Physiological potential of soybean seeds treated with thiamethoxam and submitted to storage

doi:10.4236/as.2013.411A003

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Vol.4, No.11 A, 19-25 (2013) Agricultural Sciences

http://dx.doi.org/10.4236/as.2013.411A003

Physiological potential of soybean seeds treated

with thiamethoxam and submitted to storage

Lilian Gomes de Moraes Dan1*, Alessandro Lucca Braccini1,

Alberto Leão de Lemos Barroso2, Hugo de Almeida Dan1, Gleberson Guillen Piccinin1,

Juliana Marques Voroniak1

1Department of Agronomy, State University of Maringá, Maringá, Brazil;

*Corresponding Author: [email protected]

2Department of Agronomy, University of Rio Verde, Rio Verde, Brazil

Received 18 July 2013; revised 18 August 2013; accepted 23 September 2013

tion License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly

cited.

ABSTRACT

Bio-activators such as thiamethoxam have been

used to increase the productive potential of

plants, by means of metabolic and plant mor-

phology modifications. Nevertheless, very little

is known regarding the effectiveness of this in-

secticide when incorporated into lower quality

level soybean seeds, p articularly during storage.

The objective of this work was to assess the in-

fluence of thiamethoxam on the physiological

potential of soybean seeds with different initial

vigor levels during four storage periods. Three

vigor levels of cultivar Anta—low (35%), medium

(70%) and high (85%) vigor—of soybean seeds

were utilized in this experiment. The experi-

ment al de sig n was completely randomized in a 3

× 2 × 4 factorial scheme—3 levels of vigor (low,

medium and high); 2 seed treatments (with and

without thiamethoxam) and 4 storage pe rio ds (0,

30, 60 and 90 days). The analy zed v ariables were:

germination, first germination count, acceler-

ated aging test, seedling length and emergence

speed index. Thiamethoxam-treated soybean

seeds benefit the physiological potential of both

medium and high vigor seeds during storage up

to 30 days. Whereas, the use of thiamethoxam

was shown to be ineffective in improving the

physiological quality of initial low vigor seeds.

Keywords: Glycine max; Bio-Activator; Vigor;

Physiological Quality

1. INTRODUCTION

Soybean crops comprise both increased productive po-

tential and adaptability in different regions of Brazil due

to recent technological advances in the productive seg-

ment. Among the modern farming techniques, the incor-

poration of products such as fungicides and insecticides

to seeds in order to protect the plants and increase the

productive potential is becoming an increasingly wide-

spread practice in most technified agriculture in ad-

vanced countries [1].

In general, insecticides are assessed for their efficiency

on pest control; however, some products can cause ef-

fects yet unknown, modifying the metabolism and mor-

phology of the plant [2]. The discovery of the molecule

thiamethoxam (3-(2-chloro-thiazol-5-ylmethyl)-1,3,5-

oxadiazinan-4-ylidene-N-(nitro)amine) brought about new

perspectives for agriculture, mainly regarding seeds’

treatment. A number of studies have shown the efficiency

of thiamethoxam as a bio-activator, promoting incre-

ments on germination, emergence, seedling length (roots

and aerial parts) and dry phytomass. In addition, the

product can eventually improve the performance of

plants under stress, besides increasing total protein and

enzyme content levels in plants [3]. Thiamethoxam

proved to be efficient as a bio-activator, essentially for

soybean seeds [4].

According to [5], thiamethoxam positive physiological

effect is related to germination, vigor, enzymatic activity,

nutrient level, height plant, stem diameter, radicular de-

velopment, phytomass, pods’ number per plant, grain

weight and yield, providing an increment, on average of

four bags per hectare of soybean. Increased vigor, pro-

ductivity, leaf and radicular area index, a more uniform

stand, emergence uniformity and better initial develop-

ment of soybean seedlings were observed by [6].

