Vol.2, No.4, 457-464 (2011)
doi:10.4236/as.2011.24059
C
opyright © 2011 SciRes. Openly accessible at http://www.scirp.org/journal/AS/
Agricultural Scienc es
Preparation and mechanism analyses of a new corn
seed coating agent
Defang Zeng*, Xinrong Luo
School of Resource and Environ mental Engineering, Wuhan University of Technology, Hubei Key Labora tory of Mineral Resources
Processing and Environment, Wuhan, China; *Correspon ding Author:
Received 9 September 2011; revised 24 October 2011; accepted 31 October 2011.
ABSTRACT
Due to the severity of the disease and yield losses
of corn, this study had achieved a novel corn
seed coati ng ag ent whic h was prepared with poly -
saccharide, fertilizer, microelement and other
assist ants as raw materials. The dilution and the
pH value of the corn seed coating agent, were
studied. And the biologic al toxicity and potential
hazards of the agent to environmental protection
were assessed. Over the whole trial, the best
formula and optimum conditions of a new corn
seed coating agent were determined. Also re-
sults indicated that compared with the tradi-
tional one Jinong No. 4, yields of corn was in-
creased over 9.5%, but cost was reduced over
18.75%. To sum up, this agent self-made had
three primary characteristics of high yield, less
cost and friendly environment. So it had an im-
portant value of application and promotion in
main production zone of corn and can bring us
obvious economic and environmental benefits.
Keywords: Coating; Germination Rate; Inhibitory
Rate; Antifeedant Rate; High Yield; Friendly
Environment
1. INTRODUCTION
Corn is one of the major crops in China, but in recent
years, the soil-borne of corn has been aggravated year by
year, and corn yields decreased obviously. So the soil-
borne of corn has become one of the main factors of
corn yields and quality [1,2]. To protect corn from un-
derground pest, provide the source of fertilizer, and in-
crease yields of corn, seed-coating is an effective tech-
nology and has been widely applied for crop protection
[3]. Studies have shown that a seed coating is effective
in preventing and controlling mould-induced diseases
and pests causing them, promoting seedling growth, and
increasing yields [4,5].
Since the seed coating agent for rain-fed crops first in
the 1930s, the research and development of the seed
coating agent has been rapid during the past two decades
and has provided an economical approach to seed en-
hancement, especially for larger seeded agronomic and
crops [6-8]. However, the conventional toxic seed coat-
ing agent currently used such as carbofuran and thiram
unavoidably brought serious harm to the environment
and human and make the development of ecoagricultural
practices impossible [9-11]. Studies showed that only 7
milligrams of carbofuran can kill a person. In add ition, it
leaves a Long-term pollution, elimin ated difficultly from
the environment, and becomes a great hidden danger in
China’s ecological agriculture [12,13].
The dire threat of current conventional toxic seed co at-
ing agen t forces to seek for a safe and enviro n- mentally
friendly corn seed-coating agent to resolve the current
agricultural and environmental issue. After years of re-
search, the novel seed coating agent (short for LSCA)
instead of conventional toxic fungicide, was achieved
through advanced biological chelating technology, which
prepared with natural polysaccharide as the main mate-
rial and high-activity plant growth regulator as the aux-
iliary material. Compared with a conventional seed
coating (Jinong No. 4), the effect of LSCA on the resis-
tance to smut fungus, seed germination, and corn yield,
as well as its toxicity was investigated. Yields of corn
was increased over 9.5%, but cost was reduced over
18.75%. With the application of the new coating agent,
corn seed-coatin g agent has better quality and properties
such as high yield, low cost, friendly environment, and
would have significant economic and environmental
benefits.
2. MATERIALS AND METHODS
2.1. Materials
2.1.1. Major Apparatuss and Glassware
The major instruments and glassware used in present
study were electron constant speed mixer GS28B (Shang-

D. F. Zeng et al. / Agricultural Sciences 2 (2011) 457-464
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458
hai Anting Electronic Instruments Plant, Shanghai, China),
high pressure steam sterilizer Y XQ-SG46-48SA (Shang-
hai Boyi Industry Co. Ltd., Shanghai, China), constant
temperature and humidity incubator WS-01 (Hubei
Huangshi Hengfeng Medical Instrument Co., Ltd., Hubei,
China), electronic balance (FA2004, ShanghaiYuefeng
Instrument Appearance, Ltd., Shanghai, China), warm
up hygrometer STH950 (Summit, USA), biological mi-
croscope BX41-12HO2 (Olympus Corp., Japan) and petri
dishes (90 cm dia., Shanghai Yuejin Medical Treatment
Instrument Pl ant, Shanghai , China). 24 Ori fices of plate.
