Asmini Budiani
Balai Penelitian Bioteknologi Perkebunan Indonesia

Published : 46 Documents
Articles

Embriogenesis somatik langsung dan regenerasi planlet kopi arabika (Coffea arabica) dari berbagai eksplan Direct somatic embryogenesis and regeneration of arabica coffee plantlets (Coffea arabica) from different explants OKTAVIA, Fetrina; SWANTO, .; BUDIANI, Asmini
E-Journal Menara Perkebunan Vol 71, No 2: Desember 2003
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (2082.995 KB) | DOI: 10.22302/iribb.jur.mp.v71i2.161

Abstract

SummaryTissue culture technique for arabica coffeefaces some problems, mainly in plantletsregeneration from cultured explants. Theobjectives of this experiment were to examine theeffect 2,4-D and 2-ip combinations on somaticembryogenesis and regeneration of arabicacoffee from several different explants. Basalmedium used in this experiment was MS mediumwith ½ concentration of macro and micro salts.Experiment to induce primary somatic embryos(SE) was arranged in factorial randomizedcomplete design with 10 repeats. The first factorwas the type of explants, leaf, epicotyl, hipocotyland root explants. The second factor was plantgrowth regulator i.e. combination of 1  M 2,4-Dwith 5, 10, 15, 20  M and combination of 5  M2,4-D with 5, 10, 15 and 20  M 2-ip. To multiplySE, secondary SE was induced from primary SEon medium containing combination of 0.6  MIAA and 13.3; 17.8 and 22.2  M BAP.Cotyledonary SE were germinated on mediacontaining GA 3 (0, 5, 10 and 15  M), and thenregenerated on medium free of growth regulator.Plantlets with 4-5 leaf pairs were transfered intothe soil medium for acclimatization. The resultsshow that primary SE can be induced from allexplants with the highest frequency on mediumcontaining 1  M 2,4-D and 15  M 2-ip.Induction of primary SE, in leaf explant wasmore effective than other explants. Mediumcontaining 0.6  M IAA and 22.2  M BAP gavethe highest percentage of SE multiplication i.e.52.6% with average SE number of 6.25. Plantletsregeneration can be conducted by culturing SEon maturation medium free of growth regulatorfor one month followed by germinating onmedium containing GA 3 , and then culturing onmedium free of growth regulator again. Thehighest percentage of germinated embryos wasobtained after three weeks and six weekscultured in the medium containing 5  M GA 3 , i.e49% and 90.15 respectively. From total plantletsobtained, 75% of them were normal. Sixtypercents of the young plants grew well in thegreenhouse.RingkasanTeknik kultur jaringan tanaman kopi arabikamasih menghadapi beberapa kendala terutamapada tingkat regenerasi planlet dari eksplan yangdikulturkan. Penelitian ini bertujuan untukmengetahui pengaruh kombinasi 2,4-D dan 2-ipterhadap embriogenesis somatik dan regenerasikopi arabika dari berbagai eksplan. Media dasaryang digunakan adalah medium MS ½konsentrasi garam makro dan mikro. Percobaaninduksi embrio somatik (ES) primer disusunmenurut rancangan acak lengkap faktorial dengan10 ulangan. Faktor pertama adalah jenis eksplan,erdiri atas daun, epikotil, hipokotil dan akar invitro. Faktor kedua adalah zat pengatur tumbuh,yaitu kombinasi 1 M 2,4-D dengan 5, 10, 15dan 20M 2-ip, serta kombinasi 5 M 2,4-Ddengan 5, 10, 15 dan 20 M 2-ip. Untuk mem-perbanyak jumlah ES yang didapatkan, dilakukaninduksi ES sekunder dari ES primer pada mediumyang mengandung kombinasi 0,6 M IAA dan13,3; 17,8 dan 22,2 M BAP. ES fase kotiledonkemudian dikecambahkan pada medium yangmengandung GA 3 (0, 5, 10 dan 15 M) danselanjutnya diregenerasikan pada medium tanpazat pengatur tumbuh. Planlet yang mempunyai4-5 pasang daun dipindahkan ke medium tanahuntuk aklimatisasi. Hasil yang diperolehmenunjukkan bahwa ES primer dapat diinduksipada semua eksplan yang digunakan denganfrekuensi tertinggi pada medium yang me-ngandung 1 M 2,4-D dan 15 M 2-ip. InduksiES primer pada eksplan daun lebih efektifdibandingkan eksplan lainnya. Untuk per-banyakan ES, medium yang mengandung IAA0,6 M dan BAP 22,2 M memberikanpersentase tertinggi pembentukan ES sekunderyaitu 52,6% dengan rata-rata jumlah ES 6,25.Regenerasi planlet dapat dilakukan denganmengkulturkan ES pada medium maturasi tanpazat pengatur tumbuh selama satu bulan, kemudiandikecambahkan dalam medium yang mengan-dung GA 3 , dan selanjutnya dipindah ke mediumtanpa zat pengatur tumbuh kembali.Perkecambahan ES tertinggi diperoleh padamedium dengan penambahan GA 3 5 M yaitu40,9% setelah tiga minggu dan 90,1% setelahenam minggu. Dari total planlet diperoleh 75%planlet normal. Hasil aklimatisasi menunjukkanbahwa 60% bibit mampu bertahan di rumah kaca.
Purification, characterization, and bioassay of putative protease inhibitors from Hevea brasiliensis latex PUTRANTO, Riza Arief; SISWANTO, .; MULYATNI, Agustin Sri; BUDIANI, Asmini; TISTAMA, Radite
E-Journal Menara Perkebunan Vol 84, No 2 (2016): Desember 2016
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1452.514 KB) | DOI: 10.22302/iribb.jur.mp.v84i2.220

