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Biodiversitas Journal of Biological Diversity
Published by Universitas Sebelas Maret
ISSN : 1412033X     EISSN : 20854722     DOI : https://doi.org/10.13057/biodiv/d230231
Core Subject : Science, Agriculture,
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology
The Biodiversitas Journal was first published in 2000 by the Department of Biology, FMNS, Universitas Sebelas Maret, Surakarta, Indonesia, then in 2006 it was co-published by the Society for Indonesian Biodiversity and that department; since 2017 it was also hosted by Smujo. From 2003-2012 it was accredited by DGHE, Ministry of Education, R.I., since 2014 was indexed by Scopus, where DOAJ and Google Scholar was indexed first.
Arjuna Subject : Biokimia, Genetika dan Biologi Molekuler - Biokimia, Genetika dan Biologi Molekuler (Lain-Lain)
Articles 19 Documents
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Search results for , issue "vol. 27 no. 9 (2026)" : 19 Documents clear
Screening of endophytic and rhizobacterial strains based on phosphate-solubilizing and chlorpyrifos dissipation-enhancing activities for rice-peanut growth VITA RATRI CAHYANI; ANIDA HASNA NOOR FADHILAH; HADIWIYONO HADIWIYONO; SUDADI SUDADI
Biodiversitas Journal of Biological Diversity Vol. 27 No. 9 (2026)
Publisher : Society for Indonesian Biodiversity

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13057/biodiv/d270903

Abstract

Abstract. Cahyani VR, Fadhilah AHN, Hadiwiyono H, Sudadi S. 2026. Screening of endophytic and rhizobacterial strains based on phosphate-solubilizing and chlorpyrifos dissipation-enhancing activities for rice-peanut growth. Biodiversitas 27 (9): d270903. https://doi.org/10.13057/biodiv/d270903. Nutrient deficiency and insecticide toxicity remain major constraints in crop production. This study aimed to screen endophytic and rhizospheric bacteria from organic and conventional rice fields for phosphate-solubilizing activity and insecticide tolerance, and to evaluate selected multifunctional isolates for their potential to enhance chlorpyrifos dissipation and promote the growth of rice and peanut in insecticide-contaminated soil. Thirty bacterial isolates representing distinct colony morphotypes were screened in vitro; 16 solubilized inorganic phosphate, while 24 and 21 tolerated monovariant and multivariant insecticides, respectively. Five high-performing isolates were further characterized for IAA production, potential nitrogen fixation, and non-pathogenicity. One additional isolate, Bacillus paramycoides NEROI-2, which showed superior capabilities in previous studies, was included as a comparison isolate, resulting in six isolates evaluated in two independent greenhouse pot experiments with rice and peanut. INErO5 and INRsC5 showed strong effects in rice by increasing soil-available P, reducing soil chlorpyrifos residues, and promoting plant growth. INElC4 performed best in peanut, while INRsC3 was effective in both crops. With basal fertilizers (33.3% NK dose and rock phosphate), these four isolates produced vegetative dry weights of 64.95-92.11% and 56.77-107.48% of the 100% NPK treatment in rice and peanut, respectively, while reducing soil chlorpyrifos residues by 52.90-61.59% and 61.74-72.83%, respectively. INRsC3, INErO5, INRsC5, and INElC4 were closely related to Bacillus sp., Stenotrophomonas sp., Lysinibacillus sp., and Bacillus cereus, respectively, whereas INElO7 (Proteus mirabilis) showed the least effect on both crops. These isolates are promising candidates for biofertilizer development and bioremediation in rice-peanut systems cultivated on Alfisol; however, field experiments are required to validate these findings.
Integrative morphometric and molecular assessment of Tetragonula stingless bees (Apidae: Meliponini) at Edufarm Universitas Andalas, West Sumatra, Indonesia ELNI FATIMAH; HENNY HERWINA; DAHELMI DAHELMI; PUTRA SANTOSO; SITI SALMAH; JASMI JASMI; MUHAMMAD NAZRI JANRA; RUSDIMANSYAH RUSDIMANSYAH; ALPONSIN ALPONSIN; YANDRIZAL YANDRIZAL
Biodiversitas Journal of Biological Diversity Vol. 27 No. 9 (2026)
Publisher : Society for Indonesian Biodiversity

