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Screening of Rhizobacteria for Plant Growth Promotion and Their Tolerance to Drought Stress RAHAYU FITRIANI WANGSA PUTRIE; ARIS TRI WAHYUDI; ABDJAD ASIH NAWANGSIH; EDI HUSEN
Microbiology Indonesia Vol. 7 No. 3 (2013): September 2013
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (271.581 KB) | DOI: 10.5454/mi.7.3.2

Abstract

Rhizobacteria have been known for their capability as plant growth promoter through some mechanisms, directly and indirectly. The purpose of this research to screen rhizobacteria of Bacillus spp. and Pseudomonas spp. for their drought tolerance as plant growth promoter of maize (Zea mays). Screening of rhizobacteria as growth promoter of 47 isolates of Bacillus CR and 34 isolates of  Pseudomonas CRB resulted 24 and 9 isolates were able to stimulate the growth of maize sprouts, respectively. Further screening of those growth promoter of the rhizobacterial isolates to drought tolerance resulted 7 isolates of Bacillus CR and 6 isolates of Pseudomonas CRB that were able to grow on medium with osmotic pressure -1 and -2 MPa, respectively. Potential rhizobacterial isolates of growth promoter and drought tolerance were tested for antagonist mechanisms which aims to determine ability to live together in one carrier medium if to be made formulation. Both non antagonist rhizobacterial isolates were evaluated for their potential in producing exopolysaccharide (EPS) revealing that CRB 19 and CR 90 exhibited the highest activity of EPS production up to 0.346 mg mL-1 on medium with -2.0 MPa and 0.107 mg mL-1 on medium with -0.73 MPa, respectively. Based on 16S rRNA sequence analysis, it revealed CRB 19 and CR 90 had highest similarities to Pseudomonas aeruginosa strain B2 and Brevibacillus brevis B33, respectively. Those growth promoter and drought tolerant of Bacillus CR and Pseudomonas CRB had potency to be developed as inoculants in dry land agriculture.
Peningkatan laju pertumbuhan benih ikan gurame (Osphronemus goramy Lac.) yang direndam dalam air yang mengandung hormon pertumbuhan ikan mas [Growth enhancement of Osphronemus goramy Lac.juvenile immersed in water containing recombinant Cyprinus carpio growth hormone] Irmawati Irmawati; Alimuddin Alimuddin; Muhammad Zairin Jr; Muhammad Agus Suprayudi; Aris Tri Wahyudi
Jurnal Iktiologi Indonesia Vol 12 No 1 (2012): Juni 2012
Publisher : Masyarakat Iktiologi Indonesia (Indonesian Ichthyological Society)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.32491/jii.v12i1.126

