Environmental and Toxicology Management 1 (2021) 9-22

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Environmental and Toxicology Management

journal homepage: www.journal2.unusa.ac.id/etm

A review of application of natural products as fungicides for chili

Khoirul Ngibad1,* , Afidatul Muadifah2 , Lailatul Jannah Triarini1 , Laily Rizki Amalia1 , Novita Karel Damayanti1
1 Study Program of Medical Laboratory Technology, Faculty of Health Sciences, Universitas Maarif Hasyim Latif, Sidoarjo, Indonesia
2 Study Program of Pharmacy, Sekolah Tinggi Ilmu Kesehatan Karya putra Bangsa, Tulungagung, Indonesia

Abstract

Anthracnose disease in chillies is a serious problem for farmers. So far, synthetic fungicides have been used as solution for the treat-
ment of this disease. However, the side effects of synthetic fungicides to public health and environment raised awareness on alternative
fungicides derived from natural resources. This paper aims to review plants that are potential as an alternative to fungicides for chili
plantation, fabrication of test solutions, in vitro and in vivo fungicide test. Many plants were investigated as alternatives to plant-based
fungicide. The utilization of leaves as samples including rhizomes, roots, tubers, weevils, seeds, fruit, flowers and other parts of the
plant. The extract fabrication method used as a fungicide test include: maceration method, gradual fractionation method, and de-
coction method. The maceration method is the method most widely used to extract fungicidal active compounds from plants. Some
studies that carried out in vitro tests were unable to compare with synthetic fungicides so it was not possible to determine their ef-
fectiveness for plant-based fungicide for chillies when compared to synthetic fungicides. In vitro extract of 80% alcohol and 10%/60%
n-hexane of pacar cina (Aglaia odorata L.) leaves can be compared with the performance of propineb 0.2%. In addition, the 60% and
70% kirinyuh (Chromolaena odorata L.) leaf extracts were also able to match Acrobat 0.2% performance in vitro. Based on the in vivo
test, suren (Toona sureni Merr) leaf extract and nut bulbs can be used as an alternative to vegetable / natural fungicides to help over-
come the problem of anthracnose in chilies.

Keywords :
Chili anthracnose disease, in vitro test, in vivo test, natural fungicide

1 Introduction
Anthracnose is a red chili plant disease caused by 2 types
One of the goals of the Sustainable development goals (SDGs) of fungi, namely: Colletotrichum capsici and Colletotrichum
is to achieve food security and declare as sustainable agriculture. gloeosporiodes. Colletotrichum capsisi population is fewer than
Chili is one of the food commodities whose production must be in- Colletotrichum gloeosporiodes. The Colletotrichum capsica fun-
creased in order to realize food security in Indonesia. Every year, gus attacks ripen chilies that are reddish in color, while Col-
there are increased in demand for chilies which is in line with the letotrichum gloeosporiodes which has 2 strains, namely: the R
growth in population and the development of food industry that strain which only attacks ripe red chilies and the G strain which can
require the chilies as raw material (Subagyono et al., 2010). In attack all parts of the plant, including mature red chillies and those
addition, there is always increase in the price of chili in particu- that are still unripe and green. These two types of pathogens are
lar month due to low productivity of chili harvest. The decrease seed-borne diseases because they are able to survive in the seeds
in chili productivity can be caused by pests and plant diseases for a long time to form acervulus (Piay et al., 2010).
(Warisno Dahana, 2018). The pests attack the plants and causes The use of chemical or synthetic pesticides is the most
chilies suffered severe damage and crop failure. The pests that can common control. Some examples of synthetic pesticides in-
attack chili plants include: peach aphids, thrips pests, mites, fruit cluded: Pyraclostrobin, Azoxystrobin, Picoxystrobin, Difenocona-
fly pests, and fruit borer pests. On the other hand, chili plant dis- zole, Thiophanate-methyl, Mancozeb (Gao et al., 2017), Metalaxyl-
eases include: anthracnose, phytophthora rot, fusarium wilt, cer- M (Esyanti et al., 2020), Orion 72 WP, Indofil Z-78 WP, Metarial
cospora leaf spot, bacterial wilt, yellow virus, mosaic disease (Piay 72 WP, Proven 250 EC, Folicure 5 EC, Propicon 250 EC, Fuji one
et al., 2010). Therefore, control of plant pest organisms must be 40 EC, Flowin HT, dan Winner 250EC (Naznin et al., 2016). The
done in order to increase the production of chilies (Badan Pusat negative impact of usage chemical / synthetic fungicides continu-
Statistik Republik Indonesia, 2019). ously includes: 1) polluting / damaging the environment, 2) caus-
ing residues on plants thus endanger health, and 3) causing resis-
*Corresponding Author. tance on pathogens (Amelia et al., 2020). Therefore, to overcome
Email Address : [email protected] the negative impact of usage synthetic fungicides, plant-based /
https://doi.org/10.33086/etm.v1i2.2022 natural fungicides can be used. The advantages of natural fungi-
Received from 20 March 2021; cides include: 1) relatively more environmentally friendly and safe
Received in revised from 30 April 2021; for humans because they are made from natural materials that are
Accepted 30 April 2021; easily biodegradable, and 2) cheaper, easy to obtained and easy to
Available online 22 May 2021; applied.