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13057/biodiv/d270904

Abstract

Abstract. Fatimah E, Herwina H, Dahelmi, Santoso P, Salmah S, Jasmi, Janra MN, Rusdimansyah, Alponsin, Yandrizal. 2026. Integrative morphometric and molecular assessment of Tetragonula stingless bees (Apidae: Meliponini) at Edufarm Universitas Andalas, West Sumatra, Indonesia. Biodiversitas 27 (9): d270904. https://doi.org/10.13057/biodiv/d270904. Several stingless bee species of the genus Tetragonula are morphologically similar, making accurate identification difficult using morphology alone. This study characterized morphometric variation and provided preliminary molecular support for the identification of five Tetragonula taxa from Edufarm, Universitas Andalas, West Sumatra, Indonesia, using morphometric and mitochondrial DNA markers. Fifty worker bees were measured for 34 morphometric characters, while one representative per taxon was analyzed using cytochrome C Oxidase subunit I (COI) and 16S rRNA genes. Principal Component Analysis (PCA) showed that PC1 explained 93.10% of the total variance (eigenvalue = 31.654), indicating overall body size as the main source of morphometric variation. Hierarchical clustering showed consistent phenetic differentiation among taxa, and PERMANOVA confirmed significant multivariate differences (F₄,₄₅ = 253.37, R² = 0.9575, p = 0.001). PERMDISP showed no significant differences in within-group dispersion (F₄,₄₅ = 2.019, p = 0.116), indicating that PERMANOVA mainly reflected differences among group centroids. COI analysis provided preliminary molecular support for several taxa. Tetragonula collina showed low pairwise distances to conspecific references (0.020-0.028), whereas T. fuscobalteata showed marked divergence (approximately 0.139-0.299). Tetragonula minangkabau, T. minangkabau forma darek, and T. testaceitarsis had relatively low COI distances and formed a closely related group, although their relationships could not be resolved from single-specimen sampling. The 16S rRNA marker showed more conservative divergence and supported the identification of several taxa, while the placement of T. collina remained unresolved. This study also provides new COI sequences for T. minangkabau, T. minangkabau forma darek, and T. testaceitarsis, and the first reported 16S rRNA sequence for T. testaceitarsis, providing a useful molecular reference baseline for future taxonomic and biodiversity studies in West Sumatra, Indonesia.
Intertidal gastropod biodiversity associated with shrimp aquaculture effluents exposure in Lombok and Sumbawa Islands, Indonesia EDWIN JEFRI; JAMALUDDIN JOMPA; NADIARTI NURDIN KADIR; NURLIAH BUHARI
Biodiversitas Journal of Biological Diversity Vol. 27 No. 9 (2026)
Publisher : Society for Indonesian Biodiversity

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13057/biodiv/d270912

Abstract

Abstract. Jefri E, Jompa J, Kadir NN, Buhari N. 2026. Intertidal gastropod biodiversity associated with shrimp aquaculture effluents exposure in Lombok and Sumbawa Islands, Indonesia. Biodiversitas 27 (9): d270912. https://doi.org/10.13057/biodiv/d270912. Shrimp aquaculture has expanded rapidly across the Indonesian archipelago, raising concerns regarding its potential influence on adjacent intertidal ecosystems. This study assessed the biodiversity and community structure of intertidal gastropods at coastal sites associated with shrimp aquaculture effluent exposure on Lombok and Sumbawa Islands, West Nusa Tenggara, Indonesia. Gastropods were sampled using 1 × 1 m² quadrats along 100 m transects; abundance, diversity, dominance, and evenness indices were calculated, and generalized linear models (GLMs) and Correspondence Analysis (CA) evaluated spatial patterns in community metrics and family composition. A total of 124 species belonging to 63 genera and 31 families were recorded. Cerithiidae and Nassariidae were the most species-rich families (14 species each), followed by Cypraeidae (10) and Pisaniidae (9). Species richness ranged from 17 at SKT to 56 at SBL. The highest and most seasonally stable mean diversity occurred at SBL, whereas PMK and PTN exhibited lower diversity and higher dominance, consistent with assemblage simplification in effluent-associated environments. GLM analyses revealed significant location effects on diversity (Wald χ²: 56.484, df: 3, p<0.001) and dominance (Wald χ²: 17.598, df: 3, p<0.001); seasonal effects were not significant. CA confirmed significant family-location associations (χ²: 1186.835, df: 120, p<0.001), with PMK most compositionally isolated (Dimension 1 contribution: 55.9%) and high spatial turnover across stations (mean Bray-Curtis: 0.885±0.109). These findings highlight substantial spatial variation in gastropod assemblages and support the use of intertidal gastropods as bioindicators for ecological monitoring in coastal ecosystems associated with aquaculture development.
Sodium alginate-microencapsulation of Bacillus cereus MH1 reduced chemical fertilizer dependency in maize (Zea mays) NUR AQIELA KAMARUDIN; NUR MAIZATUL IDAYU OTHMAN; DARIUS EL PEBRIAN; AZHAN HASHIM @ ISMAIL; WAN ZURAIDA WAN MOHD ZAIN; NOR HAFIZAH ZAKARIA; ALI TAN KEE ZUAN; NURUL MAYZAITUL AZWA JAMALUDIN
Biodiversitas Journal of Biological Diversity Vol. 27 No. 9 (2026)
Publisher : Society for Indonesian Biodiversity