2.1.2. Major Chemicals and Reagents
The major chemicals and reagents used in present study
were polysaccharide (PC, self-made in lab), film-form-
ing agent, polyethylene glyco (self-made in lab), the ag-
riculture regulates, acetic acid, carbamide, natrium hy-
droxydatum, violet pigment (Guangdong shantou Ming
frideric food additive Co., LTD), sand, The conventional
corn seed coating agent Jino ng No. 4 ® (Hubei Provincial
Seed Group Co., Ltd., China) (Prefecture Seed Science
and Technology Development, Ltd., Henan, China), Ar-
tificial feed, Smut fungus (North Coating Agents Factory,
Tianjin, China), Corn seeds (China Jilin Seed Co.).
2.1.3. Experiment al Animals
Cutworm (the consist ent growt h) (provi ded by Hei long-
jiang Agriculture University, Sprague-Dawley rats (20
males, 20 females; 200 ± 12 g) (Animal Testing Center
of Tongji Medical College of Huazhong University of
Science and Technology).
2.2. Experiment Methods
The experiment stepwise development and technique
route of corn seed coating agent was showed in the Fig-
ure 1. According to three principles of cheapness, envi-
ronmental protection, high efficiency, a number of the
recipes of the corn seed coating agents were designed to
prevent seedling and increasing yields. Once all the ma-
jor performance parameters of agents made in lab exceed
those of the traditional one Jinong No. 4, we optimized
further on the basis of experiment to get the best recipe,
then carried the field text, to determine the best formula
of agent with a high cost performance and environmental
friendship.
2.2.1. Comparative Experiment of the Corn See d
Coating Agent in the Laborato r y
As stated by the manufacturer of Seafresh Chitosan
(Lab) Co. Ltd., China, the natural polysaccharide with a
molecular weight of from 20 to 1500 KD and deacetyla-
tion degree of from 80% to 90%, was prepared as: 1
wt% acetic acid to dissolve polysaccharide completely to
a final concentration 1wt% under stirring at 25˚C for
approximately 4 h.
The optimal coating formulation was determined th-
rough orthogonal test. Aqueous solutions of fertilizer,
microelement, ethylene glycol, film former and crocuses
red were prepared respectively at a certain concentration.
After mixing every component completely at room tem-
perature.
Concrete experimental process are as follows:
2.2.2. Test of Seed Germination
Comparative experiment of the corn seed coating
agent in the laboratory was as follows: the same corn
seeds were coated with the new seed coating agents
(A-E) and the conventional one (Jinong No. 4) respec-
tively, and then put in the constant temperature and hu-
midity incubator under the same condition of the con-
stant temperature 28˚C ± 1˚C and the constant humidity
85%. Experimental results of the germination tendency
and germination rate should be observed and compared
every day. Each treatment was replicated 3 times.
Design and preparat ion
of corn seed coating
agent
Primary comparative
tests in the lab Analysis o f re sultsInhibito ry rate
Germination rate
Ge rm inat ing tendency
tendency
Confirm t he best
formula in the lab
Field tests
Confirm t he fin al
formula of the new
agent
Ana lysis of results
Cost
Yields
Toxicity
p
ollution
Antifeedant rate
Figure 1. The experiment stepwise development and technique route of corn seed coating agent.

D. F. Zeng et al. / Agricultural Sciences 2 (2011) 457-464
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459459
According to the seed test procedures methods of in-
ternational seed association (ISTA) [14], germinating
tendency (GT) was calculated on the 4th day and germi-
nation rate (GR)was calculated on the 7th day. The cal-
culation formulas were as follow:
GT (%) = (G3/S) × 100 (1)
GR (%) = (G7/S) × 100 (2)
where G3 is the number of germinated seeds on the third
day; G7 is the number of germinated seeds on the sev-
enth day; S is the number of total seeds investigated.
2.2.3. Antibacterial Experiment in the Lab
According to microbiological test requirements for
inhibitory rate trial, the Setosphaeria turcica blocked 5
mm were inoculated to agar which was sterilized with a
high-pressure steam and mixed with different agents,
water as to control respectively, and then cultivated at
25˚C for 2 days [15]. The tests were repeated 3 times
and recorded the diameter using cross measuring method.