Abstract

Lateks yang menyerupai cairan susu putih diperoleh dari penyadapan kulit batang tanaman karet (Hevea brasiliensis). Lateks merupakan sitoplasma dari jaringan pembuluh bernama latisifer yang didalamnya terkandung berbagai macam komponen, termasuk protein-protein penting. Berbagai jenis enzim yang memiliki fungsi terkait pertahanan tanaman dari serangan patogen dan pelukaan telah berhasil dideteksi di dalam lateks, di antaranya protease inhibitor (PI). Protease inhibitor memiliki aktivitas senyawa antifungi sehingga berpotensi untuk  dimanfaatkan sebagai biofungisida. Pada penelitian ini, protease  inhibitor putatif yang berasal dari serum B (lutoid) lateks tanaman karet telah berhasil diisolasi menggunakan teknik Ion Exchange Chroma-tography. Dari total 70 fraksi protein yang diekstrak dari kolom, hanya 26 fraksi yang menunjukkan kadar protein yang terukur. Kandungan protease inhibitor putatif yang di-peroleh berkisar antara 0,0067 hingga 0,022 mL/g serum B dari hasil 3 fraksi terpilih. Aktivitas penghambatan terhadap empat enzim protease (subtilisin A, tripsin, α-kimotripsin, dan papain) menunjukkan karakteristik protease inhibitor putatif tersebut sebagai serine dan/atau cysteine inhibitor protease dengan persentase hambatan di atas 15% terhadap protease target. Hasil SDS-PAGE memperlihatkan pemisahan protein dominan yang diperkirakan merupakan protease inhibitor putatif dengan berat molekul sebesar 21,5 kDa. Uji bioassay aktivitas antifungi secara in vitro dari protease inhibitor memperlihatkan penghambatan pertumbuhan miselium dari fungi Ganoderma boninense, Sclerotium sp., dan Rigidosporus lignosus. [Kata kunci : protease inhibitor, Hevea brasiliensis, lateks, serum B, ion exchange chromatography]AbstractLatex, a milky white liquid, is the main product from rubber tree (Hevea brasiliensis). Latex is the cytoplasm of complex cellular networks named laticifers in which it contains many different components, including important proteins. Various types of enzymes carrying functions associated with plant defense against pathogen and wounding have been detected in latex in which one of these enzymes is protease inhibitor (PI). Plant protease inhibitor has tremendous potential as an antifungal agent which can be developed as biofungicide. In this work, protease inhibitors from B-serum (lutoid) of rubber tree latex were isolated and purified using Ion Exchange Chromatography (IEC) technique. Of the total 70 fractions of proteins extracted from the columns, only 26 fractions showed measurable levels of protein. The concentration of obtained putative protease inhibitors (three fractions of IEC) ranged from 0.007 to 0.022 mL/g B-serum. Inhibitory activity against four protease enzymes (subtilisin A, trypsin, α-chymotrypsin, and papain) showed the characteristics of Hevea putative protease inhibitors from B-serum as serine and/or cysteine protease inhibitors with more than 15% inhibitory activity of target protease. Based on SDS-PAGE visualization, the molecular weight of dominant protein considered as Hevea putative protease inhibitors was 21.5 kDa. In vitro bioassay test of antifungal activity for Hevea putative protease inhibitors showed reduced mycelium growth of Ganoderma boninense, Sclerotium sp., and Rigidosporus lignosus.[Keywords: protease inhibitor, Hevea brasiliensis, latex, B-serum, ion exchange chromatography]
Dinamika populasi Trichoderma harzianum DT38 pada campuran arang hayati tandan kosong kelapa sawit (TKKS) dan gambut Population dinamic of Trichoderma harzianum DT38 on mixture of empty fruit bunches of oil palm (EFBOP) biochar and peat KRESNAWATY, Irma; SUHADA, Sayhas; BUDIANI, Asmini; DARMONO, T W
E-Journal Menara Perkebunan Vol 80, No 1: Juni 2012
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (281.329 KB) | DOI: 10.22302/iribb.jur.mp.v80i1.45

