Page 1 of 8
European Journal of Applied Sciences – Vol. 11, No. 5
Publication Date: October 25, 2023
DOI:10.14738/aivp.115.15778
Prasetyo, J., Aeny, T. N., Rahmadanti, T. P., & Efri. (2023). Intergrated Control of Corn Downy Mildew Using Trichoderma
Asperellum and Curcuma Aeruginosa. European Journal of Applied Sciences, Vol - 11(5). 393-400.
Services for Science and Education – United Kingdom
Intergrated Control of Corn Downy Mildew Using Trichoderma
Asperellum and Curcuma Aeruginosa
Joko Prasetyo
Department of Agrotechnology,
Faculty of Agriculture, University of Lampung
Titik Nur Aeny
Department of Agrotechnology,
Faculty of Agriculture, University of Lampung
Tita Prenti Rahmadanti
Department of Agrotechnology,
Faculty of Agriculture, University of Lampung
Efri
Department of Agrotechnology,
Faculty of Agriculture, University of Lampung
ABSTRACT
This study aims to determine the effect of the spore density of Trichoderma
asperellum and the concentration of Black Curcuma extract and their interaction
on the intensity of downy mildew and the growth of maize. This research was
conducted at the Integrated Field Laboratory of the Faculty of Agriculture and the
Laboratory of Plant Diseases, Department of Agrotechnology, Faculty of
Agriculture, University of Lampung from March 2019 to May 2019. The design used
in this experiment was a randomized block design, which was arranged in factorial
experiment. The first factor was the treatment of T. asperellum which consists of 3
levels, namely without T. asperellum (T0), T. asperellum with a spore density of 106
spores / mL (T1), and T. asperellum with a density of 108 spores / mL (T2), applied
in rhizosphere. The second factor was the concentration of Black Curcuma
(Curcuma aeruginosa) extract which consists of 4 levels, namely 0% (F0), 20% (F1),
40% (F2) and 60% (F3), applied at the growing point of corn. The variables
observed were incubation period, disease incidence, disease severity, plant height
and stover weight. The observational data obtained were analyzed using analysis of
variance and then tested by the LSD test at the 5% level. The results showed that
the treatment of T. asperellum density of 108 spores / mL can reduce the incidence
and severity of downy mildew. Treatment of T. asperellum of the density of 106
spores / mL can increase the dry weight of plant shoots and T. aspereellum density
of 108 spores / mL can increase the dry weight of plant roots. Treatment of Black
Curcuma extract with a concentration of 40% can reduce disease incidence, disease
severity and increase stover weight. The best density of T. asperellum in
suppressing downy mildew disease was the density of 108 spores / mL and the best
Page 2 of 8
Services for Science and Education – United Kingdom 394
European Journal of Applied Sciences (EJAS) Vol. 11, Issue 5, October-2023
concentration of Black Curcuma extract in controlling downy mildew, namely a
concentration of 40%.
Keywords: Peronosclerospora maydis., Curcuma aeruginosa, Tricchoderma asperellum,
spore density, and concentration
INTRODUCTION
Corn (Zea mays L.) is a source of carbohydrates in addition to rice and wheat. In addition, corn
can also be used as animal feed and industrial raw materials (food, beverage, chemical, and
pharmaceutical). Corn is used as an industrial raw material that will provide added value for
farming in this commodity. Corn can also be processed into cooking oil, margarine, food
formulas, and pharmaceutical industries (Najiyati and Danarti, 2000; Ranum et al., 2014).
In Indonesia, corn is a secondary commodity that deserves to be one of the leading agribusiness
commodities. Maize production in Indonesia, especially in Lampung Province, has decreased.
According to the Central Statistics Agency (2016), maize production in Lampung Province in
2010 reached 2,126,571 tonnes, in 2011 and 2012 maize production reached 1,817,906 tonnes
and 1,760,275 tonnes, in 2013 maize production reached 1,760,278, while in 2014 and 2015,
maize production experienced a decline in production, reaching 1,719,386 tons and 1,502,800
tons respectively.
One of the reasons for the decline in maize production in Lampung Province is plant pests.
According to some studies (Semangun, 2004; Soenartiningsih and Talanca, 2010), downy
mildew in maize can reduce yields over 80%. The disease is caused by Peronoslerospora spp.