Thiamethoxam used in soybean seed treatment, ac-

celerates germination and induces further development

L. G. de Moraes Dan et al. / Agricultural Sciences 4 (2013) 19-25

of embryonic axis, minimizing the negative effects in

situations such as presence of aluminum, salinity and

water deficit. This acceleration of germination is due to

the stimulation of peroxidase activity, preventing oxida-

tive stress [7], thus, reducing the time of crop establish-

ment in field, and therefore, lessening the harmful effects

of either weed competition or essential nutrients present

in soil [2].

Nevertheless, it is equally important to consider that

the effectiveness of thiamethoxam treatment may depend,

among a number of factors, on seed vigor at the time of

product application. According to [8], the utilization of

biologically active compounds such as bio-activators can

either cease or reduce the impact of adverse factors on

seeds’ quality and performance. Moreover, one of the

greatest limitations of soybean farming is unavailability

of high-vigor seeds at the sowing time [9]. Additionally,

little is known about the bioactive effect of thiameth-

oxam applied to initial low- or medium-vigor soybean

seeds.

Another factor to be considered is that soybean seeds

usually present reduction in physiological quality during

long term storage [10], even under favorable conditions.

In Brazil, the preservation of soybean seeds’ quality dur-

ing storage is among the largest obstacles for seed indus-

try. According to [11], seed quality maintenance until

sowing time ensures the increased emergence potential

of seedlings in field, resulting in productivity gains. Dur-

ing storage, seed quality can either remain close to the

initial level or reduce to a level which makes the seed

unavailable for sowing. This fact is related to several

circumstances, such as environmental conditions during

seed production, occurrence of pests, oil content in seeds,

water content, mechanical damages over the course of

processing and packaging, temperature and relative hu-

midity of the air during storage [12-14], besides chemical

treatment of seeds using insecticides [15,16].

Thus, thiamethoxam proved to be effective as a bio-

activator on soybean seeds’ treatment, implying the ne-

cessity to assess its effectiveness regarding its use in

lower physiological quality level seeds, particularly dur-

ing storage.

In this scope, this present study had the objective to

assess the influence of thiamethoxam on physiological

potential of soybean seeds with different initial vigor

levels in four periods of storage.

2. MATERIAL AND METHODS

2.1. Experiment Installation and Conduction

This experiment was conducted at the State University

of Maringá (UEM), Maringá, Paraná State, and assess-

ments on physiological quality of seeds were carried out

at the Laboratory of Seed Technology at the Applied Ag-

riculture Research Nucleus (NUPAGRI), Center for Ap-

plied Agricultural Research of UEM.

Soybean seeds of cultivar Anta 82 RR from Adriana’s

Seeds—maturity group 7.4 and semi-determinate growth

habit—were used in this study. Tetrazolium test was used

for this cultivar [17] to select lots with three levels of ini-

tial vigor: low (35%), medium (70%) and high (85%).

The experimental design was completely randomized in

a 3 × 2 × 4 factorial scheme—3 levels of vigor (low, me-

dium and high); 2 seed treatments (with and without

thiamethoxam) and 4 storage periods (0, 30, 60 and 90

days).

Chemical treatment of soybean seeds was carried out

using thiamethoxam (Cruiser® 350 FS), at a dose of 2 mL

per kg of seeds.

Syrup (product + distilled water) was applied using a

graduated pipette at the bottom of a transparent plastic

bag and spread across its walls up to 15 cm high. The

volume of syrup was 0.3 L 100 kg−1 of seeds. The wit-

ness received distilled water on the same volume of

syrup with the insecticide. The quality of treated seeds

was assessed at the following times: 0, 30, 60 and 90

days after soybean seed treatment.

2.2. Assessed Features

The following tests were carried out both at the labo-

ratory of seeds and greenhouse using trays containing

washed and sterile sand, and then utilized to assess the

physiological quality of soybean seeds.

2.2.1. Germination Test

Four subsamples of 50 seeds were used for each lot

and replicate use; they were then placed for germination

between three-leaf germination paper imbibed in distilled

water in a ratio of 2.5 times as much the paper dry mass.

They were then rolled up and placed in a Mangelsdorf

germinator kept at a constant temperature of 25˚C.