The following formula was used to calculate the inhibi-
tory rate of corn seed coating agents.
IR (%) = [(D0 – D)/D0] × 100 (3)
where IR is Inhibitory rate; D0 is colonies diameter in
the control petri dish, and D is the colonies diameter in
the test petri dish.
Once all the major performance parameters of agents
made in lab exceed those of the traditional one, then we
optimized further on the basis of experiment to get the
best recipe, named after LSCA, repeated the aboved and
stability experiment.
2.2.4. Antifeedant Test of Pests in the Lab
According to the guideline for laboratory bioassay of
pesticides, the antifeeding effect of LSCA was studied
with the artificial mixed feeding method The process of
the antifeeding test was as follows: a certain amount of
artificial feed mixed a certain amount of LSCA, the
proportion (w/w) were 5%, 10%, 15%, and an untreated
group (CK) as control was designed. Then the feeds
above and the consistent growth cutworms were put into
the Orifices plates. Cutworms were feeded in the incu-
bator at a temperature of 25˚C ± 1˚C, and relative hu-
midity of 75% - 85%. After 48 h the remaining feeds
(control and treated) were weighted, the test was valid
only the mortality of larvae within 5%. Every text repli-
cated three times the formula is as follows [16]:
AR (%) = [(W0 – W)/W0] × 100 (4)
where AR is Antifeedant rate; W0 is the weight of con-
trol feed and W is the weight of feed treated by corn
seed coating agent.
2.2.5. The Security Test
According to the toxicological test methods of pesti-
cides for registration, the toxic effect of LSCA and Jing
No. 4 were studied with the LD50 (the median lethal dose)
method. In the study, the rats were fasted overnight, then
treated at various do sages of LSCA and Jinong No. 4, b y
oral ingestion and skin infection respectively. The toxic
symptoms of rats and the median lethal dose (LD50)
were recorded after 14 days. According to the results of
the toxicity test, the difference of toxicity LSCA and
Jinong No. 4 was estimated.
2.2.6. Field Trials
Through the above formula and the experimental re-
sults, we calculated the germinating tendency and ger-
mination rate, inhibitory rate, antifeedant rate etc., and
finally the best formulation was evaluated in the field.
Field trial was carried out to verify further whether the
efficacy of (LSCA) in the laboratory was stabilized, and
the effect on the yield was higher than that of the tradi-
tional one. In this test, the method of seed treatment was
the same as that in the laboratory experiment, the corn
seeds were coated with LSCA and Jinong No. 4 in the
proportion of 1:50 (w/w).The experiment was conducted
at Hubei Provincial Seed Group Company. The experi-
ments were designed as a randomized block, and each
treatment was replicated three times.
2.2.7. Statistic Alanalysis
Analyses of variance (ANOVA) and Duncan means
comparisontest with asignificance levelof 0.05 were ap-
plied.
3. RESULTS AND DIS CUSSION
3.1. Results
3.1.1. Primary Experiment Results in the
Laboratory
The primary purpose was to select the best environ-
mentally friendly corn seed coating agent, the price-
performance ratio of which preponderated over that of
Jinong No. 4. Primary experiment results in the labora-
tory were showed in Tab le 1, corn seed coating agents
A-H were prepared-self, the traditional one Jinong No. 4,
CK uncoated seed as control.
It can be seen from Ta ble 1, different agents had dif-
ferent influences on corn seedling growth, and the main
indexes of B-E exceeded over Jinong No. 4. It can be
seen that E had a most effect on germinating tendency,
germination rate, inhibitory rate, and enhanced by 9.4%,
4.0%, 8.1% respectively compared to Jinong No. 4. E
had a positive promotional effect on seed germination

D. F. Zeng et al. / Agricultural Sciences 2 (2011) 457-464
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460
and seedling growth. So E was the best formulation,
named after LSCA.
It can be seen from Ta b l e 2, these results were stabi-
lized and consistent with those in Table 1.
These results showed that seed coating agents played
a significant role in improving germinating tendency,
germination rate and inhibitory rate, but th e efficiency of
LSCA was obviously superior to Jinong No. 4.
3.1.2. Optimization Experiments in the
Laboratory
Based on the best formula of the seed coating agent
LSCA, changing two main factors such as pH and the dilu-
tion to confirm the optimum conditions of seed coating
agent, so that the new agent can achieve the best effect
and improve the price-performance ratio.