The downy mildew control that is usually done at this time is the use of synthetic fungicide, it
is still the main choice of farmers. The problem that arises is the presence of resistant
pathogenic variants (Burhanuddin, 2009; Ginting et al., 2020; Gisi and Sierotzki, 2008).
Therefore, it is necessary to find other effective control alternatives. One of the biological agents
used to suppress plant diseases is Trichoderma sp. The fungus Trichoderma sp. reported to be
effective in suppressing several plant pathogens such as Fusarium sp., Pythium sp., Sclerotium
rolfsii, Phytophthora sp., and R. Solani (Suharna, 2003 in Ilyas, 2006). T. asperellum can reduce
the incidence of downy mildew in corn (Prasetyo et al., 2021). Black Curcuma contains
saponins, flavonoids, bitter substances, tannins and polyphenols as well as essential oils from
0.3 to 2% (Sari and Erba, 2016) which can function as antibacterial., antifungal, antimicrobial
so that it has the potential to control the attack of these pathogens. Based on the description
above, it is necessary to test the effect of the extract of Black Curcuma (Curcuma aeruginosa.
Roxb) and Trichoderma sp. against downy mildew (P. maydis.). This study aims to determine
the effect of the spore density of T. asperellum. and the concentration of Black Curcuma extract
and their interaction on the intensity of downy mildew and the growth of maize.
MATERIALS AND METHODS
Place and Time
This research was conducted from March 2019 to May 2019 at the Integrated Field Laboratory
and Protection Laboratory Plants, Department of Agrotechnology, Faculty of Agriculture,
University of Lampung.
Page 3 of 8
395
Prasetyo, J., Aeny, T. N., Rahmadanti, T. P., & Efri. (2023). Intergrated Control of Corn Downy Mildew Using Trichoderma Asperellum and Curcuma
Aeruginosa. European Journal of Applied Sciences, Vol - 11(5). 393-400.
URL: http://dx.doi.org/10.14738/aivp.115.15778
Treatment Arrangement
The design used in this experiment was a randomized block design (RBD) which was arranged
in a factorial with 3 blocks as replications. The first factor is the treatment of Trichoderma sp.
which consists of 3 levels, namely without T. asperellum. (T0), T. asperellum with a spore
density of 106 spores / ml (T1), and T. asperellum with a density of 108 spores / ml (T2). The
second factor is the concentration of Black Curcum aextract which consists of 4 levels namely
0% (F0), 20% (F1), 40% (F2) and 60% (F3).
Treatment Implementation
The implementation of this research includes the preparation of planting media, planting corn,
preparation of botanical fungicides, molecular identification of isolate Trichoderma sp.
obtained from Polinela, multiplication of T. asperellum, application of T. asperellum, making
conidia suspension of P. maydis., and inoculation of P. maydis.
Observation Variable
The parameters observed were the incubation period, the intensity of downy mildew and the
growth of maize. Observation of downy mildew disease consisted of observing disease
incidence and disease severity. Observation of plant growth consists of observations of plant
height and dry weight of plant parts.
Incubation Period
The incubation period is observed daily until the end of the observation. The incubation period
is the time from the time of inoculation to the appearance of symptoms.
The Incidence of Downy Mildew (DM)
The incidence of DM is calculated using the following modified formula of Ginting (2013):
TP = n / N X 100%
Information:
• TP: Disease incidence (%)
• n: Number of plants attacked
• N: The number of all plants observed
DM Severity
DM severity was calculated using a disease score or scale consisting of 5 levels of attack, namely
mild score, moderate, severe, very severe, or healthy leaf table 1
Table 1: The categories of DM that can be used are a scale consisting of 5 categories as
follows:
SCORE INFORMATION
0 There are no symptoms
1 Symptoms occur up to ≤ 10% of the leaf
2 Symptoms occur in> 10% to ≤ 25% of the leaves
3 Symptoms occur in ≥25% to ≤ 50% of the leaves
4 Symptoms occur in ≥ 50% or dead leaves
Page 4 of 8
Services for Science and Education – United Kingdom 396
European Journal of Applied Sciences (EJAS) Vol. 11, Issue 5, October-2023
PP = (∑ (nxv)) / (NxV) X 100%
Information:
• PP = severity of disease (%)
• n = Number of leaves with a certain score
• v = The score of each category of attack
• N = Number of leaves observed (sample)
• V = highest score or scale
Plant Height
The height of the maize plants was measured from the soil surface to the tip of the highest leaf
of the plant using a tape measure. Measurement carried out starting from 1, 2, 3, 4, and 5 wap
(weeks after planting).
