The Effect of Papaya Leaf Extract (Carica papaya L.) on the Mortality Rate of Spodoptera litura Fabricius Larvae and the Level of Damage to Soybean Leaves in Malang, Indonesia: A Greenhouse Simulation

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INTRODUCTION

Nowadays, soybeans have become an important food commodity in Indonesia after rice and corn (Sulistyo & Inayati, 2016). Soybeans have a high protein content. Soy protein is one source of vegetable protein that is important for health, which can reduce the risk of cancer and reduce risk factors for cardiovascular disease and other chronic diseases (Qin, Wang, & Luo, 2022).

Soybeans are used to make tofu and tempeh;

as a result, the need for soybeans is very high.

Indonesia’s annual soybean demand is around 2.4 million tons, 83.7% for tempeh and tofu production (Padjung, Syam’un, & Kasim, 2021).

Soybean productivity in Indonesia is still not able to meet national needs. Domestic soybean production can only meet 35% of the demand (Ningrum, Irianto, & Riptanti, 2018). One of the causes of this condition is the presence of pests in soybean cultivation, especially in the tropics (Krisnawati & Adie, 2019). One of the pests that attack soybean plants is the larvae of S. litura Fabricius (Fattah, Sjam, Daud, & Dewi, 2018). The larvae attack soybean leaves and pods. The attack of S. litura Fabricius larvae on soybean plants can reduce production by up to 68% (Bayu, Krisnawati,

& Adie, 2018).

ARTICLE INFO Keywords:

Leaf damage intensity Papaya leaf extract S. litura Fabricius larvae Soybean plant

Article History:

Received: April 2, 2022 Accepted: December 15, 2022

*⁾ Corresponding author:

E-mail: sofia.ery.fmipa@um.ac.id

ABSTRACT

Spodoptera litura Fabricius pest control using botanical pesticide from papaya leaves extract (PLE) is an alternative to substitute chemical pesticides. The study aimed to determine the levels of PLE’s activity and evaluate the effectiveness of PLE from different altitudes (low and middle lands) and cultivars (Thailand and Indonesian) against the attack of S. litura Fabricius larvae. An experimental study with a randomized block design (n=5) was conducted from December 2020 until April 2021.

Each replication used 100 soybean plants and sprayed with 0% or 20%

PLE of Indonesian purple cultivar low land (UR-20%), Indonesian purple cultivar middle land (US-20%), Thailand cultivar low land (TR-20%), and Thailand cultivar middle land (TS-20%), respectively and observed at 72 hours after treatment. The results demonstrate that the middle lands contained active compounds slightly higher than the lowlands.

The damaged soybean leaves and intensity of soybean leaf damage are highest in control (77% and 63%) and significantly different from the treatment group, 53-60% and 29-41%, respectively. The mortality of S. litura Fabricius larvae in the control group was 11%, and in the treatment group, 47-63%. PLE from different altitudes and cultivars reduce the intensity of soybean leaf damage by S. litura Fabricius larvae.

ISSN: 0126-0537Accredited First Grade by Ministry of Research, Technology and Higher Education of The Republic of Indonesia, Decree No: 30/E/KPT/2018

Cite this as: Rahayu, S. E., Leksono, A. S., Gama, Z. P., & Tarno, H. (2023). The effect of papaya leaf extract (Carica papaya L.) on the mortality rate of Spodoptera litura fabricius larvae and the level of damage to soybean leaves in Malang, Indonesia: a greenhouse simulation. AGRIVITA Journal of Agricultural Science, 45(1), 20–30. http://doi.

org/10.17503/agrivita.v45i1.3745v41i0.3635

The Effect of Papaya Leaf Extract (Carica papaya L.) on the Mortality Rate of Spodoptera litura Fabricius Larvae and the Level of Damage to Soybean Leaves in Malang, Indonesia: A Greenhouse Simulation

Sofia Ery Rahayu^1*), Amin Setyo Leksono²⁾, Zulfaidah Penata Gama²⁾ and Hagus Tarno³⁾

1) Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Negeri Malang, Malang 65145, East Java, Indonesia

2) Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Brawijaya, Malang 65145, East Java, Indonesia

3) Department of Plant and Disease, Faculty of Agriculture, Universitas Brawijaya, Malang 65145, East Java, Indonesia

