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Jurnal Bioteknologi & Biosains Indonesia (JBBI)
Published by Badan Pengkajian dan Penerapan Teknologi
ISSN : 24422606     EISSN : 2548611X     DOI : -
Core Subject : Science, Agriculture,
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Immunology & microbiology
JBBI, Indonesian Journal of Biotechnology & Bioscience, is published twice annually and provide scientific publication medium for researchers, engineers, practitioners, academicians, and observers in the field related to biotechnology and bioscience. This journal accepts original papers, review articles, case studies, and short communications. The articles published are peer-reviewed by no less than two referees and cover various biotechnology subjects related to the field of agriculture, industry, health, environment, as well as life sciences in general. Initiated at the then Biotech Centre, the journal is published by the Laboratory for Biotechnology, the Agency for the Assessment and Application of Technology, BPPT.
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Front Cover JBBI Vol 3, No 2, December 2016 Catur Sriherwanto
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 3 No. 2 (2016): December 2016
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (900.646 KB) | DOI: 10.29122/jbbi.v3i2.1512

Abstract

Appendix JBBI Vol 3, No 2, December 2016: Keyword Index and Author Index Catur Sriherwanto
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 3 No. 2 (2016): December 2016
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (349.522 KB) | DOI: 10.29122/jbbi.v3i2.1513

Abstract

Appendix JBBI Vol 3, No 1, June 2016: Keyword Index and Author Index Catur Sriherwanto
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 3 No. 1 (2016): June 2016
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (350.169 KB) | DOI: 10.29122/jbbi.v3i1.1514

Abstract

OPTIMASI PROSES UNTUK EKSPRESI GEN ENDOGLUKANASE DARI Bacillus sp. RP1 OLEH Escherichia coli BL21 (DE3)/ egc Hans Victor; Maelita Ramdani Moeis
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 5 No. 1 (2018): June 2018
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1284.125 KB) | DOI: 10.29122/jbbi.v5i1.1769

Abstract

Process Optimization for Endoglucanase Gene Expression Derived from Bacillus sp. RP1 by Escherichia coli BL21 (DE3)/egcABSTRACTCellulases are one of the most used enzymes in industrial processes. In an effort to increase production, industries have developed strategies such as isolating new cellulase producing strains, genetic engineering and process optimization since the last 50 years. One endoglucanase producing strain, Bacillus sp. RP1 was isolated from hot springs. The ribosome binding site and coding sequence of the endoglucanase gene (egc) from Bacillus sp. RP1 was cloned into pGEM-T Easy. The recombinant plasmid was used to transform E. coli BL21 (DE3). Cloning was followed by process optimization. Medium composition was selected using Plackett-Burman design. The medium components tested were rice hull, molasses, ammonium chloride, urea and fishmeal. Rice hull and molasses were found to be the factors most influencing enzyme activity and dry cell weight, respectively. The next step involved Box-Behnken method and response surface methodology to optimize the responses against molasses concentration, rice hull concentration and fermentation time. The concentration intervals used to test were 1%, 5.5% and 10% while the fermentation time used were 24, 36 and 48 hours. The conditions which optimized both enzyme activity and dry cell weight were 7.45% molasses, 6.45% rice hull and 39.52 hours of fermentation.Keywords: Bacillus sp. RP1, E. coli BL21 (DE3), egc, Endoglucanase, optimization ABSTRAKSelulase adalah salah satu enzim yang banyak dimanfaatkan dalam berbagai industri. Sebagai upaya untuk memenuhi kebutuhan, 50 tahun terakhir dikembangkan beberapa strategi untuk meningkatkan produksi selulase yang mencakup rekayasa genetika dan optimasi proses. Karena itu, dilakukan kloning gen egc dan RBS yang berasal dari Bacillus sp. RP1 yang diisolasi dari sumber air panas ke dalam vektor pGEM-T Easy. E. coli BL21 (DE3) ditransformasikan dengan vektor yang mengandung gen egc tersebut. Setelah kloning, optimasi proses berupa desain medium turut dilakukan untuk mengoptimalkan ekspresi gen egc. Desain medium diawali dengan seleksi komposisi medium menggunakan metode Plackett-Burman. Komponen medium yang diuji adalah kulit beras, molase, amonium klorida, urea dan tepung ikan. Kulit beras dan molase diperoleh sebagai bahan yang paling berpengaruh terhadap aktivitas enzim dan berat kering sel. Tahap selanjutnya melibatkan metode statistik Box-Behnken dan metodologi respons permukaan yang bertujuan mengoptimalkan respons aktivitas enzim dan berat kering sel terhadap konsentrasi molase, konsentrasi kulit beras dan lama fermentasi. Konsentrasi yang diuji adalah 1%, 5,5% dan 10%, sedangkan lama fermentasi yang diuji adalah 24, 36 dan 48 jam. Konsentrasi optimal molase adalah 7,45% dan konsentrasi optimal kulit beras adalah 6,45% dengan lama fermentasi optimal 39,52 jam.Kata Kunci: Bacillus sp. RP1, E. coli BL21 (DE3), egc, Endoglukanase, optimasi
GROWTH OF NILE TILAPIA (Oreochormis niloticus) FRY FED WITH COCONUT TESTA-CASSAVA BAGASSE MIXED SUBSTRATE FERMENTED USING Rhizopus oryzae Yudha Wali Pradana; Catur Sriherwanto; Etyn Yunita; Imam Suja’i
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 4 No. 1 (2017): June 2017
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (874.114 KB) | DOI: 10.29122/jbbi.v4i1.1799