9
 Some plants that have the potential to be used as natural both in vitro and in vivo. The researchers only used one plant type
pesticides include: tembelekan/cherry pie (Lantana camara), separately to determine its potential as natural fungicide. Only
jarak tintir/coral plant (Jatropha multifida), pacar cina/chinese few researchers have combined 2 plants, for example: mixture of
rice (Aglaia odorata L.), mengkudu/noni (Morinda citrifolia L.), betel and tobacco leaf (Oktarina et al., 2017), (Anjani, 2018), (Nur
mimba/neem (Azadirachta indica A. Juss.), kenikir/compositae Rohmah, 2017) and mixture of kenikir/compositae (Cosmos cau-
(Cosmos caudatus Kunth.), sirih/betel (Piper batle L.), awar-awar datus Kunth.) and betel (Maimunah et al., 2019).
(Ficus septica) and others. Basically, natural pesticides do not In general, fungicide test methods used in many studies are di-
only come from plants, but also from bacteria, viruses, and fungi vided into 2 categories, namely: 1) in vitro and 2) in vivo. There
(Novizan, 2002). The purpose of this paper is to review: 1) plants are researchers who only focus on using in vitro test methods or in
that have the potential as an alternative natural fungicide for chili, vivo test methods. In addition, the researchers also used both test
2) fabrication of solution for in vitro and in vivo test, 3) in vitro test methods in combination . In the in vitro test method, many types
as fungicide for chili, and 4) in vivo test as fungicide for chili. of fungi that cause Anthracnose disease in chilies are used, for ex-
ample: Collectotrichum capsici, Colletotrichum gloesporioides and
Colletotrichum acutacum mushrooms. Several parameters that
2 Potential plants as alternative can be observed in the in vitro test include: percentage of inhi-
bition, diameter of fungal colony growth, zone of inhibition, spore
fungicide for chili growth, spore germination and percentage of spore density. On the
other hand, in the in vivo test more parameters can be observed
Many plants have been investigated on the potential as an alter- which include: anthracnose disease severity, intensity of fungal at-
natives to plant-based / natural fungicides for chili. Table 1 shows tack on chilies, percentage of disease incidence, effectiveness of
the names and parts of the plant and the method tested for fungi- fungicides, diameter of chili spots, incubation period of fungi in
cide. The part of plants that is widely used in research on finding chilies, plant height, number of fruit and the weight of the chilies.
alternative natural fungicides is the leaves. Few studies have used In this in vivo test, the success of the research is strongly influenced
parts of rhizomes, roots, tubers, weevils, seeds, fruit, flowers or all by environmental factors, for example: temperature, humidity and
parts of a plant (combination of flowers, leaves, stems, roots, and rainfall (Suwastini et al., 2020).
seeds). Betel leaf is a part of the plant that has been investigated

Table 1 Plants that have the potential as alternative fungicide for chili

Name of Plant Scientific Name Part of Plant Test Method for Reference
Fungicide
Umbi Teki Cyperus rotundus L. Leaves In vivo (Sihite et al., 2020)
Urang Aring Eclipta alba (L.) Hassk - In vitro (Andreas et al., 2018)
Ketepeng Cina Cassia alata Linnaeus All parts of plant In vitro (Arneti & Sulyanti,
2017)
Forest Betel Piper aduncum L. Leaves In vitro and In vivo (Elfina, 2015)
Fragrant Lemon- Cymbopogon nardus L. Leaves In vitro and In vivo (Elfina et al., 2016)
grass
Tobacco Nicotiana tabacum L. Leaves In vitro (Isman Duila, 2017)
Mixture of Betel Piper betle L. dan Nicotina tobacum Leaves In vitro and In vivo (Oktarina et al., 2017)
and Tobacco
Kunyit Curcuma longa sensu Val Rhizome In vitro and In vivo (Sari et al., 2020a)
Temu Putih Curcuma zedoaria (Berg.) Roscoe
Temu Hitam Curcuma aeruginosa Roxb
Putri Malu Mimosa pudica L. Root In vitro and In vivo (Septianing Ratri,
2017)
Soursop - Leaves In vivo (Zulkipli et al., 2018)
Betel - Leaves In vivo (Zulkipli et al., 2018)
Papaya - Leaves In vivo (Zulkipli et al., 2018)
Garlic - Tubers In vivo (Zulkipli et al., 2018)
Jarak Tintir Jatropha multifida Leaves In vivo (Suwastini et al.,
2020)
Tembelekan Lantana camara H. suaveolens (L.) Poit Leaves In vitro (Chatri & Mansyur-
- din, 2015)
Karamunting Melastoma malabathricum L. Leaves In vitro and In vivo (Suyanti et al., 2020)
Purun Tikus Eleoharis dulcis
Kirinyuh Chromolaena odorata L
Noni Morinda citrifolia Leaves In vivo (Marsuni, 2020)
Betel Piper batle L. Leaves In vivo (Juniar Dwi Cahya,
2019)
Tagetes Tagetes erecta Leaves In vitro (Satryawibowo et al.,
2015)
Suren Toona sureni Merr. Leaves In vivo (Andriyani et al.,
2020)
In vitro (Andriyani & Pur-
wantisari, 2019)
Betel Piper betle Leaves In vitro and In vivo (Trisnawati, D., Nu-
groho, L. P. E., 2019)

10
Fragrant Lemon- Cymbopogon nardus L. Leaves In vitro and In vivo (Syabana et al., 2015)
grass
Papaya Carica papaya Linnaeus Leaves In vitro (Liswarni & Edri-
wilya, 2020)
Pacar Cina Aglaia odorata L. Leaves In vitro and In vivo (Efri et al., 2017)
Neem Azadirachta indica A. Juss Leaves In vivo (Aziziy et al., 2020)
Kepok Banana - Hump
Noni Morinda citrifolia L. Leaves In vitro (Anggreini et al.,
2016)
Neem Azadirachta indica A. Juss.

