METABOLITES ACTIVITY OF ENDOPHYTIC STREPTOMYCES SP. IPBCC. B.15.1539 FROM TINOSPORA CRISPA L. MIERS: ALPHA-GLUCOSIDASE INHIBITOR AND ANTI-HYPERGLYCEMIC IN MICE

Objective: This research work aimed to assess the capability of endophytic Streptomyces sp. IPBCC. b.15.1539 isolated from Tinospora crispa inproducing α-glucosidase inhibitor compound and examined the effect of its ethyl acetate extract containing α glucosidase inhibitor in lowering blood glucose in streptozotozin mice.Methods: Streptomyces sp. IPBCC. b.15.1539 was grown in a bioreactor filled with International Streptomyces Project 2 medium, for 5, 10, 15, and20 days, and assayed for its in vitro α-glucosidase inhibitory activity. The ethyl acetate extract was examined for its IC50Results: The crude extract produced value. An in vivo experimentwas set up using thirty mice, which were divided into five treatment groups: (a) acarbose (0.03 mg/30 g body weight) used as a positive control, (b) placebo used as a negative control, (c-e) treatment groups were treated with ethyl acetate extract at 0.036 mg/30 g body weight (P1), 0.36 mg/30 gbody weight (P2), 0.036 mg/30 g body weight (P3).98.5% α-glucosidase inhibitory activity, with 15,6 biomass after 10 days of production. The ethyl acetateextract at a concentration of 1000 µg/ml produced 96.08% α-glucosidase inhibitory activity, while acarbose at the same concentration gave 97.46%inhibition. The IC50 for the ethyl acetate extractConclusions: The was 0.047 µg/ml, while for acarbose was 0.003 µg/ml. The ethyl acetate extract applied as the P1treatment group lowered blood glucose levels in streptozotozin mice by 26%. α-glucosidase inhibitor of Streptomyces sp. IPBCC. b.15.1539 of T. crispa has the potency as an antidiabetic agent for type 2 DMtherapy

COMMUNITY STRUCTURES OF ENDOPHYTIC ACTINOBACTERIA FROM MEDICINAL PLANT CENTELLA ASIATICA L. URBAN-BASED ON METAGENOMIC APPROACH

Objective: This study aimed to assess the community structure of actinobacteria in rhizosphere and endophyte of a medicinal plant, Centella asiatica, based on a metagenomic approach using Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) of 16S rRNA gene.Methods: Total genomic DNA was extracted from the rhizosphere and plant tissue followed by PCR amplification of actinobacterial 16S rRNA gene using nested PCR. The community structure of actinobacteria was analyzed using the DGGE techniques on polyacrylamide gels. PCR products of excised bands result from polyacrylamide gel were sequenced and analyzed by bioinformatics software to construct a phylogenetic tree.Results: The results of separation in DGGE gel showed 16 major bands from rhizosphere and plant tissue. The bands distribution pattern showed that the community of actinobacteria in the plant tissue was slightly more diverse than rhizosphere, although it is not significantly different based on Shannon-Wiener analysis. The BLAST. N analysis showed that 7 bands related to Streptomycetaceae (83-100%), 5 bands related to Micromonosporaceae (99-100%), 1 bands related to Gordoniaceae (99%) and 3 bands still belonged to unculturable (87-99%). There were 6 genera under those 3 families, i.e. Streptomyces, Micromonospora, Verrucosispora, Actinoplanes, Couchioplanes, and Gordonia. The percentage of strain similarity comparison to the database showed that there were 4 bands with<97% maximum identity which may be related to novel endophytic actinobacteria in C. asiatica.Conclusion: Diversity of endophytic actinobacteria based on a metagenomic approach using 16S rRNA gene-targeted PCR-DGGE analysis was found associated with C. asiatica. Several of them may have potency as novel actinobacteria and can be further explored for their medicinal function.Keywords: Centella asiatica, PCR-DGGE, Endophytic Actinobacteria, Metagenomic, 16S rRN

