Indonesian Medicinal Plants Research Articles

Thermostability and free radical scavenging activity of Annona muricata(L.) leaf extract in antiaging cream

Annona muricata is an Indonesian medicinal plant commonly used as an anticancer agent. The finding of its strong antioxidant activity leads to the development of cosmetic products, particularly as antiaging cream. Formulation may influence thermostability and free radical scavenging activity of A.muricata leaf extract. The purpose of this study was to investigate the thermostability and free radical scavenging activity of A.muricata leaf extract incorporated in Gattefosse Skin Cashmere cream. Accelerated stability study utilizing climatic chamber at a temperature of 40oC ± 2oC and relative humidity of 75% ± 5% for 20 days of storage was used to evaluate thermostability. 1,1-diphenyl-2-picrylhydrazil (DPPH) using spectrophotometer was used for free radical scavenging activity test. The result from physical properties including appearance of antiaging cream, pH, viscosity and rheological properties of each formula showed that all of the formulas were stable at a temperature of 40oC ± 2oC and relative humidity of 75% ± 5% for 20 days. Free radical scavenging activity using DPPH showed high antioxidant activities from the A.muricata leaf extract in the Gattefosse Skin Cashmere cream. We concluded that A. muricata leaf extract could be developed into natural antiaging cosmetic product.

Allelopathic Potency and an Active Substance from Anredera cordifolia (Tenore) Steenis.

Anredera cordifolia (Tenore) Steenis is widely planted as an ornamental and medicinal plant in Indonesia. On the other hand, in some other countries this plant is classified as a noxious weed. As a harmful weed, A. cordifolia is reported to have the ability to smother all native vegetation, collapse canopies of tall trees, cultivate as a ground cover and disrupt native seedling development. There is no available information about the involvement of any allelochemicals from A. cordifolia related to these issues. The present study evaluated the allelopathic effect by isolating and identifying the allelopathic substance from A. cordifolia leaf extract. The allelopathic potency of A. cordifolia was determined by a series of bioassays of shoot and root growth on some selected test plants. Separation and purification of the active substances was achieved through several chromatography processes. Finally, the substances with allelopathic activity were identified through high-resolution electrospray ionization mass spectrometry (HRESIMS) analysis and determined by the specific rotation of compound, proton and carbon NMR spectroscopies. The results show that A. cordifolia possesses allelopathic properties which affect other plant species. The isolated compound from the plant material, 3-hydroxy-alpha-ionone, may contribute to the allelopathic effects of A. cordifolia.

Effect of Sodium Carboxymethylcellulose and Sorbitol on Anti-Peptic Ulcer Activity of Anredera cordifolia Leaves Extract

Anredera cordifolia or binahong is one of the Indonesian medicinal plants that is used to treat peptic ulcer. The purpose of this study was to evaluate the effect of the addition of sodium carboxymethylcellulose (CMC) and sorbitol on anti-peptic ulcer activity of A. cordifolia leaves extracts in male Wistar rats. The plants were extracted using decoction method and freeze dried. Three liquid formulas were used i.e., i) a combination of sodium CMC and sorbitol; ii) only sorbitol; iii) extract only. The rats were divided into 6 groups, i.e., positive control (sucralfate 35 mg/kg body weight); negative control (80% ethanol); normal control; and 3 formulas. After the administration of the liquid formula, all groups, except normal control, were given 80% ethanol (l5 ml/kg body weight) to induce peptic ulcer. Antipeptic ulcer activity was evaluated using direct observation on rats gastric mucosa, and histopathology assessment. The result showed that the strongest anti-peptic ulcer was shown by sorbitol only (96.95% inhibition), followed by the combination of sodium CMC and sorbitol (92.68% inhibition). The formula which only contained extract showed only 31.70% inhibition. Statistical analysis showed significant differences between formula 1 and 2 with negative controls. In conclusion, A. cordifolia leaves extract with the addition of sorbitol showed the strongest anti-peptic ulcer activity. Keyword: Anredera cordifolia, peptic ulcer, suspense, Wistar rat.