Abstract

This study was aimed toenhance the growth of the giant gourami juve-nile by immersion with recombinant Cyprinus carpio growth hormone (rCcGH). Recombinant growth hormone was expressed by plasmid pCold-I/CcGHin E. coli BL21 (DE3). The inclusion bodies were isolated from E. coli using lyso-zyme and centrifugation method. Immersion with water containing 0.9% NaCl and 0.01% bovine serum albumin and inclusion bodies with different doses of rCcGH, 5 mgL-1 (C1), 15 mgL-1 (C3), and 30 mg L-1 (C6) was performed for 1 hour on weekly basis for the first four weeks of experimental period. Fish were subjected to salinity shock of 30 ppt NaCl for 2 minutes before it was transferred into the water containing rCcGH. Controls fish were without immersion and salinity shock (control), immersed in 30 ppt NaCl for 2 minutes (SS), immersed in 0.01% BSA (BSA), and immersed in medium containing BSA and 30 mg L-1 inclusion bodies of pCold without rCcGH (pCold).The giant gourami ju-venile treated with 30 mg L-1 rCcGH were 72.90% heavier and 21.04% longer (p<0.05) than the control, and 43.07% heavier and 14.64% longer (p<0.05) than the pCold. No significant difference in survival rate was obtained between treatments and controls. Biomass of fish treated 30 mgL-1 rCcGH was the highest among others. The rCcGH was able to promote soluble protein synthesis and lipid utilization as energy sources to spare protein (protein sparing effect). Thus, immersion with rCcGH could be applied to enhance the growth of giant gourami juvenile. AbstrakPenelitian ini bertujuan untuk memacu pertumbuhan benih ikan gurame melalui perendaman dengan hormon pertumbuhan rekombinan ikan mas (rCcGH). Hormon pertumbuhan rekombinan diekspresikan oleh plasmid pCold-I/CcGH di dalam Escherichia coli BL21(DE3). Badan inklusi diisolasi dari E. coli menggunakan lisozim dan metode sentrifugasi. Perendaman dilakukan selama 1 jam di dalam air yang mengandung 0,9% NaCl, 0,01% albumin serum sapi (BSA), dan badan inklusi rCcGH pada dosis5 mg L-1 (C1),15 mg L-1 (C3), dan 30mg L-1 (C6),sekali seminggu pada 4 minggu pertama. Ikan diberi kejutan salinitas 30 ppt NaCl selama dua menit sebelum ikan dipindahkan ke dalam air yang mengandung rCcGH.Sebagai kontrol ialah: benih ikan gurame tanpa perendaman (kontrol), benih ikan gurame yang diberi kejutan salinitas (SS), benih ikan gurame yang direndam di dalam air media yang mengandung BSA (BSA), dan benih ikan gurame yang direndam di dalam air yang mengandung BSA dan protein pCold-I tanpa rCcGH (pCold). Setelah tujuh minggu pemeliharaan, kelompok ikan yang direndam dalam air yang mengandung 30 mg L"1 rCcGH,72,90% bobot lebih berat dan 21,04% badan lebih panjang di-bandingkan dengan kontrol serta 43,07% bobot lebih berat dan 14,64% badan lebih panjang dibandingkan dengan pCold (p>0,05). Kelangsungan hidup antar perlakuan dan kontrol tidak berbeda nyata. Biomassa kelompok ikan yang direndam dengan 30 mg L-1 adalah yang tertinggi dibandingkan perlakuan lainnya. GH rekombinan ikan mas meningkatkan sintesis protein terlarut dan pemanfaatan lipid sebagai sumber energi sehingga mengoptimalkan pemanfaatan protein untuk pertumbuhan (protein sparing effect). Dengan demikian, perendaman dengan rCcGH dapat diaplikasikan untuk memacu pertumbuhan benih ikan gurame.
Identification of nifD and nifH Genes of Methanotrophic Bacteria from Rice Field Ari Fina Bintarti; Iman Rusmana; Aris Tri Wahyudi
ANNALES BOGORIENSES Vol 18, No 2 (2014): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (879.425 KB) | DOI: 10.14203/ann.bogor.2014.v18.n2.13-25

Abstract

Metanotrophic bacteria have ability to oxidize methane and fix atmospheric nitrogen, hence the bacteria has an important role as a nitrogen source provider on wetland area like rice fields. Nitrogen fixation process is catalyzed by the nitrogenase enzyme complex, encoded by nifD and nifH genes. However, characteristic of these genes from indigenous-methanotrophic bacteria still poorly understood. Hence, nifD and nifH genes of methanotrophic bacteria isolated from rice fields in Indonesia (BGM3, BGM9, SS1, SS3, SS10, ST18, SP3 and INP4) were identified and characterized. Detection of nifH and nifD genes was conducted by polymerase chain reaction (PCR) amplification. nifH and nifD gene sequences were analyzed using BLAST-X and phylogenetic trees were constructed using Neighbour Joining method. Based on nifH sequences analysis, SS1 closely related to Beijerinckia mobilis and SS3, SS10, ST 18 closely related to Beijerinckia indica subsp. indica ATCC 9039, while, BGM3, INP4, and BGM9 related to nifH of uncultured nitrogen-fixing bacterium. In other hand, sequence analysis of nifD gene showed that SS1, SS3, SS10, ST 18 closely related to B. indica subsp. indica ATCC 9039 and BGM3, BGM9, INP4 closely related to Xanthobacter autotrophicus Py2. Identification by 16 SrRNA indicated that SS1, SS3, SS10, and ST18 had closeness to Beijerinckia sp. P310-1, while INP4 closely related to Xanthobacter sp. M5C24.
Konstruksi Mutan Pseudomonas sp. untuk Meningkatkan Produksi Indole Acetic Acid (IAA) melalui Mutagenesis dengan Transposon Aris Tri Wahyudi; Mutiha Panjaitan; Nisa Rachmania
Majalah Ilmiah Biologi BIOSFERA: A Scientific Journal Vol 26, No 3 (2009)
Publisher : Fakultas Biologi | Universitas Jenderal Soedirman