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The control of S. litura Fabricius pests on soybean plantations uses chemical pesticides. Using chemical pesticides can immediately overcome these pest attacks, but there are detrimental effects on humans and the environment. Chemical pesticides are used more often, and the dosage exceeds the dose (Ilhamiyah, Salamiah, Samharinto, & Halim, 2019). Using chemical insecticides causes losses because it can cause death for non-target insects, toxicity to consumers, and resistance to insect pests (Hernández-Lambraño, Caballero-Gallardo, &

Olivero-Varbel, 2014; Kuppusamy, Dhamodharan,

& Jayakumar, 2016). At this time, the development in agricultural research is the discovery and development of alternative pesticides derived from nature that are less toxic and biodegradable. Using plant materials as pesticides can attack pests on plants (Adesina, Ileke, Yallappa, & Ofuya, 2016;

Meena, Lal, & Swaminathan, 2018).

The basis for using plants as pesticides is the content of active compounds in plants. The function of active compounds for plants is as a response to pest attacks and pathogenic infections. These compounds include phenols, flavonoids, alkaloids, and tannins (Su et al., 2018). Indonesia has many plants that have the potential to be used as botanical pesticides. One of these plants is papaya (Carica papaya L.). Papaya plants have many cultivars, including Thailand and Indonesian purple cultivars.

Both papaya plant cultivars are found in low and middle lands. The results of the phytochemical screening of active compounds from the two cultivars contain compounds classified as phenols, flavonoids, and alkaloids (Rahayu, Leksono, Gama,

& Tarno, 2020) and contain tannins and saponins (Ikeyi, Ogbonna, & Eze, 2013).

Several researchers have carried out the use of PLE as a natural pesticide. Tatun, Vajarasathira, Tungjitwitayakul, & Sakurai (2014) examined the effect of PLE on the growth and development of Tribolium castaneum, and Aihetasham, Rasib, Hasan, & Bodlah (2017) researched termites. Until now, there has been no research using PLE of Thailand and Indonesian purple cultivars against the primary pest of soybean plants, namely larvae of S. litura Fabricius. For this reason, this study focused on finding the effectiveness of using PLE on soybean plants against the attack of S. litura Fabricius larvae. In addition, it measures the levels of active compounds contained in PLE. Furthermore, the implications of this study are to help explore the

potential of papaya leaves of the two cultivars as plant-based pesticides.

MATERIALS AND METHODS Sample Collection and Extraction

The papaya leaves used for the extract consisted of Thailand and Indonesian Purple cultivars taken from lowland (Nganjuk) and medium plains (Malang) during December 2020. Papaya leaves were washed, dried at room temperature, and baked at 50^°C for three days. Dried papaya leaves are crushed into powder form. The papaya leaf powder was then extracted using methanol as the solvent using the maceration method. PLE was screened for phytochemicals using LCMS to determine the content of active compounds.

The extraction was done in the Plant Physiology Laboratory, Department of Biology, Universitas Negeri Malang, in December 2020.

Soybean Plant Preparation

Soybean seeds of the var. Grobogan were sown in polybags with soil and humus medium. Each polybag was given two seeds, and soybean plants with good growth were selected at the treatment time. The soybean plants used for treatment were 17 days old and had 5-6 leaves.

Treatment

This research activity was carried out in a greenhouse from December 2020 to April 2021 in the greenhouse of the Department of Biology, Universitas Negeri Malang. The independent variables in this study were PLE (concentration 20%) and control (0%), while the dependent variables consisted of percentage leaf damage and larvae mortality. The application of a 20% concentration of PLE was made based on the results of the antifeedant test carried out previously by Rahayu, Leksono, Gama, & Tarno (2020). This study adopted a completely random sampling, which consists of the four treatments as follows: lowland Indonesian purple cultivar (UR-20%), middle-land Indonesian purple cultivar (US-20%), lowland Thailand cultivar (TR-20%), middle-land Thailand cultivar (TS-20%) and a control (aquades). Each treatment and control were sprayed on 20 soybean plants. The treatments were replicated five times. One 3^rd instar larva of S.

litura Fabricius was put on each plant. The soybean plants were watered every day. Leaf damage and mortality of S. litura Fabricius larvae were carried out 72 hours after the application of spraying PLE.

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The total number of leaves obtained was 2500 pieces taken randomly from 500 soybeans for all treatments and replicates. Leaves are separated between damaged and undamaged conditions.