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13057/biodiv/d270905

Abstract

Abstract. Kamarudin NA, Othman NMI, El Pebrian D, Ismail AH@, Zain WZWM, Zakaria NH, Zuan ATK, Jamaludin NMA. 2026. Sodium alginate-microencapsulation of Bacillus cereus MH1 reduced chemical fertilizer dependency in maize (Zea mays). Biodiversitas 27 (9): d270905. https://doi.org/10.13057/biodiv/d270905. Maize (Zea mays) relies heavily on chemical inputs, which may harm soil health and the environment. This study evaluated the effectiveness of sodium alginate-encapsulated Bacillus cereus (isolate MH1) (Accession No. NR_074540.1) as a fertilizer for reducing chemical fertilizer dependency in maize cultivation. The experiment was conducted using a Completely Randomized Design (CRD) with four treatments and three replications: unfertilized control (T0), a 100% NPK fertilizer (5 g NPK 15:15:15) (T1), encapsulated PGPR alone (5 g) (T2), and an Integrated Nutrient Management (INM) treatment consisting of (2.5 g NPK + 5 g encapsulated PGPR) (T3). The encapsulated biofertilizer contained approximately 1.0 x 108 CFU/g viable cells. Plant growth, yield performance and soil nutrient concentration status were evaluated over three months. Results showed that T3 significantly improved chlorophyll content (47.33), fresh root weight (312.33 g), root length (37.40 cm), yield weight (240.93 g), cob diameter (11.30 cm), calcium concentration (8644 mg/kg), and magnesium concentration (3024.67 mg/kg) compared with the control at p<0.05. The integrated treatment (T3) produced maize growth and yield comparable to the full chemical fertilizer treatment (T1), indicating that an integrated treatment of reduced NPK and encapsulated PGPR can achieve results similar to the full rate of NPK fertilizer. These findings demonstrate that sodium alginate-encapsulated B. cereus MH1 has strong potential as a sustainable biofertilizer and provides a practical framework for integrating microbial biotechnologies into sustainable agriculture, aligning with the Sustainable Development Goal (SDG) of life on land. However, the absence of a 2.5 g NPK-alone treatment in this experimental design limits the ability to isolate the specific contribution of PGPR to integrated management in T3.
Summer diet composition and burrow-centered activity of Athene cunicularia in a Peruvian coastal wetland MELANIE B. LOZADA-VILLANUEVA; LUIS N. VILLEGAS-PAREDES; CÉSAR R. LUQUE-FERNÁNDEZ
Biodiversitas Journal of Biological Diversity Vol. 27 No. 9 (2026)
Publisher : Society for Indonesian Biodiversity

