Influence of fungicides and vesicular arbuscular mycorrhiza on growth and nutrient balance of soybean by used DRIS equation

doi:10.4236/as.2012.35089

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Vol.3, No.5, 738-744 (2012) Agricultural Sciences

http://dx.doi.org/10.4236/as.2012.35089

Influence of fungicides and vesicular arbuscular

mycorrhiza on growth and nutrient balance of

soybean by used DRIS equation

Dalshad A. Darwesh1*, Kadija K. Mustafa2

1Department of Environmental Sciences, College of Science, University of Salahaddin, Hawler, Iraq;

*Corresponding Author: [email protected]

2Department of Biology, College of Science Education, University of Salahaddin, Hawler, Iraq

Received 13 June 2012; revised 20 July 2012; accepted 1 August 2012

ABSTRACT

A pot experiment was conducted to study the

effect of mycorrhiza, fungicides in difference

concentration and there combination on growth

and nutrients balance index of soya bean. Tow

AM treatments including with and with out my-

corrhiza and tow fungicides (parasmid and ant-

arcol) each of them including four concentration

(0.0, 0.25, 0.5 and 1 kg·donum−1) were tested in

factorial completely randomized design in the

three replication. The results indicates that the

higher value of total dry matter weight was re-

corded from combination treatment (F1M1C1)

which attained 11.09 gm·pot−1, while the lower

value 2.25 gm·pot−1 was produced by combina-

tion treatment (F2M2C3), however the same com-

bination treatments was showed that the nitro-

gen, phosphorus, magnesium and iron in the

shoot tissues in the mycorrhizal plant with lower

concentration of fungicides were significantly

greater (P < 0.01) compared to control and higher

concentration treatments. The higher concen-

tration of the above mentioned nutrients (28.33,

8.36, 6.34 mg·g−1, 251.00 mg·Kg−1) was recorded.

The result of nutrient index and nutrient balance

index revealed that the lower NBI (56.18) was

recorded in combination treatments (F1M1C1),

while the higher NBI (2033.81) was produced

from combination treatments (F2M2C3), moreover

the results shows significant negative correla-

tion ship between NBI and total dry matter

weight (r = −0.63*) .