3.1.3. Influence of pH Value of Seed Coating
Agent
Influence of pH value (the range of 4.0 - 7.0) of seed
coating agent was shown in Table 3.
As can be seen in Table 3, the best effects of germina-
tion tendency, germination rate and inhibitory rate were
observed at pH 6.0. From 4.0 to 6.0, the higher the pH
value was, the higher main parameters, and they reached
the maximum at pH 6.0. When pH was above 6.0, with
the increasing of the pH value, they declined. Therefore
the best pH value of corn seed coating agent was 6.0.
Table 1. The preliminary experi mental re sult s of th e seed co at in g
agents in the laboratory.
Treatments GT (%) GR (%) IR (%)
A 72.5c 86.4b 80.3c
B 77.2b 89.8b 83.7c
C 80.2a 92.4a 86.8b
D 83.6a 94.3a 89.5b
E 85.3a 93.6a 92.3a
F 80.5a 92.7a 91.2a
G 77.4b 90.1a 88.5b
H 75.6bc 88.4b 85.8b
Jinong No. 4 76.9b 88.7b 84.2c
CK 68.5d 76.4c 0
Averages in column separated by Duncan’s new multiple range test, 5%
level. A-H: seed coating agents self-made; Jinong No. 4: a conventional
seed coating agent; CK: blank control. GT: Germinating tendency; GR:
Germination rate; IR: Inhibitory rate.
Table 2. The preliminary experimental results of the seed
coating agents in the laboratory.
Treatments GT (%) GR (%) IR (%)
LSCA 88.6b 94.8a 93.6a
Jinong No.4 79.5d 87.1b 85.6bc
CK 78.6d 85.5bc -
These results are consistent with those reported by
Jun-ang [17] that the optimal pH range was between 5.3
and 6.0 for the highest antifungal activity. This increase
in pH was due to the increased consumption of H+ ions
in solution by the protonation of the free amine groups
of polysaccharide [18]. When the pH value was 6.0, the
protective film on the surface of the seeds combined
with seed coat more tightly and did not change various
functions of seed inner, so when the pH value was 6.0,
the film had better permeability which can guarantee
plants supplyin g ample oxygen an d water, and creating a
beneficial condition for the better growth of seeding.
3.1.4. Influence of the Dilution of Seed Coating
Agent
Results of the experiment in the lab showed different
proportion of agent and seeds had different influence on
corn seedling growth in Table 4.
It can be seen from Table 4, the effect on sprout poten -
tial, germination rate of 1:30, 1:40 was lower than that of
CK, especially at 1:30, the agent restrained the growth of
seedings, showing obviously a series of signs of hide-
bound symptoms such as shorter seedling and stem thin.
When at 1:50, 1:60, 1:70, the main parameters of LSCA
was higher3 times than that of CK and the growing and
leaf color of seedings were consistent. Therefore, 1:50
was the best proportion. When at 1:50, the germination
Table 3. The influence of pH on the main performance pa-
rameters of seed coating agent.
pH GT (%) GR (%) IR (%)
4.0 74.5d 82.5c 83.6c
4.5 78.9d 86.5b 86.3b
5.0 82.6c 89.9b 88.5b
5.5 86.6b 91.7a 91.5a
6.0 88.6b 93.7a 93.6a
6.5 85.6bc 91.4a 92.9a
7.0 82.6c 90.6a 90.2a
Averages in column separated by Duncan’s new multiple range test, 5%
level.
Ta ble 4. Compare of germination energy and germination rate
in different proportion of seed.
The dilution of LSCA GT (%) GR (%) IR (%)
1:30 84.9c 89.9b 89.5b
1:40 86.6b 91.7a 91.3a
1:50 88.6b 94.8a 92.9a
1:60 87.5b 92.6a 90.5a
1:70 85.6bc 90.6ab 87.6b
CK 82.6c 86.5b -
Averages in column separated by Duncan’s new multiple range test, 5%
level.

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461461
rate was up to 9 4.8%, which displayed obviously to pro-
mote sprout and growth of seedings. It can be explained
that when the proportion was 1:50, the protective film
provides the favorable conditions for seed germination,
growth, and plays significant role in maintaining bacte-
ricidal and insecticidal effect.