Kenikir Cosmos caudatus Kunth. Leaves In vitro and In vivo (Amelia et al., 2020)
Babadotan Ageratum conyzoides Leaves In vitro (Wulandari et al.,
2015)
Awar-awar Ficus septica Leaves In vitro and In vivo (Sudirga, 2018)
Mixture of Betel Piper betle L. dan Nicotina tobacum Leaves In vitro and In vivo (Anjani, 2018)
and Tobacco
Gelinggang Cassia alata L. Leaves In vivo (Supriati et al., 2016)
Jarak Pagar Jatropha curcas L. Seed In vitro and In vivo (Lestari et al., 2020)
Mixture of Betel Piper betle L. dan Nicotiana tabaccum L. Leaves In vitro and In vivo (Nur Rohmah, 2017)
and Tobacco
Binahong Anredera cordifolia Leaves In vitro and In vivo (Yulia et al., 2019)
Neem Azadirachta indica A. Juss. Leaves In vitro and In vivo (Ali et al., 2012)
Noni Morinda citrifolia L. Fruits In vitro and In vivo (Ali et al., 2013)
Betel Piper betle L Leaves In vivo (Damiri, 2011)
Noni M. citrifolia Fruits In vitro (Septiana et al., 2013)
Betel Piper betle L. Leaves In vitro (Ningtyas et al., 2013)
Babadotan Ageratum conyzoide
Noni Morinda citrifolia Leaves, Flowers, In vivo (Efri, 2010)
and Fruits
Jarak Jatropha curcas L. Leaves In vivo (Wanda et al., 2014)
Mimba Azadirachta indica
Betel Piper betle L. Leaves In vivo (Wati et al., 2014)
Babadotan Ageratum conyzoides L.
Babadotan A. conyzoides - In vivo (Gusmarini et al.,
2014)
Siam C. odorata -
Reed I. cylindrica -
Teki C. rotundus -
Camplong Callophyllum inophyllum Fruits In vitro (Sholehah, 2012)
Patchouli Oil - - In vitro and In vivo (Sakerebau & Wahyu,
2013)
Ubi Ungu Ipomoea batatas Leaves In vitro and In vivo (Saputri & Utami,
2020)
Mixture of Cosmos caudatus dan Piper betle Leaves In vitro (Maimunah et al.,
Kenikir and Betel 2019)
Putri Malu Mimosa pudica Leaves In vivo (Eviyanti, 2020)
Kirinyuh Euphatorium odoratum L. Leaves In vitro and In vivo (Indrawati, 2021)
Cinnamon Cinnamomum burmannii Leaves In vitro (Darmadi et al., 2021)
Neem Azardiachta indica Leaves In vitro (Rahman et al., 2019)
Garlic Allium sativum Rhizome In vitro (Rahman et al., 2019)
Zinger Zingiber officinale Rhizome
Termaric Curcuma longa Rhizome
Tulsi Oscimum sanctum Linn. Leaves
Mahogoni Swietenia mahogoni Leaves
Mehendi Leaves

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 Table 2 Methods of extract preparation for in vitro and in vivo test

Plant Method Solvent Sample : Sol- Result Reference

vent (w/v)
Binahong Leaf Maceration for 1 x 24 hours then 90% 1:4 Sticky (Yulia et al., 2019)
concentrated using rotary evapora- methanol
tor
Banana Hump The sample was sieved with size of 100% 6 : 10 Condensed (Tobing & Mulyan-
and Mimba 25 mesh and macerated separately methanol extract ingsih, 2020)
Leaf for 1 x 24 hours then concentrated
with rotary evaporator to 250 mL
Jarak Pagar Seed Maceration for 48 hours then con- 96% ethanol 1 : 3 and 1 : 2 - (Lestari et al., 2020)
centrated with rotary evaporator
Mixture of Be- The sample was separately dried Distilled 1:1 - (Oktarina et al., 2017)
tel and Tobacco and then crushed by adding dis- water
Leaf tilled water and then filtered
Leaf of Pasang Leaf powder is macerated for 2 x 24 96% ethanol 1:4 - (Suyanti et al., 2020)
Surut Weeds hours then concentrated with ro-
tary evaporator at a temperature of
40 - 70 °C and followed by evapo-
ration process with water bath at
temperature of 50 - 60 °C
Awar Awar Leaf Maceration for 72 hours then evap- Methanol 1 : 10 - (Sudirga, 2018)
orated with rotary evaporator
Babadotan Leaf Graded fractionation Water, 1 : 10 - (Wulandari et al.,
methanol, 2015)
ethyl acetate,
and n-hexane
Noni Leaves Maceration for 3 x 24 hours then Methanol 1:3 Condensed (Nurul et al., 2020)
and Fruit concentrated with rotary evapora- extract
tor and waterbath at temperature of
60 °C
Kenikir Leaf Maceration for 2 x 24 hours: the first 96% ethanol 1 : 3 and 1 : 2 - (Amelia et al., 2020)
soaking for 6 hours, then stirring
it then leaving it for 18 hours then
concentrating it with rotary evapo-
rator and then concentrating again
with a waterbath at temperature of
40 °C
Noni and neem Using simple fractionation tool Water 1:5 - (Anggreini et al.,
leaf 2016)
Pacar Cina The sample was added with ster- Distilled 1 : 20 - (Arneti & Sulyanti,
Leaf Graded ile distilled water, blended until water 2017)
fractionation smooth and put in sterile erlen-
Distilled water, meyer and covered with aluminum
alcohol, and foil. extract was heated until
n-hexane - - boilling and then filtered
(Efri et al., 2017)
Ketepeng Cina
Fragrant The sample was heated in water at Water - Concentrated (Syabana et al., 2015)
Lemongrass 90 °C for 30 minutes then concen- extract
Leaf trated on rotary evaporator
Suren Leaf Maceration for 1x24 hours then 70% ethanol 1:3 Pure extract (Andriyani & Pur-
concentrated with rotary evapora- wantisari, 2019)
tor
Tagetes Leaf Using simple fractionation tool Water, - - (Satryawibowo, 2015)
methanol,
ethyl acetate,
and n-hexane
Putri Malu Root The maceration and then evapo- Ethanol 9 : 10 Condensed (Eviyanti, 2020)
rated with rotary evaporator extract
Jarak Tintir and The sample was separately ex- Water 1:5 Dry extract (Suwastini et al.,
Tembelekan tracted using simple fractionation 2020)
tool and then evaporated with
rotary evaporator
Neem, Betel, Maceration for 3 days and stirring 3 96% ethanol 1:5 Concentrated (Sitompul, 2017)
and Clove Leaf times a day then concentrated with extract
rotary evaporator