Abstract

Abstract Biochar offers option for managing land as a source  of carbon and soil conditioner. The ability of biochar in  increasing soil  fertility associates with its ability to retain water, reduce soil acidity, and keep the availability of essential nutrients for plant thus increasing crop produc-tivity, and reducing the risk of soil erosion. Biochar is also substance to provide a  suitable environment for the growth of beneficial microbes, including an isolate of  Trichoderma harzianum used in this study, that has been proven capable in stimulating plant growth and suppressing soil borne diseases. The purpose of this research was to determine the in vitro compatibility of T. harzianum DT38, Indonesia Biotech-nology Research Institute for Estate Crop (IBRIEC) collection, in mixtures of  EFBPO biochar and peat during 28 days. This research was performed in completely randomized design with single factor, comprising of five formulas: 1) 100% EFBOP biochar (K1), 2) 100% peat (K2), 3) Mixture of EFBOP biochar and peat = 1 : 4 (F1), 4) Mixture of EFBOP biochar and peat=  1 : 8 (F2), dan 5) Mixture of EFBOP biochar and peat=  1 : 12 (F3). The colony forming units were determined after storage to express the amount of fungal propaguls in each mixture. The results was analized using one-way ANOVA test and Duncan Test. Result showed that the total of  T. harzianum DT38 propaguls was not significantly difference among five mixture preparations tested during 0 and 7 days storage. The total propaguls were insignificantly difference between F1 and K2, and also between  F2 and F3in 14, 21 and 28 days incubation. Peat addition on biochar increased the total of  T. harzianum DT38 propaguls during 28 days incubation. The total propaguls which are remain high in F1, F2 and F3 formula up to 28 days storage indicating that the mixtures suitable for microbe media and biofertilizer formula.Abstrak Penggunaan arang hayati (biochar) merupakan alternatif pengelolaan tanah terutama sebagai penyedia karbon dan pembenah tanah.  Kemampuan biochar dalam meningkatkan kesuburan tanah berhubungan dengan kemampuannya untuk menahan air, mengurangi keasaman tanah, menjaga keter-sediaan nutrien yang penting bagi tanaman sehingga mening-katkan  produktivitas  tanaman, serta mengurangi resiko erosi  tanah. Arang hayati  juga berperan dalam menyediakan ling-kungan yang   cocok   untuk   pertumbuhan   mikroba,  ter-masuk isolat Trichoderma harzianum yang digunakan dalam penelitian ini dan teruji mampu meningkatkan pertumbuhan tanaman dan mengendalikan penyakit tular tanah. Tujuan penelitian ini untuk mengetahui kompatibilitas T. harzianum DT38 koleksi BPBPI pada bahan pembawa berupa campuran biochar tandan kosong kelapa sawit (TKKS) dan gambut selama penyimpanan 28 hari secara in vitro. Penelitian ini menggunakan rancangan acak lengkap (RAL) untuk menguji lima perlakuan, yaitu : 1) 100% biochar TKKS (K1), 2) 100% gambut (K2), 3) Campuran biochar TKKS dan gambut 1 : 4 (F1), 4) Campuran biochar TKKS dan gambut 1 : 8 (F2), dan 5) Campuran biochar TKKS dan gambut 1 : 12 (F3). Hasil pengamatan pada penyimpanan 0 dan 7 hari menunjukkan bahwa jumlah propagul T. harzianum DT38 dari berbagai formula tidak berbeda nyata.  Jumlah propagul antara formula F1 dan K2, serta F2 dan F3 tidak berbeda nyata pada penyim-panan 14, 21 dan 28 hari.  Penambahan gambut pada biochar TKKS dapat meningkatkan jumlah propagul T. harzianum DT 38 selama penyimpanan 28 hari secara in vitro.  Jumlah propagul T. harzianum DT38 pada media F1, F2 dan F3 selama penyimpanan 28 hari masih memenuhi jumlah minimal propagul dalam bahan pembawa yang menunjukkan bahwa media ini sesuai untuk pertumbuhan mikroba dan berpotensi sebagai formula pupuk hayati.
Nucleotide sequence of cryIA gene cloned from Btk isolate of Bacillus thuringiensis and comparison with cryIA(c) gene from B. thuringiensis subsp. kenyae Sekuen nukleotida gen cryIA dari B.thuringiensis isolat Btk dibandingkan dengan gen crylA(c) dari B. thuringiensis subsp. Kenyae BUDIANI, Asmini; SANTOSO, Djoko
E-Journal Menara Perkebunan Vol 68, No 1: Juni 2000
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (54.059 KB) | DOI: 10.22302/iribb.jur.mp.v68i1.134