Keywords: VA-Mycorrhi za ; DRIS; Nutrients

Balance Index

1. INTRODUCTION

Mycorrhizal symbioses are essential components of

approximately 80% of plant systems, and the beneficial

role that mycorrhizal fungi play in agricultural production

is well recognized [1]. Although the use of pesticides,

herbicides and fungicides is fundamental for cultivation,

there is now a growing body of evidence of their effects

and impact on plant growth and on microorganisms such

as mycorrhizal fungi [2,3]. Systemic fungicides applied to

soil around plants can be absorbed by roots and translo-

cated to other parts of the plant. These kinds of fungicides

are persistent in the plant and their action on mycorrhizal

fungi can either be against their vegetative or their re-

production structures [4]. Benomyl is among the most

frequently used systemic fungicides against pathogenic

fungi of cereals and oil seed plants [5]. It has been found

to negatively affect mycorrhizal symbioses by delaying or

preventing the formation of AM symbiosis between fungi

and roots and by decreasing plant P-uptake [6]. Benomyl

suppresses mycelial growth by preventing nuclear divi-

sion. In fact, this fungicide inhibits mitosis by blocking

the formation of microtubules when chromosomes are

separated. Combined use of multiple types of fungicide

can amplify the negative effects on mycorrhiza. When the

soil was treated with the two fungicides in combination,

mycorrhiza did not appear at all on the roots. This dem-

onstrates that fungicides used against different soil borne

pathogens can have significant side effects on mycorrhiza

in soil, the extent of which may be dependent on the type

of fungicide [7]. The results of [8] show that benomyl

inhibited fungal activity of both internal and external

hyphae at the low application level (1 mg·g–1 soil) corre-

sponding to the recommended field dose. Propiconazole

decreased the activity of the external hyphae at the low

application level (0.21 mg·g–1 soil) but did not affect the

internal activity at any application level. Treating seeds

with different chemicals such as fungicides to ensure their

higher germination is a common practice before planting

[9]. However such treatments may influence soil micro-

bial population including arbascular mycorrhizal fungi

D. A. Dar wesh, K. K. Mustafa / Agricultural Sciences 3 (2012) 738-744 739

developing a beneficial symbiosis association with most

of the terrestrial plants including crops plants. Consid-

eration of the actions and interaction between plants and

soil microorganisms including arbascular mycorrhiza,

influenced by different parameters such as chemicals in

the field can be very useful for the development of fa-

vorable strategies, which can enhance the efficiency of

agricultural production although using different chemical

including fungicides may effect AM activities, which are

of great significance for the production and health of

ecosystems [7]. Accordingly, it can very pertinent to

evaluate how different fungicides may effect the activities

of VM in the soil and how AM can interactively adjust the

unfavorable effect of fungicides on soil microbial popu-

lation. This is because AM have some very great abilities,

including alleviating the effects of different soil stresses

on plant growth [10], and also controlling the adverse

effects of soil pathogens [11]. Soy bean is one of the most

important legume crops for human nutrition and has been

traditionally cultivated in saline soils in arid and semi-arid

regions, the agronomical importance of soybean (Glycine

max L.) is linked to its high protein content (25% - 29%)

[12]. Diagnosis and Recommendation Integrated System

(DRIS) is a method to evaluate a plant nutritional status

that uses a comparison of the leaf tissue nutrient concen-

tration ratio of the nutrient pairs with norms from a high

yielding treatment. The first step to implement DRIS or

any other foliar diagnostic system is the establishment of

the standard values or norms ,thus the DRIS norms and

their standard deviation and coefficient of variance for N,

P, Ca, Mg, K, Fe, and total dry matter ratio were calcu-

lated from the high yielding treatments ,that is to say

DRIS norms were established locally depending on the

nutrient concentration and their ratio from the high

yielding plants (treatment) from these different set of

conditions are similar. After the establishment of DRIS

norms, we calculate the index of each nutrient that

ranges from negative to positive values. The negative

index values indicate that the nutrient levels are below

the optimum. Consequently, the more negative index, the

more deficient the nutrient, similarly a positive index

indicates that the nutrient levels are above the optimum,

and the more positive index the more excessive the nu-

trient that is relative to normal, and the DRIS index is

equal to zero indicating that the nutrient is at optimum

levels, but there are authors that do not consider a nutri-

ent deficiency or excessive when the DRIS indices are

negative or positives and near to zero [13]. The DRIS

also computes an overall index, which is the sum of the

absolute values of the nutrient index, called nutrient bal-

ance index, the smaller the absolute sum of al DRIS in-

dices, the lesser the imbalance among nutrient. Little

information exists on the effect of current soybean seed

applied fungicides on colonization of vesicular arbuscular

mycorrhiza in soy beans. Also the information on the

effect of fungicide and mycorrhiza on nutrient balance by

using the DRIS equation are rare. Thus the objects of this

study are to evaluate the effect of some fungicides on

mycorrhiza activities and yield of soybean, also to inves-

tigate the combination effect of fungicide and mycorrhiza

on nutrient balance in soybean.

2. MATERIALS AND METHODS

The research was conducted in the green house of

College of Science Education, University of Salahaddin/

Erbil. The experiment was a factorial on the basis of

completely randomized design including two factors and

three replication the first factors involved AM treatment

and without AM treatment, the second factors was two

fungicides (Brasamid and Antracol) including four con-

centration (0.0, 0.25, 0.5 and 1 kg·donum−1) and their

combination on growth, nutrient status and nutrient bal-

ance in soybean. Hence 16 experimental treatments were

tested in each replicates making the total of 48 pots .The

soil was dispensed in to plastic pots (7 kg soil per pots) the

5 gm of vesicular arbuscular mycorrhizal inoculum were

distributed and the sterilized soy bean seeds (6 seeds)

were planted in each pots after germination thinned to 4

plants. Plant irrigated with tap water to maintain soil

moisture near field capacity by using weight method. Leaf

samples were collected as described by [14] and washed

thoroughly with dilute acid (0.2 N HCl) and pure water

to remove residues. The samples were dried at 65˚C and

then wet digested in H2O2 and H2SO4 acid (1/1, v/v)

mixture [15]. The N content was measured by the micro-

Kjeldahl digestion method of [16], the P content was

determined by the molybdenum blue colorimetric me-

thod of [17], Ca, Mg and Fe were determined by atomic

absorption, the flame photometric method of [18] was

used to determined K. The plants were harvested before

flowering stage and oven dry to obtain total dry weight.

