A O RTICLE

3830
Advances in Environmental Biology, 5(13): 3830-3835, 2011
ISSN 1995-0756
This is a refereed journal and all articles are professionally screened and reviewed
ORIGINAL ARTICLE
Effect of Trichoderma spp and Compost on Seedling Emergence and Early Growth of
Corn (Zea mays L.)
1
Mohammad Yazdani, 2Abbas Ghanbari-Malidarreh and 3Hossein Bagheri
1
PhD Student, Department of Agriculture ,Tabriz Branch, Islamic Azad University,Tabriz,Iran.
Department of Agronomy, Jouybar Branch, Islamic Azad University, Jouybar, Iran
3
Department of Agriculture ,Chaloos Branch, Islamic Azad University, Chaloos, Iran. 2
Mohammad Yazdani, Abbas Ghanbari - Malidarreh and Hossein Bagheri; Effect of Trichoderma spp
and Compost on Seedling Emergence and Early Growth of Corn (Zea mays L.).
ABSTRACT
Effects of Trichoderma spp and different rate of compost on corn germination and early growth seedling
were investigated. An experiment was conducted at glasshouse in Sari Agricultural and Natural Resources
Sciences University. The pot experiment was laid out factorial based on completely randomized design with
four replications. Three seedbed materials consisted of field soil as check, compost (20 Mg.ha-1) and (20 Mg.ha1
) and two species of Trichoderma (T. harzianum and T. viride) with a check were the treatments. Results show
that application of compost with 20 Mg.ha-1, emergence of the seedlings (Percentage and Rate Emergence) a
9.1% and 6.6% increased compared to the other treatments respectively. Seedling vigor increased by higher
dosage of Trichoderma spp in the compost (20 Mg.ha-1) treatment. Dry weight of corn grown in the compost
were significantly affected by the application of T, harzianum and T, viride that is the dosage main effect was
statistically significant. Furthermore, application of two fungi with compost simultaneously increase seedling
emergence compared to check. However, Trichoderma and compost combination had additive effect in on
seedling emergence and early growth of corn.
Key words: Trichoderma, Compost, Seedling emergence, Corn.
Introduction
The use of bio-fertilizers is the key to
sustainable agriculture [17]. Bio-fertilizers being
essential components of organic farming play vital
role in maintaining long term soil fertility and
sustainability by fixing atmospheric nitrogen,
mobilizing fixed macro and micro nutrients or
convert insoluble P in the soil into forms available to
plants, there by increases their efficiency and
availability [23,4,7]. Fungal biofertilizers help to
enhance crop yield and promote sustainable
agricultural production and are safe to the
environment. Fungal biofertilizers have advantages
in terms of nutrient supply, soil quality and crop
growth and yield. Development in the effectiveness
of fungal species for formulation as biofertilizers
should be considered. Applying beneficial
microorganisms to seed during the priming process is
commercially
realistic,
as
microorganism
suspensions can easily be incorporated into the water
used for seed priming. It is equally important that the
microorganisms remain viable and can colonies the
developing roots and rhizosphere in order to continue
improving plant growth and to potentially control
disease. Seed-applied microorganisms have the
potential to become established in the rhizosphere of
plants, as they may transfer onto the developing root
as it emerges from the seed [11,13]. Even small
improvements in plant growth and health can result
in significant economic benefits.
Different species of Trichoderma have the
potential to control soil borne plant pathogens more
effectively than chemicals [12]. Use of these fungi is
not as harmful to the environment as chemical
pesticides. They can also compete with other
microorganisms; for example, they compete for key
exudates from seeds that stimulate the germination of
propagules of plant-pathogenic fungi in soil and,
more generally, compete with soil microorganisms
for nutrients and/or space. Furthermore, they inhibit
or degrade pectinases and other enzymes that are
essential for plant-pathogenic fungi, to penetrate leaf
surfaces. In vitro studies have shown that
micronutrients and insoluble phosphates became
soluble and available, therefore useful to the roots
interacting with T. harzianum in the root zone.
