18530-20805-1-PB

Journal of Natural Sciences Research www.iiste.org
ISSN 2224-3186 (Paper) ISSN 2225-0921 (Online)
Vol.4, No.23, 2014
139
Virulence of Russian Wheat Aphid, Diuraphis Noxia (Kurdjumov)
(Homoptera: Aphididae) Populations in Kenya
Watson A. Ngenya1*
J.N. Malinga1
E.A. Masinde1
I.M. Tabu2
M.Cakir3
1.Kenya Agricultural research institute, Njoro, Private Bag, Njoro, Kenya
2.Department of Crops, horticulture and Soils, Egerton University, P.O. Box 536 Egerton, Kenya
3.Murdoc University, Perth. Australia
*
Correspondence: watsonngenya24@gmail.com
Abstract
The Russian wheat aphid (RWA) Diuraphis noxia (Kurdjumov) is a serious pest of wheat in Kenya.
Development and use of RWA resistant wheat (Triticum aestivum L.) varieties, has been constrained by RWA
populations evolving with differential virulence to given resistant host plants. To fully exploit host plant
resistance (HPR) in management of RWA, local populations of RWA have to be evaluated for differential
virulence and biotypes in order to develop and deploy cultivars that exhibit cross biotype resistance. A study was
conducted at KARI-Njoro to characterize virulence of RWA populations from the endemic areas (Eldoret, Mau
Narok, Njoro and Egerton) in Kenya. A factorial experiment in randomized complete block design replicated
three times was set up to evaluate seedling resistance to RWA with variety and aphid collection source as main
factors in the screen house. Five adult RWA aphids from each of the four collection locations were used to infest
four host genotypes; PI624933-1 containing Dn4 gene, 2414-11-2 containing Dn7 gene, KRWA9 which
contains an unknown Dn gene and a susceptible check, K.KWALE, for 28 days to determine virulence of the
RWA aphids to seedlings of the four wheat genotypes in the greenhouse. Data was recorded on damage scores,
plant height, plant height reduction, shoot biomass and biomass reduction of test plants 28 days after infestation.
Results of an analysis of variance of these plant parameters show that Egerton population was more virulent than
populations selected from other areas as it caused more damage on resistant lines.
Keywords: Diuraphis noxia, biotype, virulence, RWA, Wheat.
Introduction
Diuraphis noxia (Kurdjumov) (Homoptera Aphididae) is a new invasive pest of wheat, barley and other small
grains in Kenya. Since its discovery in Kenya in the mid 1990’s, the aphid has become an important pest of
barley, Hordeum vulgare L., and wheat, Triticum aestivum L. D. noxia feeding significantly reduces chlorophyll
and carotenoid content in susceptible plants. This chlorophyll reduction has an effect on plant yield (Heng-moss
et al, 2003). Plant resistance has been considered an especially useful way to control D. noxia, because the
aphid’s habit of feeding within rolled wheat leaves may limit the effectiveness of contact insecticides and some
natural enemies (Burd et al., ;Brewer et al. 2005) besides, using for aphid management is cheaper and has no
negative effects on the environment. The importance of host plant resistance in an integrated pest management
program is however limited by development of virulent biotypes. Insect biotypes are infraspecific classifications
based on biological rather than morphological characteristics, they are generally morphologically
indistinguishable. Insect biotypes have been described as members of an insect species with similar genetic
composition for a biological attribute (Saxena and Burrion, 1987) where as Diehl and Bush, (1984) described
biotypes based on survival and development on a particular host and by host preference for feeding, oviposition
or both. Insect biotypes including Diuraphis noxia biotypes have traditionally been described by their ability to
damage crops that express host plant resistance. (Puterka et al., 1992). As virulent RWA biotypes with superior
fitness replace the previously avirulent RWA biotype populations, the result is breakdown of resistance and
ineffective control of RWA. Virulent biotypes of RWA have been reported in the USA (Haley et al 2004), South
Africa (Tolmay, 2007), Asia (Dolatti et al, 2005), Europe, (Basky, 2003) and South Amarica (Smith et al, 2004).