3.1.5. Antifeedant Effect of Seed Coatings
Results of antifeeding test showed that LSCA had an
excellent antifeedant effect on selective antifeedant rate
by observing experimental process and analysing of
weight. According to the observation, the artificial feed
containing LSCA was rarely bitten, while the artificial
feed without LSCA (CK) was nearly eaten up. From
Table 5, it showed when the proportion of LSCA was
15%, 10%, 5% on cutworm, the antifeedant rates were
respectively 87.4%, 70.9% and 48.3%. All of those in-
dicated the antifeedant effect of LSCA was increased
with increasing dosage of LSCA.
3.1.6. Results of Acute Toxicity Test
Results of acute toxicity test showed there were sig-
nificant differences in the median lethal dose (LD50) be-
tween LSCA and Jinong No. 4 (Table 6).
In the acute oral toxicity test, rats infected with a high
dosage of Jinong No. 4 exhibited toxic symptoms after 3 -
6 min such as systemic muscle spasm, salivation, and
even convulsions, and sticky nasal and ocular secretions
in the 12 hours before death. The rats infected with low
dosages still showed a sign of slight muscle spasm.
However, the rats infected with LSCA didn’t exhibit the
previously mentioned symptoms and exposured to a low
level. The datas showed that the acute oral LD50 and
acute dermal LD50 in female rats were 607 mg·kg–1 and
2106 mg·kg–1, respectively. In the acute skin toxicity test,
the rats were observed consecutively for two weeks
when they were infected with the toxicant By safety ex-
periment, we see that the acute toxicity of Jinong No. 4
was significantly higher (about 2 times) than that of
LSCA. Therefore, the novel seed-coating agent LSCA
was safe for human and meets the requirements of the
environment protection during usage and disposal.
3.1.7. The Comparative Result of Field Trials
The comparative result was shown in Table 7.
Results of field trial showed that LSCA noticeably
improved obviously the main performance indices such
as the germination index, seedling rate (Ta b l e 7 ), when
comparing with Jinong No. 4, the germination rate and
per ha yield of LSCA were increased by 5.6% and 9.5%
respectively; when compared to the uncoated corn seed
(CK), and they were enhanced by 6.0% and 13.6% re-
spectively, while the cost was declined by 18.75%. It had
indicated that the germination energy and germination
rate in the laboratory were consistent with the trend of
increasing yield in field experiment.
Table 5. The experimental result of antifeedant effect of the seed coating agents.
Treatment Dosage of LSCA (%) Weig h t o f AF (g) Weight of AF in 48 h (g) Feeding volume (g) AF (%)
15 5.488 5.193 0.294 87.4a
10 5.693 4.966 0.677 70.9b
LSCA
5 6.267 4.963 1.204 48.3c
CK - 5.550 3.213 2.327 -
Averages in column separated by Duncan’s new multiple range test, 5% level. AF means artificial feed; LSCA means the novel seed coating ag ent; CK means
the blank control.
Table 6. The result of the security test in rats.
Results of acute toxicity test
Experimental Groups
Aot LD50/mg·kg–1 TC Ast LD50/mg·kg –1 TC
798 (Male) Slight 2239 (Male) Slight
LSCA 607 (Female) Slight 2106 (Fema l e) Slight
243 (Male) M ode rate 1792 (Male) Moderate
Jinong No.4 127 (Female) Moderate 1554 (Female) Moderate
Acute oral toxicity grading scale: 1) Slightly toxic: LD50 > 500 mg·kg–1; 2) Moderately toxic: 50 < LD50 < 500 mg·kg–1g; 3) Highly toxic: LD50 < 50 mg·kg–1.
Acute dermal toxicity grading scale: 1) Slightly toxic: LD50 > 2000 mg·kg–1; 2) Moderately toxic: 200 < LD50 < 2000 mg·kg–1; 3) Highly toxic: LD50 < 200
mg·kg–1.

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Table 7. The investigation results of the field trial.
Measurement Indicators LSCA Jinong No. 4CK
Germination index 11.2 10.8 10.2
Seedling rate (%) 96.4 92.5 91.9.
The length of function l e a f (cm) 96 84 83
The thick of stem (cm) 3.6 2.7 2.5
The rate of infect ed plant (%) 0 5.3 25.6
Yield (kg·ha–1) 986 892 852
Cost (US＄·kg–1) 1.3 1.6
Toxicity Slight Moderate
Averages in column separated by Duncan’s new multiple range test, 5%
level.