Species in the filamentous fungal genus Trichoderma
are of great economic importance as sources of
enzymes and antibiotics; plant growth promoters;
Corresponding Author
Mohammad Yazdani, Department of Agriculture, Tabriz Branch, Islamic Azad University,
Tabriz, Iran.
E-mail: [email protected]
3831
Adv. Environ. Biol., 5(13): 3830-3835, 2011
degraders of xenobiotics, and most importantly, as
commercial biofungicides. Different mechanisms
have been suggested as being responsible for their
biocontrol activity, which include competition for
space and nutrients, secretion of chitinolytic
enzymes, mycoparasitism and production of
inhibitory compound. Trichoderma ability to attack
other fungi (plant pathogens) mycelia, Trichoderma
hyphens growing surround of other fungi mycelia,
penetrate and feed from them. In addition to the
above, various species of Trichoderma were also
effective in the promotion of growth and yield in
various crops. The ability of Trichoderma to
recognize and parasitize phytopathogenic fungi in the
rhizosphere has been ascribed to several complex
mechanisms, such as nutrient competition, antibiosis,
mycoparasitism, induction of systemic resistance,
and increased plant-nutrient availability [11,5,19,9].
Compost and compost extracts applied to soil
improve its quality by altering the chemical and
physical properties, increase organic matter content,
water holding capacity, overall diversity of microbes,
provide macro- and micronutrients essential for plant
growth and suppress diseases which indirectly
contribute to plant growth enhancement [7]. Corn
(Zea mays L.) is one of the main crops and is
commonly employed in the human diet in its natural
form as sweet corn or as a sub product in bread, flour
and dough, among others. In addition Corn suffers
from various fungal diseases in its entire growing
period from germination of seeds to the mature plant
stage. Seedling emergence is an important trait that
can limit commercialization of sweet corn hybrids. A
rate of seedling emergence and leaf appearance is
important in developing a corn crop with earlier
canopy closure and better seasonal light interception.
The aim of this study was to effect of Trichoderma
spp and compost on germination and early growth of
corn. Therefore, this study was design to find out the
effect of different Trichoderma strains and compost
on the percentage emergence, rate emergence and
seedling parameters of corn seeds in glasshouse
conditions.
2.2. Trichoderma Spp and Inoculation:
This work investigated the application of two
selected beneficial microorganisms (T. harzianum
and T. viride) to corn seed and their subsequent
survival and establishment in the rhizosphere once
the seed was planted. Briefly this involved storing
agar plugs taken from the periphery of an actively
growing colony in a 10% (v:v) aqueous solution of
glycerol in straw ampoules. Fungi recovered from
this system were subsequently grown on potato
dextrose agar (PDA; Merck) [21] slopes for storage
for 7 days at 30 °C in a termostated incubator
chamber with air circulation. This period showed to
be sufficient for fungi sporulation. The substrates
used were wheat bran. Wheat bran showed to be the
most suitable substrate to produce Trichoderma
spores for all strains that were evaluated. The
substrates were sterilized at 121 °C for 15 min in a
Phoenix autoclave model AV 50 and cooled down to
room temperature before the inoculation, which was
done until 24 h after sterilization. The dosages of two
funguses applied were 5 and 10g per pot. Control
plants were also available. A control plot in each
block was left untreated and only water without the
antagonistic fungus was provided. Corn seeds were
sown in plastic pots on July 6, 2009. Availability of
water in the soil plays an important role in
facilitating establishment and effectiveness of
Trichoderma in the soil, Therefore irrigation was
done so on a regular basis.
2.3. Observations Recorded:
Percentage emergence and rate emergence was
recorded on the15th day after sowing, shoot length
and total weight and seed vigor on the 45 the day
after sowing were recorded. Seedling vigor index
was recorded after 15 days. Vigor index for each
treatment was determined using the following
formula developed by Abdul-Baki and Anderson [1].
Seedling Vigor = [root length + shoot length] ×
percentages of germination.
Materials and Methods
The plants were removed 45 day after sowing,
and roots were washed using slow running water to
remove soil particles and organic debris. The dry
mass of shoot and root samples, root length and
shoot length was determined after drying in an oven
at 60 °C with forced air.