Community of Soil Actinobacteria in PTPN VI Oil Palm Plantation Jambi (Sumatra, Indonesia) Based on Amplicon Sequencing of 16S rRNA Gene

In Sumatra, Indonesia, increased oil palm production encourages land expansion for oil palm plantations. And soil Actinobacteria have a potential role in agriculture and plantations ecosystems. The use of fertilizer and herbicide affects soil microbial diversity, including Actinobacteria. This research analyzed and investigated the community composition and diversity of Actinobacteria in soils of oil palm plantations in Jambi Sumatra. Amplicon-based analysis of the 16S rRNA gene (V3-V4 hypervariable region) was used to amplify actinobacterial full-length 16S sequences. The V3-V4 actinobacterial specific 16S rRNA gene sequencing was done using Next-Generation Sequencing. This study confirmed that actinobacterial specific 16S rRNA gene primer could amplify the actinobacterial 16S rRNA gene. Frankiales dominated the community composition of soilborne Actinobacteria. The diversity and community composition of soilborne Actinobacteria were not significantly affected by the interaction between fertilization and weed treatments. Furthermore, the use of NPK fertilizer significantly affected the abundance of Kineosporiales, whose abundance increased with the increasing concentration of NPK fertilizer. The interaction between fertilization and weeding treatments in the oil palm plantations has no impact on soil Actinobacteria's community composition and diversity

Pelapisan Benih dengan Aktinobakteri untuk Meningkatkan Pertumbuhan Tanaman Padi: Actinobacterial Seed Coating for Promoting Rice Plant Growth

The need for rice continues to increase along with the increase in population. Efforts to increase rice production is generally carried out through proper and balanced fertilization. Other than that, plant growth-promoting actinobacterial (PGPB) inoculants can be used as an alternative solution. This study aimed to examine the effect of the application of actinobacterial seed dressing and NPK fertilizers on the growth of rice plants grown in a glass house.  A randomized block design with three factors was conducted, consisting of actinobacterial seed dressing (added and not added), type of carrier (zeolite, peat, and combinations) and doses of NPK fertilizer (0 g/pot, 0.375 g/pot, and 0.75 g/pot). The addition of actinobacteria consortium, peat-zeolite combination 1:3, and NPK fertilizer at a dose of 0.75 g/pot (A2C3P3) gave consistent results in increasing the average yield of rice vegetative and reproductive parameters observed in the glass house. The A2C3P3 treatment had a significant effect on the number of tillers, width of flag leaf, dry weight of roots and shoots of rice observed at 10 WAP compared to other treatment combinations. The actinobacterial seed coating plays a pivotal role in supporting rice plant growth

Lipase Activity of Endophytic Actinobacteria from Medicinal Plants

Endophytic bacteria are known to reside within host plant tissue without giving a harmfull effect. The endophytes may play an important role, as they may produce similar bioactive compounds as produced by the host plant. Various medicinal plants have long been used to cure diseases. Traditionally, leaves extract of Guazuma ulmifolia, Psidium guajava, or the rhizome of Curcuma xanthorrhiza can be used to treat disease, e.g. hyperlipidemic. The mechanism can be through lipase activity, where the lipase catalyzes the hydrolysis of triacylglycerol to fatty acids and acylglycerol. The objective of this research was to assess potency of endophytic bacteria as anti-hyperlipidemic compounds producer through their lipase activity. Sixty nine endophytic bacteria which comprised of 22, 27 and 20 isolates were isolated from the leaves of G. ulmifolia, P. guajava, and the rhizome of C. xanthorrhiza, respectively. Eight out of the 69 isolates showed lipase activity, and the two selected isolates, i.e. DPG 3(2) and AJB 4(4) were considered as good lipase producers. The highest specific lipase activity of DPG 3(2) isolate was observed for 0.874 units per mg at 38 h, whereas AJB 4(4) isolates had the specific lipase activity at 1.139 units per mg after 72 h observation. These data indicate that the two selected isolates have the potency as antihyperlipidemic compounds producer through their lipase activity