Genetic studies of Kenyan populations of D.noxia demonstrated that there is limited genetic variability among
the biotypes under study (Maling’a et al., 2007). Kiplagat, (2005) however found that some Kenyan populations
of D. noxia were virulent to wheat containing Dn4 gene for D noxia resistance. Malinga, 2007 also found that
despite the Kenyan populations showing little genetic variation, the same populations had variations in
development and population growth. In this study our primary goal was to characterize virulence of Kenyan
biotypes of RWA on selected resistant genotypes of wheat with a view to identify the most appropriate
genotypes to incorporate into Kenyan breeding programmes in order to develop a variety of wheat that is
resistant to RWA populations in Kenya.
Materials and Methods
The study was conducted at KARI-Njoro located in the lower highlands (LH3), at an altitude of 2166 meters
above sea level.

Vol.4, No.23, 2014
140
Plant material
Wheat genotype Kkwale, KRWA9, PI624933-1 and 2414-11-2 2414-11-2 were used. In our earlier
experiment Kkwle had been found susceptible and the other three wheat line were resistant (results not
publiched).
RWA Populations
RWA populations were collected from the endemic areas (Eldoret, Mau Narok, Njoro and Egerton) in Kenya
during 2010. Kenya Pasa (susceptible) wheat, used as the rearing plant, was grown in a sterilized potting mixture
composed of forest soil, sand and manure ratio 3:1:1. The potting mixture was amended with 50 kg/ha equivalent
of DAP. 10 seeds each of Kenya Pasa (K. Pasa) were planted 2.5 cm deep in the potting mixture in a 1L plastic
pot and the pots placed in a water bath in an insect rearing box to keep the emerging seedlings clean from aphid
contamination in the greenhouse. When rearing plants reached Zadocks et al. (1974) growth stage 12, a single
adult female RWA was settled in the leaf whorl using a fine hair brush. The plants were watered regularly by
replenishing water in the water bath after every three days so that the seedlings were not water stressed.
Environmental conditions were 18±20
C with a photoperiod of 12:12(L:D) h. The aphid was allowed to
multiply freely to form a colony. The insect rearing boxes were kept a minimum 10m from each other to
eliminate accidental contamination of clones by mixing. The four established clones above were used to test for
variation in the ability to cause feeding damage to four wheat genotypes.
Screening protocol
The experiment was a factorial experiment in randomized complete block design replicated three times. Wheat
genotype and Diuraphis noxia (origin) collection point were the two factors. Wheat genotype had four levels
(Kkwale, KRWA9, PI624933-1 and 2414-11-2 2414-11-2 ) whereas RWA origin had five levels (Egerton,
Eldoret, Mau Narok, Njoro and a non infested control). The experiment was conducted in a screen house during
the period September 2010 to January 2011.
Two seeds of the test material were planted in a potting mixture as described earlier in a 1L pot. After
emergence, at Zadocks et al. (1974) growth stage 10, they were thinned to leave one seedling per pot. The single
seedling was infested with five aphids placed in the leaf whorl using a fine hair brush at growth stage12 (Zadoks
et al., 1974). The infested seedlings were then caged using polyester mesh supported on wires and the aphids left
to multiply and feed on the test plants for 28 days. Three seeds were planted per pot and later thinned to two
seedlings per pot. Water was supplied regularly by filling the water bath until the soil in the pots was ascertained
to be wet by visual examination. Scoring for overall plant damage was done at 7 days, 14 days, 21 days and 28
days post inoculation, the damage to the test plants were qualitatively evaluated using a 1-9 scale(Table, i) where
1-resistant; 9-susceptible (Maling’a, 2007; Tolmay, 1999)
Traits measured
Plant height was measured 28 days after infestation (DAI). The plants were then cut at the soil surface and
weighed to determine fresh weight. The sampled plant were dried at 1050
C for 48hrs, and weighed to determine
above ground biomass. Proportional height, fresh weight and dry weight was determined using the formular
∗ 100……………………………………………………Equation 1
DWT- Proportional reduction
Dc- Value measured on non infested control plant
Dt- Value measured on infested plant
. Temperatures range during the duration of the experiment was between 18–28°C
Statistical analysis
Analysis of variance (ANOVA) was performed on collected data using Genstat (,
Significant differences in treatment means were separated using Least significant difference (LSD) at α= 0.05
level of significance.
Results and Discussion
There were significant differences in damage level on wheat genotypes at different duration of RWA infestation.