Abstract

Ringkasan Perakitan tanaman perkebunan yang toleran terhadap serangga hama dapat ditempuh melalui rekayasa genetika menggunakan gen cry. Gen cryIA merupakan gen cry yang paling banyak dipelajari di antara gen cry lainnya. Berdasarkan homology sekuen dan spesifisitas protein yang disandinya terhadap serangga sasaran, gen ini telah diklasifikasikan menjadi 10 subklas. Tulisan ini melaporkan hasil sekuensing (ragmen gen cryIA penyandi domain toksin yang diisolasi dengan teknik PCR dari Bacillus thuringiensis isolat Btk dan diklon menggunakan vektor pGEM­T. Untuk menentukan sekuen gen cryIA yang berukuran sekitar 2 kb tersebut, dilakukan kons­truksi satu seri mutan terdelesi searah dari ujung 5' menggunakan kit Erase-a-Base-System. Tiga DNA gen cryIA mutan dengan tingkat delesi yang sesuai dan satu nonmutan dipilih untuk sekuensing DNAnya. Sekuensing dilakukan dari satu arah menggunakan primer universal SP6 pada alat ABI 377A automatic DNA sequencer. Sekuen lengkap dari gen cryIA diperoleh dengan cara meng­gabungkan sekuen ketiga mutan dengan sekuen dari gen cryIA nonmutan secara manual. Untuk konfirmasi sekuen ujung 3', dilakukan sekuensing dari arah lainnya menggunakan primer universal T7. Sekuen lengkap dari fragmen tersebut mengandung 2021 nukleotida dan menyandi protein dengan 673 asam amino. Dibandingkan dengan sekuen gen crylA(c) dari B. thuringiensis subsp. kenyae, terlihat adanya sepuluh mutasi titik masing-masing pada nukleotida ke 444, 477, 1089, 1092, 1098, 1242, 1566, 1869, 1906 dan 1961. Tujuh mutasi titik pada nukleotida ke 444, 477, 1089, 1092, 1242, 1566, dan 1869 tidak merubah asam amino, sedangkan tiga mutasi lainnya mengakibatkan perubahan asam amino, yaitu pada nukleotida ke 1098 (kodon ke 366, yang menyebabkan perubahan dari Phe menjadi Leu), nukleotida ke 1906 (kodon ke 636, yang mengubah Val menjadi Leu) dan pada nukleotida ke 1961(kodon ke 654, yang mengubah Cys menjadi Tyr).Summary Estate crops tolerant to pests can be devel­opment through genetic engineering using cry gene. CryIA is the best studied among cry genes. Based on the sequence homology and specificity of their encoded proteins to the, targeted insect, these genes have been classified into 10 sub­classes. This paper reports sequencing of cryIA gene fragment en-coding toxin domain isolated from Btk isolates of Bacillus thuringiensis using PCR technique and cloned with pGEM-T vector. To determine the full sequence of the 2-kb gene fragment, a series of mutants uni-directionally deleted at the 5'-end were constructed. Mutation was done using Erase-a Base-System kit. Three DNA mutants with appropriate degree of deletion and the un-mutated DNA were chosen for sequencing. Sequencing was conducted from one direction with SP6 universal primer using the ABI 377A automatic DNA sequencer. The full sequence of cryIA fragment was assembled manually using the sequences of DNA mutants and the non-mutant cryIA fragment. To confirm the sequence of the 3'-end, sequencing from the other direction was performed using the T7 universal primer.The completed sequence of the fragment contains 2021 nucleotides encoding a protein of 673 amino acids. Compares to the sequence of cryIA(c) from B. thuringiensis subsp. kenyae, it was shown that there were ten point mutations (nucleotides of 444, 477, 1089, 1092, 1098, 1242, 1566, 1869, 1906 and 1961), sevent of them (nucleotides of 444, 477, 1089, 1092, 1242, 1566 and 1869) were identified as silent mutations, while the other three substituted the amino acids, which are at the nucleotide 1089 (codon 366, substitution of Leu for Phe), nucleotide 1906 (codon 636, substitution of Leu for Val), and nucleotide 1961 (codon 654, substitution of Tyr for Cys).
Karakterisasi gen penyandi lipase dari kapang Rhizopus oryzae dan Absidia corymbifera Characterization of gene encoding lipase from fungus Rhizopus oryzae and Absidia corymbifera PUTRANTO, Riza A; SANTOSO, Djoko; TRI-PANJI, .; SUHARYANTO, .; BUDIANI, Asmini
E-Journal Menara Perkebunan Vol 74, No 1: Juni 2006
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (471.091 KB) | DOI: 10.22302/iribb.jur.mp.v74i1.118

