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Hayati Minarsih
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INDONESIA
Menara Perkebunan
Published by Pusat Penelitian Kelapa Sawit
ISSN : 01259318     EISSN : 18583768     DOI : -
Core Subject : Agriculture,
Agriculture, Biological Sciences & Forestry
Menara Perkebunan as a communication medium for research in estate crops published articles covering original research result on the pre- and post-harvest biotechnology of estate crops. The contents of the articles should be directed for solving the problems of production and/or processing of estate crops of smallholder, private plantations and state-owned estates, based on the three dedications of plantation. Analyses of innovative research methods and techniques in biotechnology, which are important for advancing agricultural research. Critical scientific reviews of research result in agricultural and estate biotechnology.
Arjuna Subject : -
Articles 3 Documents
 1 
Search results for , issue "Vol 73, No 2: Desember 2005" : 3 Documents clear
Konstruksi pustaka cDNA dari daun klon karet AVROS 2037 yang diinfeksi patogen Corynespora cassiicola Construction of a cDNA library from leaf of AVROS 2037 rubber clone infected by Corynespora cassiicola pathogen . NURHAIMI-HARIS; Hajrial ASWIDINNOOR; Antonius SUWANTO; Maggy T. SUHARTONO; Nurita TORUAN-MATHIUS; Agus PURWANTARA
E-Journal Menara Perkebunan Vol 73, No 2: Desember 2005
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (952.615 KB) | DOI: 10.22302/iribb.jur.mp.v73i2.156

Abstract

SummaryConstruction of cDNA library derived fromtranscripts made under certain condition is animportant first step to understand diseaseresistant mechanisms. To identify rubber genesor transcripts involved in defense responsetoward Corynespora cassiicola, cDNA librarywas constructed using rubber clone AVROS2037, one of resistant clone to this pathogen.cDNA library was constructed based on thestrategy of leaves infection using conidia, withthe assumption that transcript expression relatedto defense response would be induced bypathogen infection. RNA was isolated from leavesthree days after inoculation with conidia ofC. cassiicola. Steps involved in the cDNA libraryconstruction were RNA isolation, mRNApurification, cDNA synthesis, vector modifcation,cDNA insert ligation, plasmid transformation andclone verifications. Each gram of leaf producedapproximately 300  g RNA, and 0.25% of themwas mRNA. The mRNA was used to synthesizedcDNA. Ligation of cDNA and modified vectorwas facilitated by restriction enzyme SfiI. Theconstructs were transformed into the E. coliDH5 competent cells. A total of 8000 colonieswere produced. Random examination of 270colonies showed that approximately 93% of thesecolonies carried plasmid vector with DNA insertsize of 200 – 2000 bp, with average size of 500 –800 bp. cDNA library construction of rubberleaves from AVROS 2037 clone as well as somenecessary modification steps are presented in thispaper.RingkasanKonstruksi pustaka cDNA yang me-ngandung transkrip yang diekspresikan dalamkondisi tertentu merupakan tahap awal yangsangat penting dalam berbagai studi biologi.Untuk mengidentifikasi gen karet atau transkripyang berperan dalam respons pertahanan tanamankaret terhadap Corynespora cassiicola, pustakacDNA dibuat dengan menggunakan daun klonAVROS 2037 yang merupakan salah satu klonresisten terhadap patogen tersebut. PustakacDNA dibuat berdasarkan strategi menginfeksidaun dengan konidia C. cassiicola denganpertimbangan bahwa ekspresi transkrip yangberperan dalam respons pertahanan akandiinduksi oleh adanya infeksi patogen. Dengandemikian pustaka cDNA yang dibuat diharapkanmengandung gen atau bagian gen yang ber-hubungan dengan respons pertahanan. RNAdiisolasi dari daun setelah daun diinokulasiselama tiga hari dengan konidia C. cassiicola.Beberapa tahapan telah dilakukan, dimulaidengan isolasi RNA, pemurnian mRNA, sintesiscDNA, modifikasi vektor kloning, ligasi fragmencDNA utas ganda dengan vektor kloning sertatransformasi hasil ligasi ke bakteri Escherichiacoli DH5 kompeten. Dari setiap gram jaringandaun berhasil diisolasi RNA sekitar 300 g, dandari jumlah tersebut sekitar 0,25% mRNA dapatdiisolasi. mRNA yang diisolasi digunakan untuksintesis cDNA. cDNA dipotong dengan enzimrestriksi SfiI dan diligasi ke vektor plasmid yangdimodifikasi dengan menyisipkan situs enzimSfiI. cDNA-vektor rekombinan ditransformasi kedalam sel bakteri E. coli DH5 kompeten meng-gunakan metode standar. Transformasi konstrukini menghasilkan 8.000 koloni. Pengujian PCRterhadap 270 koloni yang dipilih secara acakmengindikasikan bahwa sekitar 93% kolonitersebut membawa cDNA sisipan dengan ukuranfragmen cDNA yang menyisip berkisar antara200 sampai 2000 bp. cDNA sisipan terbanyakterdapat pada ukuran antara 500 – 800 bp. Dalamtulisan ini dibahas tahap demi tahap proses yangdilakukan untuk membuat pustaka cDNA asaldaun karet klon AVROS 2037 serta beberapamodifikasi yang diperlukan.
Biokonversi minyak sawit kasar menggunakan desaturase amobil sistem curah pada skala semipilot Bioconversion of crude palm oil using immobilized desaturase in batch system at semi pilot scale . TRI-PANJI; . SUHARYANTO; . GUNAWAN; Khaswar SYAMSU
E-Journal Menara Perkebunan Vol 73, No 2: Desember 2005
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (3113.418 KB) | DOI: 10.22302/iribb.jur.mp.v73i2.154