All wheat genotypes developed damage symptoms associated with RWA infestation as early as 7 days post
infestation. (Figure 1).
Wheat genotype varied significantly in plant damage resulting from RWA biotypes on all days when
plant damage was assessed. As expected, wheat K. Kwale was generally susceptible throughout the period while

Vol.4, No.23, 2014
141
wheat KRWA9 was moderately resistant. Wheat PI624933-1 PI624933-1 and 2414-11-2 were generally
moderately resistant to RWA attack (Figure 1).
Aphid biotype caused significant damage on wheat genotypes. The biotypes also varied in their
virulence on wheat genotypes (Figure 2). At 7 days post infestation, all aphid biotypes caused plant damage with
Egerton, Eldoret and Njoro biotypes causing the most damage. Irrespective of wheat genotype, Mau Narok
biotype caused the least damage on infested wheat. Njoro Egerton and Eldoret biotypes emerge as the most
virulent biotypes 21 days after infestation and are significantly different from Mau Narok biotype,
There was significant interaction of RWA population and Wheat genotype on four instances when
damage score was assessed indicating that populations varied in virulence depending on wheat genotype (Figure
3 and 4). The degree of damage also depended on the duration of infestation for some genotypesAll RWA
populations significantly damage wheat but the degree of damage now depends on the RWA population infesting
the wheat and the duration under which wheat remains infested.
There were no significant differences among RWA populations on PI624933-1 and 2414-11-2 2414-
11-2 at 14, 21 and 28 days of RWA infestation. The two wheat genotypes were moderately resistant to all RWA
populations that were tested. Significant differences were noted among populations on KRWA9 during the entire
period of RWA infestation. The genotype was clearly susceptible to all populations of RWA after 7 and 14 days
of infestation with Njoro and Eldoret populations being the most virulent during this period. Mau narok
population was the least virulent to KRWA9 during the entire period of evaluation.KRWA9 has significantly
lower mean damage score 21 and 28 days after infestation. There was significant variation in the reduction of
seedling growth attributed to Diuraphis noxia populations. Percent reduction of plant height varied significantly
across wheat genotypes. PI624933-1 and 2414-11-2 2414-11-2 were the tallest plants followed by KKWALE
and KRWA9 28 days after infestation with RWA populations (Table ii). There was significant reduction in
wheat shoot fresh and dry weight 28 days after infestation of bread wheat with Diuraphis noxia. PI624933-1 and
2414-11-2 2414-11-2 had the lowest height, and shoot weight reductions, an indication that these two
genotypes are better at tolerating RWA infestation at seedling stage. Differences among individual wheat
genotypes were expected and arise from feeding by RWA that causes infested plants to become stunted and
genotype characteristics in relation to infestation.
RWA populations varied significantly in their effect on, % leaf reduction, and % height reduction 28
days after infestation. (Table iii). This is an indication that all RWA populations cause plant stunting when they
infest wheat plants, however the percent reduction differs significantly among populations.
Egerton population caused the highest % reduction in plant height followed by Eldoret and Njoro
population respectively. Mau Narok population caused the least reduction in plant height among RWA
populations.
There were significant differences in % reduction in shoot fresh weight of test plants,(Table iii) Test
plants infested with Egerton population had the least shoot fresh weight and the population had the highest %
fresh shoot weight reduction among RWA populations.
There were significant two way interactions between host genotype and aphid population in some
growth measurements of wheat seedlings. Significant interaction was observed in plant height reduction on
wheat genotype Krwa9 (Table IV). Significant interaction was also observed in % reduction in plant height, %
reduction in shoot fresh weight, and % reduction in shoot dry weight on KKwale, PI624933-1PI624933-1 , and
2414-11-2 2414-11-2 , (Tables iv and v).
Table, viii shows Pearsons’ correlation coefficients among parameters of wheat infested with RWA and
RWA damage scores at 21, and 28 days in the greenhouse. Significant positive associations were observed
between damage score and plant height reduction, percent shoot fresh weight reduction and shoot dry weight
reduction. Allowing RWA to infest wheat results in stunted growth and reduced straw weight. A significant
negative relationship exists between damage scores and plant height, shoot fresh weight and shoot dry weight.
Discussion
Symptoms of Russian wheat aphid damage started manifesting in all wheat genotypes 7 days after infestation.