Anti-Bacterial and Anti-Fungal Activities from Macaranga bancana Leaves Extract

Macaranga bancana is one of the Indonesian medicinal plants that is empirically used to treat infectious disease. The aim of this study was to evaluate anti-bacterial and anti-fungal activities of M. bancana against human pathogenic microbes, i.e., Escherichia coli, Staphylacoccus aureus, and Candida albicans. The dried leaves of M. bancana were extracted using cold extraction method with n-hexane, dichloromethane, ethylacetate, methanol, and ethanol solvents. The evaluation of anti-microbial activity from these extracts at the concentration of 1000 µg/ml was conducted using microdilution method. Amoxicillin, cephadroxil, and ketoconazole were used as positive controls. All extracts showed 100% inhibitory activity against S. aureus, with the exception of ethanol extract which was 72.8%. Nevertheless, methanol and ethanol extracts showed 100% inhibitory activity against E. coli, while the lowest activity was shown by n-hexane extract ( 45.2%). All extracts exhibited 100% inhibitory activity against C. albicans. In conclusion, leaves extract of M. bancana is a potential source of anti-bacterial and anti-fungal agents. Keywords: anti-bacterial, anti-fungal, M. bancana, microdilution

EFFICACY OF JAVANESE TURMERIC ETHANOL EXTRACT IN ERADICATING STREPTOCOCCUS SANGUINIS AND PORPHYROMONAS GINGIVALIS BIOFILM

Objective: Javanese turmeric is an Indonesian native medicinal plant with antibacterial activities. This study aimed to analyze the efficacy of identifiedJavanese turmeric ethanol extract (IJTEE) in eradicating Streptococcus sanguinis and Porphyromonas gingivalis biofilms.Methods: Biofilm assay: Single and combination biofilms formed at different phases were exposed to IJTEE in 0.5–25% concentrations for 1 h. Thepercentage of eradication was tested using the microtetrazolium assay.Results: The efficacy of IJTEE in eradicating the biofilm was equal to that of chlorhexidine against the early phase of biofilm formation. IJTEE is moreeffective against S. sanguinis biofilm formation than against P. gingivalis biofilm formation.Conclusion: IJTEE can eradicate S. sanguinis and P. gingivalis biofilms.

THE POTENCY OF JAVANESE TURMERIC (CURCUMA XANTHORRHIZA ROXB.) ETHANOL EXTRACT TO ERADICATE WILD STRAIN CANDIDA ALBICANS BIOFILM

Objective: The pathogenic yeast Candida albicans forms biofilm to increase its resistance toward antifungal agents. Javanese turmeric is an Indonesianmedicinal plant reported to have antifungal effects due to the active component, xanthorrhizol. The objective of this study was to measure the in vitropotential of Javanese turmeric ethanol extract to eradicate C. albicans biofilm.Methods: C. albicans was exposed to Javanese turmeric ethanol extract for 1 h during biofilm formation phases. MTT assay was used to test thepercentage of biofilm eradication.Results: The minimum inhibitory concentration and minimum fungicidal concentration of Javanese turmeric ethanol extract against planktonicC. albicans were 15%. The minimum biofilm eradication concentration (MBEC50) was 25% in the early phase and 15% in the intermediate andmaturation phases.Conclusions: Javanese turmeric ethanol extract is effective at eradicating clinical isolate of C. albicans biofilm.

Ion wind from yield corona discharge and its application to drying of Turmeric slices (Curcuma domestica Val)

Turmeric (Curcuma domestica Val) is a native Indonesian medicinal plant. Drying sliced turmeric is an important process to get the basic herbal ingredient of a durable herb. The purpose of this study was to apply the ion wind of corona discharge to dry the turmeric slices and its characterization.The corona discharge is generated by using a pin-three of ring electrode which is subjected to a high voltage DC with positive pin electrode polarity and a negative concentric three-ring electrode. The ion will flow from the pin electrode to the three-ring electrode and a portion will be forwarded through the gap between three concentric rings. The drying of the turmeric slices is carried out at distance between electrodes which is constant. It is placed just below the concentric three-ring electrode and the distance between electrode is 2 mm. The drying of turmeric slices is time-varying 5 to 65 minutes with a time interval of 5 minutes. The study result of drying of turmeric slices is obtained drying rate and mass shrinkage is inversely proportional to a drying time. While the efficiency of drying is inversely proportional to the drying rate.