Stover Dry Weight
Observation of dry weight of stover was calculated as 43 days after planting. The dry weight of
stover is weighed by removing the corn plant from the planting medium and then cleaning it
from clinging dirt such as soil. Furthermore, the stover is cut into pieces, separating the roots,
stems and leaves, then put in a different envelope according to the part of the plant; and then
ovened with temperature 80 ̊C for 4 days until stover weight has been constant.
Data Analysis
The data obtained were analyzed for variance, homogeneity of variance was tested by Barlett's
test, data additivity was tested by Tukey's test. If the assumptions are fulfilled, then it is
followed by analysis of variance. The difference in the mean treatment value was tested with
the least significant difference test (LSD) at the level of confidence α=5%.
RESULTS AND DISCUSSION
Based on the observations, it was shown that the spore density treatment of T. asperellum and
the concentration of Black Curcuma has no significant effect on incubation period and plant
height. However, it had a real effect on incidence and severity of DM and dry weight of plant
stover. Early symptoms of downy mildew in maize can be seen on the 14th day after inoculation.
The initial symptom of downy mildew was the presence of white lines parallel to the leaf bones.
Furthermore, the symptoms of chlorosis appear all over the leaf surface. On the upper and
lower surfaces of the maize leaves when seen in the morning there were white conidia like flour.
This is in line with the opinion of Semangun (2004), symptoms in maize plants begin with
chlorotic spots appearing that extend parallel to the leaf bones with a border which is clear and
there is mass like flour under the surface of the leaves.
Disease Incidence and Severity of DM
Based on the analysis of the various treatments of T. asperellum can reduce disease incidence
at 21 days after insulation and disease severity at 35 days after inoculation (Tables 2 and 3).
The result of further test showed that the treatment of T. asperellum with a density of 108 spores
/ mL can reduce the incidence of DM compared with controls. This is in line with the opinion of
Sutama et al. (2010) and Prasetyo et al.(2021), the fungus T. asperellum has the ability to trigger
Page 5 of 8
397
Prasetyo, J., Aeny, T. N., Rahmadanti, T. P., & Efri. (2023). Intergrated Control of Corn Downy Mildew Using Trichoderma Asperellum and Curcuma
Aeruginosa. European Journal of Applied Sciences, Vol - 11(5). 393-400.
URL: http://dx.doi.org/10.14738/aivp.115.15778
a number of peroxidase enzymes in plants, where these enzymes play a major role in
suppressing downy mildew. Furthermore, the peri oxidase enzymes produced by plants play a
role in strengthening plant cell walls so that plants are able to inhibit infection by pathogens
that cause disease.
Table 2: Disease incidence (%) in various treatments of spore density of T. asperellum.
and the concentration of Black Curcuma extract
Treatment The incidence of DM (%) in
21 DAI
Original Data Trans Data (√(x+0,5))
The spore density of T. asperellum.
0 11,11 1,46 a
106 5,56 1,36 ab
108 1,39 1,30 b
LSD 5 % 0,12
Concentration of Black Curcuma extract (%)
0 12,96 1,49 a
20 9,26 1,44 a
40 0,00 1,26 b
60 1,85 1,30 b
LSD 5 % 0,14
Note: The values in the same column followed by the same letter are not significantly different (LSD Test 5%). DAI:
days after inoculation
Table 3: DM severity (%) in various treatments, the spore density of T. asperellum and
the concentration of Black Curcuma extract
Treatments Disease severity (%) pada
21 DAI 35 DAI
Original
Data
Trans Data
(√(x+0,5))
Original
Data
Trans Data
(√(x+0,5))
The spore density of T. asperellum.
0 1,21 1,34 a 12,34 1,52 a
106 1,44 1,32 a 11,35 1,48 a
108 0,29 1,28 a 2,15 1,33 b
BNT 5 % 0,06 0,14
Concentration of black curcuma
0 2,77 1,38 a 13,81 1,53 a
20 1,15 1,33 a 10,51 1,48 a
40 0,00 1,26 b 4,35 1,35 a
60 0,00 1,27 b 5,77 1,41 a
LSD 5 % 0,07 0,16
Note: The values in the same column followed by the same letter are not significantly different (LSD Test 5%). DAI:
days after inoculation.