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 Curcuma spp. Maceration Methanol - Condensed (Sari et al., 2020b)
Rhizome extract
Urang aring Maceration for 3 x 24 hours then Ethanol 1:4 Concentrated (Andreas et al., 2018)
evaporated with rotary evaporator extract
at temperature of 40 °C
Shallot and Gar- Extraction and then extracted in Distilled 1:1 Crude extract (Sittisart et al., 2017)
lic centrifuge and filtered water
Carica papaya Maceration for 24 hours then the Ethanol 1 : 8, 1 : 10, 1 : 12 - (Chávez-Quintal et
L. Cv. Leaf and extract was filtered and centrifuged al., 2011)
Maradol Seed then evaporated with rotary evapo-
rator
Noni Leaf Using multilevel extraction Water, al- 1:5 Dry extract (Putra, 2017)
cohol, ethyl
acetate
Galangal Rhi- Maceration extraction for 24 hours 95% ethanol - Condensed (Harianto, 2018)
zome, Clove then evaporated by rotary evapora- extract
Leaf, and Ban- tor
gun Bangun

3 Preparation of extract the active polar fungicide compound which is polar. Decreasing
the level of polarity starting from methanol, ethyl acetate, and n-
The preparation of test solutions for chilies fungicide was sum- hexane solvents is expected to be able to separate the active fungi-
marized in Table 2. In general, the method of extracts preparation cide compounds based on their polarity level.
used can be classified into 3 types, namely: 1) maceration method,
2) graded fractionation method, and 3) decoction method.
3.3 Decoction method
3.1 Maceration method Decoction method has been used to extract the fungicidal ac-
tive compounds found in betel leaf. Samples were boiled in wa-
This method are most widely used to extract active compounds ter with ratio of 1: 1 for 1 hour. The extract are filtered and steril-
from certain plants for fungicide. Plants were prepared in the pow- ized using autoclave at temperature of 121 °C to obtain sterile be-
der or flour form are added with a solvent and then are soaked tel leaf extract (Trisnawati et al., 2019). The boiling process of the
for a designated time. The filtrate is separated from the dregs and Cassia alata Linnaeus sample which was blended with water was
the maceration process can be continued with new solvent until carried out for 15 minutes (Arneti Sulyanti, 2017). This decoction
color filtrate is clear. The filtrate is concentrated using rotary evap- method is rarely used because it is feared that the active fungici-
orator with temperature control according to the type of solvent dal compounds present in the sample could be damaged by heat
used until concentrated extract is free solvent (K Ngibad, 2019), treatment.
(Khoirul Ngibad, 2019), (Wibowo et al., 2019). The solvents used in
the extract preparation for test fungicide on chilies, including: wa-
ter solvent (ultrapure water) and organic solvents (90% methanol, 4 In vitro test as fungicide for chili
methanol, 70% ethanol, 96% ethanol, ethanol, ethyl acetate, and
n-hexane). The usage of solvents in the maceration process is ex- The review results of research related to in vitro fungicide test
pected to be able to extract the large fraction of possible fungicidal are summarized in Table 3. The concentration of the test solution
active compounds. In addition, there are differences in the ratio of was carried out in various ways. For example, the concentration
sample weight and volume of solvent used between researchers, of mixture of betel leaf and tobacco extract with concentration of
ranging from 1: 1 to 1: 10. The greater the ratio of solvent volume 30% was made by mixing 7 ml of PDA (Potato Dextrosa Agar) and
and sample weight will maximize the extract or active fungicidal 3 ml of mixture of betel and tobacco extracts (Nur Rohmah, 2017).
compound produced. However, it is necessary to pay attention to Another technique was found in preparation of kenikir leaf extract
the effectiveness of usage the solvent volume. test solution which is done by mixing the extract with Tween 80
as emulsifier with ratio of 1: 1 (w / v) and diluted using sterile dis-
3.2 Stratified fractionation method tilled water to get concentration of 5%, 10%, 15%, and 20% (Amelia
et al., 2020). In other cases, the suren concentrated extract was as-
Practices of the graded fractionation method have been carried sumed to be 100% concentration then the concentrated extract was
out, for example: the fine powder of Chinese henna leaves was diluted using distilled water into several concentrations (25%, 50%,
fractionated in stages using filter made of various sizes of par- and 75%) (Andriyani et al., 2020). Besides water, methanol was also
alon to form funnel containing activated charcoal as filter and used as solvent to make test solution for the Curcuma sp. rhizome
adsorption of nonpolar compounds. The liquid-liquid solvent with concentration of 4-12 ppm (Sari et al., 2020b).
extraction method used cold distilled water. Then, it was fol- The synthetic fungicide control used by several researchers in
lowed by solution of alcohol or n-hexane with concentrations of in vitro tests included: propineb 70%, 0.2% propineb, azoxistrobin,
10, 20, 30, 40, 50, 60, 70, 80, and 90%, respectively (Efri et al., diphenoconazole, benomyl, anthracol, 0.2% acrobat and 0.2% car-
2017). Then, babadotan/goatweed (Ageratum conyzoide) leaf pow- bendazim. The usage of synthetic fungicide controls is very use-
der was placed into simple fractionation tool, then the filtered ful as comparison against the plants being studied. Many stud-
residue was collected and air-dried. The filtrate or crude extract ies do not use synthetic fungicide controls so that the potential
was added with methanol solvent then was collected and air-dried of these plants is less known when compared to synthetic fungi-
to obtain the methanol fraction of the babadotan leaf extract. In cide controls. On the other hand, the most widely used fungi for
the same way, to get ethyl acetate and n-hexane extract (Wulan- in vitro tests are Colletotrichum capsici and then Colletotrichum
dari et al., 2015). The water solvent is expected to be able to extract gloeosporioides.