Evaluations on the percentage of normal seedlings were

carried out both on the fifth (first count) and eighth (final

count) days, according to the established criteria of the

Seed Analysis Rules [18]. First count was considered as

an indicative of vigor and final count as the total seeds

germination percentage.

2.2.2. Accelerated Aging T est

Plastic boxes (gerbox) provided with horizontal

stainless steel screen fixed at the median position were

used in this experiment. 40 mL of distilled water (in or-

der to get approximately 100% of Relative Humidity)

was added to the bottom of each gerbox, and seeds were

uniformly distributed on the screen in order to cover its

surface, forming a single layer. Subsequently, the boxes,

together with the seeds, were sealed and stored in a Wa-

ter Jacket Incubator and kept at a constant temperature of

L. G. de Moraes Dan et al. / Agricultural Sciences 4 (2013) 19-25 21

41˚C for 48 hours [19]. After this period, seeds were

taken for germination under the same conditions de-

scribed previously on germination test.

2.2.3. Emergence speed index

It was carried out from sowing in trays containg 8 kg

of washed and sterile sand in 4 replicates of 50 seeds for

each sample. Test was conducted at the greenhouse and

emerged seedlings were counted daily between the on-

sets (five days after installation) of emergence until the

numerical stabilization of the counts (ten days after in-

stallation). Emergence speed index calculation was ac-

complished in accordance with [20], using the formula

proposed by [21].

2.2.4. Seedling Length

Five samples of 20 seeds from each treatment were

distributed in germination paper rolls imbibed with dis-

tilled water in a 2.5:1 ratio (mL distilled water per dry

paper mass in grams) and kept into a germination cham-

ber at 25˚C for five days [20]. A line was longitudinally

traced in the upper third upon the humidified paper towel,

where seeds were placed pointing the micropyle down-

ward. Seedlings length which was considered normal [18]

was determined at the end of the fifth day using a milli-

meter-gauged ruler.

2.3. Statistical Analysis

Variables which characterize the physiological quality

of seeds were all submitted to variance analysis

(ANOVA) using the Statistical Analysis System-SISVAR.

Regression analysis was utilized to verify polynomial

adjustment for dependent variables according to storage

periods, at 5% probability [22].

3. RESULTS AND DISCUSSIONS

3.1. Germination Assessment

Significant interaction was shown for both treatment

of seeds and periods of storage (Figure 1) by assessing

the germination test results. For high and medium-vigor

seeds, submitted or not to thiamethoxam treatment, re-

ductions in germination performance over storage were

similar. Nevertheless, taking into account the study ac-

complished by [7], thiamethoxam, used for seed treat-

ment accelerated soybean germination, whose effect was

pronounced when twice-fold dose of that recommended

for the crop was utilized (4 mL·kg−1 of seed).

Furthermore, at 90-day storage, germination percent-

ages were higher to 80% in both seed treatments assessed,

thus being within the trading-accepted standards of soy-

bean seeds in Brazil [23]. In thiamethoxam-treated soy-

bean seeds any germination and vigor differences were

found [24], as well as [25] who observed no germination

Figure 1. Polynomial regression for soybean seeds germina-

tion percentage with different levels of initial vigor, treated (T)

and not treated (NT) with thiamethoxam in four periods of

storage.

effects on bean seeds.

The lack of thiamethoxam positive effects on germina-

tive performance in this current work may be on the ba-

sis of the recommended dose used for the crop (2

mL· k g −1 of seed). However, the choice for using the

recommended dose in this study was mainly not only due

to economic viability but also to environmental and eco-

logical aspects related to chemical dose increments.

For low vigor thiamethoxam-treated seeds (Figure 1),

47% germination was observed in the onset of storage,

whereas for non-treated seeds, germination level was

38%. This fact shows the positive action of thiameth-

oxam treatment in low-vigor seeds, when not stored.

Nevertheless, the effect of storage is negative for both

treated seeds and for those which received no chemical

treatment. These results are in accordance with [16] who

observed that soybean seeds treated or not with thia-

methoxam showed germination reduction over a 45-day

storage.