Abstract

SummaryIncreasing unsaturation level of crude oilpalm (CPO) could be carried out by usingdesaturase enzyme of Absidia corymbifera. Thisbiocatalyst could also produce polyunsaturatedfatty acids (PUFA) such as gamma linolenic acidthat beneficial for healthy oil. The objective ofthis research was to determine the optimumcontact time and ratio of immobilized desaturaseenzyme-substrate in batch system at semi pilotscale (5,000-15,000 mL). Desaturase wasextracted from A. corymbifera biomass andimmobilized on activated zeolite (3-6 mm).Immobilized enzymes were then used forbioconversion process in batch system by mixingthe enzyme with CPO in a bottle placedhorizontally then rotated using a rotator machineat room temperature (25-30 o C). The resultshowed that optimum contact time with ratioimmobilized enzyme-substrate 1:1; 1:2; and 1:3were 30, 40, and 50 min resulted in increasingiodine number 2.84; 3.94; and 4.46 g I 2 /100 gCPO, respectively. An optimum enzyme-subtrateratio was achieved at 1:2, resulted in increasingof iodine number 9-11 g I 2 /100 g CPO, productrecovery of 17,000 mL (21 batches) up to 18 hours. It was detected that active desaturasesduring CPO bioconversion were  6 ,  9 , and  12 desaturases as shown by the increase of oleic(4.5%), linoleic (0,85%) and linolenic acids(60.7%).RingkasanPeningkatan ketidakjenuhan minyak sawitkasar (crude palm oil, CPO) dapat dilakukandengan enzim desaturase Absidia corymbifera.Biokatalis ini juga mampu menghasilkan asamlemak tidak jenuh majemuk (polyunsaturatedfatty acids, PUFA) yang bermanfaat untukkesehatan seperti asam gamma linolenat (GLA).Tujuan penelitian adalah menetapkan waktukontak dan nisbah enzim desaturase amobil-substrat optimum dalam sistem curah pada skalasemipilot (5.000-15.000 mL). Desaturase di-ekstraksi dari biomassa A. corymbifera dandiamobilisasi pada zeolite (3-6 mm) yang telahdiaktivasi. Enzim amobil kemudian digunakanuntuk proses biokonversi dalam sistem curahdengan cara mencampurkan dengan CPO dalambotol yang diletakkan secara horizontal kemudiandiputar dengan mesin rotator pada suhu ruang(25-30 o C). Hasil penelitian menunjukkan bahwawaktu kontak optimum enzim desaturase-substratdengan nisbah 1:1; 1:2; dan 1:3 adalah 30, 40,dan 50 menit dan menghasilkan peningkatanbilangan iod berturut-turut sebesar 2,84; 3,94;dan 4,46 g I 2 /100 g CPO. Nisbah enzim-substratoptimum dalam proses biokonversi CPO adalah1:2 yang menghasilkan peningkatan bilangan iod9-11 g I 2 /100 g CPO dan perolehan produk17.000 mL (21 kali curah) selama 18 jampemakaian. Penelitian juga dapat mendeteksibahwa desaturase yang aktif selama prosesbiokonversi CPO adalah  6 ,  9 , dan  12desaturase yang ditunjukkan oleh peningkatanasam oleat (4,5%), linoleat (0,85%) dan linolenat(60,7%).
The development of somatic embryos of sago palm (Metroxylon sagu Rottb.) on solid media *) Perkembangan embrio somatik tanaman sagu (Metroxylon sagu Rottb.) pada medium padat Imron RIYADI; J.S. TAHARDI TAHARDI; . SUMARYONO
E-Journal Menara Perkebunan Vol 73, No 2: Desember 2005
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (297.957 KB) | DOI: 10.22302/iribb.jur.mp.v73i2.155