13
 Table 3 In vitro fungicide test on chili test

Plant Concentration Synthetic Type of Test Results Reference

Test Solution Fungicide Mushroom
Control
Jarak Pagar 10 - 40% - Collectotrichum The percentage of inhibition of fun- (Lestari et al., 2020)
Seed capsici gal mycelium (%): 18.90 – 31.08
Betel and To- 30% with con- - Colletotrichum Percentage of Colletotrichum sp (Nur Rohmah, 2017)
bacco centration ratio sp colony inhibition (%): 0.56 - 30.44
(1: 1), (1: 2), (2:
1), (1: 3) (3: 1)
Betel and To- 30% with ratio - Colletotrichum Average of inhibition power (%): (Anjani, 2018)
bacco of 3: 1 sp 45.08 - 15.68
Average of spore density (106spores
/ mL): 15.8 - 35.2
Babadotan - Propineb 70% Colletotrichum Colony diameter of C. capsici (Wulandari et al.,
Seed 1 g/L capsici Water extract: 99.28% 2015)
Methanol extract: 76.81%
Ethyl acetate extract: 137.20%
N-hexane extract: 85.27%
Propineb 70%: 0%
Number of spores
Extract water: 5.33 × 106
Methanol extract: 2.34 × 106
Ethyl acetate extract: 7.78 × 106
N-hexane extract: 4.90 × 106
70% propineb: 0.00 × 106
Noni Leaves 5% - Colletotrichum Percentage of inhibition (%) (Nurul et al., 2020)
and Fruit capsici Leaves: 2.27 Fruits: 2.78
Amount of conidium (conid-
ium/mL) Leaves: 1.44
Fruits: 1.42
Kenikir Leaf 5 – 20% - Colletotrichum Percentage of inhibition (%) : 10.76 (Amelia et al., 2020)
sp – 41.12
Pacar Cina Aquades extract Propineb Colletotrichum Growth diameter of C. capsici on (Efri et al., 2017)
Leaf and 10 - 90% al- 0,2% capsici day 7 (cm): 5.85 - 1.00 Spore density
cohol extract of C. capsici: 15.77 - 0.88
Aquades extract Colletotrichum Growth diameter of C. capsici on
and 10 - 90% n- capsici day 7 (cm): 2.16 - 0.5 Spore density
hexane extract of C. capsici: 4.8 – 0
Papaya Leaf 1–5% - Colletotrichum Colony area and effectiveness: 41.32 (Arneti & Sulyanti,
gloespori- - 20.11 and 6.13% - 64.04% Mush- 2017)
oides room wet weight: 4.69 - 45.16%
Mushroom dry weight: 8.33 - 54.16%
Number of conidia: 27.5 - 82.5%
Suren Leaf 10 – 30% - Colletotrichum Percentage of inhibition of C. cap- (Andriyani & Pur-
capsici sici colony diameter: 45.49 - 62.74 wantisari, 2019)
Tagetes Leaf Water, Propineb 70% Colletotrichum Percentage of colony diameter: (Satryawibowo, 2015)
methanol, capsici 55.18 - 92.47%
ethyl acetate, Percentage of Spore density: 7.20 -
and n – hexane 36.88
extracts)
Pasang Surut Extract of purun Azoxistrobin, Colletotrichum The percentage of inhibition: 6.99 - (Suyanti et al., 2020)
Weeds tikus, extract dipheno- sp 79.54
of kirinyuh, conazole, and
and extract of benomyl
karamunting
Essential Young leaves 0.5 - Colletotricuhm Percentage of inhibition (%): 50 - 65 (Chatri & Mansyur-
oil (Hyptis – 2.5% gloeosporoides din, 2015)
suaveolens L Mature leaves
) 0.5 - 2.5%)
Putri Malu 30 – 90% - Colletotrichum Percentage of inhibitory power (%): (Septianing Ratri,
sp 14.36 - 28.01 2017)
The average of spore density (106
spores / ml): 19.11 - 4.44
Curcuma spp 4 – 12 ppm - Colletotrichum The average of colony growth diam- (Sari et al., 2020b)
capsici eter (cm): 1.50 - 0.58

14
 Betel and To- Biorational - Colletotrichum Percentage of inhibition of fungal (Oktarina et al., 2017)
bacco Leaf extract: (1:1), capsici colonies (%): 0.56 - 30.44
(1:2), (2:1), (1:3),
(3:1)
Average of spore density (106
spores/ ml): 31.9 - 7.6
Tobacco 25 – 100% - Colletotrichum Percentage of inhibition (%): 6.56 - (Isman Duila, 2017)
sp 33.78
Average of spore density (106 spores
/ ml): 19.57 - 100
Flour of fra- 50 – 250 g/l - Colletotrichum Percentage of inhibitory power (%): (Elfina et al., 2016)
grant lemon- capsici 17.47 - 34.43
grass
Ketepeng 5% - Colletotrichum Percentage of inhibitory power (%) (Arneti & Sulyanti,
Cina gloeospori- Root: 11.96 2017)
oides Seeds: 17.21
Interest: 30.68
Young leaves: 38.40
Stems: 56.37
Old leaves: 64.30
Urang Aring 5 - 25% Antracol Colletotrichum Number of mushroom colonies on (Andreas et al., 2018)
sp day 7: 125.75 - 72.75
Diameter of mushroom colonies on
day 7: (1.254 - 1.38)
Banana 15 – 45% - Colletotrichum Percentage of inhibition zone for (Tobing & Mulyan-
Hump and capsici fungal colonies: 10.76 - 6.58 ingsih, 2020)
Mimba Leaf
Cinamon leaf 0,5 - 1,50% - Colletotrichum Percentage of inhibition (%): 17 - (Darmadi et al., 2021)
Extract capsici 100%
Percentage of spore density (%): 51
100
Leaf Extract of 1 – 5% - Colletotrichum Colony diameter (mm): 29.72 - 81.39 (Sudirga et al., 2014)
Ficus septica acutacum
Percentage of spore density
(105 spora / ml): 63.21 - 99.11
Percentage of spore density (105
spora / ml): 63.21 - 99.11
Kirinyuh Leaf 10 – 70% Acrobat 0,2% Colletotrichum Average percentage of fungal (Indrawati, 2021)
Extract capsici colony growth (%): 100.00 - 0.00
Fungal inhibition zone on day 10:
1.59 - 1.10
Mansoa alli- 1 – 5% - Colletotrichum Colony diameter (mm): 63.25 - (Sudirga et al., 2019)
acea Extract acutacum 17.00
Spore growth (105 spores / mL): 4.19
-0
Spore germination (105 spores /
mL): 2.59 - 0
Akar Putri 25 - 100% - Colletotrichum Percentage of inhibition zone di- (Eviyanti, 2020)
Malu sp ameter (%): (70 - 10)
Neem Seed Neem oil, garlic Carbendazim Colletotrichum Percentage of inhibition: 74.77- (Musakhan &
Kernel Extract bulb extract, 0,2% capsici 68.75 Zacharia, 2017)
combinate
application of
neem, garlic,
ginger, onion
plant extract,
and neem sed
kernel extract
(NSKE)
Purple Sweet 5 – 40% - Fusarium sp Average percentage of inhibitory (Saputri & Utami,
Potato power (%): 56.7 - 76.6 2020)