Maling’a, (2007) also found that symptoms of damage started to manifest in both resistant and susceptible plant
entries as early as seven days after first infestation, thus for effective management, control of the Russian wheat
aphid should start early to minimize yield losses due to aphid damage. Though all populations caused damage on
wheat genotypes, some populations caused significantly more damage. Njoro populations caused the severest
damage symptoms though mau narok biotype cauld cause severe damage symptoms only on 2414-11-2 2414-
11-2 indicating a possible resistance breaking variant of RWA in Kenya. This findings concur with Kiplagat,
(2005) who found that Nakuru RWA populations were more virulent compared to RWA populations from other
wheat growing regions. All four Russian wheat aphid populations established from single female aphids
collected from the various wheat growing regions in Kenya were virulent to Kenya Kwale which does not have
any Dn resistance gene. K.Kwale is a highly popular variety in Kenya grown by almost 60% of wheat farmers

Vol.4, No.23, 2014
142
even though it is susceptible to Diuraphis noxia. The variety would form a good background in any breeding
program to breed wheat resistant to RWA. The other resistant introductions had varying damage ratings
depending on RWA population used to infest the resistant line and the duration of infestation. The line RWA9
had high damage values in the first and second week and this could be due to the line having been used for a
long time in breeding programs in Kenya to develop RWA resistant wheat and the aphid populations could be
acclimatized to the resistance in RWA9. Jyoti and Michaud (2005) reported high damage values for Trego
infested with USA RWA biotype 1 because the biotype had been acclimatized on the variety. 20 generations of
the aphid had been raised on the variety and were acclimatised to it. This indicates that when a resistant source is
exposed to an aphid population for a long time, resistance may begin to break down as the the aphid becomes
acclimatized on the variety developing novel strategies for neutralizing resistance factors in the variety. This is
an indication that a resistant variety cannot be grown indefinitely as a control measure, therefore the process of
searching for new sources of resistance should be continuous. Botha et al., (2005) also postulated that several
defense strategies could be employed in wheat defenses including systemic acquired resistance. KRWA9
responds to RWA attack by becoming moderately resistant over time, this could be due to the fact that it may
take time for the plant to build up defence factors from the first time of aphid infestation when the plant
recognisez proteins in the aphid saliva during the initial sampling before settling. It also indicates that the Dn
resistance gene in KRWA9 may be responsible for systemic acquired resistance observed in KRWA9. The two
resistant lines, PI624933-1 and 2414-11-2 were generally resistant and had low damage ratings for all RWA
populations except that PI624933-1 had uncharacteristically high damage score when infested with Mau Narok
RWA population. There is a high likelihood of the Mau Narok RWA population having evolved into a virulent
resistance breaking biotype and therefore an indication that PI624933-1 would not be a suitable resistant parent
in a breeding program to develop RWA resistant wheat in that region. It also could mean that RWA population
in Mau Narok will need to be managed using resistant materials that contain more than one resistant gene and
therefore have different modes of resistance.
There was also some reductions in some of the growth parameters measured such as plant height due to
RWA infestation. Plant damage due to Diuraphis noxia is associated with developmental, morphological,
physiological and biochemical processes in the host plant. Reduction in biomass of wheat plants infested with
Diuraphis noxia was also reported by Ni and Quinsenberry, (2006). The greater the reduction, the more virulent
the population.
Russian wheat aphid populations in Kenya have been evaluated for biotype development (Malinga et al.,2007).
However this study has clearly indicated that there are at least two possible biotypes based on the result of the
study.
Conclusion
The Russian wheat aphid populations tested showed distinct differences in how they affected damage and growth
of resistant and susceptible wheat entries. Njoro and Egerton populations emerge as the most virulent
populations whereas Mau Narok population emerges as the least virulent of all populations tested. This shows
that there are at least two distinct RWA biotypes in Kenya. Among the wheat genotypes tested, PI624933-1 and
2414-11-2 performed more uniformly in plant damage scores, had the least reduction in height, shoot fresh
weight and shoot dry weight reduction and generally exhibited moderate resistance to the RWA populations
tested.
Recommendations
Tolerance to RWA should be incorporated into breeding programs in order to slow down biotype development
by reducing selection pressure. Symptoms of RWA damage are manifested as early as seven days after
infestation. Therefore, management of the pest should start as soon as RWA is seen in the field.