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (226.596 KB) | DOI: 10.20884/1.mib.2009.26.3.158

Abstract

Pseudomonas sp. is one of bacterial groups having ability to promote plant growth and health. Of a hundred successfully isolated Pseudomonas sp. from soybean rhyzosphere, 98 were found to produce indole acetic acid (IAA) ranging from 0.33 to 16.02 ppm. These isolates are bacilli, motile, Gram negative, and showing positive oxidase assay. One of them, i.e. Pseudomonas sp. CRB17, can promote plant growth by means of significant stimulation of primary root length and lateral root number. This isolate was then subject to mutagenesis using transposon Mini-Tn5Km1 to increase IAA production. Mutagenesis was done by conjugation between E coli S17-1 (l pir) carrying transposon mini-Tn5Km1 (donor) and Pseudomonas sp. (recipient), resulting in conjugation frequency of approximately 3.1 x 10-5 cell per recipient. The resulted CRB17 mutants were then tested for their ability to produce IAA, one of which showed an increment of IAA production up to 77.5%, while some others showed no significant change or even had a reduction to 55.3%.  Sequence analyisis of 16S rRNA gene of Pseudomonas sp. CRB17 indicated that it has a high homology with that of Pseudomonas plecoglossicida (identical value of 99%). The results recommends that mutagenesis using transposon can be applied to increase IAA production, especially in Pseudomonas sp. CRB17.
Abundance of Culturable Bacteria Isolated from Maize Rhizosphere Soil Using Four Different Culture Media ERNIN HIDAYATI; ARIS TRI WAHYUDI; ANTONIUS SUWANTO; RAHAYU WIDYASTUTI
Microbiology Indonesia Vol. 8 No. 1 (2014): March 2014
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (708.386 KB) | DOI: 10.5454/mi.8.1.5

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Composition and abundance of culturable bacteria of four soil samples (Ktr50II, D50II, G50II, and A50II) were analyzed. The soil samples were collected from maize rhizosphere that planted in dryland Lombok Island. Each soil sample give different growth performance of maize in greenhouse experiment. This study was to investigate the relation of maize growth performance with culturable bacterial community of their rhizosphere and the effect of culture media on number of bacterial isolates recovery. The rhizosphere bacteria were cultured and isolated on commercial media (SEA) and non commercial modification media (NAln, NAln-SE and NAln- RE). Thirty four strains rhizosphere bacteria were obtained from four maize rhizophere soil samples. D50II is the soil sample that caused the better growth performance to the maize, contrary to Ktr50II. D50II has significantly highest number of culturable bacterial types, while significantly lowest on Ktr50II. In D50II, at least 17 bacterial isolates contributed to better growth performance in maize and have relative abundance of dominant isolate not more than 35.34%. In comparing the rhizosphere bacteria recovered using different culture media, bacteria cultivated on SEA have different growth characteristic compared with bacteria cultivated on NAln, NAln SE and NAln-RE. Six bacterial isolates showed antagonistic ability when grew on SEA but not in all of three media. Compared with commercial media, non commercial modification media can increase total isolates recovery about 70.6%.
16S rRNA-based Metagenomic Analysis of Endophytic Actinomycetes Diversity from Tinospora crispa L. Miers MONA PRIMANITA; ARIS TRI WAHYUDI; YULIN LESTARI
Microbiology Indonesia Vol. 9 No. 1 (2015): March 2015
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1339.777 KB) | DOI: 10.5454/mi.9.1.4