Abstract

Utilization of agroindustrial byproduct as cheap raw materials for aquafeed was hampered by its poor nutritional value as well as high antinutrition content which could be overcome through fermentation. Coconut testa (CT) and cassava bagasse (CB) were mixed, and fermented using Rhizopus oryzae for preparing aquafeed. Subsequent feeding test was carried out on tilapias (Oreochormis niloticus L.) using 5 feeding treatments: one unfermented feed (commercial feed 100%), and the other four feeds produced by fermentation using substrate mixture of CT and CB in 4 different ratios, namely 100%:0%, 75%:25%, 50%:50%, and 25%:75%. Feeding 100% commercial feed (true protein 22.64% and crude fibre 14.67%) showed the best results on the fish growth with body weight gain of 3.96 g and feed conversion ratio of 8.63. Meanwhile, feeding fermented feeds (true protein 7.96-20.27% and crude fiber 14.14-18.47%) resulted in body weight gain in the range of 2.22 to 2.75 g with feed conversion ratio of 10.89 to 13.62. Thus, the fermented feeds promoted growth in tested tilapias albeit less optimally than commercial feed did.Keywords: Rhizopus oryzae, Oreochromis niloticus, coconut testa, cassava bagasse, fermentation ABSTRAKPenggunaan hasil samping agroindustri sebagai bahan pakan murah ikan terkendala rendahnya nutrisi dan tingginya antinutrisi yang dapat diatasi melalui fermentasi. Dalam penelitian ini, kulit daging buah kelapa (KK) dan onggok singkong (OS) dicampur dengan perbandingan tertentu, lalu difermentasi menggunakan Rhizopus oryzae untuk pakan ikan. Uji pemberian pakan dilakukan terhadap ikan nila (Oreochormis niloticus L.) dengan 5 perlakuan: satu perlakuan pakan tanpa fermentasi (pakan komersial 100%), dan empat perlakuan pakan fermentasi substrat campuran KK dan OS dengan 4 perbandingan yang berbeda, yakni 100%:0%, 75%:25%, 50%:50%, dan 25%:75%. Pemberian pakan komersial 100% (protein sejati 22,64% dan serat kasar 14,67%) memperlihatkan hasil terbaik pada pertumbuhan ikan nila dengan pertambahan bobot badan 3,96 g dan rasio konversi pakan 8,63. Sebaliknya, pemberian pakan fermentasi (protein sejati berkisar 7,96-20,27% dan serat kasar 14,14-18,47%) menghasilkan pertambahan bobot badan ikan pada kisaran 2,22-2,75 g dengan rasio konversi pakan 10,89-13,62. Dengan demikian pakan fermentasi tersebut mendorong pertumbuhan ikan nila namun kurang optimal dibandingkan pakan komersial.Kata Kunci: Rhizopus oryzae, Oreochromis niloticus, kulit daging buah kelapa, onggok, fermentasi
PENINGKATAN PRODUKSI SEFALOSPORIN C DARI Acremonium chrysogenum CB2/11/1.10.6 DENGAN OPTIMASI MEDIA MENGGUNAKAN METODE RESPON PERMUKAAN Erwahyuni Prabandari; Dyah Noor Hidayati; Diana Dewi; Eni Dwi Islamiati; Khaswar Syamsu
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 4 No. 1 (2017): June 2017
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (816.471 KB) | DOI: 10.29122/jbbi.v4i1.1808