2.1. Soil Properties

Some physical and chemical characteristics of the soil

under study were as follows: texture: sandy loam; CaCO3

150 g·kg−1; pH (saturated paste) 7.7; EC 0.6 dS·m−1; or-

ganic matter 1.7 gm·kg−1, NH4OAc-extractable K 1.49

meq·100 g−1; exchangeable Ca 11.0 meq·100 g−1; Mg 5

meq·100 g−1; total N 0.44 mg·kg−1; NaHCO3-extractable

P 5.6 mg·kg−1 and DTPA-extractable Fe, as 0.009 mg·kg−1,

respectively. Chemical soil analysis was conducted as

described in [19].

2.2. Calculation of DRIS Indices

The DRIS indices were calculated by using the fol-

owing index equations by [20]: l

D. A. Dar wesh, K. K. Mustafa / Agricultural Sciences 3 (2012) 738-744

740











fNP fNK fNCa fNMg fNFe fNDM

N Indexn

 

;











fNP fNKfNCafNMg fNFe fNDM

P Indexn



;











fNK fPK fKCa fKMg fKFefKDM

K Indexn

 

;















fNCafPCafKCafCa MgfCa FefCa DM

Ca Indexn

 

;















f NMgf PMgf KMgf CaMgf MgFef MgDM

Mg Indexn

 

;















fNFe fPFe fKFe fCaFe fMgFefFeDM

Fe Indexn

 

;















f NDMf PDMf KDMfCaDMf MgDMf FeDM

DM Indexn

 

.



x/y

fXYXYxy1*100k CVif XYxy 

creased the plant growth compared to control .However

the higher concentration of F2 Antracol significantly de-

pressed the total dry weight in comparison with F1

Prasamid concentration in both mycorrhizal and non-

mycorrhizal plants as shown in (Table 4). The higher

total dry matter weight (8.43 gm·pot−1) was recorded in

case application of 0.25 kg·donum−1 fungicides, whereas

the lower value 4.01 gm·pot−1 was recorded in treatment

C3 which include the application of 1 kg·donum−1 of fun-

gicides, the total dry matter of plants was not signifi-

cantly affected by both fungicides, but in general the

Prasamid give higher dry matter than those treated by

Antracol. Soybean shoot treated or not with mycorrhiza

and difference concentration of fungicides are given in

Tables 1-4. The nitrogen, phosphorus, magnesium and

iron in the shoot tissues in the mycorrhizal plant with

lower concentration of fungicides were significantly

greater (P < 0.01) compared to control and higher con-

centration treatments. The higher concentration of the

above mentioned nutrients (28.33, 8.36, 6.34 mg·g−1,

251.00 mg·Kg−1) was recorded in the factorial treatment

(F1M1C1). The application both fungicides with or with-

out mycorrhiza had no significant effect on the Ca2+ and

K+ content in shoot. Fungicide addition in different con-

centration significantly decrease the nitrogen, phosphorus,

magnesium and iron contents of shoot, except Ca2+ and

K+ not effect by fungicides and their concentration. The

higher concentration of above nutrient was recorded in

case of application of (0.25 kg·donum−1) fungicides.



x/y

fX Y1xyXY*100k/CVif x yXY

X and Y = nutrients (N: nitrogen, P: phosphorus: po-

tassium, Mg: magnesium, Ca: calcium, Fe: iron);

N = number of ratios;

x/y mean for X/Y, in reference population (high-yield-

ing treatment, Norm).

CV(x/y) = coefficient of variation for x/y, in reference

population (high-yielding group).

K = sensitivity coefficient (1).

2.3. Statistical Analysis

The experiment was designed in completely random-

ized design with 3 replications (factorial CRD). The ex-

perimental data were analyzed by ANOVA and differ-

ences between the treatment means were separated by

Adj.LSD Test [21].