English

Javanese ginseng (Talinum paniculatum) is an herb and one of the traditional medicinal plants in Indonesia which accumulate saponin in the root organ. However, slow root growth hampers the accumulation of the compound in this plant. Therefore, in vitro culture of adventitious roots offers an alternative way to increase the production of saponin. Furthermore, this study was performed to investigate the effect of methyl jasmonate (MeJA) and salicylic acid (SA) on the growth and saponin content in adventitious root of Javanese ginseng. Adventitious root culture was induced on solid Murashige and Skoog (MS) medium supplemented by 10 µM IBA (indole-3-butyric acid). Subsequently, adventitious roots were subcultured into a half strength MS liquid medium with the addition of different concentrations of MeJA or SA and incubated for 5, 10 and 15 days. Despite inhibition on the root growth, saponin production increased by 1.5 and 1.3 fold upon elicitation with 0.2 mM MeJA and SA for 15 days, respectively. Thus, MeJA and SA elicitations regulate saponin biosynthesis in adventitious root culture of Javanese ginseng as a time and dose-dependent manner. Key words: Adventitious root, elicitation, Javanese ginseng, saponin, Talinum paniculatum.

Pengaruh Ekstrak Bunga Kembang Sepatu (Hibiscus Rosa-Sinensis Linn.) terhadap Epididimis, Prostat dan Vesikula Seminalis

Hibiscus rosa-sinensis Linn. merupakan salah satu tanaman obat di Indonesia yang digunakan sebagai kontrasepsi pria. Bunga kembang sepatu mengandung flavonoid yang dapat menurunkan testosteron. Tujuan penelitian ini untuk mengetahui pengaruh ekstrak bunga kembang sepatu terhadap berat epididimis, tebal epitel epididimis cauda, berat prostat dan vesikula seminalis tikus jantan. Penelitian ini merupakan penelitian eksperimental dengan menggunakan Rancangan Acak Lengkap (RAL). Sampel dalam penelitian ini menggunakan 25 tikus jantan strain Sprague Dawley® usia 60-70 hari dengan berat badan 200-210 gram. Dosis perlakuan 200 mg/kgBB, 300 mg/kgBB, 400 mg/kgBB, 500 mg/kgBB selama 30 hari. Data disajikan dalam bentuk tabel serta narasi untuk menginterpretasikan data tersebut.Hasil penelitian menunjukan adanya penurunan berat epididimis pada kelompok perlakuan dibanding kelompok kontrol (p=0,000), terjadi juga penyusutan tebal epitel epididimis cauda (p=0,021), penurunan berat prostat dan vesikula seminalis antara kelompok perlakuan dibanding kelompok kontrol (p=0,001). Dapat disimpulkan bahwa ada pengaruh ekstrak bunga kembang sepatu tehadap epididimis, prostat dan vesikula seminalis. Pengaruh tersebut kemungkinan disebabkan kandungan bahan kimia yang ada di dalam bunga kembang sepatu yang bersifat antifertilitas.

Antihyperlipidemic Activity of Ethanol Extract Mindi’s Leaves (Melia azedarach Linn.) in Male Wistar Rats Induced Propiltiouracil

Mindi’s leaves (Melia azedarach Linn.) is an Indonesian medicinal plant that used as traditional medicine. Mindi leaves contains some secondary metabolites which have potency to decreased total cholesterol and LDL level. The study purposed to know antihyperlipidemic effect of ethanol extract of mindi’s leaves (Melia azedarach Linn.) seen from total cholesterol, and LDL in male albino rats. Male wistar albino rats were divided into 5 groups, negative control group (sodium CMC 0.5%), positive control group (simvastatin 0.193 mg/200 gBW), group I (ethanol extract mindi’s leaves with dosage 300 mg/200 gBW), group II (ethanol extract mindi’s leaves with dosage 600 mg/200 gBW), and group III (ethanol extract mindi’s leaves with dosage 1200 mg/200 gBW). The rats were given high-fat supplement and propylthiouracil for 15 days to increase cholesterol, and the extract was given for the next 15 days. Average cholesterol level, LDL, and body weight after induction was 90.28 mg/dL, 31.09 mg/dL, and 222.32 g. The result showed ethanol extract mindi’s leaves could decreased total cholesterol level and LDL level with % decreased in total cholesterol (%PDTC) and % decreased in LDL (%PDLDL) of group I is 37.78% and 35.57%, group II is 45.99% and 40.39%, and for group III is 56.29% and 52.42%. The result showed that ethanol extract of mindi’s leaves has antihyperlipidemic activity and significantly different from negative control (p>0.05). Based on the percentage relation of decreased total cholesterol and LDL levels to dose, then the effective dose 50 (ED50) value of ethanol extract mindi’s leave is 869 mg/200 gBW for total cholesterol and for LDL reduction level is 1086.84 mg/200 gBW