Abstract

SummaryLipase is a group of enzymes which catalyze fat hydrolysis. Lipase is recently used to produce diacylglycerol (DAG) from triacylglycerol (TAG). Lipase  can be used to produce healthy oil. Having a rich biodiversity, Indonesia has the opportunity to produce lipase using indigenous microbes, such as molds. This research aimed to detect  LIPASE  gene on several strains of molds employing PCR technique. Genomic DNAs were isolated from four strains of molds (M. sitophila, R. oryzae, R. microsporus, and A. corymbifera). Heterologous primers for LIPASE  were designed based on the conserved region of 12 LIPASE  sequences accessed from GenBank and used to amplify the genomic DNA resulted in a 466 bp fragmen. BLAST analysis showed that the bands of DNAs have high homology with common lipase protein in several strains of  Rhizopus.Ringkasan Lipase merupakan kelompok enzim yang berfungsi sebagai biokatalis hidrolisis lemak. Lipase banyak digunakan untuk konversi triasilgliserol (TAG) menjadi diasilgliserol (DAG). Penggunaan lipase penting untuk produksi minyak sehat (healthy oil). Indonesia dengan keanekaragaman hayati tinggi berpeluang besar   mengembangkan   produksi   lipase   dari mikroba lokal, salah satunya adalah kapang. Deteksi gen merupakan langkah awal dalam upaya peningkatan produksi lipase melalui rekayasa genetika. DNA genomik empat galur kapang (M. sitophila, R. oryzae, R. microsporus, dan A. corymbifera) telah berhasil diisolasi. Sepasang primer heterologous telah berhasil dirancang berdasarkan daerah terkonservasi 12 sekuen gen LIPASE dari GenBank. Amplikon DNA yang diperoleh pada PCR menggunakan pasangan primer RLP memiliki panjang 466 bp. Analisis BLAST memperlihatkan bahwa amplikon PCR memiliki homologi yang tinggi dengan protein LIPASE  beberapa galur Rhizopus. 
Aplikasi biokaolin untuk perlindungan buah kakao dari serangan PBK, Helopeltis spp. dan Phytophthora palmivora Application of biokaolin in protecting cocoa pod from cocoa pod borer, Helopeltis spp. and Phytophthora palmivora infestation KRESNAWATY, Irma; BUDIANI, Asmini; WAHAB, Abdul; DARMONO, T W
E-Journal Menara Perkebunan Vol 78, No 1: Juni 2010
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1384.025 KB) | DOI: 10.22302/iribb.jur.mp.v78i1.77