3. RESULTS

The effects of fungicides applications in different

concentration on plant growth of mycorrhizal and non-

mycorrhizal plants are presented in Tables 1-4. Plant

colonized by mycorrhiza grow well and were significantly

larger (P < 0.01) than non inoculated .The higher value of

total dry matter weight was recorded from combination

treatment (F1M1C1) which attained 11.09 gm·pot−1, while

the lower value 2.25 gm·pot−1 was produced by combi-

nation treatment (F2M2C3). This stimulatory effect of VA

mycorrhiza on total dry matter weight was maintained

with application lower concentration of both fungicides.

In contrast the higher concentration of fungicides de-

The results that are presented in Table 5 show the sig-

nificant effect of mycorrhiza, fungicides, concentration of

fungicides and their combination on nutrient index and

nutrient balance index in soyban. The lower NBI (56.18) e

OPEN ACCESS

D. A. Dar wesh, K. K. Mustafa / Agricultural Sciences 3 (2012) 738-744 741

Table 1. Combination effect of mycorrhiza, fungicides and their concentration on total dry weight and nutrient content of soya bean.

Nutrients concentration

Treatments Nitrogen

(mg/gm)

Phosphorus

(mg/gm)

Calcium

(mg/gm)

Magnesium

(mg/gm)

Potassium

(mg/gm)

Iron

(mg/kg)

Total dry

weight

(gm/pot)

F1M1C0 8.50 5.79 22.50 3.09 26.55 91.33 4.15

F1M1C1 28.33 8.36 35.33 6.34 16.94 251.00 11.09

F1M1C2 19.67 5.36 58.66 1.92 22.28 119.33 6.26

F1M1C3 18.00 5.17 33.00 2.27 23.35 115.00 5.11

F1M2C0 17.83 5.26 47.00 2.51 21.21 128.00 6.20

F1M2C1 16.66 5.51 23.66 3.67 18.01 119.00 7.28

F1M2C2 13.16 5.59 30.66 1.92 28.68 113.33 5.65

F1M2C3 15.50 5.26 35.33 2.27 22.28 71.00 3.29

F2M1C0 12.00 5.69 30.66 2.74 23.35 90.33 5.62

F2M1C1 22.55 7.28 40.00 4.51 22.28 151.33 7.35

F2M1C2 17.83 5.35 35.33 2.27 20.14 130.33 5.70

F2M1C3 16.66 4.52 40.00 2.51 27.61 121.00 5.39

F2M2C0 10.83 3.92 33.00 2.86 27.61 84.33 4.02

F2M2C1 8.50 4.30 35.33 2.74 19.08 124.33 8.02

F2M2C2 12.00 4.36 37.66 2.62 26.55 113.00 5.27

F2M2C3 7.33 3.46 35.33 2.39 22.28 55.67 2.25

Adj. LSD (1%) 9.31 3.48 5.86 16.15 1.96 52.24 2.25

M1 = with mycorrhiza; F1 = Brasamid; F2 = Antraco; M2 = without mycorrhiza; C0 = 0; C1 = 0.25 Kg·donum−1; C2 = 0.5 kg·donum−1; C3= 1 kg·donum−1.

Table 2. Effect of mycorrhiza inoculum on total dry weight and nutrients content of soya bean.

Nutrients concentration

Treatments Nitrogen

(mg/gm)

Phosphorus

(mg/gm)

Calcium

(mg/gm)

Magnesium

(mg/gm)

Potassium

(mg/gm)

Iron

(mg/kg)

Total dry

weight

(gm/pot)

M1 17.95 8.91 36.94 3.20 22.82 131.71 6.33

M2 12.73 7.06 34.75 2.62 23.22 99.08 5.24

Adj. LSD (1%) 3.19 1.19 5.53 0.67 2.01 17.90 0.77

Table 3. Effect of fungicides on total dry weight and nutrients content of soybean.

Nutrients concentration

Treatments Nitrogen

(mg/gm)

Phosphorus

(mg/gm)

Calcium

(mg/gm)

Magnesium

(mg/gm)

Potassium

(mg/gm)

Iron

(mg/kg)

Total dry

weight

(gm/pot)

F1 17.21 8.68 35.77 3.20 22.42 124.00 6.13

F2 13.47 7.29 35.92 2.62 23.62 106.79 5.45

Adj. LSD (1%) 3.19 1.19 5.53 0.67 2.01 17.90 0.77

D. A. Dar wesh, K. K. Mustafa / Agricultural Sciences 3 (2012) 738-744

742

Table 4. Effect of different concentration of fungicides on total dry weight and nutrients content of soybean.