Abstract

Cephalosporin is a β-lactam antibiotic produced by Acremonium chrysogenum using submerged fermentation. Carbon and nitrogen are the most influential medium ingredients for cephalosporin formation. The purpose of this study was to obtain the best composition of media for cephalosporin C production. Response surface methodology was used for production optimization. The results showed that molasses of 70 g/Lwas the best carbon source, while the best nitrogen source was the combination of corn steep liquor, urea and ammonium sulphate. DL-methionine, carbon, and nitrogen source significantly affected  the production of cephalosporin C. The mathematically modelled optimization showed that the highest production of cephalosporin C (3876 mg/L) was obtained using medium composition of 68.28 g/L molasses, 71.61 g/L nitrogen, and 0.4 g/L DL-methionine. Laboratory verification using the same medium composition produced 3696 mg/L of cephalosporin C, being 4.65% different from the mathematically optimized results. Medium optimization increased the cephalosprin C production which was 1.48 times higher than that using the previous medium, where the maximum production was only 2487 mg / L.Keywords: Carbon,  cephalosporin C, cultivation medium, nitrogen, A. chrysogenum ABSTRAKSefalosporin C adalah golongan antibiotik β-lactam yang dihasilkan Acremonium chrysogenum melalui fermentasi cair. Komponen yang sangat berpengaruh terhadap produksi sefalosporin C adalah sumber karbon dan nitrogen. Penelitian ini bertujuan mendapatkan komposisi media terbaik untuk produksi sefalosporin C. Optimasi dilakukan menggunakan metode respon permukaan. Hasil menunjukkan bahwa molases 70 g/L adalah sumber karbon terbaik dan kombinasi corn steep liquor, urea dan ammonium sulfat adalah sumber nitrogen terbaik. DL-methionin, sumber karbon, dan nitrogen berpengaruh nyata terhadap produksi sefalosporin C. Optimasi menggunakan model matematika menunjukkan produksi sefalosporin C tertinggi (3876 mg/L) yang diperoleh dengan komposisi media 68,28 g/L molases, 71,61 g/L nitrogen, dan  0,4 g/L DL-methionin. Verfikasi di laboratorium menggunakan komposisi media yang sama menghasilkan sefalosporin C sebesar 3696 mg/L, berbeda 4,65% dibanding dengan hasil optimasi matematis. Optimasi media mampu meningkatkan produksi sefalosprin C sebesar 1,48 kali dibanding media yang digunakan sebelumnya, dimana maksimal hanya menghasilkan 2487 mg/L.Kata kunci: Karbon, sefalosporin C, media kultivasi, nitrogen, A. chrysogenum
PRODUKSI REKOMBINAN SEFALOSPORIN ASILASE SEBAGAI BIOKATALIS UNTUK PRODUKSI ASAM 7-AMINOSEFALOSPORANAT Bima Wedana Isdiyono; Dudi Hardianto; Fransiskus Xaverius Ivan
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 4 No. 1 (2017): June 2017
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1087.024 KB) | DOI: 10.29122/jbbi.v4i1.2059