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13057/biodiv/d270901

Abstract

Abstract. Lozada-Villanueva MB, Villegas-Paredes LN, Luque-Fernández CR. 2026. Summer diet composition and burrow-centered activity of Athene cunicularia in a Peruvian coastal wetland. Biodiversitas 27 (9): d270901. https://doi.org/10.13057/biodiv/d270901. This study assessed summer diet composition and burrow-centered activity of Athene cunicularia in a coastal wetland of southern Peru. A total of 502 pellets were collected from three active burrows between December 2022 and March 2023. Diet composition was quantified using Relative Numerical Frequency (RNF), representing the numerical contribution of each prey category, and frequency of occurrence in pellets (FAE), representing its recurrence among pellets. Trophic diversity, niche overlap, and multivariate diet composition were explored among burrow-month units, whereas camera-trap and direct-observation records were used descriptively to characterize burrow-centered activity. These complementary approaches allowed dietary patterns and activity observations to be evaluated within the same local ecological and seasonal context. This study identified 6031 prey items, of which 95.94% were invertebrates. Orthoptera and Coleoptera numerically dominated the diet and were recurrent across burrows and months. Under the available sampling design, analyses did not detect statistically supported differences in trophic diversity among burrows or months, while trophic overlap values ranged from 0.47 to 0.88. Feeding records occurred mainly during nocturnal and crepuscular periods, whereas vigilance, preening, and burrow maintenance were more frequently documented during daytime. These findings provide a local summer baseline consistent with the trophic flexibility of A. cunicularia and document its burrow-centered activity in a heterogeneous coastal wetland, providing useful ecological information for future comparative studies of this species in similar coastal environments.
The discovery of dendrophylliid scleractinian corals from Sempu Island, East Java, Indonesia OKTIYAS MUZAKY LUTHFI; DEWA GEDE RAKA WIADNYA; M. AZUREA BAHRI LUTHFI; ANDIK ISDIANTO; ADE YAMINDAGO; FERNAN P. TUPAS; SITI NURTAHIRAH JAAFAR
Biodiversitas Journal of Biological Diversity Vol. 27 No. 9 (2026)
Publisher : Society for Indonesian Biodiversity

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13057/biodiv/d270902

Abstract

Abstract. Luthfi OM, Wiadnya DGR, Luthfi MAB, Isdianto A, Yamindago A, Tupas FP, Jaafar SN. 2026. The discovery of dendrophylliid scleractinian corals from Sempu Island, East Java, Indonesia. Biodiversitas 27 (9): d270902. https://doi.org/10.13057/biodiv/d270902. This study reports the first documented locality records of two dendrophylliid scleractinian corals, Duncanopsammia peltata and Turbinaria mesenterina, from the Sempu Island Nature Reserve and adjacent coastal waters, East Java, Indonesia. Previous coral surveys conducted within this protected area did not document either species, despite their widespread distribution throughout the Indo-Pacific region. Field surveys were conducted in November 2021 and July 2024 at seven reef-flat and reef-slope sites. Standardized searches were performed along 100-m transects covering approximately 200 m² per site, while snorkeling was used on the shallow reef flat at Kondang Merak Beach. Species identification was based on non-destructive in situ observations, scaled underwater photographs, and diagnostic skeletal characters, including colony growth form, corallite morphology, corallite diameter, septal arrangement, corallite density, and live colony coloration. A single juvenile colony of D. peltata was recorded on the shallow reef flat of Kondang Merak Beach, where it exhibited an encrusting form, tubular corallites, and a conspicuous dome-shaped columella. T. mesenterina was observed on sediment-influenced reef slopes, exhibiting encrusting, foliose, and vase-shaped growth forms with densely packed unifacial corallites. Visible sediment accumulation on several colonies further characterized the habitat conditions in which T. mesenterina occurred. Because no colonies or skeletal fragments were collected, identification relied exclusively on photographic and field-based morphological evidence. These records expand the documented coral inventory of the Sempu Island Nature Reserve and provide updated morphological documentation and baseline information to support field identification, biodiversity assessment, and future monitoring of dendrophylliid corals in tropical reef ecosystems.
Optimization of thermohalostable α-amylase production from a marine extremophile Geobacillus sp. PLS 41 using response surface methodology and enzymatic characterization VIVI MARDINA; FEBRIANI FEBRIANI; NURDIN SAIDI; SUKMAWAN F. SANTOSA; TEUKU M. IQBALSYAH
Biodiversitas Journal of Biological Diversity Vol. 27 No. 9 (2026)
Publisher : Society for Indonesian Biodiversity