The composition and diversity of plant species in pasak bumi’s (Eurycoma longifolia) habitat in Batang Lubu Sutam forest, North Sumatra, Indonesia

Susilowati A, Rachmat HH, Elfiati D, Hasibuan MH. 2019. The composition and diversity of plant species in pasak bumi’s (Eurycoma longifolia) habitat in Batang Lubu Sutam forest, North Sumatra, Indonesia. Biodiversitas 20: 413-418. Pasak bumi (Eurycoma longifolia Jack) is one of the most popular medicinal plants in Indonesia. Currently, E. longifolia is being over-exploited due to its potential and popularity as herbal medicine and its high value in the market. Therefore, the study on the population structure of the species and habitat characterization is required to ensure successfulness of conservation of this species. The study was carried out in lowland forest, located in Limited Production Forest within the Register Number 40, situated administratively in Papaso Village, Sub-District of Batang Lubu Sutam-Padang Lawas, North Sumatra, Indonesia. Batang Lubu Sutam forest is known as a source of pasak bumi material in North Sumatra. Every year tons of pasak bumi are collected from this forest and exported to Malaysia and surrounding countries. Due to illegal harvesting for its roots, pasak bumi has been seriously depleted in some areas in Batang Lubu Sutam forest. Therefore, current data on its population are needed for conservation effort of Batang Lubu Sutam's pasak bumi. The objectives of this study were to assess the structure of forest where pasak bumi grows and to determine the correct management for sustainability the future stocks. Vegetation analysis was carried out purposively in four transect belts, each consisting of five quadrats or plots. The results showed that the Importance Value Index of pasak bumi at seedling, sapling, pole and tree stages was 21.26%, 9.147%, 0, and 0 respectively. The Shannon Diversity Index (H) ranged between 3.050 and 3.220 and was inside its common value distribution. The evenness index was high, more than 0.85, at all stages of plant growth. The disappearance of pasak bumi at pole and tree stages indicates threatening condition for the sustainability of pasak bumi in the study sites because of excessive harvesting.

Morphological characters, flowering and seed germination of the Indonesian medicinal plant Orthosiphon aristatus

Febjislami S, Kurniawati A, Melati M, Wahyu Y. 2019. Morphological characters, flowering and seed germination of the Indonesian medicinal plant Orthosiphon aristatus. Biodiversitas 20: 328-337. Orthosiphon aristatus (Blume) Miq is a popular medicinal plant in Southeast Asia. The morphological variation of O. aristatus is narrow and information on flowering and seed germination is limited. This study aimed to determine the morphological characters, flowering and seed germination of O. aristatus. The study was conducted on 19 accessions (ex situ collections) of O. aristatus from West, Central and East Java. It was found that the differences in morphological and flowering characters were mainly based on shape and color. The dominant stem color is strong yellowish green mixed with deep purplish pink in different proportions. The dominant leaf shape was medium elliptic. O. aristatus flower has three kinds of colors: purple, intermediate and white (the most common color). O. aristatus has heterostyled flower with a long-styled morph. The stigma has two shapes: open and close (the dominant shape). The open and close stigma was found in Tuban accession, the open stigma was found in Pamekasan accession. O. aristatus accessions have a narrow diversity of 84% similarity rate. O. aristatus started flowering at 4-5 weeks after planting and blooming 2-3 weeks later. O. aristatus seeds begin to germinate on the fourth day and take about eight days to grow from seedling to become complete sprouts. Count I ranged from day 5-6 and count II on days 10-11. Tuban accession has the highest germination rate of 72.97%. The results of this research can be considered as new information, regarding information about the diversity of morphological, flowering characters and the ability of O. aristatus seed to germinate.

Antimicrobial activity of essential oil from Indonesian medicinal plants against food-borne pathogens

The aim of the study was to evaluate antimicrobial activity of essential oil from local plants or plants cultivated in Indonesia against three food-borne pathogens, Salmonella Thypimurium, Bacillus cereus and Staphylococcus aureus. A total 6 essential oil extracted from ginger (Zingiber officinale), turmeric (Curcuma longa), candlenut (Aleurites moluccanus), lemongrass (Cymbopogon citratus), clove bud (Syzygium aromaticum), and galangal (Alpinia galanga). By using disc diffusion assay, the highest antimicrobial activity against S. Tyhimurium, B. cereus and S. aureus was shown by clove bud oil (3.67 ± 0.58 mm), lemongrass oil (3.58 ± 0.14 mm) and clove bud oil (5.25 ± 0.75 mm), respectively. By using Minimum Inhibitory Concentration (MIC), the concentration of essential oil to inhibit the growth of S. Tyhimurium, B. cereus and S. aureus was found on clove bud oil for 0.39 %, lemongrass oil for 1.56 % and clove bud oil for 0.78 %, respectively. Out of the essential oil tested, clove bud oil and lemongrass oil showed promising antibacterial activity against S. Thypimurium, B. cereus and S. aureus.