3.2. First Count of Germination Test

Significant interaction between seed treatment and

storage period (Figure 2) was observed at the polyno-

mial regression assessment for normal seedling percent-

age in first germination count. In high-vigor soybean

seeds, treatment using thiamethoxam and the witness

showed normal seedling percentage of 0.15% and 0.14%,

respectively in each day’s storage. Thus, demonstrating

similarity to that reduction of physiological performance

of initial high-vigor seeds among treatments during stor-

age.

In both medium- and low-vigor seeds (Figure 2), re-

ductions on normal seedling percentage in the first count

were 0.10% and 0.17%, respectively for each day’s stor-

L. G. de Moraes Dan et al. / Agricultural Sciences 4 (2013) 19-25

Figure 2. Polynomial regression for normal seedling percent-

age in the first count of soybean seed germination with dif-

ferent initial vigor levels, treated (T) and not treated (NT) with

thiamethoxam in four periods of storage.

age for thiamethoxam-treated seeds. Whereas, for the

witnesses (no chemical treatment), vigor decrements

were 0.16% (medium vigor) and 0.21% (low vigor) for

each day’s storage. These results support thiamethoxam

beneficial effects on vigor maintenance of soybean seeds

with initial quality ranging from intermediate to low

during storage [26], when working with cotton crops,

concluded that higher increments are reached by using

thiamethoxam in lower physiological quality seeds, par-

ticularly in vigor tests. Thiamethoxam promotes soybean

seed expression related to both enzyme synthesis and

activity by modifying amino acids of phytohormone pre-

cursors, resulting in an increase of germination, vigor

and radicular growth [27].

Nevertheless, at day 90 of storage, only initial high-

vigor seeds showed normal seedling percentage over

80%, despite the reduction observed along with storage

period increment.

3.3. Vigor Assessment in the Accelerate

Aging Test

Results on accelerated aging test (Figure 3) showed

that in initial high-vigor thiamethoxam-treated seeds,

decrement was more intense in normal seedling percent-

age during storage, ranging from 89.8% at the beginning

of storage to 57.4% after 90-day storage. On the other

hand, assessment carried out right after thiamethoxam

application (period zero) in seeds, percentages of 89%

for vigor and 83% for treatment without the insecticide

were observed. These results show that the positive ef-

fects on initial high-vigor soybean seeds is limited to

short term storage when submitted to stress conditions in

the accelerated aging test.

Figure 3. Polynomial regression for normal seedling percent-

age after accelerated aging test of soybean seeds with different

initial vigor levels, treated (T) and not treated (NT) with thia-

methoxam in four periods of storage.

In medium-vigor seeds, thiamethoxam incorporation

via seed treatment presented positive action during stor-

age because angular coefficient indicated 0.16% reduc-

tion on seed vigor for each day’s storage. In contrast, for

the witness, seed vigor decrease after accelerated aging

test was 0.19% in each day of storage (Figure 3).

Accelerated aging test is one of the most used for

physiological potential assessment of several species

[28]. This test has as key principle lead to a considerable

increase on seed deterioration when exposed to high lev-

els of temperature and relative air humidity, which are

considered preponderant environmental factors for both

intensity and speed deterioration. It is notorious that

stress conditions induce reactive oxygen species (ROS),

which damage biomolecules, leading to cellular death.

However, there are antioxidative enzymes such as su-

peroxide dismutase and peroxidase which are responsible

for ROS elimination. According to [7], thiamethoxam

speeds up seed germination under stress conditions by

eliminating the ROS generated. Thiamethoxam moves

through plant cells and activates various physiological

reactions, such as the expression of functional proteins

related to defense mechanisms of plants against stress

factors, for instance, water deficit, high temperatures,

toxic effects, just to name a few [4]. Thus, the influence

of thiamethoxam on these defense mechanisms and on

antioxidative enzyme activities may have played a role

against adverse conditions of high temperature and in-

creased relative humidity imposed by the aging test, and

therefore, having attenuated the physiological potential

decrement of soybean seeds during storage.