Furthermore, the leaves whose condition was damaged due to being eaten by S. litura larvae were grouped according to the reference by Ortega, All, Boerma, & Parrot (2016) (Fig. 1).

The number of damaged leaves was counted and divided by the number of leaves observed (100) and then multiplied by 100 to obtain a percentage.

The intensity of the damaged leaves is obtained from the number of damaged leaves multiplied by

the intensity scale and then divided by the number of observed leaves multiplied by the largest scale of damaged leaves according to the following formula.

... (1) Where: I = attack intensity, n₁= number of damaged leaves on the scale v₁, v₁ = leaf scale value, N = number of leaves observed, Z = the highest scale value of damaged leaves; Broken leaf scale value (v₁): 0 = no damaged leaves, 1 = leaf damage >

0-20%, 3 = leaf damage > 20-40%, 5 = leaf damage

> 40-60%, 7 = leaf damage > 60-80%, 9 = leaf damage > 80-100%.

Fig. 1. The example for the category of damages soybean leaves which attacked by pest.

Table 1. The concentration of ten active compounds in papaya leaf methanol extract.

No. Active Compounds Kind of active compound

The concentration of active compounds (µg/ml) in papaya leaf methanol extract

Thailand Indonesian purple

Low land Middle land Low land Middle land

1 Gallic acid Phenolic 16.798 17.282 17.228 18.152

2 Kaempferol Flavonoid 9.560 10.011 9.961 10.823

3 Quercetin Flavonoid 11.028 11.487 11.435 12.313

4 Benzyl glucosinolate 12.526 12.997 12.945 13.845

5 Luteolin 7 glucoside Flavonoid 9.644 10.095 10.044 10.906

6 Isoquercitrin Flavonoid 8.415 8.839 8.791 9.600

7 Dehydrocarpaine II Alkaloid 5.517 5.925 6.222 7.002

8 Dehydrocarpaine I Alkaloid 4.967 5.363 5.652 6.410

9 Carpaine Alkaloid 9.600 10.058 10.007 10.880

10 Pseudocarpaine Alkaloid 9.335 9.825 9.770 10.705

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Data Analysis

Data on the concentration of active compounds in PLE and the average number of damaged soybean leaves based on the level of damage were analyzed descriptively. Data on the percentage of broken soybean leaves, the intensity of damage to soybean leaves, and mortality of S.

litura Fabricius larvae were analyzed using analysis of variance (ANOVA) and continued with Duncan’s follow-up test at a 5% confidence level.

RESULTS AND DISCUSSION

Papaya Leaf Extract Phytochemical Screening The results of phytochemical screening of PLE using LCMS have been carried out and found the content of active compounds. The number of active compounds in the Thailand cultivar methanolic PLE was 60 compounds, and in the Indonesian purple cultivar was 62. Ten detected compounds had the extract’s highest concentration (Table 1). These compounds are phenols, flavonoids, alkaloids, and benzyl glucosinolates. PLE from the medium plains had a slightly higher concentration of each active compound than the PLE from the lowlands.

Likewise, the concentration of active compounds in the Indonesian purple cultivar was somewhat higher than in the Thailand cultivar. In line with our results, the previous study reported that the total phenolic and flavonoids of Taxus walliciana increased following the altitude increment (Adhikari, Joshi, Singh, & Pandey, 2020).

PLE of Thailand and Indonesian purple cultivars from different regions (middle and lowlands)

contain active compounds belonging to the phenol, flavonoid, alkaloid, and benzyl glucosinolate groups.

The concentration of each active compound in papaya leaves from the middle lands was slightly higher than from the lowlands. The difference in the data from the concentration measurement of the active compound is due to differences in habitat conditions and the altitude of the papaya plant habitat. It appears that the active compounds produced by plants are the response of plants to different environmental factors such as light, temperature, humidity, and soil fertility. The type of compound and its quantity depend on the species and geographical conditions (Isah, 2019; Lopes et al., 2018; Yang et al., 2018). The number of active compounds of the two cultivars was different. Still, for the active compound with the highest percentage composition, it was shown that there was a similarity in compounds. The highest bioactive compounds in PLE were carpaine (alkaloid), quercetin, and kaempferol (flavonoid) (Sudi et al., 2022). Alkaloid has a bitter taste which may reduce the appetite of the larva. Further, flavonoids disturb enzyme activity, which may interrupt the insect’s development and reduce its eating palatability (Marques et al., 2015).