Identification of 4-4’-(1-methylethylidene)-bisphenol from an Endophytic Fungus Cladosporium oxysporum derived from Aglaia odorata

<p>Endophytic fungi has an economic potential as raw material for biologically active compounds<em>. Cladosp</em><em>o</em><em>rium oxysporum</em><em> </em>is one of the endophytic fungi isolated from Indonesian medicinal plant <em>Aglaia odorata </em>Lour (Local name: Pacarcina). This planth has been used for fever, cough, diarrhea, inflammation and injury. In our previous study, the ethyl acetate extract and several fractions of the extract of <em>C. oxysporum </em>showed antimicrobial activity against <em>Candida albicans, Escherichia coli</em>, and <em>Staphylococcus aureus</em>. The objective of the currents study is to investigate the chemical constituent of the active fraction.<strong> </strong>Purification of the metabolite was achieved by using column chromatography followed by preparative thin layer chromatography. Identification of the metabolite was conducted by using TLC densitometry, GC-FID and GC-MS. Compound 1 was isolated from fraction 12. The purity of this compound was determined by 2D-TLC and GC-FID. The UV-Vis profile of compound 1 indicates a phenolic compound. Further analysis by using GC-MS shows one peak at a retention time (Rt) of 23.80 minutes, predicted as 4-4’-(1-methylethylidene)-bisphenol. The chemical constituent of the sub fraction 12.2.7 (fraction <strong>a</strong>) is identified as 4-4’-(1-methylethylidene)-bisphenol.</p>

Development of 10 single-copy nuclear DNA markers for Euchresta horsfieldii (Fabaceae), a rare medicinal plant.

Premise of the Study Euchresta horsfieldii (Fabaceae) is a rare and endangered medicinal plant in Indonesia with restricted distribution. Single‐copy nuclear DNA (scnDNA) markers were developed for this species to facilitate further investigation of genetic diversity and population structure.Methods and ResultsWe performed RNA‐Seq and de novo assembly of the transcriptome. Ten primer sets were developed for E. horsfieldii, all of which also amplified in E. japonica and E. tubulosa.ConclusionsThese scnDNA markers will be an important resource for the study of genetic diversity and population structure of E. horsfieldii and other species in the genus Euchresta.

Active Compounds and Antimalaria Properties of some Medicinal Plants in Indonesia – A Review

Active Compounds and Antimalaria Properties of some Medicinal Plants in Indonesia – A Review

Diversity of Indonesian Medicinal Plant in The lowland Forest, Bodogol and Its Surrounding of Mount Gede-Pangrango National Park, West Java

Ethnobotanical research of medicinal plant was carried out in the lowland forest at Bodogol, Mount Gede Pangrango National Park, West Java. The study site is inhabited by the Sundanese community in several villages of surrounding National Park. The used methods of research were ethno–directed sampling, open–ended interviews with the local community, and a ‘walk in the woods’ in four permanent forest plots. A total of 93 medicinal plants were recorded in the Bodogol lowland forest and its surroundings, belonging to 78 genera and 50 families. Those species are used for treatment of 32 different diseases. The species is most frequently used for post-pregnancy. Three of the recorded species namely Alstonia scholaris, Cinnamomum sintoc, and Fibraurea tinctoria are endangered plants. The community rarely collects medicinal plants from the forest; they prefer to collect from around the village or from cultivated plants.

