with BABA on
Weight of Shoots
Soil drenching with the test concentrations of BABA
significantly (p = 0.05) affected shoot weights of Meloi-
dogyne infected tomato plants (Table 6). The average
Sfw and Sdw was grater, 20.43 and 2.34 g in 5 mM
treatment compared with 15.11 and 1.19 g in control.
Grater shoot weights were observed when BABA treat-
ments exceeded nematode inoculation by 3 days , 20.91,
2.65 compared with 13.11, 0.70 and 18.61, 1.52 g when
BABA treatment was at or 7 days after nematode inocu-
lation respectively. The greatest average Sfw and Sdw,
24.32 and 4.22 g were recorded for 5mM application 3
days before nematode inoculation while the lowest Sdw,
0.43 g and Sfw, 11.75 g for the1mM at the time of nema-
tode inoculation and the untreated control plants respec-
tively.
5. DISCUSSION
Results of this work indicated that foliage as well as
soil drench applicatio n of BABA induced systemic resis-
tance against Meloigogyne spp in susceptible tomato
plants. This was manifested by the reduction of galls on
roots and numbers of J2 in roots of BABA treated plants
compared with those in BABA untreated but nematode
Table 6. Effect of soil drenching of BABA on shoot weights of
tomato, Solanum lycopersicum L. infected with Meloidogyne
spp 30 days after nematode inoculation.
Shoot weight (g)
Nematode inocu l ation
3 days beforeDuring 7 days after
BABA
Conc.
(mM)
DryFreshDryFresh Dry Fresh
Mean
Dry Mean
Fresh
5 4.2224.321.0916.47 1.71 20.50 2.3420.43
2.52.7622.920.7511.96 1.64 20.90 1.7118.59
1 1.9020.510.4312.25 1.45 15.39 1.2616.05
Control1.7315.890.5311.75 1.30 17.67 1.1915.11
Mean2.6520.910.7013.11 1.52 18.61
LSD (P = 0.05) Fresh weight: Conc. = 2.50*, Traet. = 2.16*, Inter. = 4.33*.
LSD (P = 0.05) Dry weight: Conc. = 0.319*,Treat. = 0.2763*, Inter. =
0.5252*. Each number is a mean of three replicates and two plant each.
*Indicate significant deferens. Nematode inoculums (nematode infested soil
+ peat moss, 1:1) were added when plants were 4 - 5 true leaves. . BABA =
β,amino butyric acid.
Copyright © 2013 SciRes. OPEN ACCESS
S. S. Mutar, F. A. Fattah / Agricultural Sciences 4 (2013) 608-613
612
infected plants. The growth of the nematode infected
plants by decreasing root weight and increasing shoot
weight of BABA treated tomato plants. These findings
also support previous reports indicating that treatments
with β,amino butyric acid reduced root knot disease
through decreased penetration of J2, gall number on
roots and nematode development [4,11,12]. Root weight
increase in heavy Meloidogyne spp infection was previ-
ously reported and thought to be caused due to biomass
accumulations in infected roots [16]. Meloigogyne spp
infection is known to have negative effects on water and
nutrient elements as well as photosynthesis [17]. It was
reported that M. incognita infection caused biomass ac-
cumulation in roots and this is controlled by the effi-
ciency of the pathogen in capturing the light energy and
directing it in favor of the pathogen or the infected host
[18]. Because of the relative large size of females of
Meloigogyne spp and its ability to produce large number
of eggs, it requires large amount of energy. Beside this
energy requirement, these pathogens caused obvious
distortion in xylem vessels, swellings of root cells and
formation of giant feeding cells which alter root normal
functions. Treatments of nematode infected plants with
the high concentration of BABA produced more shoots
compared with untreated nematode infected plant. This is
mainly due to the fact that Meloidogyne infections em-
bed photosynthesis and chlorophyll synthesis which ne-
gatively influences plant growth [17]. It was clear from
the results of this study that treatments with BABA was
more effective when applied before nematode inocula-
tion, which is probably due to the mechanism of induce
resistance of this chemical inducer. The mechanism of
induce resistance to Meloidogyne in tomato by BABA is
not fully understood. It was believed that treatments with
this inducer render roots less attractive to J2 through al-
tered plant nutrient assimilation or render plant cell walls
harder to penetrate by J2 or that giant cells were smaller
or not able to provide enough nutrients for the develop-
ing nematodes [11]. Treatments with BABA were re-
ported to increase levels of salicylic acid (SA) and patho-
genesis related proteins (PRP) [19], and enzymes like
catalase (CAT), polyphenoloxidase (PPO) and guaiacol
peroxidase (GPOX) [4,7] and phenol compounds [20].
BABA was also reported to induce the accumulations of
PPO, GPOX, H2O2, CAT and phenols in M.javanica in-
fected cucumber roots [21].
Results of this research work clearly indicate and in-
dorse previous reports on the effectiveness of β,amino-
butyric acid to the induced systemic resistance as envi-
ronmentally sound alternative for the control of Meloi-
dogyne sp p on t omato.
6. ACKNOWLEDGEMENTS
The authors would like to thank Mr. Dhulfiqar Layth (Graduate stu-
dent, Dept., of Plant Protection, College of Agriculture,Univ. of Bagh-
dad, Baghdad, Iraq) for his valuable assistance.
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