Effect of S. litura Larvae Attack on Soybean Plants The effectiveness of PLE of Thailand and Indonesian purple cultivars was evaluated by spraying the PLE with a concentration of 20% on soybean plants in the vegetative phase. The next stage is placing the third instar S. litura Fabricius larvae, on the soybean plants. Observations found that S. litura Fabricius larvae feed on soybean leaves (Fig. 2).

Remarks: Mean values within a column followed by the different letters are significantly different at p < 0.05 and vice versa according to Duncan’s Multiple Range Test; US = Indonesian purple cultivar from middle land, TS = Thailand cultivar from middle land, UR = Indonesian purple cultivar from low land, TS = Thailand cultivar from low land, K = control, 20% = the concentration used in the study.

Fig. 2. Percentage of soybean leaves damaged by S. litura larvae attack.

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The larvae of S. litura Fabricius eat soybean leaves; as a result, the soybean leaves become hollow, even until the soybean leaves are reduced by about 50%. The highest percentage of soybean leaf damage occurred in the control group (77.2%), while in the soybean leaf sprayed with PLE, the rate of leaf damage was lower than in the control group (53.2-60.2%). Among all treatments, the spraying with PLE of the Indonesian purple cultivar from the medium plains resulted in the lowest percentage of soybean leaf damage.

S. litura Fabricius larvae consumed both the treated and untreated soybean leaves. In the control group, leaf damage did not exceed 100 percent. We assumed it was caused by the active chemicals present in soybean leaves. Soybean leaves contain flavonoids that serve as a defensive mechanism against herbivore attacks (da Silva et al., 2021). Soybean leaves contain rutin, a flavonoid that causes the larvae to avoid soybean leaves. The phytochemical analysis of Thailand and Indonesian purple papaya leaf extracts reveals the presence of rutin (Rahayu, Leksono, Gama, & Tarno, 2020).

In the treatment group, where soybean leaves were sprayed with papaya leaf extract, soybean leaf damage was significantly less than in the control group. It is possible that the rutin found in soybean leaves and papaya leaf extract possesses antinutritional effect on insects because it decreases the palatability of plant tissue to herbivores.

PLE of Thailand and Indonesian purple cultivars evaluation on soybean leaf damage by S. litura Fabricius larvae has been carried out.

The results showed that the attack of S. litura Fabricius larvae on soybean plants sprayed with PLE was quite high. TR-20% and TS-20% resulted in 59.80% and 57.4%, respectively. Meanwhile, UR- 20% and US-20% resulted in 60.2% and 53.20%, respectively. Even in the control group, soybean leaf damage reached 70%. Soybean plants with leaf damage, according to de Freitas Bueno, R. de Freitas Bueno, A., Moscardi, Parra, & Hoffmann- Campo (2011), plants can still survive until the flowering phase if soybean leaf damage reaches 35%. However, if soybean leaf damage occurs during seed development, it decreases soybean yield. In this research, leaf damage is observed only in the vegetative phase of soybean plants, so further studies are needed until the generative stage.

Soybean plants are susceptible to leaf loss.

The higher the level of defoliation, the lower the

yield. A leaf defoliation rate of 100% can result in a production loss of up to 80% (da S. Glier et al., 2015). In this study, the effect on soybean yield was unknown because the observation of damage to soybean leaves was only during the vegetative phase. Therefore, the response of soybean plants in the generative phase to S. litura Fabricius attack after being sprayed with PLE needs to be studied further in the laboratory and field tests.

Soybean leaves that were damaged by S. litura Fabricius larvae were then grouped according to the level of damage. The level of leaf damage was separated into six groups based on the percentage of damage. The distribution of the damaged soybean leaves for each criterion of the level of damage is presented in Fig. 3.

The data in Fig. 2 shows that in each treatment group, the single concentration of PLE used at 20% demonstrated was the most number in the 5% damage category and was less at the level of damage category 30-50%. The damage category, which PLE improved, suggested that PLE could protect soybean leaves from S. litura Fabricius attack. This condition indicates that the larvae searching for food will move from one leaf to another soybean leaf. If the larvae of S. litura Fabricius have found the desired soybean leaf, the leaf will continue to be eaten so that the area of the soybean leaf decreases.