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AbstractMain constraints of cacao cultivation are infestations of cocoa pod borer (Conopomorpha cramerella (Snellen), Helopeltis spp., and cocoa pod rot disease (palmivora). So far there is no technology that could efficiently controlthese important pests. This research was aimed to develop environmentally friendly new technology to protect pod surfaces of cacao. The experiment was performed in heavily infested cacao plantation in Konawe, South-East Sulawesi.The use of kaolin particle film enriched with entomopathogenic microbe was contrasted againts the use of currently recommended plastic sleeve. Cacao pods were sprayed at one week and two week intervals. The observed parameters were the number of pods infested with cocoa pod borer, pod rot and Helopeltis spp. at 4th - 14th weeks after first spray. From the observation, weekly biokaolin application showed the highest amount pods free from cocoa pod borer (33.97 %), followed by biweekly application (27.96 %), and plastic sleeving (19 %). Ten weeks after first spray, cocoa pod borer incidence was significantly reduced especially in weekly application. The percentage of pods free from pod rot were 81.92 %, 62.96 %, and 72.20 % for weekly spray, biweekly spray, and plastic sleeving, respectively. Pods being kept for 12 weeks in plastic sleeve endured the highest intensity of pod rot incidence. Biweekly biokaolin treatment was better in handling Helopeltis spp. attack. Besides reducing infestation of the main pests and diseases, biokaolin application also reduced the incidence of cherelle wilt to almost 40%. Those results gave the great expectation that biokaolin usage would significantly increase cacao yield, resulting in the increase of cacao farmer income. AbstrakKendala utama dalam upaya budidaya kakao adalah adanya serangan hama penggerek buah kakao (PBK), Conopomorpha cramerella (Snellen) dan hama kepik Helopeltis spp., serta serangan patogen penyebab busuk buah (Phythophtora palmivora). Sampai saat ini belum tersedia teknologi yang secara efisien mengendalikan ketiga-tiganya sekaligus. Peneltian ini bertujuan untuk mengembangkan teknologi yang ramah lingkungan untuk melindungi permukaan buah kakao. Percobaan dilakukan pada perkebunan kakao dengan tingkat serangan yang berat di Konawe, Sulawesi Tenggara. Dalam penelitian ini, penggunaan lapisan partikel kaolin yang diperkaya dengan mikroba entomopatogenik dibandingkan efektifitasnya dengan penyarungan menggunakan kantung plastik yang direkomendasikan selama ini. Buah kakao disemprot setiap interval satu minggu dan dua minggu sekali. Parameter yang diamati adalah jumlah buah terserang PBK, jumlah serangan busuk buah dan jumlah serangan Helopeltis spp. pada minggu keempat sampai dengan minggu ke-14 setelah penyemprotan pertama. Hasil pengamatan menunjukkan bahwa persentase tertinggi (33,97 %) buah kakao yang terbebas dari serangan PBK diperoleh pada plot dengan penyemprotan biokaolin setiap minggu, diikuti dengan penyemprotan setiap dua minggu (27,96 %), dan penyelubungan dengan kantung plastik (19,00 %). Pada minggu ke- 10 setelah penyemprotan pertama terjadi penurunan intensitas serangan PBK secara signifikan khususnya pada perlakuan setiap minggu. Persentase buah kakao yang terbebas dari penyakit busuk buah 81,92 %, 62,96 %, dan 72,20 %, secara berturutan untuk perlakuan penyemprotan setiap satu minggu,setiap dua minggu, dan penyarungan plastik. Pada minggu ke-12 buah kakao yang diberi perlakuan penyarungan mengalami peningkatan serangan busuk buah paling tinggi dibandingkan dengan perlakuan lainnya. Perlakuan penyemprotan setiap dua minggu memberikan perlindungan terbaik dari serangan Helopeltis spp. Hasil ini memberikan harapan besar bahwa aplikasi biokaolin sangat berpotensi meningkatan hasil panen petani kakao sehingga akan meningkatkan pendapatan petani.
Aktivitas antibakteri ekstrak kulit buah kakao (Theobroma cacao L.) terhadap Escherichia coli, Bacillus subtilis, dan Staphylococcus aureus MULYATNI, Agustin Sri; BUDIANI, Asmini; TANIWIRYONO, Darmono
E-Journal Menara Perkebunan Vol 80, No 2: Desember 2012
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (118.287 KB) | DOI: 10.22302/iribb.jur.mp.v80i2.39

Abstract

AbstractCocoa (Theobroma cacao L.), one of the most important export commodities from Indonesia, is widely planted with current total area of 1.6 million Ha, producing 500.000 metric tons of dry bean  in 2011 . At the time of harvest, instead of seed approximately the same volume cacao husk is produced. The aim of the study was to assess the potential of cocoa husk extract as an antibacterial against Escherichia coli, Bacillus subtilis, and Staphylococcus aureus, and to determine the minimum inhibitory concentration (MIC) of cocoa husk extract to the three test bacteria. Extraction of cocoa husk conducted by maceration method using ethanol 96%. Analysis of antibacterial activity was done by paper disc diffusion method. Completely Randomized Design of single factor presentage that is extract concentration of 0; 1; 2; 4; 8; 16; 32; and 64% (g/mL) with three replicans were applied.The results showed that the extract of cocoa pod husk has antibacterial activity against S. aureus, B. subtilis, and E. coli with the MIC are 8% (g/ mL), 16% (g/ mL), and 32% (g/ mL) respectively.AbstrakKakao (Theobroma cacao L.), salah satu komoditi ekspor terpenting Indonesia, ditanam secara luas dengan total luasan 1,6 juta Ha, menghasilkan 500.000 ton biji kering pada tahun 2011. Di samping biji sebagai hasil utama, pada saat panen juga dihasilkan kulit buah dengan volume yang hampir sama dengan biji. Penelitian ini bertujuan untuk mengkaji potensi ekstrak kulit buah kakao sebagai antibakteri terhadap Escherichia coli, Bacillus subtilis, dan Staphylococcus aureusserta menentukan konsentrasi hambat minimum (KHM) ekstrak kulit buah kakao terhadap ketiga bakteri uji. Ekstraksi kulit buah kakao dilakukan dengan metode Maserasi menggunakan pelarut etanol 96%. Analisis aktivitas antibakteri dilakukan dengan metode difusi cakram  kertas. Penelitian  ini  menggunakan  Rancangan Acak  Lengkap (RAL) dengan faktor tunggal  konsen-trasi ekstrak, yaitu 0; 1; 2; 4; 8; 16; 32; dan 64% (g/mL), masing-masing dengan tiga kali ulangan. Hasil penelitian menunjukkan bahwa ekstrak kulit buah kakao berpotensi sebagai antibakteri terhadap S. aureus, B. subtilis dan  E. coli, dengan KHM berturut-turut adalah 8% (g/mL), 16% (g/mL), dan 32% (g/mL).
Confirmation of Transgenic Robusta Coffee (Coffea canephora) Transformed by Chitinase-encoding Gene and Its Propagation Through Somatic Embryogenesis ., Priyono; Budiani, Asmini; Mawardi, Surip
Pelita Perkebunan (Coffee and Cocoa Research Journal) Vol 21, No 2 (2005)
Publisher : Indonesian Coffee and Cocoa Research Institute