Based on the results of the analysis of the various treatments of black curcuma, it can reduce
the incidence of disease at 21 DAI and disease severity at 35 DAI. In the treatment of Black
Curcuma extract concentrations of 40% and 60% can reduce the incidence and the severity of
Page 6 of 8
Services for Science and Education – United Kingdom 398
European Journal of Applied Sciences (EJAS) Vol. 11, Issue 5, October-2023
DM (Tables 2 and 3). This is because Black Curcuma contains saponins, tannins and essential
oils from 0.3% -2% (Syamsul Hidayat and Hutapea, 1991 in Sari and Erba, 2016). As stated by
Sekarsari et al. (2013), vegetable extracts that contain active ingredients that can inhibit and
damage microorganism cells, one of the active ingredients is essential oil compounds such as
terpenes and aromatic compounds. In the opinion of Koul et al. (2008), essential oil compounds
are able to suppress the growth of pathogenic fungi by damaging the pathogenic cell walls so
that important components such as proteins leave the cells and the cells die. According to Yendi
(2015), tannin compounds have the ability to interfere with the process of forming fungal cell
walls by inhibiting chitin synthesis in these fungal cells. Saponin compounds have properties
that can inhibit bacterial growth. The mechanism of the compound in pressing is by reducing
the surface tension of cell so that it will increase the permeability or leakage of cells and cause
intracellular compounds will come out (Nuria, et al., 2009).
Dry Weight of Roots and Plant Canopy
Based on the analysis of the various treatments, T. asperellum significantly effect on root dry
weight and plant canopy. Based on further tests, the concentration of Black Curcuma extract
treatment 40% can increase the dry weight of maize stover.
Treatment of T. asperellum the density of 108 spores / mL significantly increased root dry
weight (Table 4). This is in line with the opinion of Vargas et al (2009), that the application of
T. asperellum in maize plants can strengthen plant roots, increase plant growth and provides
protection against downy mildew infection.
Treatment of the spore density of T. asperellum 106 spores / mL significantly increased the dry
weight of maize plant canopy (Table 4). This is because the increase in plant dry weight is
thought to be related to the ability of T. asperellum in producing the auxin hormone, namely
IAA (Indole Asetic Acid). The IAA hormone has the ability to increase lateral root growth,
multiply shoots and increase biomass plants from shoots in Arabidopsis plants (Cornejo et al.,
2009).
Table 4: Weight of root and shoot dry stover (g) under various treatments of spore
density of T. asperellum and the concentration of Black Curcuma extract
Treatment / ml and Concentration
(%) Observation
Treatment / ml and Concentration (%) Observation
Root Weight (g) Head Weight (g) Root Weight (g) Head Weight (g)
The spore density of T. asperellum 0 4,32 b 24,36 b
10 6 5,27 ab 28,01 a
10 8 6,08 a 27,19 ab
LSD 5 % 1,05 2,87
Concentration of Black Curcuma
0 4,84 a 25,51 b
20 4,87 a 26,43 ab
40 5,92 a 29,44 a
60 5,29 a 24,07 b
LSD 5 % - 2,87
Note: The values in the same column followed by the same letter are not significantly different (LSD Test 5%).
Page 7 of 8
399
Prasetyo, J., Aeny, T. N., Rahmadanti, T. P., & Efri. (2023). Intergrated Control of Corn Downy Mildew Using Trichoderma Asperellum and Curcuma
Aeruginosa. European Journal of Applied Sciences, Vol - 11(5). 393-400.
URL: http://dx.doi.org/10.14738/aivp.115.15778
CONCLUSION
Based on the research results it can be concluded that the treatment of T. asperellum with a
density of 108 spores / mL can reduce the incidence and severity of DM and increase the dry
weight of corn crop stover. Black Curcuma extract with a concentration of 40% can reduce the
incidence and the severity of DM and increase the dry weight of the corn plant canopy. There
was no interaction between T. asperellum spore density and concentration of black curcuma
extract in controlling downy mildew and maize growth
ACKNOWLEDGMENT
This research was funded by an institutional research grant program at the University of
Lampung, Indonesia. Thanks, are also conveyed to the Plant Clinic, Plant Disease laboratory,
Faculty of Agriculture and LTSIT, University of Lampung, where this research was carried out
References
Akarchariya, N., S. Sirilun, J. Julsrigival, S. Chansakaowa, 2017. Chemical profiling and antimicrobial activity of
essensial oil from Curcuma aeruginosa Roxb. Roxb., Curcuma glans K. Larsen and J. Mood and Curuma Cf.