Nutrients concentration

Treatments Nitrogen

(mg/gm)

Phosphorus

(mg/gm)

Calcium

(mg/gm)

Magnesium

(mg/gm)

Potassium

(mg/gm)

Iron

(mg/kg)

Total dry

weight

(gm/Pot)

C0 12.29 7.75 33.29 2.80 24.68 96.50 4.99

C1 19.01 9.54 33.58 4.31 19.08 159.42 8.43

C2 15.67 7.75 40.58 2.18 24.42 117.00 5.72

C3 14.38 6.90 35.92 2.36 23.88 88.67 4.01

Adj. LSD (1%) 5.43 2.03 9.41 1.14 3.42 30.45 1.31

Table 5. Combination effect of mycorrhiza, fungicide and their concentration on nutrient index and NBI of soybean.

Nutrients Index

Treatments

N Index P Index Ca Index Mg IndexK Index Fe Index TDW Index NBI TDW (gm/pot)

F1M1C0 −387.74 20.44 −76.91 55.02 553.10 −91.53 −72.38 1257.13 4.15

F1M1C1 −4.78 −8.39 −4.86 28.09 −5.10 −1.72 −3.24 56.18 11.09

F1M1C2 −95.27 −12.04 121.88 −30.52 158.15 −92.37 −49.83 560.05 6.26

F1M1C3 127.04 7.07 −146.91 −16.56 165.07 −40.39 −95.31 598.36 5.11

F1M2C0 −85.03 1.51 118.81 2.28 166.15 −166.87 −36.86 577.51 6.20

F1M2C1 38.08 16.09 −173.15 −4.97 149.42 −49.40 23.92 455.04 7.28

F1M2C2 −196.06 −9.55 −65.35 −35.94 362.09 2.26 −57.45 728.70 5.65

F1M2C3 −43.08 44.05 66.67 6.21 288.24 −198.05 −164.04 810.35 3.29

F2M1C0 −195.35 −55.84 14.11 31.96 338.22 −111.90 −21.20 768.57 5.62

F2M1C1 −420.80 −70.37 205.67 11.03 303.19 −53.22 24.49 1088.77 7.35

F2M1C2 −32.69 −5.35 −14.48 −6.29 149.43 −46.15 −44.46 298.85 5.70

F2M1C3 −154.26 −62.08 11.42 −6.12 270.14 30.04 −89.15 623.22 5.39

F2M2C0 −299.44 −68.70 79.68 41.24 489.21 −139.68 −102.31 1220.27 4.02

F2M2C1 −438.04 −57.93 186.00 29.50 253.05 −29.75 57.16 1051.42 8.02

F2M2C2 −280.39 −58.11 95.55 14.59 365.44 −78.99 −58.09 951.17 5.27

F2M2C3 −511.02 −57.62 377.42 75.34 564.15 −216.96 −231.31 2033.81 2.25

was recorded in combination treatments (F1M1C1). While

the higher NBI (2033.81) was produced from combination

treatments (F2M2C3) also the nutrient index in the same

table indicate that inoculation with mycorrhiza fungus

changed the order of nutrients requirements in other word

the presence of endophyte improved the balance of ele-

ments, as shown by the lowest sum of indices for the

mycorrhiza soya bean. Thus the result revealed that the N

index, P index, Ca index, K index Fe index and TDW

index reduced from (−85.03, 1.51, 118.81, 166.15, 36.86

and −166.87) in non-mycorrhizal plant to (−4.78, −8.39,

−4.86, −5.10, −1.72 and −3.24) in mycorrhizal plant,

moreover the result show the application of higher con-

centration of fungicides led to increasing the nutrients

imbalance in soy bean.