Characterization of Xylanase Streptomyces spp. SKK1-8

Streptomyces spp. SKK1-8 producing xylanase was isolated from soil sample from Sukabumi West Java. The xylanase have an optimum condition at pH 6 and 50 0C. Addition of 5 mM Cu2+ decreased the xylanase activity up to about 77%, whereas not by other cations. The xylanase was stable at 3 0C for 48 hours, and the enzyme half lifetime was 1 hour 45 minute at 50 0C. This xylanase showed the highest activity on oatspelt xylan, and their molecular masses were estimated approximately 16.80, 15.21, and 13.86 kDa. HPLC analysis showed that xylosa and arabinosa were the main hydrolytic product of birchwood xylan. Key words: xilanase, Streptomyces spp., characterization, zymogram and SDS-PAGE, stabilit

Metagenomic of Actinomycetes Based on 16S rRNA and nifH Genes in Soil and Roots of Four Indonesian Rice Cultivars Using PCR-DGGE

The research was conducted to study the metagenomic of actinomycetes based on 16S ribosomal RNA (rRNA) and bacterial nifH genes in soil and roots of four rice cultivars. The denaturing gradient gel electrophoresis profile based on 16S rRNA gene showed that the diversity of actinomycetes in roots was higher than soil samples. The profile also showed that the diversity of actinomycetes was similar in four varieties of rice plant and three types of agroecosystem. The profile was partially sequenced and compared to GenBank database indicating their identity with closely related microbes. The blast results showed that 17 bands were closely related ranging from 93% to 100% of maximum identity with five genera of actinomycetes, which is Geodermatophilus, Actinokineospora, Actinoplanes, Streptomyces and Kocuria. Our study found that Streptomyces species in soil and roots of rice plants were more varied than other genera, with a dominance of Streptomyces alboniger and Streptomyces acidiscabies in almost all the samples. Bacterial community analyses based on nifH gene denaturing gradient gel electrophoresis showed that diversity of bacteria in soils which have nifH gene was higher than that in rice plant roots. The profile also showed that the diversity of those bacteria was similar in four varieties of rice plant and three types of agroecosystem. Five bands were closely related with nifH gene from uncultured bacterium clone J50, uncultured bacterium clone clod-38, and uncultured bacterium clone BG2.37 with maximum identity 99%, 98%, and 92%, respectively. The diversity analysis based on 16S rRNA gene differed from nifH gene and may not correlate with each other. The findings indicated the diversity of actinomycetes and several bacterial genomes analyzed here have an ability to fix nitrogen in soil and roots of rice plant

Potential Pseudomonas Isolated from Soybean Rhizosphere as Biocontrol against Soilborne Phytopathogenic Fungi

Plants are liable to be attacked by soilborne fungal pathogens which are responsible to reduce plant growth and losses in yield. In Indonesia, indigenous soybeans’ rhizobacteria such as antifungal producing Pseudomonas sp. have not many been reported yet. Therefore, the potential of the Pseudomonas sp. as biocontrol agent should be deeply explored. The aim of this study was to screen the indigenous soybeans’ rhizobacteria Pseudomonas sp. that possessing biocontrol characters against soilborne mainly i.e. Sclerotium rolfsii, Fusarium oxysporum, and Rhizoctonia solani, in vitro and in planta. Eleven isolates identified Pseudomonas sp. CRB numbered by CRB-3, CRB-16, CRB-17, CRB-31, CRB-44, CRB-75, CRB-80, CRB-86, CRB-102, CRB-109, and CRB-112 were affirmed to be candidates of biocontrol agents toward the soilborne fungal pathogens. Pseudomonas sp. CRB inhibited growth of the pathogenic fungi approximately 11.1-60.0% in vitro. Among of them, 7 isolates were also produced siderophore, 2 isolates produced chitinase, and 4 isolates produced hydrogen cyanide. Seed coating with the Pseudomonas sp. CRB accomplished disease suppression in planta about 14.3-100% in sterile soil condition and 5.2-52.6% in non sterile soil condition. Consistency in high performance more than 30% of disease suppression in non sterile soil condition suggested that 5 isolates i.e. CRB-16, CRB-44, CRB-86, CRB-102, and CRB-109 isolates have great promising to be developed as biocontrol agents of soilborne pathogenic fungi