Based on the level of damage to soybean leaves, it was found that the number of damaged soybean leaves with a damage level of 5% was the most in all treatments. These conditions indicated that the larvae of S. litura Fabricius, in search of food, were very active. The activity of the larvae was due to the presence of trichomes on the surface of the soybean leaves, which interfered with their movement. As a result, S. litura Fabricius larvae, in search of food, will move from one leaf to another.

The function of trichomes for plants is to protect themselves from pests.

The trichome density of each soybean cultivar is different. This study uses soybean varieties Grobogan. The study’s results (Fattah, Sjam, Daud,

& Dewi, 2018) showed that the thickness of soybean leaf trichomes of the Grobogan variety was 50.80 cm^2, and the density was the highest compared to the Anjasmoro and Argomulyo varieties. The presence of trichomes on soybean leaves will interfere with the movement, feeding mechanism, and oviposition of S. litura Fabricius larvae. Trichomes in leaves

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also help to boost plant resistance to diseases. Plant resistance increases as trichome density increases (Gupt et al., 2021). Another study that examined the relationship between the larval preference of S.

frugiperda and trichome density in some cultivated plants was carried out by Sotelo-Cardona, Chuang,

Lin, Chiang, & Ramasamy (2021). The results showed that the larvae did not like tomato leaves because there were many trichomes on the surface of the leaves. Tactile stimulation in the form of trichomes on the leaf surface also affects the selection of host plants.

Remarks: US = Indonesian purple cultivar from middle land, TS = Thailand cultivar from middle land, UR = Indonesian purple cultivar from low land, TS = Thailand cultivar from low land, K = control.

Fig. 3. Distribution of damaged soybean leaves based on the level of damage.

Fig. 4. The intensity of soybean leaf damage due to S. litura larvae attack.

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Soybean Leaf Damage Intensity

The leaves of soybean plants in the vegetative phase of the treatment and control groups in the study were damaged. The results of the analysis of the intensity of damage to soybean leaves are presented in Fig. 4. Statistical analysis showed that the intensity of damage to soybean leaves in the control group had the highest damage intensity (63.68%) and significantly differed from the treatment group (29- 41%). The lowest intensity of damage to soybean leaves was given to PLE of the Indonesian purple cultivar from the temperate plains. However, Duncan’s test results showed that the intensity of damage to soybean leaves sprayed with PLE from different cultivars from different altitudes was not significantly different. The results showed that soybean leaves damage intensity on TR-20%, TS-20%, UR-20% and US-20% resulting in 38.78%, 34.70%, 41.70%, and 29.6%, respectively. The soybean leaves damage intensity in the control groups reached 63.68%.

Our current result demonstrated that PLE from the middle land protects the soybean leaves better than other PLE. Another study reported that the intensity of the soybean leaves was different in four places, ranging from 8-61-10.18% (Fattah, Sjam, Daud,

& Dewi, 2018). The previous results demonstrated that botanical pesticides are still inferior to chemical insecticides (Indiati, 2014; Kushram, Yadu, Sahu, Kulmitra, & Kumar, 2017). The botanical pesticides consisted of garlic and green chili resulting in 30.51-

63.56% of S. litura Fabricius attack reduction, while the chemical insecticides Triazophos 40EC resulted in 68.64% attack reduction (Kushram, Yadu, Sahu, Kulmitra, & Kumar, 2017). The decline of S. litura Fabricius population is indicated by the reduction of S. litura Fabricius attack on the soybean plant.

However, the different results of our present study might be assumed from the different bioactive compounds contained in the plants.

The intensity of damage to soybean leaves in the control group reached 63%, and on soybean, leaves sprayed with PLE, 29-41%. In comparison, according to Fattah, Ilyas, & Rauf (2020), the intensity of damage to soybean leaves in the vegetative phase was highest (41.45%) and occurred in monoculture plantings. The difference in the results of the power of damage in the two studies was due to different environmental conditions. The level of damage caused by pest attacks varies greatly depending on the friction and climatic conditions of the area (Fattah, Ilyas,

& Rauf, 2020). The research conducted by the author was carried out in a greenhouse condition, while Fattah, Ilyas, & Rauf (2020) carried it out on soybean farms. The environmental conditions of soybean plants on agricultural land are volatile, in contrast to greenhouse conditions. In addition, in the greenhouse conditions, the area for S. litura Fabricius larvae to find food is limited, resulting in a higher intensity of damage to soybean leaves.

Fig. 5. S. litura larva mortality after treatment.