2.1. Experimental Design:
An experiment was conducted at research farm
of Sari Agricultural Sciences and Natural Resources
University (Latitude 42.36 N, longitude 13.53E and
16 m above mean sea level), Iran during 2009.
Experiment laid out as factorial plot based on
randomized complete design. Three replicates were
set up for each treatment. Three seedbed materials
consisted of field soil as check, compost (20 Mg.ha-1)
and (40 Mg.ha-1) and two species of Trichoderma
with a check were the treatments. The compost was
applied two weeks before sowing and mixed
thoroughly with the soil of each pot.
2.4. Statistical Analysis:
Data were subjected to ANOVA using the SAS
statistical software package using GLM, [18] and
Duncan's multiple range test was performed to
compare the treatment means. The level of statistical
significant was accepted as P<0.05.
3832
Adv. Environ. Biol., 5(13): 3830-3835, 2011
case of T. harzianum and T. viride to enhanced seed
germination root and shoot length [9] as well as
increasing the frequency of healthy plants, and
boosting yield [12]. Root colonization by
Trichoderma spp therefore induces significant
changes in the plant metabolic machinery. Root
colonization by these fungi, also frequently enhances
root growth and development, crop productivity,
resistance to abiotic stresses and the uptake and use
of nutrients [11].
Results and Discussion
The effect of two Trichoderm strains (T.
harzianum and T. viride) and compost on seed
percentage and rate emergence and seedling
parameters of corn in glasshouse conditions, the
results are presented in table 1. Statistical analysis
showed significant differences in treatments at P ≤
0.05 levels.
3.1. Seedling Emergence:
3.2. Seedling Vigor:
The emergence of the seedlings (Percentage and
Rate Emergence) a 9.1% and 6.6% increased in
compost with 20 Mg.ha-1, compared to the other
treatments respectively (Table1). The increase in
emergence of the seedlings corn in compost
treatment, especially in 20 Mg.ha-1 rates, provides
further support to the hypothesis that increase of
organic matter in soil, increased Seedling emergence
[4,7]. Compost contains other essential nutrients,
such as P and K, and minor nutrients which will
stimulate plant emergence. Therefore, Nutrient-rich
organic amendments by compost can improve soil
physical and chemical properties, increasing water
holding capacity and nutrient availability, and
promoting seedling establishment [7]. Zaller reported
that compost amendments increased emergence,
elongation and biomass allocation (root: shoot ratio)
of seedlings of tomato. Additionally, compost
contains humic acids. Humic acids are molecules that
regulate many process of plant development included
macro and micro nutrients adsorption and they
stimulate plant growth. Seedlings are an important
stage in the life cycle of plants and their successful
recruitment is important for plant population
dynamics and ultimately to community development,
structure and sustainability [6]. At the same time,
seedlings are the most vulnerable part of the life
cycle of plants, being subject to a plethora of abiotic
and biotic factors that may affect their emergence,
survivorship and establishment [15]. In addition,
Compost based suppression of wide range of major
soil borne diseases has been demonstrated in the last
decade as a promising option [17]. Treatment of
compost in the 40 Mg.ha-1 rates reduced the
emergence rate of seedling. However, compost in the
20 Mg.ha-1 rates did not reduce this rate, in the same
conditions. Probably due to the high salinity and
heavy metal concentrations of this treatment (40
Mg.ha-1), which could have unfavorable affected the
emergence rate of corn seedling.
Our results showed that, all Trichoderma strains
were found effective to enhance the emergence
percentage compared to control (Figure1). However
among the five treatments, two Trichoderma strains
(T. harzianum and viride) in 10g per pot exhibited
significantly enhancement of percentage emergence,
rate emergence, total weight and seed vigor in corn
seeds. The lowest vigor index was recorded in
control (Table 1). Studies have been confirmed in
When soil was amended with compost (20
Mg.ha-1 and 40 Mg.ha-1), a 33.6% and 14.2%
increase in Seedling vigor was observed up to 15
days after sowing respectively. The highest increase
in the seedling vigor of corn was also from compost
(20 Mg.ha-1) treatment. Interestingly, however, in
some other characteristics studied such as dry weight
and percentage emergence the control plants (without
compost treatment) were with the superior figures.