Xanthorrhiza Roxb. Collecten in Thailand. Asian Paccific Journal of Tropical biomedicine 7(10): 881-885
Burhanuddin. 2009. Metalaxyl fungicide was not effective in suppressing downy mildew (Peronosclerospora
maydis) in West Kalimantan and alternative control. Proceedings of the National Seminar.
Central Bureau of Statistics. 2016. Indonesian sweet corn production data for 2015-2016. BPS. Jakarta Cahyani,
Sri Susan.
Cornejo, H.A.C., L.M. Rodriguez, C.C. Penagos, and J.L. Bucio. 2009. Trichoderma virens a plant benefical fungus,
enhances biomass production and promotes lateral root growth through an auxin-dependent mechanism in
arabidopsiss. Plant Physiology. 14 (9): 1579-1592.
Ginting C, Prasetyo J, Dirmawati SR, Ivayani, Timotiwu PB, Maryono T, Widyastuti, Chafiska DIR, Asyifa A,
Setyowati E, Pasaribu AHZ. 2020. Identification of maize downey mildew pathogen in Lampung. Ann Res Rev Biol
35 (7): 23-35. DOI: 10.9734/ARRB/2020/v35i730244
Gisi U, Sierotzki H. 2008. Fungicide’s mode of action and resistance in downy mildews. Eur J Plant Pathol 122:
157-167. DOI: 10.1007/978- 1-4020-8973-2_12.
Ilyas, M. 2006. Isolation and identification of molds in the rhizosphere niches of plants in
Koul, O., Walia, S and Dhawalia, G.S. 2008. Essential oil as green pesticides potential and constrains. Biopestic. Int
4 (1): 63-84.
Mutis Mountain Nature Reserve area, East Nusa Tenggara. Journal of Biodiversity. 5 (3): 216-220.
Najiyati, S., and Danarti. 2000. Palawija, Farming Cultivation and Analysis Edition 10. Self-help spreader. Jakarta.
Nuria, M.C., Faizatun, A and Sumantri. 2009. Antibacterial Activity Test of Jatropha curcas (Jatropha curcas L.)
Leaf Ethanol Extract against Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, and Salmonella
typhi ATCC 1408. Journal of Agricultural Sciences. 5 (2) .26-37.
Ranum P, Pena-Rosas JP, Garcia-Casal MN. 2014. Global maize production, utilization, and consumption. Ann N Y
Acad Sci 1312 (2014): 105-112. DOI:10.1111/nyas.12396.
Page 8 of 8
Services for Science and Education – United Kingdom 400
European Journal of Applied Sciences (EJAS) Vol. 11, Issue 5, October-2023
Sari, A. M and Cikta, E.V. 2016. Extract flavonoids from Black Curcuma (Curcuma aeruginosa Roxb) and
application to transparent soap. Convertible Journal 5 (1): 17-23.
Semangun, H. 2004. Food Plant Diseases in Indonesia. Gajah Mada University Press. Yogyakarta. 429 p.
Sekarsari, R.A, Prasetyo, J., Maryono, T. 2013. The effect of several vegetable fungicides on the incidence of
downy mildew in maize (Zea mays saccharata). Tropical Agrotech Journal. 1 (1): 98 - 101.
Soenartiningsih and Talanca A. 2010. The spread of downy mildew (Peronosclerospora maydis) in corn in Kediri
district in: Saenong S, Daud D, Amin N (eds). Prosiding Seminar Ilmiah Dan Pertemuan Tahunan XX PEI-PFI
Komda Sulsel. Balai Penelitian Tanaman Serealia Maros. Maros, 5 November 2008. [Indonesian]
Sutama, K, Ratih, S, Maryono. T and Ginting, C. 2015. Effect of Paenibacillus polymyxa and the fungus
Trichoderma sp. against downy mildew (Peronosclerospora maydis (Rac) Shaw) in corn plants. Agrotech
Journal. 13 (2): 199-203.
Vargas, W. A, Mandawe, J.C and Kenerly, C.M. 2009. Plant derived sucrose is a key element in the symbiotic
association with Trrichoderma virens and Maize plants. Journal of Plant Physiol 151: 792-808.
Yendi, T.P, Efri and Prasetyo, J. 2015. The effect of extracts from several zingiberaceae family plants on
anthracnose disease in bananas. Tropical Agrotech Journal. 3 (2): 231-235.