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S. litura Fabricius larvae introduced into soybean plants died. The mortality of S. litura Fabricius larvae was observed 72 hours after applying PLE on soybean plants. The mean mortality of S. litura larvae is presented in Fig. 5. The highest mortality percentage of S. litura larvae Fabricius sequentially from the treatment was US-20% (43%), TS-20% (40%), US-20% (40%), and TR-20%

(37%). The results of Duncan’s test showed that on soybean leaves sprayed with PLE of Thailand and Indonesian purple cultivars from different heights, the mortality of S. litura Fabricius larvae was not significantly different (37-43%). On the other hand, the mortality of S. litura Fabricius larvae was lowest (11%) in the control group and significantly different from the treatment group sprayed with PLE.

Insects use their sensory organs to determine the host plant. The eyes and sense of smell are sensory organs that play a function. Adult insects utilize their eyes to identify colors. The sense of smell detects volatile substances and is important in both the adult and larval stages (Cao et al., 2022).

Another factor that influenced the behavior of S.

litura Fabricius larvae in eating soybean leaves in this study was the presence of PLE of Thailand and Indonesian purple cultivars, which were sprayed on the surface of soybean leaves. Even the larvae of S.

litura Fabricius that were infested on soybean plants died. The mortality percentage of S. litura Fabricius larvae in the treatment group was much higher than in the control group. PLE contains various active compounds. The results of phytochemical screening using LCMS PLE contain alkaloids, flavonoids, phenols, and compounds. The active compounds in PLE will enter the larvae body and soybean leaves.

The active compounds such as alkaloids, phenols, flavonoids, and benzyl glucosinolates will affect the metabolism of S. litura Fabricius larvae. The alkaloid compounds in PLE include dehydrocarpaine, carpaine, and pseudocarpaine compounds. These compounds have toxic properties and cause a bitter taste (Matsuura

& Fett-Neto, 2015). The activity of alkaloid compounds in the body of S. litura Fabricius larvae also causes malnutrition and interferes with their growth (Maulina, Sumitro, Amin, & Lestari, 2018). In addition, alkaloids also inhibit -amylase and protease enzymes (Zou et al., 2018). As a result, the need for protein and carbohydrate compounds during larval growth is disrupted. Flavonoid compounds

function as insecticides, have antifeedant properties (Lingaturai, Vendan, Paulraj, & Ignacimuthu, 2011), and affect pest behavior (Mierziak, Kostyn,

& Kulma, 2014). Flavonoid compounds interfere with the enzyme reaction process in the larval body and reduce the palatability of plant tissues (Marques et al., 2015). Phenol compounds will be oxidized to hydrogen peroxide so that the larvae will have problems digesting food, and the cells in the midgut of the larvae will be damaged (Rashwan &

Hammad, 2020). Benzyl glucosinolate compounds are effective insecticides (Saran & Choudhary, 2013). The research results by Rahayu, Leksono, Gama, & Tarno (2020) also showed that the purple PLE of Thailand and Indonesian cultivars had antifeedant properties. As a result, the larvae would avoid soybean leaves sprayed with PLE.

The presence of active compounds in PLE provides a synergistic effect on S. litura Fabricius larvae. In addition, the presence of trichomes on the surface of soybean leaves disturbed the feeding activity of S. litura Fabricius larvae. Consequently, the larvae were malnourished and eventually died.

Based on the study’s results, it was shown that the PLE of Thailand and Indonesian purple cultivars with a concentration of 20% could be developed as a natural pesticide to prevent S. litura Fabricius larvae attack on soybean plants.

CONCLUSION

In conclusion, the PLE of the Indonesian purple cultivar from the middle land with 20%

concentration resulted in the lowest of the leave damage intensity caused by S. litura Fabricius attack (29.56%). At the same time, the highest intensity of leave damage, 63.68%. Additionally, the mortality data demonstrated that the PLE of the Indonesian purple cultivar from the middle land with 20% concentration resulted in 43% S. litura Fabricius larva death. In comparison, the control group only showed 11% S. litura Fabricius larva death. PLE from Thailand and Indonesian purple cultivars might be a promising natural agent to combat S. litura Fabricius attack by protecting the leaves from damage and increasing the mortality rate of S. litura Fabricius larvae.

ACKNOWLEDGEMENT

The authors would like to thank Universitas Negeri Malang for facilitating this research.

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