Effects of Trichoderma spp dosages (10g per pot) on
seedling vigor were significant (Table1). In the
present experiment, Seedling vigor increased by
higher dosage of Trichoderma spp in the compost
(20 Mg.ha-1) treatment (Figure2). [7] found greater
fungal species diversity under organic cultivation
than under conventional cultivation with inorganic
fertilizer. Enhanced plant vigor has also observed
following application of Trichoderma species to
other crops. Evidence in the literature suggests that
disease control activity or plant growth promotion
can be achieved when beneficial microorganisms are
present above 1*105 per gram of seed, root or soil
and thus this was the target application rate for seed
in this work.
Promotion of growth by Trichoderma spp. is a
result of increased root area allowing the roots to
explore larger volumes of soil to access nutrients,
and increased solubility of insoluble compounds as
well as increased availability of micronutrients
[15,10]. Seed vigor, an important agronomic trait
defined as the potential to produce vigorous
seedlings. This characteristic of Seed is a measure of
the quality of seed, and involves the viability of the
seed, the germination percentage, germination rate
and the strength of the seedlings produced [15]. In
theory, seed vigor may influence crop yield through
both indirect and direct effects. The indirect effects
include those on percentage emergence and time
from sowing to emergence. These influence yields by
altering
plant
population
density,
spatial
arrangement, and crop duration. Direct effects on
subsequent plant performance are more difficult to
discern. A number of different approaches to testing
the hypothesis that seed quality affects subsequent
plant performance (implicit in some definitions of
vigor) are illustrated. The results show that it is
possible to detect such effects in some circumstances
[8].
38333
Adv. Enviroon. Biol., 5(13): 3830-3835,
3
20111
3.3. Dry Weight:
W
i
in dryy
On daay 45, these plaants exhibited increase
weight in the compost treatment
t
especially with 200
Mg.ha-1 The
T lowest rate of compost was
w effective att
increasing dry weight of seedling corn. Dry weight off
wn in the compost
c
were significantlyy
corn grow
affected by
y the applicatiion of T, harzzianum and T,
viride that is the dosage main effect was
w statisticallyy
d
(10g) of Trichodermaa
significant. The highest dosage
spp. was main
m effect on the
t biomass in comparison too
the conttrol treatmennt without Trichoderma.
Inoculation
n of Trichodderma speciess significantlyy
increased of
o 15% T. viridae to 12% T. harzianum inn
dry matterr seedling (fig
gure3) of cornn as comparedd
with untreaated plants. Freesh weight, shooot length, dryy
weight of cucumber seeedlings as weell as seedlingg
weight of corn were inccreased signifi
ficantly by thee
applicationn of T. harzia
anum and T. viride
v
[23,24]..
Positive effects of Tricchoderma spp.. are not limiteed
to the above, many speecies of Trichoderma promoteed
ment of seed
dlings of cropps
growth and developm
[22,10,5]].
In our
o experimennts, response of seedlings to
Trichodeerma applicatiion varied in that while thhe
increasedd growth respoonse at signifi
ficant levels was
w
recorded
d for some orgaans of seedlinggs such an effeect
was not significant inn others. These effects weere
n due to nuttrient solubilizzation or to thhe
clearly not
control of plant pathogenic microorganism
m
ms.
ng to Altintas and
a Bal [5], thhe application of
Accordin
Trichodeerma spp increeased the grow
wth of seedlinng
corn wiith induced resistance, changes in thhe
microflorral compositioon on roots, en
nhanced nutrieent
uptake, including buut not limiteed to nitrogeen,
enhancedd solubilization of soil nutrrients, enhanceed
root deveelopment, incrreased root haiir formation annd
deeper roooting.
m and T. viridae annd different rate off compost on germ
mination and early growth of corn.