3.2. Discussion
3.2.1. The Mechanism of Insect Prevention
The most important difference between LSCA and Ji-
nong No. 4 was LSCA used PC as the main ingredient,
which can improve plant pest and disease resistance ca-
pability, and promote plant’s growth, crop yield and
quality [19,20]. When the plant is attacked by pest, PC
as an inducer stimulates plants to produce a series of
defense reactions and produce secondary structure (e.g.
periderm, corpus callosum, glue sluggish body, invade
fill body, excise, halo, etc.) and some functional mole-
cules (e.g. plants defend element, phenolic compounds,
duration of related proteins, pathogens, enzyme inhibi-
tors, etc.), which are inexistent before stimulating. These
substances directly or indirectly suppress chemical sen-
sors of insect mouthpart, and block the nerve conduction,
make its information transfer of feeding behavior inter-
rupted, or causing abnormal discharge from animals to
get correct taste of information and not to eat [21-24 ]. So
LSCA controls pests through the approach of repelling
pests and enhancing the immunity of seeds, instead of
poisoning pests. Finally pest lose taste function and re-
fuse feeding reaction.
In addition, if PC is used in advance, plants will mis-
take pest coming so that they can produce resistant ma-
terial (phenylalanine solution ammonia enzyme (PAL),
peroxidase (PO) and polyphenol oxidase (PPO), phy-
toalexin etc, protein related to the disease such as chiti-
nase, and beta arene glucan enzyme etc.), and thus act th e
genes of plant resistance and make plant produce system
antibody, which can keep a high level for a long time to
repel pests [25-27]. PC as an inducer u sing in the earlier
stage crop growth is better than used in the later period.
The inducible resistance function of PC is like to hit
the precaution for plants, which makes plants less ill or
no, thus promote plant’s growth, crop yield and quality.
3.2.2. The Mechanism of Yield Increase
The field experiment results showed that seeds coated
with PC had better resistance to smut fungus and higher
germination percentage and higher crop yields than the
control. The first main reason is the mentioned-above
mechanism of insect prevention, and another is that PC
can enhance the resistance against smut fungus and has a
repellent effect to the pests in the soil, so that the plants
are safe from threats and damage by disease and have a
positive seedling growth and the fruits of harvest [28-
30].
The second reason contributes to PC of LSCA, which
has the ability of forming a dense protective film on the
surface layer of seeds. The film increases the brightness
of the coated seeds. In this case, the fungicide was glued
on the surface of the seeds, it is an advantage, since it
allows the continuous contact of the fungicide with the
seed surface and increased the resistance of the fungicide
to be removed by the wind or rain. Thus, the agent was
acting not only as antimicrobial coating itself but also as
a carrier system of agrochemicals [31,32].
The film has a good selective permeability which can
prevent oxygen from entering the film, restrict loss of
CO2, and maintain a high concentration of CO2 in the
film [33-35], so as to restrain the seed’s respiration,
maintain the seed moistu re and absorb the soil moisture,
and cut off excessive soil moisture to prevent the seed
from corrupting. In short the film can provide a good
growth environment for seeds germination and growth
and make the internal nutrient consumption of seeds fall
to the lowest possible level [36-38]. It is just that this
layer of natural poly mer membranes plays a vital role on
corn yield and antifeedant effect.
The third reason contributes to the corn yield increase
may be the effect of the trace elements and fertilizer
contained in LSCA. The trace fertilizer can increases the
activity of an enzyme and provide abundant nutrient
sources for germ gr owing.
4. CONCLUSIONS
The formula was as follows: 1% PC 56%, sodium hy-
droxide 5%, trace fertilizers 22%, trace element 1.5%,
active ingredients 1%, filmogen 4%, plant growth regu-
lator 2%, water 6.5%, natural pigment 2%. In the proc-
ess of preparation, the pH of the agent was adjusted to
6.0, and the dilution was 1:50. The results showed that a
novel formulation had increased the yield by about
13.6%, 9.5% over that of CK and Jinong No. 4, respec-
tively. Furthermore, the cost of LSCA was 18.75% less
and the toxicity was lower than that of Jinong No. 4. So
it is safe, non-polluting, and has significant economic
and environmental benefits.

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463463
5. ACKNOWLEDGEMENTS
We express the appreciation to Wuhan Science and Technology Bu-
reau of China for financial support (No. 201120922303). We also th an ks
the State Key Laboratory of Crop Biology of Shandong Agricultural
University. A special acknowledgement is given to Wuhan University
of Technology for experimental conditi ons an d techn ical s upport.
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