Abstract

Production of Cephalosporin Acylase Recombinant as Biocatalyst for 7-Aminocephalosporanic Acid Production7-aminocephalosporanic acid (7-ACA) is a precursor for the production of semisynthetic cephalosporin derivatives. The enzymatic 7-ACA production can use two-stage and one-step enzymatic methods. Two-stage enzymatic method uses D-amino acid oxidase (DAAO) enzyme to produce glutaryl-7-aminocephalosporanic acid (GL-7-ACA) in the first stage and glutaryl-7-aminocephalosporanic acid acylase to produce 7-ACA in the second stage. The one-stage enzymatic method using cephalosporin acylase (CPC acylase) converts the CPC to 7-ACA directly. The aim of this research was to produce recombinant CPC acylase in Escherichia coli BL21(DE3). Transformantion culture E. coli BL21(DE3) was induced with concentrations of IPTG 0; 0.25; 0.5; 0.75; 1; 2 mM for 5 hours. The induction time of IPTG was determined at 0, 1, 2, 3, 4, and 5 hours. The results showed that CPC acylase produced by E. coli BL21(DE3) with optimum condition of CPC acylase production was 0.5 mM IPTG and optimal induction time of IPTG was 5 hours.Keywords: Cephalosporin, cephalosporin acylase, 7-ACA, protein expression, Escherichia coli BL21(DE3) ABSTRAKAsam 7-aminosefalosporanat (7-ACA) merupakan prekursor untuk produksi turunan sefalosporin semisintetik. Produksi 7-ACA secara enzimatik dapat menggunakan metode dua tahap dan satu tahap enzimatik. Metode enzimatik secara dua tahap menggunakan enzim asam D-amino oksidase (DAAO) untuk menghasilkan asam glutaril-7-aminosefalosporinat (GL-7-ACA) pada tahap pertama dan menggunakan asam glutaril-7-aminosefalosporinat asilase untuk menghasilkan 7-ACA pada tahap kedua. Metode enzimatik satu tahap dengan sefalosporin asilase (CPC asilase) mengubah CPC menjadi 7-ACA secara langsung. Tujuan penelitian adalah memproduksi rekombinan CPC asilase di dalam sel Escherichia coli BL21(DE3). Kultur Transforman E. coli BL21(DE3) diinduksi dengan konsentrasi IPTG 0; 0,25; 0,5; 0,75; 1; 2 mM selama 5 jam. Waktu induksi IPTG ditentukan pada 0, 1, 2, 3, 4 dan 5 jam. Hasil penelitian menunjukan bahwa CPC asilase diproduksi oleh E. coli BL21(DE3) dengan kondisi optimal produksi CPC asilase adalah konsentrasi IPTG 0,5 mM dan waktu induksi IPTG optimal adalah 5 jam.
PENGARUH KOMBINASI BAKTERI ASAM LAKTAT TERHADAP PERUBAHAN KARAKTERISTIK NUTRISI SUSU KERBAU Rofiq Sunaryanto
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 4 No. 1 (2017): June 2017
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (573.366 KB) | DOI: 10.29122/jbbi.v4i1.2064

