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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.
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Articles 5 Documents
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Search results for , issue "Vol 68, No 2: Desember 2000" : 5 Documents clear
Transformation of Coffee arabica using chitinase gene and regeneration of planlets from transformed-zygotic embryos Transformasi Coffea arabica menggunakan gen kitinase dan regenerasi planlet dari embrio zigotik-transforman Asmini BUDIANI; T CHAIDAMSARI; . PRIYONO; S MAWARDI; . SISWANTO
E-Journal Menara Perkebunan Vol 68, No 2: Desember 2000
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (279.03 KB) | DOI: 10.22302/iribb.jur.mp.v68i2.138

Abstract

RingkasanRekayasa genetika kopi arabika tahan penyakit cendawan dapat dilakukan dengan  cara memasukkan gen kitinase (gen chi) ke dalam genom tanaman tersebut. Penelitian ini bertujuan untuk mengintroduksikan gen chi pada  kopi arabika serta meregenerasi eksplan yang ditransformasi menjadi plantlet. Gen chi disubkloning dari pBS G11 ke dalam plasmid pCAMBIA2301. Melalui Agrobacterium tumefaciens, plasmid rekombinan pCAMBIA2301/35s-chi kemudian dimasukkan ke dalam eksplan daun dan embrio zigotik kopi arabika. Eksplan daun transforman ditumbuhkan pada media seleksi yang mengandung kanamisin untuk induksi kalus embriogenik . Beberapa kombinasi 2,4-D dan dicamba serta kinetin, BAP dan 2-iP diuji kemampuannya untuk menginduksi terbentuknya kalus embriogenik. Embrio zigotik transforman ditumbuhkan pada media MS modifikasi yang mengandung kanamisin. Hasil penelitian menunjukkan bahwa perbedaan tipe sitokinin dan kombinasinya dengan 2,4-D atau dicamba. menyebabkan terjadinya variasi  persentase pembentukan kalus embriogenik tahan kanamisin.  Penambahan 100 mg/L kanamisin dalam media seleksi cukup efektif untuk menghambat pertumbuhan eksplan daun nontransforman. Persentase tertinggi induksi kalus embriogenik pada eksplan daun non transforman maupun transforman diperoleh pada media yang mengandung 5 mM 2,4- D dengan 5 mM of kinetin atau 5 mg/L dicamba dengan 5 mM BAP. Sedangkan dalam media dengan penambahan 5 mM kinetin, 100 mg/L asam sitrat dan 100 ppm asam askorbat, jumlah eksplan yang membentuk kalus mencapai optimum pada konsentrasi 0 dan 1 ppm dicamba untuk eksplan transforman dan 10 mg/L dicamba untuk non transforman. Pada eksplan embrio zigotik transforman, peningkatan konsentrasi kanamisin dari 100 mg/L hingga 500 ppm menurunkan persentase pengecambahan embrio dari 80.5 % menjadi 49%, persentase perakaran, dari 34 % menjadi 16%, jumlah akar, panjang akar dan tinggi tunas dari 7 mm menjadi 4 mm.  Pada semua perlakuan kanamisin, embrio zigotik non transforman tidak membentuk akar dan pada umur kultur yang sama tunas yang dihasilkan lebih pendek dibandingkan dengan embrio-zigotik transforman. Hasil tersebut membuktikan bahwa gen ketahanan terhadap kanamisin (NPTII) telah terinsersi dan terekspresi dengan baik pada plantlet kopi arabika yang berasal dari eksplan embrio-zigotik transforman. Karena gen chi  dikonstruksi dalam satu vektor dengan NPTII, maka diharapkan gen tersebut juga telah terinsersi ke dalam genom tanaman kopi.SummaryGenetic engineering of arabica coffee resistant to fungal diseases might be done by introducing a chitinase-encoding gene (chi) into genome of this plant. This research was aimed to introduce chi construct into arabica coffee and regenerate plantlets from the transformed explants. The chi gene was previously subcloned from pBS G11 into pCAMBIA2301 plasmid. With Agrobacterium tumefaciens,the recombinant plasmid pCAMBIA2301/35s-chi was then introduced into leaf and zygotic embryos explants of arabica coffee. The transformed leaf explants were cultured on the selection media containing kanamycin in the presence of several combinations of 2,4-D and dicamba with kinetin, BAP and 2-iP to induce the formation of embryogenic callus. The transformed zygotic embryos were cultured on the media of modified MS containing  kanamycin. The results showed that the several types of cytokinin used in combination with 2,4-D or dicamba caused the percentage of kanamycin resistant-embryogenic calli was varied. The addition of 100 mg/L kanamycin in the selection media was effective for inhibiting the  growth of untransformed explants. Among the several combinations of auxin and cytokinin tested, the highest percentage of embryogenisis for untransformed and transformed leaf explants  were achieved on the media containing 5 mM 2,4-D and 5 mM kinetin or 5 mg/L dicamba and 5 mM BAP. However in the presence of 5 mM kinetin together with antioxidants of 100 mg/L citric acid and 100 mg/L ascorbic acid, the explants calluses was optimum at 0 - 1 mg/L dicamba for transformed explants and 10 mg/L dicamba for untransformed explants. In the explants of transformed-zygotic embryos, increasing kanamycin from 100 mg/L up to 500 mg/L decreases the percentage of embryo germination from 80.5 % to 49%, rooted-shoots from 34 % to 16%, number of roots, root length and shoot length from 7 mm to 4 mm.  At all the kanamycin treatments, root was not developed from the untransformed-zygotic embryos and the lenght of shoots were shorter compared to the transformed-zygotic embryos. This result demonstrates that the kanamycin-resistant gene (NPTII) has been inserted and well expressed in the plantlets of arabica coffee derived from transformed-zygotic embryos. Since the chi gene was constructed in one vector with NPTII, this gene might also been inserted in the genome of coffee. 
Overexpression of chitinase gene with a GC-rich synthetic enhancer in tobacco plant (Nicotiana tabacum L.) Overekspresi gen kitinase dengan enhancer sintetis kaya GC pada tanaman tembakau (Nicotiana tabacum L.) D SANTOSO; T CHAIDAMSARI; A BUDIANI; H MINARSIH; S DWI UTOMO; . SISWANTO
E-Journal Menara Perkebunan Vol 68, No 2: Desember 2000
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (264.272 KB) | DOI: 10.22302/iribb.jur.mp.v68i2.139