Acknowledgement
This study was funded by Grain Development Research Corporation (Austarlia) through Kenya agricultural
research institute (KARI). I thank the entire staff KARI-Njoro for being supportive during the study and Dr.
Vicky Tolmay for giving invaluable advice on damage scoring. I would also like to thank Prof Mehmet Cakir of
Murdoc University (Perth, Australia) for His invaluable advice and contribution towards completion of the study.
Finally, the staff at Crops Horticulture and Soils department, Egerton University for support and advice during
the study.
REFERENCES
Basky, Z. 2003. Biotypic and pest status differences between Hungarian and south African populations of
Russian wheat aphid, Diuraphis noxia (Kurdjumov) (Homoptera: Aphididae). Pest Management Science,
59:1152-1158.

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Botha, A.M.,Youchun, L. and Lapitan, N.L.V.2005. Cereal host interactions with Russian wheat aphid:A review.
Journal of plant interactions, 1(4):211-222.
Brewer. M.J., Noma, T. and Elliot, N. 2005. hymenopteran paracitoids and dipteran predators of the invasive
aphid Diuraphis noxia after enemy introductions:temporal variation and implication for future aphid
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Castro, AM et al…
Diehl, S.R. and Bush, G.L. 1984. An evolutionary and applied perspective of insect biotypes. Annual Review of
Entomology, 29:474-504.
Dolatti, L. Ghareyazie, B.Moharramipour, S., Noori-Daloii. 2005. Evidence for regional diversity and host
adaptation in Iranian populations of the Russian wheat aphid. Entomologia axperimentalis et applicata,
114:171-180.
Harley, S.D., Peairs, F.B., Walker, C.B., Rudolf, J.B. and Randolf, T.L. 2004. The occurrence of a new Russian
Wheat Aphid bitype in Colorado. Crop Science, 44:1589-1592
Heng-Moss, T.M., Ni, X.,Macedo,T., Markwell, J.P., Baxendale, F.P., Quisenberry, S.S.,and Tolmay, V.2003.
Comparison of chlorophyll and carotenoid concentrations among russian wheat aphid
(Homoptera:Aphididae)-infested wheat isolines. Journal of Economic Entomology, 96:475-481
Kiplagat, O. 2005. The Russian wheat aphid (Diuraphis noxia Mord.): Damage on Kenyan wheat (Triticum
aestivum L.) varieties and possible control through resistance breeding. PhD Thesis. Wagengen
University. Netherlands.
Malinga, J.N. 2007. Studies on Russian wheat aphid, Diuraphis noxia (Kurdjiumov)(Homoptera:Aphididae) with
special emphasis to biotypes and host plant resistance in bread wheat (Triticum aestivum L.). PhD.
Thesis. Egerton University.
Malinga, J.N., Kinyua, M.G., Kamau, A.W., Wanjama, J.K., Awalla, J.O. and Pathak, R.S. 2007. Biotypic and
genetic variation within tropical populations of Russian wheat aphid, Diuraphis noxia
(Kurdjiumov)(Homoptera:Aphididae) in Kenya. Journal of Entomology, 4(5):350-361.
Ni, X. and Quisenberry, S.S. 2006. Diuraphis noxia and Rhopalosiphum padi (Homiptera: Aphididae)
interactions and their injury on resistant and susceptible cereal seedlings. Journal of Economic
Entomology, 99:551-558
Puterka, G.J., Burd, J.D., and Burton, R.L. 1992. Biotypic variation in a worldwide collection of Russian wheat
aphid (Homoptera: Aphididae). Journal of Economic Entomology, 85:1497-1506.
Saxena, R.C and Burrion, A.A. 1987. Biotypes of agricultural crops. Insect Science and its Application, 8:453-
458.
Smith, C.M., Belay, T., Stauffer, C., Stary, P., Kubeckova, I., and Starkey, S. 2004.Identification of Russian
wheat aphid (Homoptera:Aphididae) biotypes virulent to the Dn4 resistance gene. Journal of Economic
Entomology, 97:1112-1117.
Tolmay, V.L., van der Westhuizen, M.C. and van Deventer, C.S. 1999. A six week screening method for
mechanisms of host plant resistance to Diuraphis noxia in wheat accessions. Euphytica, 107: 79-89.