Abstract

SummarySago palm (Metroxylon sagu Rottb.) isusually propagated vegetatively by suckers.However, the limited availability of uniformsuckers is a major obstacle in the establishmentof cultivated sago plantations. Tissue culture hasthe potential for large-scale mass clonalpropagation of superior genotypes of sago palm.In vitro culture of sago palm has been establishedthrough somatic embryogenesis. Embryogeniccallus derived from shoot apical tissue of youngsuckers was cultured on a modified Murashigeand Skoog (MMS) medium containing 30 g/Lsucrose, 2 g/L Gelrite, 1 g/L activated charcoal,5.0 mg/L 2,4-D, and 0.1 mg/L kinetin to inducesomatic embryos. Callus clumps formed somaticembryos within four weeks. In the subsequentculture, approximately 0.3 g initial globularcallus grown on MMS medium containing 1.0mg/L kinetin, 0.01 mg/L ABA and 0.1 mg/L GA 3produced 140 to 200 somatic embryos at differentdevelopmental stages four weeks later. All stagesof developing embryos with different sizesand colors were present at any one time ofculture. Secondary (repetitive) somatic embryo-genesis was also found in the culture.Transferring of the mature stage of somaticembryos to solid media with half-strength macro salts and with sucrose at concentration of 20 or 30 g/L without growth regulators led to the development of normal plantlets.RingkasanTanaman sagu (Metroxylon sagu Rottb.)biasanya diperbanyak secara vegetatif dengantunas anakan. Namun, terbatasnya ketersediaantunas anakan yang seragam merupakanhambatan utama dalam pembukaan perkebunansagu. Teknologi kultur jaringan mempunyaipotensi untuk perbanyakan klonal tanaman saguunggul dalam skala besar. Kultur in vitrotanaman sagu telah dikembangkan melaluiembriogenesis somatik. Kalus embriogenik yangberasal dari eksplan pucuk tunas anakandikulturkan pada medium modifikasi Murashigedan Skoog (MMS) dengan sukrosa 30 g/L,Gelrite 2 g/L, arang aktif 1 g/L, 2,4-D 5 mg/Ldan kinetin 0,1 mg/L untuk menginduksi embriosomatik. Kalus membentuk embrio somatikdalam waktu empat minggu. Dalam kulturberikutnya, dari kurang-lebih 0,3 g embrio faseglobuler yang dikulturkan pada medium MMSdengan kinetin 1,0 mg/L, ABA 0,01 mg/L danGA 3 0,1 mg/L menghasilkan 140 sampai 200embrio somatik dengan fase perkembangan yangberbeda-beda. Embrio somatik dalam semuafase perkembangan dengan ukuran dan warnayang berbeda-beda ditemukan setiap saat dalamkultur. Di samping itu, embriogenesis somatiksekunder (berulang) juga terjadi dalam kultursagu. Embrio somatik fase dewasa biladipindah ke medium padat dengan garam makrosetengah konsentrasi dan sukrosa padakonsentrasi 20 atau 30 g/L tanpa zat pengaturtumbuh akan menjadi planlet normal.

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