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13057/biodiv/d270919

Abstract

Abstract. Mardina V, Febriani, Saidi N, Santosa SF, Iqbalsyah TM. 2026. Optimization of thermohalostable α-amylase production from a marine extremophile Geobacillus sp. PLS 41 using response surface methodology and enzymatic characterization. Biodiversitas 27 (9): d270919. https://doi.org/10.13057/biodiv/d270919. α-Amylases are vital in many industries, but most conventional forms denature under the high temperatures and salinity common in industrial processes. This study optimized α-amylase production by the marine thermophilic bacterium Geobacillus sp. PLS 41 using Response Surface Methodology (RSM), followed by enzyme purification and biochemical characterization. Ammonium sulfate precipitation and DEAE Sepharose ion-exchange chromatography were used for enzyme purification. The central composite design evaluated starch concentration, NaCl concentration, temperature, and incubation time. The validated optimum conditions were 2.5% (w/v) starch, 2.5% (w/v) NaCl, 65°C, and 36 h, yielding 38.53±3.00 U/mL, approximately 3.6-fold higher than the initial condition. Ammonium sulfate fractionation at 40-60% saturation produced the highest specific activity of 42.6 U/mg, while DEAE Sepharose chromatography increased the specific activity to 206.6 U/mg, corresponding to 12.07-fold purification and 1.0% yield. The purified enzyme indicated an apparent molecular mass of approximately 63 kDa. Maximum activity occurred at 65°C and pH 7.0. Activity increased to 139.5% in 3 M NaCl, while thermal preincubation retained 104.9% activity after 100 min and reached 122.0% after 80 min. Under 3 M NaCl, residual activity increased to 171.1% after 100 min. Fe²⁺, Mn²⁺, and K⁺ enhanced activity, whereas EDTA and PMSF reduced it. Overall, the enzyme demonstrates strong thermo- and salt-tolerance under laboratory conditions, supporting further evaluation for detergent, starch-processing, kinetic, and scale-up applications.
Diversity, distribution, and environmental conditions of Caulerpa spp. in an eastern Indonesian intertidal zone ALFRED G. O. KASE; FANNY IRIANY GINZEL; YOHANES MERRYANTO; DONNY MERCYS BESSIE; WILSON L. TISERA; JAMES NGGINAK; ARDY ANDREAS BERNARD KASE; MATEUS SALVADOR
Biodiversitas Journal of Biological Diversity Vol. 27 No. 9 (2026)
Publisher : Society for Indonesian Biodiversity

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13057/biodiv/d270908

Abstract

Abstract. Kase AGO, Ginzel FI, Merryanto Y, Bessie DM, Tisera WL, Ngginak J, Kase AAB, Salvador M. 2026. Diversity, distribution, and environmental conditions of Caulerpa spp. in an eastern Indonesian intertidal zone. Biodiversitas 27 (9): d270908. https://doi.org/10.13057/biodiv/d270908. Caulerpa is a green macroalgal genus widely distributed in tropical coastal ecosystems and plays important ecological roles in shallow marine habitats. However, information on its distribution patterns and diversity in eastern Indonesian intertidal ecosystems remains limited. This study aimed to analyze the distribution patterns, diversity, and environmental conditions of Caulerpa spp. in the intertidal zone of Tablolong Beach, Kupang District, East Nusa Tenggara, Indonesia. Field sampling was conducted from August to September 2023 using transect-quadrat methods across four sampling sites. Morphological identification was based on thallus characteristics, while spatial distribution patterns and species diversity were analyzed using the Morisita index (Id) and Shannon-Wiener diversity index (H’). Environmental parameters including temperature, salinity, pH, and dissolved oxygen were also measured. Three species were identified, namely Caulerpa racemosa, Caulerpa lentillifera, and Caulerpa sertularioides, occurring predominantly in sandy intertidal habitats with varying degrees of seagrass association. All species exhibited clustered distribution patterns (Id>1), indicating aggregation within specific microhabitats. The Shannon-Wiener diversity index (H′<1) indicated low diversity across all sampling sites. C. lentillifera showed the highest mean percentage composition among the observed species. These findings suggest that localized habitat characteristics and environmental context may contribute to observed variation in spatial distribution and species composition of Caulerpa in tropical intertidal ecosystems. This study provides baseline ecological data for understanding Caulerpa communities and may support biodiversity conservation and sustainable coastal management in eastern Indonesia.
Compatibility of Microplitis manilae with reduced concentration microbial biopesticides under pre- and post-parasitization laboratory conditions NURSYIFA NADA HARIYADI; RULY ANWAR; NINA MARYANA
Biodiversitas Journal of Biological Diversity Vol. 27 No. 9 (2026)
Publisher : Society for Indonesian Biodiversity