Tolmay, V.L., 2007. Genetic variability for Russian wheat aphid, Diuraphis noxia resistance in south African
genotypes. PhD Thesis. University of the Free State. Republic of South Africa
Zadoks, J.C., Chang, T.T. and Konzak, C.F. 1974. A decimal code for the growth stages of cereals. Weed
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Tables and figures
Tables and figures
Table I. Effect of wheat genotype on growth of selected bread wheat seedlings infested with Diuraphis noxia
populations for 28 days.
HOST GENOTYPE
% PLANT
HEIGHT
REDUCTION
% FRESH SHOOT
WEIGHT
REDUCTION
%DRY SHOOT WEIGHT
REDUCTION
KRWA9 36.3 40.76 11.72
KWALE 25.3 25.85 7.43
PI 624933 6.8 17.40 4.38
2414-11-2 7.2 15.67 6.36
LSD 1.8 2.2 2.2
%CV 12.7 12.2 41.0

Vol.4, No.23, 2014
144
Table II. . Effect of rwa (Diuraphis noxia) populations on growth of bread wheat seedlings after for 28 days of
infestation
RWA POPULATION
% Plant
height
reduction
%Fresh shoot weight
reduction
%Dry shoot weight
reduction
UNINFESTED CONTROL 0.0 0.0 0.0
EGERTON 26.9 36.4 10.0
ELDORET 24.6 29.6 10.2
MAU NAROK 20.1 28.5 8.3
NJORO 22.9 30.1 8.9
LSD 2.0 2.5 2.5
CV 12.7 12.2 41.0
Table III. Effect of rwa (Diuraphis noxia) population on krwa9 and 2414-11-2 2414-11-2 wheat genotype
seedling height, shoot fresh weight and dry weight reduction 28 days after infestation
KRWA9 2414-11-2
RWA
Population
% Plant
height
reduction
%Fresh
shoot
weight
reduction
% Dry
shoot
weight
reduction
% Plant
height
reduction
%Fresh
shoot
weight
reduction
% Dry
shoot
weight
reduction
UNINFESTED 0.0 0.0 0.0 0.0 0.0 0.0
EGERTON 53.3 52.3 15.0 15.8 25.6 7.5
ELDORET 40.9 50.4 14.8 7.1 13.4 9.5
MAU NAROK 47.1 50.8 14.7 5.2 25.3 7.5
NJORO 40.1 50.2 14.1 7.9 14.0 7.4
LSD 4.9 3.1 4.9 4.5 4.7 4.7
CV 7.2 4.1 22.3 32.9 15.8 38.8
Table IV. Effect of RWA (Diuraphis noxia) population on kwale and PI624933-1 wheat genotype seedling
height, shoot fresh weight and dry weight reduction 28 days after infestation
KWALE PI624933-1
RWA
Population
% Plant
height
reduction
%Fresh
shoot
weight
reduction
% Dry
shoot
weight
reduction
% Plant
height
reduction
%Fresh
shoot
weight
reduction
% Dry
shoot
weight
reduction
UNINFESTED 0.0 0.0 0.0 0.0 0.0 0.0
EGERTON 29.1 36.2 9.8 9.5 31.5 7.8
ELDORET 42.6 32.6 11.2 7.7 21.9 5.3
MAU NAROK 23.1 23.5 6.4 4.9 14.5 4.6
NJORO 31.4 36.9 9.7 12.0 19.2 4.2
LSD 4.5 4.8 4.8 3.2 2.9 3.5
CV 9.5 34.2 34.2 24.7 8.8 42.6

Vol.4, No.23, 2014
145
Figure 1. Mean plant damage values on wheat at various days after infestation with Diuraphis noxia.
Figure 2. Mean plant damage score at varying days of wheat infestation with Diuraphis noxia biotypes.

Vol.4, No.23, 2014
146
Figure 3Level of plant damage caused by different aphid (Diuraphis noxia) populations on KRWA9 and 2414-
11-2 2414-11-2 wheat genotypes at varying times of infestation
Figure 4. Level of Plant damage caused by different Diuraphis noxia aphid populations on PI624933-1 and
Kwale wheat genotype at varying times of infestation

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