Table 1: Effeect of T. harzianum
Treatmentss
Rate Emergennce
Perrcentage Emergencce
Seedlinng vigor
-1
Compost (220 Mg.ha )
5.61 a
96.00 a
57.9 a
Compost (440 Mg.ha-1)
5.02 b
94.1 ab
44.8 b
Control
5.14 b
90.00 b
38.4 c
Trichoderm
ma spp
Control
4.82 b
88.8 b
37.96 c
T. harzianuum5g
5.47 a
93.33 b
46.94 b
T. harzianuum10g
5.43 a
97.77 a
51.63 a
T. viridae5
5g
5.33 a
91.22 ab
45.52 b
T. viridae1
10g
5.54 a
95.66 a
53.43 a
Levels of siggnificant:* P< %5, ** P<%1, NS = not
n significant.
Fig. 1: Efffect of Trichodeerma spp and different
d
rate oof compost on rate
r emergencee seedling.
d
rate oof compost on seedling
s
vigor of corn.
Fig. 2: Efffect of Trichodeerma spp and different
Dry Weight
180.6 a
163.6 b
147.2 c
159.0 d
170.2 bc
175.5 ab
164.7 dc
179.6 a
38334
Adv. Enviroon. Biol., 5(13): 3830-3835,
3
20111
d
rate oof compost on dry
d weight of seedling
s
corn.
Fig. 3: Efffect of Trichodeerma spp and different
5. Conclussion:
mpost with 200
In this study, appliication of com
e
off the seedlinngs increasedd
Mg.ha-1, emergence
compared to the other treatments. Seedling
S
vigorr
b higher dossage of Trichooderma spp inn
increased by
the compo
ost (20 Mg.ha-1
) treatment. Dry
D weight off
corn grow
wn in the compost
c
were significantlyy
affected by
y the applicatiion of T, harzzianum and T,
viride that is the dosage main effect was
w statisticallyy
significant. The preseent study cooncludes thatt
Trichoderm
ma species haave potential too enhance thee
seedling em
mergence in corn
c
which cann be useful too
enhance thhe emergencee percentage of
o corn seedss
besides red
ducing loses due
d to delayedd germination..
Trichoderm
ma harzianum
m and Trichooderma viridee
promoted the growth of corn plannt, increasingg
percentagee and rate emeergence and seedling
s
vigor..
However, Trichoderma and compostt combinationn
had additive effect in on
o seedling em
mergence andd
early grow
wth of corn. Further inveestigations aree
required to
o study in vivo,, effects of thesse fungi on thee
morphologgical and physiological chaaracteristics inn
corn plant and seed produuction.
Referencees
1. Abdul-Baki, A., J.D
D. Anderson, 1973. Vigorr
S
seed by multiplee
determiination of Soybean
criteriaa. Crop Sciencee, 13: 630-633.
2. Amand
da, J. Bennettt, John M. Whipps,
W
2008..
Beneficcial microorgaanism survival on seed, rootss
and in rhizosphere soil
s following application too
seed during
d
drum priming.
p
Biological Control,,
44: 349
9-361.
3. Altintas, S. and U. Bal, 2008. Effects
E
of thee
commeercial productt based on Trichodermaa
harzian
num on plant, bulb
b
and yield characteristicss
of onioon. Scientia Horticulturae, 1166: 219-222.
4. Badrelddin, S.M.S., M.
M Attia and S.A
A. Abosedera,,
2000. Field
F
assessmeent of compostts produced byy
highly effective ceellulolytic miicroorganisms..
Biologyy Fertilizer andd Soils, 32: 35--40.
5. Bal, U. and S. Alltintas, 2006. A positive sidde
effectt from Triichoderma harzianum,
h
thhe
biological control agent: Increeased yield in
vegettable crops. Journal of Environmenttal
Proteection and Ecollogy, 7(2): 3833-387.
6. Baeteen, L., H. Jacquuemyn, H. van
n Calster, E. Vaan
Beekk, R. Devlaeminck, K. Verheeyen, M. Herm
my,
2009. Low recruitm
ment across liife stages parttly
n of forest herbbs.
accouunts for the sloow colonization
Journnal Ecology. 977: 109-117.
7. Bulluuck, L.R., M. Brosius, G. Evanylo
E
and J.B
B.
Kristaino, 2002. Organic
O
and syynthetic fertiliity
amenndments influeence soil miccrobial, physiccal
and chemical properties on organic annd
conveentional farmss. Applied Sooil Ecology, 19:
147-1160.