Forhidrol, a bioactive fraction of Phaleria macrocarpa (Scheff.) Boerl., increases reverse cholesterol transport pathway by down-regulation of cholesteryl ester transfer protein activity

Phaleria macrocarpa (Scheff.) Boerl. is one of the most popular medicinal plants in Indonesia. This plant has traditionally been used to lower cholesterol and control hypertension. This study was performed to examine the effect of Forhidrol, a bioactive fraction of P. macrocarpa (Scheff.) Boerl., to increase reverse cholesterol transport through the down-regulation of cholesteryl ester transfer protein (CETP) activity in hepatocellular carcinoma cell lines. Measurement of target genes including liver X receptor (LXR), sterol regulatory element-binding protein 1 (SREBP1), scavenger receptor class B type 1, low-density lipoprotein (LDL) receptor, apolipoprotein B, CYP11B1, CYP11B2, peroxisome proliferatoractivated receptor α (PPARα), peroxisome proliferator-activated receptor δ (PPARδ) and peroxisome proliferator-activated receptor γ (PPARγ) were done using realtime polymerase chain reaction assay. Secreted CETP and apolipoprotein A-1 were analyzed using western blot. CETP activity in vitro was measured using CETP inhibitor drug screening kit and triglyceride synthesis was measured using enzyme-linked immunosorbent assay. Forhidrol was found to significantly down-regulate CETP mRNA expression, as well as total CETP and cholesteryl esters transfer activity (P<0.05). It specifically reduced transcriptional level of regulatory genes of CETP promoter including SREBP-1 and LXR. Forhidrol also significantly increased PPARδ and PPARα expression (P<0.05) and slightly repressed triglycerides synthesis. In vivo study showed elevated high-density lipoprotein (HDL) levels in rabbits after 4-week treatment of Forhidrol at a dose of 37.5 mg/1.5 kg body weight compared to placebo. Conversely, LDL, triglyceride and CETP activity were decreased. Forhidrol increased HDL levels by reducing CETP-dependent transfer of cholesterol from HDL to LDL particles. Looking at possible side effects, Forhidrol apparently acted as a safe agent without negative effect towards blood pressure. These findings suggested that Forhidrol may be further developed as a potential anti-atherogenic drug.

De Novo Transcriptome Profiling of Buasbuas (Premna pubescens. Blume)

Buasbuas is the one of the medicinal plants in Indonesia that contains bioactive compounds potential as antimicrobial, antioxidant, antidiabetes, antiinflammation and anticancer. Exploring the pathway and gene related of buasbuas bioactive compounds production has led to the renaissance of understanding buasbuas molecular mechanism database. The aim of this study was to developed data-mining framework of buasbuas to study plant specialized metabolism for phytochemical biosynthesis. Material and Methods: This project was started by collecting shoots and leaves of Buasbuas. Focus of the project was exploring the molecular mechanisms on biosynthesis phytochemical of Buasbuas. Illumina Mi-Seq Next Generation Sequencing was utilized to understand the molecular mechanisms of biosynthesis. Transcriptomes then trimmed and assembled with CLCBio genomic software. Assembled contigs then annotated towards Arabidopsis thaliana using CLC Bio genomic software. Digital Gene Expression was performed to analyze the transcriptional changes in control culture and treatment. Results: There were 5.342 unigenes that expressed only in treatment shoot cultures. Annotation with Gene Ontology showed that 57.9% (3.446) unigenes play role in Biological Process, 56.7% (3.375) unigenes play role in Cellular Components and 63.4% (3.772) unigenes play role in Molecular Functions. Annotation with Kyoto Encyclopedia of Genes and Genomes shows 853 unigenes essentially have role in 24 biological pathways. Highest process with highest unigenes involvement is biosynthesis of plant hormones and biosynthesis of alkaloids. Conclusion: This study showed that phytochemical biosynthesis in buasbuas induces level expression of several genes involved in the jasmonic acid, cytokinin, gibberellin, salicylic acid and ethylene biosynthesis pathway.

First record of entomopathogenic fungi on autumn leaf Caterpillar (Doleschallia bisaltide)

Caricature plant is one of the medicinal plants in Indonesia to cure hemorrhoids, menstruation, and others. The cultivation constraints of caricature plant is autumn leaf caterpillars (Doleschallia bisaltide). Utilization of synthetic insecticides is not allowed to avoid bioaccumulation of chemical residues. Entomopathogenic fungi is an alternative way to control D. bisaltide. The objective of the research was to obtain isolates of entomopathogenic fungi of D. bisaltide. The research conducted by two steps, which were exsploration of infecfted D. bisaltide. The second step was identification of the fungi. Exploration results of 16 pupae of D. Bisaltide were infected by fungi. Identification done by classify the mcroscopic and microscopic fungi isolate characteristic. One from five fungal isolates were entomopathogenic fungi from Verticillium genera.