Abstract

Effects of Lactic Acid Bacteria Combination on Characteristic Change of Buffalo Milk Nutrition Determination of physical and chemical characteristics on fermented milk using a mixture of Lactobacillus bulgaricus, Streptococcus thermophilus, and Bifidobacterium sp. has been conducted. Fermentation was carried out for 20 hours at 37ºC in a facultative aerobic condition. Physical characteristics were performed by comparing the physical properties before and after fermentation such as viscosity and texture, including chemical properties such as pH, acidity, protein, fat, and sugar contents. Inoculants of lactic acid bacteria were varied using a combination of lactic acid bacteria mixture. Results showed that the combination of bacteria inoculants used has no significant effect on changes in protein, fat, ash, and water contents. However, the effects on total acid, pH, sugar content, and physical appearance such as viscosity and texture were significant. The combination of L. bulgaricus, S. thermophilus and Bifidobacterium sp. (A6) produced a soft texture appearance and high viscosity. The combination of inoculant bacteria L. bulgaricus with S. thermophilus (A5) produced a texture similar to A6 but with a lower viscosity than A6.Keywords: Lactobacillus bulgaricus, Streptococcus thermophilus, Bifidobacterium sp, buffalo milk, fermentation. ABSTRAKPenelitian untuk mengetahui karakteristik fisik dan kimia hasil fermentasi susu kerbau menggunakan campuran Lactobacillus bulgaricus, Streptococcus thermophilus, dan Bifidobacterium sp telah dilakukan. Fermentasi dilakukan selama 20 jam pada suhu 37ºC secara aerobe fakultatif. Karakteristik fisika dilakukan dengan membandingkan sifat fisik seperti kekentalan dan tekstur, serta sifat kimia yang meliputi kandungan protein, lemak, gula, pH, keasaman sebelum dan sesudah fermentasi. Inokulan bakteri asam laktat divariasikan menggunakan kombinasi campuran bakteri asam laktat. Hasil penelitian menunjukkan bahwa kombinasi inokulan yang digunakan dalam proses fermentasi susu kerbau tidak berpengaruh nyata terhadap perubahan kadar protein, lemak, abu, dan air. Namun demikian berpengaruh nyata terhadap total asam, pH, kadar gula, dan penampakan fisik seperti kekentalan dan tekstur. Kombinasi inokulan L. bulgaricus, S. thermophilus dan Bifidobacterium sp. (A6) menghasilkan susu fermentasi dengan penampakan tekstur halus dan kekentalan yang lebih padat. Kombinasi bakteri L. bulgaricus dengan  S. thermophilus (A5) menghasilkan tekstur yang mirip dengan A6, namun demikian menghasilkan kekentalan yang lebih rendah dibandingkan A6.Kata kunci: Lactobacillus bulgaricus, Streptococcus thermophilus, Bifidobacterium sp, susu  kerbau, fermentasi
IMPROVING THE FUNCTION OF CRISPR-CAS9 FOR GENOME EDITING THERAPY: EDITING THE EDITOR Alva Sahiri Alexander Supit
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 4 No. 1 (2017): June 2017
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (831.041 KB) | DOI: 10.29122/jbbi.v4i1.2068

Abstract

Meningkatkan Fungsi CRISPR-Cas9 untuk Terapi Pengeditan GenPengeditan gen menjadi mudah dilakukan sejak ditemukannya clustered regularly interspaced short palindromic repeat (CRISPR) dan CRISPR-associated protein 9 (Cas9) sebagai alat untuk menyunting gen suatu organisme. Sebagian besar penyakit genetik tidak dapat disembuhkan secara kausal dengan terapi yang ada, maka pengeditan gen merupakan suatu cara yang prospektif dalam terapi medis di masa depan. Sayangnya, pengeditan gen dengan Cas9 yang ada saat ini masih memiliki banyak kelemahan, yaitu: 1) kurang spesifik, di mana RNA pemandu dapat berikatan dengan beberapa segmen pada genom manusia, sehingga memungkinkan terjadinya salah target; 2) kurang efisien, karena sekalipun telah berhasil memotong utas ganda DNA, kebanyakan penyambungan kembali akan dilakukan secara non-homology end joining (NHEJ), yang justru meningkatkan peluang terjadinya mutasi; 3) sulit disalurkan ke dalam inti sel karena berbagai sawar fisiologis maupun biokimiawi. Tulisan ini akan membahas perkembangan terkini dalam mengatasi ketiga masalah di atas. Untuk meningkatkan spesifisitas, dapat dilakukan modifikasi RNA pemandu dan struktur Cas9. Efisiensi dapat ditingkatkan dengan meningkatkan peluang terjadinya homology-directed repair dibandingkan NHEJ, sedangkan untuk meningkatkan distribusi ke dalam sel, dapat digunakan berbagai macam vektor, seperti virus dan nanopartikel. CRISPR-Cas9 merupakan area yang aktif diteliti dalam bidang biosains, dan dalam waktu dekat, diharapkan dapat dimanfaatkan dalam bidang klinik.Kata kunci: CRISPR, Cas9, efektivitas, spesifisitas, terapi genABSTRACTGene editing has become reasonably easy since the discovery of clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR-associated protein 9 (Cas9). Most genetic diseases cannot be treated causally, and currently available therapies are mainly symptom-based. To treat the etiology of genetic diseases, a firm gene editing therapy is necessary to be established. This posits Cas9-facilitated gene editing as a prospective modality to become a clinically approved therapy in the future to treat genetic disorders. However, until recently, Cas9-based genome editing is still facing several hurdles, including low specificity, low effectiveness, and difficult delivery. Currently available Cas9 nucleases are able to bind to non-specific DNA sequence and produce non-specific cleavage. The efficiency has been relatively low due to the preference of non-homologous end-joining (NHEJ) over homology-directed repair (HDR) by the host cell. Furthermore, in order to deliver Cas9 into the nucleus, multiple physiological barriers have to be overcome. This review discussed recent developments in tackling these three hurdles, ranging from designing the guide RNA using multiple bioinformatics tools, modifying Cas9 structure, as well as packaging the nuclease-guide RNA complex into viral vectors and nanoparticles. Considering the active research on this area, it is expected that CRISPR/Cas9 can be utilized as a clinical therapy in the near future.Received: 02 June 2017        Accepted: 07 July 2017        Published: 19 July 2017
PENGARUH BEBERAPA JENIS SITOKININ PADA MULTIPLIKASI TUNAS ANGGREK Vanda douglas SECARA IN VITRO Karyanti Karyanti
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 4 No. 1 (2017): June 2017
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (792.84 KB) | DOI: 10.29122/jbbi.v4i1.2200