Sponge-Associated Actinobacteria: Morphological Character and Antibacterial Activity against Pathogenic Bacteria

Sponge-associated actinobacteria may diverse and have potency to produce bioactive compounds. Diversity and antimicrobial activity of indigenous sponge-associated actinobacteria isolated from the marine ecosystem in Indonesia have not much been explored. This work aimed to assess morphological and antibacterial activity of sponge-associated actinobacteria. The morphological characteristics were examined based on their color of aerial and substrate mycelia, and pigmentation, while antibacterial activities were assayed using the antagonist technique. The selected actinobacterial isolate was identified using 16S rRNA gene. Various sponge-associated actinobacteria were successfully isolated from Hyrtios sp., Callyspongia sp., and Neofibularia sp. sponges. A total of 62 actinobacterial isolates were obtained, and each isolate showed a variety of morphological characters, which could be seen in aerial mass color, substrate mass color, and pigmentation. Actinobacterial isolates were tested against human pathogenic bacteria, i.e. Staphylococcus aureus and Methicillin-Resistant S. aureus, representing Gram-positive, and Escherichia coli EPEC K1-1 and Shigella dysenteriae, representing Gram-negative. Most of actinobacterial isolates had antimicrobial activities at least against one of pathogenic bacteria. High activity was shown by NOHa.2, isolated from Neofibularia, and HRHa.5 isolated from Hyrtios. The NOHa.2 showed the highest antimicrobial activity against S. dysenteriae, meanwhile, HRHa.5 showed antimicrobial activity against 3 of 4 tested bacterial pathogens. These data showed diversity of sponge-asccociated actinobacteria from marine ecosystem in Indonesia, and several of them have potency as source of antibacterial compound

The Existence of Endophytic Actinobacteria from Rhododendron zoelerri Revealed by Culture-Dependent and Culture-Independent Approaches

Endophytic actinobacteria from medicinal plant may play a significant role in producing bioactive compounds. The information regarding their diversity is an important.  Rhododendron are traditionally used for treating human disorders. One of the selected Rhododendron used in this study was R.  zoelleri from Papua origin, which has been conserved and grown in Cibodas Botanical Garden, West Java, Indonesia. The aim of this study was to assess the existence of endophytic actinobacteria from R. zoelleri based on a culture-dependent and their community structure based on a culture-independent approach. Culturable actinobacteria were isolated and cultured on HV medium. Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) targeting the metagenomic 16S rRNA was used to analyse the structure of the actinobacterial community. Six culturable endophytic actinobacteria (200 cfu/g fresh weight) from R. zoelleri were successfully isolated, three isolates from leaf, and the other isolates were obtained from stem. The six culturable isolates were RZP 1.3, RZP 1.1, RZP 2.2, RZPB 1.1, RZPB 7.1, RZPB 4.1. Based on their morphological characteristics, the endophytes have Streptomyces characters. The existence of Streptomyces spp. were also confirmed with molecular analysis based on 16S rRNA gene. The phylogenetic analysis based on 16S rRNA gene to the reference strains available in EzTaxon-e database showed that six isolates were closely related to S. djakartensis strains of NBRC 15409ᵀ(99.19%), S. tritolerans strains of DAS 165T(99.90%), S. coelicoflavus strains of NBRC 15399T(99.59). However, they showed differences in morphological characteristics as compared with the reference strains. The metagenomic analysis of the DGGE profile based on 16S rRNA gene showed the community structure of endophytic actinobacteria from R. zoelleri which was represented by 13 DGGE bands. The bands were closely related to Agromyces, Gordonia, Microbacterium, Micromonospora, Propionibacterium, Saccharomonospora, Streptomyces which have 93.18%-100% similarity. Based on the data, it showed diversity of endophytic actinobacteria from R. zoelleri which may be further assess for their novelty and bioprospecting