4. DISCUSSION

The data in the present study indicates that the meta-

bolic activities of the mycorrhizal fungus in infected soy

bean plants varied with treatments of soil by different

fungicides. Differences were reflected in the growth of

mycorrhizal plants, nutrients accumulation, nutrient index

and nutrients balance index. These results support previ-

ous reports on the variation in the toxicity of fungicides to

vesicular arbuscular mycorrhiza [22]. The dominance of

D. A. Dar wesh, K. K. Mustafa / Agricultural Sciences 3 (2012) 738-744 743

sinoculateted treatments with mycorrhiza for nitrogen,

phosphorus, agnesium, iron and total dry weight, this may

be due to the fact that the mycorrhiza increase the ab-

sorption of phosphorus and other essential elements re-

quired for growth these results and explanation are similar

with those reported by [23]. Application of the fungicides

Prasamid and Antracol relatively enhance the plant to

mycorrhizal inoculation these fungicides essentially eli-

minated or reduced the beneficial effect of mycorrhizal

fungus on the growth of plant, macro and micronutrients

nutrition, mycorrhizal fungi can be relatively susceptible

to certain fungicides particularly when the fungicides is

applied to the seeds or into the soil, while other fungi-

cides can stimulate mycorrhiza growth [24]. When Pra-

samid and Antracol were applied half of the field rate

recommended for root diseases control the plant dry

matter nutrients content and nutrient balance were sig-

nificantly increased compared to field rate multiple field

rate application. These results are in agreement with

those of [25] who reported stimulatory effect of lower

concentration of some fungicides on mycorrhizal root

colonization and plant dry matter. It is of interest to note

that the increases in growth of mycorrhizal plants is not

strangely parallel with the concentration of both fungi-

cides, its mean that the dray matter production and nu-

trients uptake reduce with higher rate application of fun-

gicides, this could be explained on the basis that the

fungi toxicity of these fungicides when applied at higher

concentration may be injure fungal cell at many site and

it is also possible that may inhibited a number of en-

zymes involved in nutrient assimilation. [6] reported that

the application of higher concentration of fungicides af-

fect vesicular arbuscular mycorrhizal primarily by inhi-

bition spore germination and infection processes, also

[26]. Their results have indicated that root exudates may

be the factors governing mycorrhizal symbiosis, pesti-

cides that increase root exudation may increase my-

corrhizal infection. Both experimental factors and their

interaction significantly affected soy bean nutrients con-

centration and nutrients balance index. Inoculation of

soybean plants with vesicular arbuscular mycorrhizal

fungi was associated with alterations in the contents of

nutrients in the shoots. Total nitrogen, phosphorus, mag-

nesium and iron tended to be higher in vesicular arbus-

cular mycorrhizal-colonized soybean than in non-colo-

nized plants. the total amount of mentioned nutrients

accumulation in the shoots of soybean were greater when

low concentration of fungicides applied with higher con-

centration of fungicides there was less mycorrhizal effect

on nutrient accumulation. Soya bean plants colonized by

vesicular arbuscular mycorrhiza had no significant dif-

ference in Ca and K content than control, moreover Ca

and K contents of shoot in mycorrhizal and mycorrhizal

plants increase with increasing the concentration of fun-

gicides, however the combination between mycorrhiza

and fungicides enhance the nutrients balance index in

soybean plants. The enhancement of nutrient acquisition

by mycorrhizal infection can be attributed to direct hyphal

uptake or indirect effects brought about by morphological

a physiological changes in the host roots, in addition to

that the VAM differs when subjected to different fungi-

cides and some combination of AM and fungicides may

be more efficient under certain circumstances. According

to our result vesicular arbuscular mycorrhizal inoculation

can significantly alleviate the unfavorable effects of fun-

gicides on soybean yield and nutrients balance. These all

indicated that how using chemicals, among their other

unfavorable effects, particularly on the environments can

influence the nutritional quality of the plants [10].

5. CONCLUSION

The result of this study concluded that the metabolic

activities of the mycorr hizal fungus in infected soy bean

plants varied with treatments of soil by different fungi-

cides. Differences were reflected in the growth of my-

corrhizal plants, nutrients accumulation, nutrient index

and nutrients balance index. These results support previ-

ous reports on the variation in the toxicity of fungicides to

VA mycorrhiza.

6. ACKNOWLEDGEMENTS

Grateful acknowledgement is extended to biology department. Col-

lege of Scientific Education University of Salahaddin-Hawler for pro-

viding laboratory facility.

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[1] Bethlenfalvay, G.J. and Linderman, R.G. (1992) Preface:

Mycorrhizae in sustainable agriculture. ASA Special Pub-

lication No. 54. American Society for Agriculture, Madi-

son.

[2] Ocampo, J.A. (1993) Influence of pesticides on VA my-

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