Abstract

The Effect of Several Types of Cytokinin on Shoot Multiplication of Vanda douglas Orchid In VitroThe study was aimed to determine the response of Vanda douglas orchid on shoot-multiplication media to different cytokinin concentrations in vitro. A completely randomized design experiment was employed with one factor cytokinin, in which the cytokinins used were TDZ (thidiazuron), BAP (6-Benzylaminopurine) and kinetin at the concentrations of 0, 0.5, 1, and 1.5 mg/L. The results showed that kinetin 0.5 mg/L was the best concentration for shoot formation, occuring on average at 14.88 days after planting; while TDZ 0.5 mg/L was the best concentration for increasing the height of the plant, being on average 0.53 cm. TDZ at 0.5 mg/L concentration also had positive effect on shoot and leaf formation, which resulted in the highest number of shoots and leaves. The average number of shoots was 8.00 buds, and the average number of leaves was 12.25 sheets. Keywords: Vanda douglas, thidiazuron, BAP, kinetin, shoots multiplication ABSTRAKPenelitian ini bertujuan untuk mengetahui respon anggrek Vanda douglas terhadap media perbanyakan tunas pada beberapa konsentrasi jenis sitokinin secara in vitro. Penelitian ini menggunakan Rancangan Acak Lengkap (RAL) dengan satu faktor, yaitu sitokinin. Sitokinin yang digunakan adalah TDZ (thidiazuron), BAP (6-Benzylaminopurine) dan kinetin, dengan konsentrasi 0, 0,5, 1, dan 1,5 mg/L. Hasil penelitian menunjukkan bahwa pemberian kinetin 0,5 mg/L menunjukkan hasil terbaik pada peubah waktu pembentukan tunas, dengan rata-rata 14,88 hari setelah tanam. Sedangkan konsentrasi TDZ 0,5 mg/L merupakan jenis sitokinin dan konsentrasi terbaik terhadap pertambahan tinggi tanaman, dengan rata-rata 0,53 cm. TDZ dengan konsentrasi 0,5 mg/L juga berpengaruh positif terhadap pertumbuhan tunas dan daun, dengan menghasilkan jumlah tunas tertinggi dan jumlah daun terbanyak. Rata-rata jumlah tunas adalah 8,00 tunas, dan rata-rata jumlah daun adalah 12,25 helai.Kata Kunci: Vanda douglas, thidiazuron, BAP, kinetin, perbanyakan tunasReceived: 19 June 2017        Accepted: 02 July 2017        Published: 12 July 2017

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