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Abstract

Genetic engineering of Robusta coffee resistant to fungal diseases might be done by introducing a chitinase-encoding gene into genome of this plant. This research was aimed to confirm transgenic plant of BP 308 clone Robusta coffee transformed by chi gene and to evaluate its ability for the somatic embryogenesis. Confirmation of transgenic was carried out by analysis the presence of NPTII gene as a selectable marker for Canamysin resistant using PCR technique. The somatic embryo initiation and reproduction were evaluated in 11 plant accessions. Three kinds of sucrose concentration, 20%, 30% and 40% were applied in initiation stage of somatic embryo germination. The suitability of 4 medium, namely M1 (without addition by liquid medium), M2 (addition by liquid medium contained 0.25 mg/l kinetin), M3 (addition by liquid medium contained 0.25 mg/l IAA) and M4 (addition by liquid medium contained 0.25 mg/l GA3 ) was evaluated for somatic embryo maturation. The result showed that 8 out of 10 plant accessions tested were transgenic and they could be propagated through somatic embryogenesis. The ability of transgenic plant for somatic embryo initiation, reproduction and regeneration were similar with that of nontransgenic one. Germination of somatic embryo could be improved by using 40% sucrose. Maturation of somatic embryo could be improved by addition of fresh liquid medium on the ancient gelled medium that used for somatic embryos reproduction. The best result was obtained on addition of fresh medium contained 0.25 mg/l GA 3 in which 65% of the somatic embryos developed to pre-germinate somatic embryo. Key words: Coffea canephora, transgenic plant, somatic embryogenesis.
Kloning dan karakterisasi daerah promoter gen penyandi ADP glucose pyrophosphorylase dari Metroxylon sagu rendemen pati-tinggi dan -rendah [Cloning and characterization of promoter region of ADP glucose pyrophosphorylase-encoding gene from Metroxylon sagu with high- and low-starch content] BUDIANI, Asmini; PUTRANTO, Riza Arief; MINARSIH, Hayati; RIYADI, Imron; SUMARYONO, .; ABBAS, Barahima
E-Journal Menara Perkebunan Vol 84, No 1: Oktober 2016
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (52.299 KB) | DOI: 10.22302/iribb.jur.mp.v84i1.200

Abstract

ADP-glucose pyrophosphorylase (AGPase) is one of the key enzymes in the starch biosynthesis. In many plants, the activity of this enzyme was reported to affect the yield and composition of the produced starch. This research is a part of an effort to develop molecular markers for early selection of high starch-yielding of sago palm. The purpose of the research was to isolate promoters of AGP gene and to analyze the differences in their DNA sequences between sago palm with high starch content (MsHS) and low starch content (MsLS). DNA was isolated and purified from the leaves of the two sago palm. The promoter region of AGP was amplified by Genome Walking technique. The specific primers were designed by Primer3 program based on the information of DNA sequence of AGP genes of sago palm from previous studies. Selected DNA fragments resulted from Genome Walking were isolated from the gel, cloned into E. coli, and analyzed its DNA sequence. DNA sequence analysis showed that one DNA fragment from MsHS  (± 1500 bp) and one DNA fragment from MsLS (> 2000 bp) were confirmed as a 5’ upstream of the AGP gene.  Further in silico analysis using MEME program identified various DNA motifs of cis-acting elements, which confirmed that those DNA fragment were promoter region of the gene. Preliminary analysis showed the differences in DNA sequences and motives of cis-acting elements in the promoter region of the two samples which might influence or indirectly associated with the character of the starch yield in sago palm.
Mikropropagasi planlet tebu menggunakan sistem perendaman sesaat (SPS) Micropropagation of sugarcane plantlets using temporary immersion system (TIS) MINARSIH, Hayati; RIYADI, Imron; SUMARYONO, .; BUDIANI, Asmini
E-Journal Menara Perkebunan Vol 81, No 1: Juni 2013
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (448.502 KB) | DOI: 10.22302/iribb.jur.mp.v81i1.53