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Endophytic actinomycetes associated with medicinal plants is very important as source of various bioactive compounds. The fact that more than 99% of microbes that exist in nature may have the potency but still unexplored. Published data regarding diversity of endophytic actinomycetes from T. crispa is mainly based on a culturable approach. This paper describes the first reported data regarding metagenomic analysis on the diversity of endophytic actinomycetes from T.crispa based on 16S rRNA gene using PCR-DGGE. There were some similarities amongst endophytic actinomycetes found in stems, roots, and leaves with soil actinomycetes community in the rhizosphere of T. crispa. There were a total of 21 bands found from the DGGE analysis which were interpreted using Phoretix 1D software. Diversity of actinomycetes in the stems, leaves, roots were represented by 17, 16, and 14 bands, respectively. Whereas only 10 bands represented diversity of actinomycetes in the soil rhizosphere. The 12 dominant and or different bands with 180 bp in size were molecularly sequenced. The A4 and A9 bands have 95% and 86% similarities with Williamsia and Streptomyces, respectively. These similarities were less than 97% thus may indicate novel actinomycetes. The other 10 sequenced bands have closed similarity ranging from 97-100% and they were closely related to the genus Streptomyces, Microbacterium, Amycolatopsis, Actinomadura, Actinoplanes, Actinokineospora, Kibdelosporangium, Williamsia and Kocuria. These findings indicate that diversity of actinomycetes can be found associated with T. crispa. 
Detection of Antibody to Burkholderia pseudomallei in Captive and Wild Macaquesin West Java and Bali, Indonesia VINCENTIUS ARCA TESTAMENTI; DIAH ISKANDRIATI; ARIS TRI WAHYUDI; JOKO PAMUNGKAS
Microbiology Indonesia Vol. 12 No. 1 (2018): March 2018
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (500.83 KB) | DOI: 10.5454/mi.12.1.4

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Melioidosis is an emerging zoonotic disease caused by the saprophytic Burkholderia pseudomallei, which infects human and a wide range of animal species. Melioidosis may lead to septicemia and pneumonia in human patients, which can be fatal if the patient is not treated accordingly. The disease is spread in tropical areas and is highly endemic to Southeast Asia and Northern Australia. However, melioidosis is poorly reported in Indonesia, especially in the veterinary field. This research provides serological evidence of antibodies to B. pseudomallei in both captive and wild nonhuman primates. Plasma samples were taken from a total of 390 monkeys in captivities and wild habitats in West Java and Bali, Indonesia. Enzyme-linked immunosorbent assay (ELISA) showed that the seroprevalence was 42.21% for Macaca fascicularis and 43.59% for Macaca nemestrina. Furthermore, the seroprevalence was 53.41% for captive macaques and 17.83% for wild macaques. The findings showed that exposure to B. pseudomallei happened in both captive and wild macaques. Based on this serosurveillance results, further studies such as comprehensive culture and clinical study are required to discover the clinical burden of the disease in nonhuman primates.
Optimization of Xylanase Production by Streptomyces costaricanus 45I-3 Using Various Substrates through Submerged Fermentation SIPRIYADI SIPRIYADI; ARIS TRI WAHYUDI; MAGGY THENAWIDJAYA SUHARTONO; ANJA MERYANDINI
Microbiology Indonesia Vol. 14 No. 1 (2020): March 2020
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1468.005 KB) | DOI: 10.5454/mi.14.1.5