In initial low-vigor seeds (Figure 3), the use of thia-

methoxam showed no positive effects on the improve-

ment or maintenance of seed vigor during storage for 90

days, once angular coefficient presented the same reduc-

L. G. de Moraes Dan et al. / Agricultural Sciences 4 (2013) 19-25 23

tion in both seed treatments (treated and not treated).

3.4. Emergence Speed Index Assessment

In the results of emergence speed index (ESI) (Figure

4), reductions observed upon this variable were also sig-

nificant as storage period increased in both seed treat-

ments. ESI reductions observed over the course of stor-

age, initial high- and low-vigor seeds highlight thia-

methoxam negative effect on soybean seedling emer-

gence speed derived from treated seeds and submitted to

storage. In contrast, this insecticide treatment in medium

vigor seeds showed similar effect when compared to

those of the witnesses on the ESI performance, present-

ing 0.05 reduction unit each day in which treated seeds

were stored. These results are conflicting to those found

by [4], who concluded that soybean seeds treated with

thiamethoxam showed faster germination due to the ac-

tion of the product that stimulates enzymatic activity,

providing a more uniform and better initial seedling de-

velopment.

3.5. Seedling Length Assessment

Soybean seedlings height (Figure 5) showed signifi-

cant reduction (p < 0.05) as storage period increased in

both seed treatments. In initial high- and medium-vigor

thiamethoxam-treated seeds, reductions on seedling

lengths in the order of 0.08 and 0.04, respectively, were

observed in each day of storage. In the witnesses, reduc-

tions of 0.04 and 0.03 cm were observed in each day of

storage for high- and medium-vigor seeds.

Despite the highest reduction on this response variable

during storage is determined in thiamethoxam treatment,

seedling length was 4.94 cm taller than the witness for

initial high-vigor seeds at the storage period onset (pe-

riod zero).

In medium-vigor seeds, the maximum efficiency of the

product, for this variable, enabled seedlings height to in-

crease up to 3.0 cm, comparatively to seeds with no

product applications. Thus, that the bioactive effect of

thiamethoxam in initial high- and medium vigor-seeds is

limited to a 30-day storage period is evidenced. The

thiamethoxam tendency to increase seedling length cor-

roborates with the effect observed by [24] in soybean

crop.

After a 30-day period, the natural aging process of

seeds, which occurs during storage and culminates in

physiological quality loss, negatively interferes in the

thiamethoxam action in order to increase seedling growth.

This fact can also be observed on results of seedlings

derived from initial low-vigor seeds, treated or not with

thiamethoxam, which showed the same reduction during

storage. Thus, the absence of thiamethoxam positive in-

fluence on seedling length from low-vigor seeds may be

Figure 4. Polynomial regression for the emergence speed in-

dex (ESI) of seedlings derived from soybean seeds with dif-

ferent initial vigor levels, treated (T) and not treated (NT) with

thiamethoxam in four periods of storage.

Figure 5. Polynomial regression for seedling length (cm) de-

rived from soybean seeds with different initial vigor levels,

treated (T) or not treated (NT) with thiamethoxam in four pe-

riods of storage.

due to the advanced deterioration degree which these

seeds presented, indicating that this insecticide shows no

effect as a bio-activator in low quality seeds.

An overall review of data obtained with soybean cul-

tivar Anta suggests that, for most assessments, greater

increments were reached using thiamethoxam in higher

physiological quality seeds. In addition, physiological

quality of soybean seeds was favored by thiamethoxam

treatment, providing more expressive effects right after

its application up to a 30-day storage period for treated

seeds. These characteristics of thiamethoxam, along with

the use of increased genetic and physiological quality

seeds maximize seed production capacity.

L. G. de Moraes Dan et al. / Agricultural Sciences 4 (2013) 19-25

4. CONCLUSION

Soybean seed treatment using thiamethoxam favors

the physiological potential of high- and medium-vigor

seeds during 30-day storage. In initial low-vigor seeds,

the use of thiamethoxam is inefficient in improving

physiological quality.

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