Some of the parameters used in the in vitro test include: colony Then, the colony diameter is calculated using formula (Andreas et
diameter, percentage of colony inhibition, density / number of al., 2018) :
spores, and colony area. Colony diameter is measured by mak-
ing vertical and horizontal lines perpendicular to each other at 𝐶 𝑜𝑙𝑜𝑛𝑦𝑑𝑖 𝑎𝑚𝑒𝑡 𝑒𝑟 (𝑐𝑚) = (𝐷 1 + 𝐷 2)/2 (1)
the bottom of the petri dish as vertical and horizontal diameters. With :

15
 D1 = diameter of horizontal colony With:
D2 = diameter of vertical colony C = spore density per ml of solution
Observations are made by measuring the diameter of the t = total number of spores in the sample box observed
growth of C. capsici colonies. The measurement of inhibition us- n = number of sample boxes (5 large x 16 small boxes)
ing the formula: 0.25 = correction factor for the use of a small-scale sample
box on the haemacytometer
𝐷𝐻 ( %) = (𝑎 − 𝑏)/𝑎𝑥 100% (2)
With : Colony area was measured using millimeter plotting paper by
DH = Inhibition (percent) depicting the colony area on plastic glass (Liswarni Edriwilya,
a = diameter of C. capsici colony (mm) (negative control) 2020). The plants studied as an alternative to natural fungicides
b = diameter of C. capsici colony (mm) (treatment) for chili have the ability to inhibit the growth of anthrax-causing
fungi in chilies by in vitro study, which include: Colletotrichum
Spore density was determined by taking 1 ml of spore sus- capsici, Colletotrichum gloeosporioides, and Colletotrichum acu-
pension from isolate propagation treatment. Furthermore, the tacum. However, many in vitro studies do not compare with syn-
spore density was calculated using hemocytometer that had been thetic fungicides. So, it is not possible to know the effectiveness of
dropped by the suspension under a double lens (binocular), which the performance of natural fungicides for chilies when compared
is one type of lens from a light microscope with a magnification of to synthetic fungicides. Based on Table 4, it can be seen that 80%
400 times. (Herlinda et al., 2006). The spore density was calculated alcohol extract and 10% n-hexane extract and 60% Chinese henna
using Gabriel Riyatno formula (1989) (Gabriel Riyanto, 1989): leaves is similar with the 0.2% propineb performance by in vitro
𝑡 study. In addition, the 60% and 70% kirinyuh leaf extracts were also
𝐶 = 𝑋 106 (3) able to match 0.2% acrobat performance by in vitro study.
𝑛𝑥 0, 25

Table 4 Comparison of effectiveness of natural fungicides for chilies with synthetic fungicides

Plants Synthetic Fungicides Explanation Reference

Babadotan Leaf 70% Propineb The effectiveness of the three extract frac- (Wulandari et al., 2015)
tions < propineb 70%
Pacar Cina Leaf 0.% Propineb The effectiveness of 80% alcohol extract (Efri et al., 2017)
and 10% and 60% n-hexane extract is is
comparable to 0.2% propineb
Tagetes Leaf 70% Propineb Cannot match the effect of 70% propineb (Satryawibowo, 2015)
Urang Aring Antracol Effectiveness of urang-aring < Antracol (Andreas et al., 2018)
Kirinyuh Leaf Extract 0.2% Acrobat The effectiveness of 60% and 70% kirinyuh (Indrawati, 2021)
leaf extract is is comparable to 0.2% acro-
bat
Neem Seed Kernel 0.2% Carbendazim The effectiveness of neem seed kernel ex- (Musakhan & Zacharia,
Extract tract < 0.2% carbendazim 2017)

5 In vivo test as fungicide for chili ious plants with certain concentrations as shown in Table 5. This
in vivo test was directly applied to chili plants to be treated with
In effort to find alternatives natural fungicides, the researchers natural fungicides with test conditions appropriate to the actual
focused not only on in vitro studies but also in vivo studies of var- environment in chili farm.

Table 5 In vivo fungicide test for chili

Plants Concentration Synthetic Test Parameters Test Results Reference

of Test Solu- Fungicide
tion
Neem Leaf 5 – 20% - Incubation period 9 – 12 days (Sitompul, 2017)
Extract
The severity of anthracnose 8.84 – 8.43%
Betel Leaf Ex- Incubation 12 – 19 days (Sitompul, 2017)
tract period
The severity of anthracnose 8.88 – 7.74%
Clove Leaf Extract Incubation period 14 – 14.33 days (Sitompul, 2017)
Putri Malu 30 – 90% - Percentage of disease inci- 62.5 – 0% (Eviyanti, 2018)
Root Extract dence
Spot diameter 0 mm
Incubation period 12 days
Suren Leaf Ex- 25 – 100% Mankozeb Percentage and fresh weight 91.16 – 99.68% (Andriyani et al.,
tract 1g/L of healthy cayenne pepper 2020)
affected by anthracnose
Morphometry of chili fruit Length: 7.74 – 8.54
cm