Abstract

Xylanase is an important hydrolytic enzymes with many application in several industries, but to obtain enzyme derived products is not easy. Thus, the optimization of efficient xylanases production is a great interest for biotechnological application. This study aims to determine the type of substrate, medium composition, and optimum conditions of xylanase production by S. costaricanus 45I-3. Determination of substrate type was done by growing the tested bacteria on birchwood xylan, beechwood xylan, oat spelled xylan, corn cobs xylan, and tobacco xylan substrate, meanwhile the determination of medium composition and enzyme production were done by measuring xylanase activity at various substrate concentration and replacing the carbon, nitrogen, phosphate and surfactants source. The results showed that the highest enzymatic index (EI) produced from corn cob xylan substrate at 3.60 meanwhile the second highest was beechwood xylan substrate at 2.87 EI, however this substrate is purer, thus this substrate was selected and used as xylan sources for further optimization measurement. The best xylanase activity (2.29 U/mL) obtained on eighth day after inoculation on rotary incubator at 120 rpm in 28 ºC. Arabinose as the source of carbon generate the highest activity at 3.161 U/mL meanwhile the most preferred source of phosphate is Na2HPO4 (2.37 U/mL). Both source of nitrogen i.e. nitrogen ammonium sulphate (NH4)2SO4 and yeast extract were able to produce xylanase at 2.57 and 2.36 U/mL. The addition of surfactant in production medium showed addition of SDS surfactant (0.146 U/mL) and Tween 80 (0.438 U/mL) showed a negative response by decreasing the activity. The conclusion showed that the xylanase activity was increased after optimization at various C, N, and P sources, and the use of nitrogen source (NH4)2SO4), become a more economical alternative to replacing a nitrogen source yeast extract so it can lower the production costs of xylanase enzyme.
Antifungal Substances Produced by B. subtilis Strain W3.15 Inhibit the Fusarium oxysporum and Trigger Cellular Damage Rury Eryna Putri; Nisa Rachmania Mubarik; Laksmi Ambarsari; Aris Tri Wahyudi
HAYATI Journal of Biosciences Vol. 30 No. 5 (2023): September 2023
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.30.5.843-854

Abstract

Soybean Fusarium wilt and root rot disease caused by a necrotrophic ascomycete pathogen, F. oxysporum, triggered severe damage to the plant tissues and organs and impacted heavy losses. Biocontrol agents, Bacillus subtilis, were commonly used to produce a broad spectrum of antifungal substances and were gradually used in biocontrol studies for plant disease management. Investigation and determination of the inhibiting mechanism of antifungal substance produced by B. subtilis on F. oxysporum should be done to protect the soybean plant. This study revealed that basal nutrient broth (NB) gives the best antifungal activity. The stationary phase of the bacterial growth curve was obtained on two days of cultivation and showed the maximum antifungal activity against F. oxysporum. Ethyl acetate (EA) extraction of bacterial supernatant generated crude EA extract, which showed half inhibition (IC50) at 306.42 µg/ml obtained from the dose-response regression curve. Post-treatment mycelia of F. oxysporum with bacterial extract were demonstrated as hyphal deformation followed by malondialdehyde (MDA) accumulation. Furthermore, cellular leakage on fungal cells that may be triggered by antifungal compounds from strain W3.15 occurred. Last, the related antifungal compounds were predicted to be epicatechin and benzophenone from the LC-MS/MS analysis of crude EA extract. Accordingly, the biocontrol agent B. subtilis strain W3.15 promises a strong potency for biofungicide development.
Antioxidant Properties of Active Fraction Extract Derived from Yellow-Red Pigment Produced by the Marine Sponge-Associated Bacterium Bacillus haikouensis AGS112 and Identification of Related Compounds Uci Cahlia; Rika Indri Astuti; Jun Nomura; Aris Tri Wahyudi
HAYATI Journal of Biosciences Vol. 30 No. 5 (2023): September 2023
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.30.5.874-884