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13057/biodiv/d270913

Abstract

Abstract. Hariyadi NN, Anwar R, Maryana N. 2026. Compatibility of Microplitis manilae with reduced concentration microbial biopesticides under pre- and post-parasitization laboratory conditions. Biodiversitas 27 (9): d270913. https://doi.org/10.13057/biodiv/d270913. Heavy reliance on synthetic insecticides for managing the invasive pest Spodoptera frugiperda underscores the urgency of integrating biological control agents, particularly the indigenous endoparasitoid Microplitis manilae, with microbial entomopathogens. This study evaluated the compatibility of commercial formulations of Bacillus thuringiensis and Metarhizium anisopliae, applied at half the recommended field concentrations, based on the biocontrol efficacy and pre-adult parameters of M. manilae. The experiment was conducted using a Randomized Block Design (RBD), analyzed via Linear Mixed-effects Models (LMM), in which the time cohort defined as a random effect. Based on the parasitization-rate reduction criteria set by the International Organization for Biological Control (IOBC), both biopesticides were classified as Class 1 (Harmless). However, significant sublethal effects were observed. Pre-parasitization exposure to M. anisopliae significantly prolonged the egg-larval and pupal development stages compared to the control, it also induced a strong male-biased sex ratio, reducing the female proportion to 28.60%. Conversely, a significant reduction in adult female longevity was observed following post-parasitization exposure to B. thuringiensis, with a mean longevity of 21.46 days compared to 26.84 days in the control group. Preliminary qualitative observations revealed rare physical malformations affecting the distal antennal segments of two eclosed female adults in the post-parasitization treatment. These laboratory findings provide an initial indication that post-parasitization application may be less disruptive at reduced concentrations, thereby offering a safer integration strategy in sustainable pest management programs for S. frugiperda.
Comparative phytochemical profiling of leaves from three Indonesian Vitex species using spectrophotometry, HP-TLC, and LC-HRMS SRI LULIANA; FADLINA CHANY SAPUTRI; SARINI ABDULLAH; RATNA SURYA ALWI; PUTU GITA MAYA W. MAHAYASIH; CHOO CHEE YAN; ABDUL MUN’IM
Biodiversitas Journal of Biological Diversity Vol. 27 No. 9 (2026)
Publisher : Society for Indonesian Biodiversity

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.13057/biodiv/d270906

Abstract

Abstract. Luliana S, Saputri FC, Abdullah S, Alwi RS, Mahayasih PGMW, Yan CC, Mun’im A. 2026. Comparative phytochemical profiling of leaves from three Indonesian Vitex species using spectrophotometry, HP-TLC, and LC-HRMS. Biodiversitas 27 (9): d270906. https://doi.org/10.13057/biodiv/d270906. The genus Vitex is widely used in traditional medicine, yet comparative phytochemical data integrating quantitative assays and metabolite profiling for Indonesian collections remain limited. This study compared leaf extracts from three Indonesian Vitex collections representing Vitex pinnata, Vitex negundo, and Vitex trifolia. Leaves were extracted with 70% ethanol using microwave-assisted extraction. Extraction yield, Total Phenolic Content (TPC), and Total Flavonoid Content (TFC) were determined from three independent extraction batches (n = 3), while HP-TLC and LC-HRMS were used for qualitative phytochemical profiling. V. pinnata showed the highest extraction yield (34.77±1.54%), TPC (424.31±24.48 mg GAE/g extract), and TFC (32.03±2.01 mg QE/g extract). HP-TLC revealed distinct chromatographic fingerprint patterns among the three collections. LC-HRMS yielded 79 putatively annotated features (MSI Level 2), comprising 43 annotations in V. pinnata, 28 in V. negundo, and 39 in V. trifolia. All three collections shared seven features, while 10 were shared only by V. pinnata and V. trifolia, four by V. pinnata and V. negundo, and three by V. negundo and V. trifolia; 22, 14, and 19 features were unique to the respective collections. Flavonoids were most prominent in V. pinnata, iridoid-related annotations were comparatively abundant in V. negundo, and V. trifolia showed relatively high phenolic and terpenoid annotation counts. These findings provide a preliminary comparative phytochemical reference for Indonesian Vitex materials. Because each species was represented by a single geographically distinct collection, the observed differences should be interpreted at the collection level. Multisite sampling and confirmation using authentic standards are required to establish robust species-associated markers.

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