16
 Diameter : 1.14 – 1.48
cm
Chili plant height 122.8 – 131.6 cm
The severity of anthracnose 4 – 0%
Extract of 5 – 25% Propineb The severity of anthracnose 13.00 – 17.50% (Sihite et al., 2020)
Umbi Teki
Plant height 37.56 – 34.18 cm
Number of fruit 7.5 – 3.2 fruit
Fruit weight 6.2 – 3.8 gram
Extract of 4 – 12 ppm - Symptoms of anthracnose 0.67 – 0.00 cm (Sari et al., 2020b)
Curcuma in red chili
Longa Sensu
Mycelium dry weight 80.53 – 0.00 mg
Extract of Symptoms of anthracnose 0.88 – 0.00 cm
Curcuma in red chili
aeroginosa
Mycelium dry weight 0.00 mg
Extract of Symptoms of anthracnose 0.78 – 0.00 cm
Curcuma in red chilies
zedoaria
Mycelium dry 69.00 – 0.00
weight mg
Citronella 50 – 250 g/L - When the early symptoms of 2.48 – 2.00 days (Elfina et al., 2016)
Leaf Extract anthracnose disease appear
in red chilies
The intensity of the attack to 19 - 20
C. capsici
Effectiveness and level of -11,76 until 17,65%
fungicidal ability
Awar-Awar 1 – 5% - Percentage of incidence of 41.00 – 0% (Sudirga, 2016)
Leaf Extract anthracnose in red chilies
Percentage of incidence of 41.00 – 0%
anthracnose in red chilies
Disease intensity 38.21 – 0%
Yield / amount of red chilies 0.163 – 0.587
(kg / plant)
Losing the salvaged yield of 59.51 – 88.76%
red chilies
Betel and (1:2), (1:2), - Anthracnose incidence rate 25 – 75% (Oktarina et al., 2017)
Tobacco Leaf (2:1), (1:3),
Extract (3:1)
The incubation period for 4 – 9 days
anthracnose
Betel Extract 200 – 600 - Number of fruits / plants 33 – 36 fruits (Juniar Dwi Cahya,
mL/L 2019)
Number of fruit / plot 75 – 76 fruitd
Fruit weight / plant 362.23 – 387.21 gram
Fruit weight / plot 757.80 – 777.93 gram
Number of healthy fruit / 21 – 24 fruits
plant
Number of damaged fruit / 8 – 5 fruits
plants
Percentage of healthy fruit / 88.15 – 97.46%
plot
Percentage of damaged fruit 12.24 – 2.56%
/ plot
The intensity of the plant at- 3.06 – 1.25%
tacked
Extract of Pu- 30 – 90% - Incidence of anthracnose in 62.5 – 0% (Eviyanti, 2018)
tri Malu red chilies
Incubation period 6 – 12 days
Spot diameter 6.8 – 0 mm
Betel Leaf Ex- - - Incidence of anthracnose in 43 – 15% (Trisnawati et al.,
tract chilies 2019)
Neem Leaf 15% - 45% - Average height of chili 25.41 – 22.79 cm (Tobing & Mulyan-
Extract plants ingsih, 2020)

17
 Average width of fungal 25.02 – 22.80 cm
spots
Interaction of average of 33.17 – 33.03 cm2
plant leaf area
Average weight of chilies 5.97 – 4.89 gram
Interaction of the main 36.67 – 52.22%
branches of the chili plant
Dry weight of chilies 20.79 – 15.07 gram
Wet weight of chilies 20.53 – 26.74 gram
The average of root wet 3.58 – 4.99 gram
weight
The average of root dry 0.36 – 0.49 gram
weight
Disease incidence in chilies 20.3 – 0.00%
The average of incidence of 0.52 – 2.85%
pest infestation
Banana Wee- 15 - 45% - Average height of chili 23.57 – 25.49 cm (Tobing & Mulyan-
vil Extract plants ingsih, 2020)
Average width of fungal 24.58 – 23.50 cm
spots
Interaction average leaf area 28.99 – 40.21 cm2
of plants
Average weight of chilies 5.42 – 5.83 gram
Interaction of the main 43.00 – 59.11%
branches of the chili plant
Dry weight of chilies 22.17 – 19.63 gram
Wet weight of chilies 26.81 – 22.86 gram
The average weight of the 3.98 – 4.44 gram
wet roots
The average dry weight of 0.39 – 0.45 gram
the roots
Disease incidence in chilies 1.31 – 4.17%
The average incidence of 2.94 – 1.17%
pest infestation
Neem Leaf 15 – 45% - Height of red chili plant 16.39 – 17.26 cm (Aziziy et al., 2020)
Extract
Number of red chili leaves 18.47 – 18.89 leaves
Number of productive 5.11 – 5.33
branches
Fruit weight / pPlant 46.95 – 50.03 gram
Number of pieces / plant 12.83 – 13.83 fruits
Fruit length 10.63 – 10.37 cm
Fruit diameter 1.10 – 1.12 cm
Banana 15 – 45% - Height of red chili plant 16.30 – 17.65 cm (Aziziy et al., 2020)
Weevil Mole
Extract
Number of red chili leaves 18.64 – 19.22 leaves
Number of productive 5.06 – 5.28
branches
Fruit weight / plant 44.08 – 53.20 gram
Number of pieces / plant 12.19 – 13.89 fruits
Fruit length 10.37 – 10.62 cm
Fruit diameter 1.11 – 1.12 cm
Noni Leaf Ex- 4 mL/100 mL, - Anthracnose intensity 12.75% (Marsuni, 2020)
tract 8 mL/100 mL,
12 mL/100 mL,
16 mL/100 mL
aquadest
Betel and (1:1), (1:2), - Disease incidence rate 75 – 25% (Nur Rohmah, 2017)
Tobacco (2:1), (1:3),
Extracts (3:1)
Incubation period 4 – 9 days
Spot diameter 6 – 4 mm
Kenikir Leaf 5 – 20% - Incubation period 4.40 – 5.13 days (Amelia et al., 2020)
Extract
Disease incidence 72.5 – 52.5%
Percentage of incidence of 72.5 – 87.5%
anthracnose in chilies