Abstract

Sponge-associated bacteria can produce bioactive compounds similar to the host. Here, the investigation of antioxidant properties of the yellow-red pigment produced by sponge-associated bacterium AGS112 was conducted using in vitro and in vivo analysis. The 16S rRNA gene sequence showed the sponge-associated bacterium AGS112 has the closest similarity with Bacillus haikouensis C-89 (99%). The crude pigment extract produced by Bacillus haikouensis AGS112 contained both phenolic and flavonoid. The peaks occurred at 412 nm and 664 nm, which indicated as the presence of carotenoids and flavonoids, respectively. The active fraction extract obtained through bio-autography TLC, and had more potent antioxidant activity against DPPH (2,2-diphenyl-1-picrylhydrazyl) compared to the crude pigment extract with an IC50 value of 68.62±0.59 µg/ml and 198.88±1.66 µg/ml, respectively. Moreover, the active fraction extract at the concentration of 35 µg/ml could better enhance the viability of Schizosaccharomyces pombe and achieve the highest expression of sod1 and ctt1 genes. Metabolite profiling using LC-MS analysis confirmed the active fraction extract contains eudesmin and artelastin that might be contributed as antioxidants. These results suggest that the active fraction extract of the yellow-red pigment produced by B. haikouensis AGS112 had potency to be used as candidate for natural antioxidants.
Co-Authors Abdjad Asih Nawangsih Abdjad Asih Nawangsih ABDUL MUNIF ABDUL MUNIF Akhmad Endang Zainal Hasan Alimuddin Alimuddin ALINA AKHDIYA Alina Akhdiya ANDINI PURNAWIJAYA Anja Meryandini Anja Meryandini Antonius Suwanto Ari Fina Bintarti Ari Fina Bintarti, Ari Fina ARI SUSILOWATI Aris Tjahjoleksono BRAMANTYO JATI PRASOJO Budi Tjahjono C Hanny Wijaya DIAH ISKANDRIATI Dina Aribah DINI NURDIANI Edi Husen EDI HUSEN Edi Husen EDI HUSEN Edi Husen Engelhaupt, Martin ERNIN HIDAYATI Hamim Hamim Hari KAPLI Hermawaty Abubakar Iman Rusmana Irmawati Jun Nomura LAKSMI AMBARSARI Latifah Kosim Darusman Maggy Thenawidjaja Suhartono Maggy Thenawidjaya Suhartono Marini Wijayanti Meliah, Siti Mia Setiawati MONA PRIMANITA MUHAMMAD AGUS SUPRAYUDI Muhammad Faiz Amri Muhammad Zairin Jr MUNTI YUHANA Mutiha Panjaitan Ni Putu Ratna Ayu Krishanti NI PUTU RATNA AYU KRISHANTI NISA RACHMANIA MUBARIK NURFITRIANI RINA Pamungkas, Joko Puspitasari, Esti Raden Ajie Syahbarie RAHAYU FITRIANI WANGSA PUTRIE RAHAYU WIDYASTUTI Rasti Saraswati RASTI SARASWATI Rasti Saraswati Rika Indri Astuti Rini, Adityawati Fajar Rury Eryna Putri Rustam, Yepy Hardi Sarah Asih Faulina Sarah Asih Faulina, Sarah Asih Satya, Andreas Adhi Sigit Tri Wibowo Sipriyadi Sipriyadi Sipriyadi Siska Tridesianti Siti Meliah Siti Sholekha Sri Budiarti Suryo Wiyono SUSILOWATI1 SUSILOWATI1 SYAMSUL BAHRI SYAMSUL BAHRI Tati Barus TEDJA IMAS Uci Cahlia Umi Fatmawati VINCENTIUS ARCA TESTAMENTI WAODE MUNAENI Wati, Cheppy WIDANARNI WIDANARNI Wiraswati, Sri Martina Yohanes Bernadino Putera Saju Yuli Siti Fatma YULIN LESTARI