Abstract

Ringkasan Perakitan tanaman perkebunan toleran terhadap serangan cendawan patogenik  dilakukan dengan mengoverekspresikan gen penyandi kitinase. Untuk itu elemen DNA peningkat ekspresi (enhancer E52) yang berupa oligonukleotida 52 pb dan kaya kandungan basa purin (GC) disisipkan di ujung 5’ konstruk 35S-chi. Penyisipan E52 tersebut dilakukan secara lebih terarah pada situs ganda HindIII-SalI dari MCS pCAMBIA2301. Melalui situs HindIII yang terletak tepat di ujung 3’ E52, konstruk 35S-chi kemudian disambungkan dengan E52 pada pCAMBIA tersebut. Transformasi DNA rekombinan ke dalam sel tembakau dikerjakan melalui perantaraan Agrobacterium tumefaciens LBA4404. Sel tanaman transgenik     diseleksi    dan  diregenrasi dalam media  yang    mengandung  3%  sukrosa, 0,5 mg/L diregenerasikan pada media   MS   padat  benzilaminopurin   (BAP)  dan  50 mg/L kanamisin. Pada media ini tunas transgenik    tembakau   mulai   terbentuk    setelah 5 minggu penanaman. Analisis  tingkat aktivitas enzimatis menunjukkan bahwa aktivitas kitinase pada tembakau transgenik 40 hingga 80 kali lebih tinggi daripada non-transgenik. Pengujian hibridisasi protein menggunakan antibodi anti-kitinase, dot blot dan western blot, membuktikan bahwa enhancer tersebut dapat meningkatkan ekspresi transgen chi pada tanaman tembakau.Summary Development of estate crops tolerant to pathogenic fungi is conducted by overexpressing chi gene. For this purpose, a synthetic enhancer consisting of 52 base pairs and GC-rich was inserted at immediate 5’ end of a 35S-chi cassette. Insertion of the E52 was directed at HindIII-SalI restriction sites of  the pCAMBIA2301 MCS.  With HindIII restriction site located just after the 3’ end of the E52 sequence, the 35S-chi construct was then ligated with the E52 of the pCAMBIA. Transformation of the resulting recombinant DNA into tobacco cells was mediated by Agrobacterium tumefaciens LBA4404. The transgenic cells were selected and regenerated on a solid MS medium supplemented with 3% sucrose, 0.5 mg/L benzylamino purine (BAP) and 50 mg/L kanamycin. Tobacco   shoots   were   initiated  after 5 weeks inoculation on the selection media. Enzymatic analysis demonstrated that chitinase activity of transgenic tobacco was 40 to 80 folds higher than that of the control plant. Analysis of  enzymatic activity using hybridization with anti-chitinase antibody indicated that the level of chitinase activity in the transgenic tobacco carrying the enhancer is higher than that  without enhancer. These data suggest that the enhancer improved the expression of chi transgene in tobacco. 
Development of tobacco plant cells in the presence of kanamycin at various levels for transgenesis Perkembangan sel tanaman tembakau pada kanamisin berbagai konsentrasi untuk transgenesis D SANTOSO; Ferry I CUGITO; H MINARSIH
E-Journal Menara Perkebunan Vol 68, No 2: Desember 2000
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (225.248 KB) | DOI: 10.22302/iribb.jur.mp.v68i2.140