18
 Tobacco 25 – 100% - Disease incidence 75 – 0% (Isman Duila, 2017)
Extract
Incubation period 4 – 13 days
Extract of 30% - 50% - Incubation period 2.57 – 5.4 days (Lestari et al., 2020)
Jarak Pagar
Seed
The incidence of anthrac- 65 – 85%
nose in chilies
Extract of For- 25 – 100 g/L - The appearance of early an- 3.33 – 4.25 days (Elfina et al., 2015)
est Betel Leaf thracnose symptoms
Flour
The intensity of fungal at- 18.00 – 10.00%
tack on chilies
Effectiveness and level of
fungicidal ability 14.00 – 52.00%
Soursop, - - The intensity of the attack 39%, 31%, 33%, 35% (Zulkipli et al., 2018)
Betel, Papaya,
and Garlic
Leaf Extracts
Fragrant 0.1 – 0.5% 0.2 % b/v Incubation period 5.0 – 6.0 days (Syabana et al., 2015)
Lemongrass synthetic
Leaf Extract fungicides
The intensity of the attack 13.3 – 6.6%
Reduction in weight of 13.9 – 7.4%
chilies
Average total yield 3.8 and 2.5 kg

Several parameters that are often observed in in vivo tests in- Several studies have also identified other parameters in the
clude: 1) The percentage of incidence of anthracnose disease and in vivo test, for example: fruit weight, mycelium dry weight, spot
2) The percentage of severity of anthracnose disease or the inten- diameter, yield / number of red chilies, number of fruits, effective-
sity of disease / attack. ness and level of fungicidal ability, incubation period of anthrac-
nose disease, morphometry of cayenne pepper, period incubation,
Determination of the percentage of disease occurrences is percentage and fresh weight of healthy cayenne pepper affected
done by counting the number of symptomatic chilies in each plant by anthracnose disease, when the early symptoms of anthracnose
with the following formula (Suwastini et al., 2020): disease appeared in red chilies, and the height of chili plants. The
plants studied for chilies had the effectiveness of being used as a
𝑛
𝑇𝑃 𝑥 100% (4) natural fungicide. However, many in vivo studies do not compare
𝑁
with synthetic fungicides. So, it is not possible to know the effec-
With :
tiveness of the performance of natural fungicides for chilies when
TP = Occurrence of disease (%)
compared to synthetic fungicides. Table 6 shows that suren leaf
n = Number of infected (symptomatic) fruit per plant
extract and nut bulbs can be used as alternatives to natural fungi-
N = Total number of fruits observed per plant
cides to help overcome the problem of anthracnose in chilies.
Anthracnose disease in chilli is characterized by the appear-
Table 6 Comparison of the effectiveness of natural fungicides for
ance of blackish brown spots that will expand into soft rot with
chilies with synthetic fungicides
black dots in the middle which are collection of seta and conidia of
C. capcisi fungi. The attack of C. capsici fungi begins by attaching
the spores to the fruit and then the spores will germinate. Further- Plants Synthetic Explanation Referen
more, through the fungal hyphae inject the fruit tissue and take Fungi- ce
nutrients in it so that it can interfere with metabolism and even cides
cause cell death. The more severe the disease attack, the more ex- Suren Mankozeb In general, the performance (Andriy
tensive the rotting area on the fruit will be, this is due to damage to Suren 1g/L of suren leaf extract as nat- ani et
the fruit tissue and even cell death which ultimately results in the Leaf ural fungicide > mankozeb al.,
fruit experiencing dry rot or shrinking. (Andriyani et al., 2020). Dis- Extract fungicide 2020)
ease severity is the surface area of chilies that shows symptoms of Extract Propineb The treatment of nut bulb (Sihite
disease. Disease severity can also be interpreted as the part of the of Umbi flour with concentration et al.,
plant affected by disease or the disease area of the sample plant. Teki of 5%, 15%, and 25% was 2020)
Determination of the percentage of disease severity can be calcu- comparable to the propineb
lated by the formula as follows (Suwastini et al., 2020): fungicide which is effective
Í
𝑛𝑥𝑣 in controlling anthracnose
𝑇𝑃 𝑥 100% (5) in chili plants.
𝑁 𝑥𝑉
Fragrant 0.2% In general, the performance (Syaban
With : Lemon- w/v syn- of fragrant lemongrass leaf a et al.,
KP = Disease severity (%) grass thetic extract as natural fungicide 2015)
N = The number of fruit observed per plant Leaf fungi- < 0.2% w / v synthetic fungi-
n = The number of fruits in each attack category Extract cide cide
v = Numeric value for each attack category
V = Highest score

19
6 Conclusion Agrotek Tropika, 4(1).
Anjani, A. G. (2018). Efektivitas Lama Penyimpanan Campuran Ek
This paper reviews the potential plants as an alternative to chili strak Sirih Dan Tembakau Pada Colletotrichum Sp. Penye-
fungicides, the preparation of test solutions, in vitro and in vivo bab Antraknosa Cabai. Universitas Muhammdiyah Jember.
fungicide tests. The part of the plant that is widely studied as fungi- Arneti, A., Sulyanti, E. (2017). Pengujian Ekstrak Sederhana Bagian
cide for chilies is the leaves, while the parts of the plant that are Tumbuhan Cassia Alata Linnaeus Terhadap Colletotrichum
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weevils, seeds, fruit, flowers or all parts of the plant. The methods (Journal Of Plant Protection), 1(2), 42–51.
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method, stratified fractionation method, and decoction method. gan Antraknosa Pada Pertumbuhan Cabai Merah (Capsicum
The plants studied had the ability to inhibit the growth of the Annuum L.) Setelah Aplikasi Larutan Daun Mimba Dan Mol
Colletotrichum capsici, Colletotrichum gloeosporioides, and Col- Bonggol Pisang. Jurnal Agronida, 6(1), 22–32. Badan Pusat
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n-hexane extract of Chinese henna leaves can be equal with the gan Komoditas Cabai Merah Indonesia 2019. Bps Ri/Bps-
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60% and 70% kirinyuh leaf extracts were also able to match acrobat Chatri, M., Mansyurdin, A. B. P. (2015). Potensi Minyak Atsiri Hyp
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The authors declare no known competing interests that could Darmadi, A. A. K., Suriani, N., Darmayasa, I. D. A. B. G., Bagus, I. D.
have influenced the work reported in this paper. A., Suaskara, M., Gari, N. I. M., Fudholi, A. (2021). Cinnamon
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