Abstract

bstractTo achieve Indonesian sugar self-sufficiency in2014, the national production needs to be escalatedthrough land extensification that requires a largenumbers of cane planting materials. This can be achievedby mass propagation of sugarcane through in vitroculture. Solid medium is commonly used for callusproliferation in sugarcane tissue culture. However, solidmedium is considered inefficient in terms of plantletproduction level, labour and space. The use of liquidmedium may solve the problem by allowing automationto increase plantlet production scale and uniformity.Temporary immersion system (TIS) is based on a shortperiodic immersion of explants in a liquid medium for aspecific frequency and duration. Research on in vitromass propagation of sugarcane using TIS was conductedat the Indonesian Biotechnology Research Institute forEstate Crops. Callus initiated from immature unfoldedleaves of PSJT 941 and PS 881 was cultured on liquidMS medium in TIS with different frequencies (12 and24 h) and durations (1 and 3 min) of immersion. Eachtreatment was replicated three times. The callus biomassof two elite cane varieties (PSJT 941 and PS 881)cultured in TIS for six weeks was higher (2 – 4 times fold)than that of on solid medium. The PSJT 941 varietyreached the highest calli biomass with immersion forthree min every 24 h. However, PS 881 variety reachedits highest biomass with immersion for one minute every24 h. The propagation of sugarcane using TIS culturewas proven to produce higher calli biomass up to fourfolds and to form more numbers and uniform shootscompared to the solid medium culture. The callus wassuccesfully regenerated to shoots and plantlets.AbstrakUntuk mencapai swasembada gula, perlu dilakukanpeningkatan produksi gula nasional melalui perluasanareal pertanaman tebu sehingga diperlukan bibit dalamjumlah besar. Hal tersebut dapat diatasi antara laindengan perbanyakan tebu melalui kultur in vitro. Peng-gunaan medium padat pada perbanyakan kalus tebumelalui kultur in vitro merupakan teknik yang umumdigunakan saat ini. Akan tetapi penggunaan mediumpadat dianggap kurang efisien dalam hal jumlah planletyang diproduksi, tenaga kerja dan ruang digunakan.Penggunaan medium cair dapat mengatasi kelemahantersebut dengan dimungkinkannya otomatisasi sehinggadapat meningkatkan skala produksi secara massal dankeseragaman planlet. Sistem perendaman sesaat (SPS)merupakan teknik kultur in vitro dalam medium cairmenggunakan bejana bersekat dimana kontak antaraeksplan dan medium terjadi hanya secara sesaat danperiodik. Penelitian perbanyakan massal bibit tebumelalui SPS dilakukan di Balai Penelitian BioteknologiPerkebunan Indonesia. Kalus diinisiasi dari daun meng-gulung varietas PSJT 941 dan PS 881 yang ditumbuhkanpada media MS cair dalam kultur SPS dengan frekuensiyang berbeda (12 dan 24 jam) dan lama perendaman (1dan 3 menit). Setiap perlakuan diulang tiga kali. Bobotbasah (biomassa) kalus dari dua varietas tebu (PSJT 941dan PS 881) yang ditumbuhkan dengan metode SPSsetelah enam minggu menunjukkan pening-katan yanglebih tinggi yaitu antara 2 - 4 kali lipat dibandingkandengan kontrol (media padat). Peningkatan biomassatertinggi pada varietas PSJT 941 diperoleh pada per-lakuan SPS dengan interval perendaman 24 jam dan lamaperendaman tiga menit. Sedangkan pada PS 881,peningkatan tertinggi biomassa diperoleh pada intervalperendaman 24 jam dan lama perendaman satu menit.Perbanyakan dengan metode SPS terbukti dapat mening-katkan biomassa kalus lebih dari empat kali lipat danpembentukan tunas yang lebih seragam dibandingkandengan pada media padat. Kalus yang dihasilkan dapatdiregenerasikan menjadi tunas dan planlet.