8. Dorddas, C., 2006. Foliar
F
boron appplication affeccts
lint and
a seed yield and improvess seed quality of
cotton
n grown onn calcareous soils. Nutrieent
Cycliing in Agro ecoosystems, 76(1
1): 19-28.
9. Dubeey, S.C., M. Suresha, B. Singha, 20007.
Evaluuation of Tr
Trichoderma species
s
againnst
Fusarrium oxysporuum f. sp. cicerris for integrateed
manaagement of chicckpea wilt. Bioological Controol,
40: 118-127.
10. Hanson, L.E., 20000. Reduction of Verticilliuum
wilts symptoms in cotton following
f
seeed
treatm
ment with Tricchoderma vireens. The Journnal
of Cootton Science, 44: 224- 231.
11. Harm
man, G.E., 20006. Overview of mechanism
ms
and uses
u
of Trichoderma spp. Phytopathologgy,
96: 190-194.
n, P. John, R.D
D. Tyagi, D. Prevost,
P
Satindder
12. Rojan
K. Brrar, Stephan Poouleur, R.Y. Su
urampalli, 2010.
Mycooparasitic Trichhoderma viridee as a biocontrrol
agentt against Fusarrium oxysporuum f. sp. adzuuki
and Pythium arrhhenomanes andd as a grow
wth
prom
moter of soybeaan. Crop Proteection, 29: 14521459.
13. Harm
man, G.E., 20006. Overview of mechanism
ms
and uses
u
of Trichoderma spp. Phytopathologgy,
96: 190-194.
14. Gary, E. Harmann, Charles R. Howell, Adda
Viterrbo, Ilan Cheet and Matteoo Lorito, 20004.
3835
Adv. Environ. Biol., 5(13): 3830-3835, 2011
20. Scott, S.J., R.A. Jones and W.A. Williams, 1984.
Review of data analysis methods for seed
germination. Crop Science, 24: 1192-1199.
21. Chacon, M.R., O.G. Rodriguez, T. Benitez S.
Sousa, M. Rey, A. Llobell, J.D. Jarana, 2007.
Microscopic and transcriptome analyses of early
colonization of tomato roots by Trichoderma
harzianum. International microbiology, 10: 1927.
22. Rabeendran, N. Moot, D.J. Jones, E.E. Stewart,
A. 2000. Inconsistent growth promotion of
cabbage and lettuce from Trichoderma isolates.
N.Z. Plant Prot., 53: 143-146.
23. Raviv, M., B.Z. Zaidman Kapulnik, Y., 1998.
The use of compost as a peat substitute for
organic vegetable transplants production.
Compost Sci Util., 6(1): 46-52.
24. Yedidia, I., A.K. Srivastva Y. Kapulnik I. Chet,
2001. Effects of Trichoderma harzianum on
microelement concentrations and increased
growth of cucumber plants. Plant and Soil., 235:
235-242.
Trichoderma species opportunistic, a virulent
plant symphonist (review). Microbiology, 2: 4366.
15. Leck, M.A., V.T. Parker, R.L. Simpson, (Eds.).
2008. Seedling Ecology and Evolution.
Cambridge University Press, Cambridge.
16. Michal Shoresh and Gary E. Harman, 2008. The
molecular basis of shoot responses of maize
seedlings to Trichoderma harzianum T22
inoculation of the root: A Proteomic Approach.
Plant Physiology, 147: 2147-2163.
17. Nakkeeran, S., W.G.D. Fernando, Z.A. Siddiqui,
2005. Plant growth promoting rhizobacteria
formulations and its scope in commercialization
for the management of pests and disease. In:
Siddqui, Z. A. (Ed.), PGPR: Biocontrol and
Biofertiization.
Springer,
Dordrecht,
the
Netherlands, pp: 257-296.
18. SAS. Institute, Inc., 2000. SAS/STAT Users
Guide, version 6.12. SAS Institute. Inc. Cary,
NC.
19. Singh, A., S. Srivastava, H.B. Singh, 2007. Effect
of substrates on growth and shelf life of
Trichoderma harzianum and its use in biocontrol
of diseases. Bioresource Technology, 98: 470473.