Abstract

Ringkasan Diferensiasi sel tanaman dalam proses regenerasi tanaman transgenik umumnya dilakukan bersamaan dengan proses seleksi menggunakan bahan penyeleksi. Kanamisin merupakan salah satu antibiotika yang biasa digunakan dalam proses seleksi. Dengan spektrum yang luas, kanamisin menghambat pertumbuhan sel dan mengganggu proses translasi pada saat ekspresi gen. Untuk tujuan regenerasi tanaman transgenik yang mengeks-presikan gen NPTII, konsentrasi kanamisin perlu dioptimasi sehingga cukup untuk membedakan sel yang tertransform dengan yang tidak tertransform. Penelitian ini betujuan untuk mempelajari perkembangan eksplan tembakau pada media regenerasi yang mengandung kanamisin. Kecepatan inisiasi tunas dan jumlah tunas terbentuk merupakan kriteria utama untuk evaluasi. Ekstrak protein dari eksplan yang berregenerasi dianalisa dengan SDS-PAGE untuk melihat kemungkinan keterlibatan protein tertentu dalam proses regenerasi tersebut. Hasil penelitian menunjukkan bahwa konsentrasi optimum kanamisin untuk tujuan tersebut adalah 50 mg/L, dimana tunas tembakau transgenik terinisiasi pada hari ke 25 kultur sedangkan eksplan non-transgenik hingga hari ke 56 kultur tidak mampu menginisiasi tunas. Data analisis protein menunjukkan adanya 3 protein dengan ukuran terdenaturasi relatif kecil, antara 14,5 hingga 21,5 kDa pada eksplan yang berregenerasi, namun tidak dijumpai pada ekstrak eksplan yang tidak berregenerasi. Ketiga protein ini kemungkinan terlibat dalam proses regenerasi.  Summary Plant cell differentiation toward regeneration of transgenic plants is usually conducted simultaneously with selection process in the presence of selecting agent. Kanamycin is one of antibiotics widely used as selection agent. Having a wide spectrum activity, kanamycin hinders cell growth through inhibition of translation process during gene expression. To regenerate transgenic plant expressing NPTII gene, the level of kanamycin has to be optimized so that high enough to differentiate genetically transformed cells from those untransformed. This research is aimed to investigate in vitro development of tobacco cells toward plantlet regeneration on the media supplemented with kanamycin. The rate of shoot initiation and number of shoots formed are the major criteria evaluated. Protein extracts of the regenerated explants were analyzed with SDS-PAGE to examine possible involvement of specific proteins in the regeneration process. The experimental result suggested that optimum level of kanamycin for the purpose is 50 mg/L, by which transgenic tobacco shoots were initiated at 25-day culture whereas the untransformed explants did not initiated any shoots even though after 56-day culture. Preliminary data from the protein analysis indicated the presence of relatively small size denatured proteins, between 14.5 and 21.5 kDa, likely to involved in the regeneration process.      
Extraction and characterization of humic acid from plantation’s solid organic waste composts Ekstraksi dan karakterisasi asam humat dari kompos limbah padat organik perkebunan LAKSMITA P.SANTI P SANTI; D H GOENADI; H WIDIASTUTI; N MARDIANA; . ISROI
E-Journal Menara Perkebunan Vol 68, No 2: Desember 2000
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (503.074 KB) | DOI: 10.22302/iribb.jur.mp.v68i2.141

Abstract

Ringkasan Kompos dari limbah padat organik (LPO) perkebunan memiliki kandungan asam humat yang relatif tinggi.  Namun, metode ekstraksi dan karakteristik asam humat asal kompos tersebut belum banyak diteliti.   Oleh karena itu suatu rangkaian penelitian dilakukan dengan tujuan memperoleh paket teknologi ekstraksi dan menetapkan karakteristik asam humat asal kompos tandan kosong kelapa sawit (TKKS), kulit buah kakao (KBK), dan sisa pangkasan teh (SPT). Pengomposan dilakukan melalui tahapan pengumpulan limbah organik padat perkebunan, pencacahan, pencampuran dengan bioaktivator, inkubasi dan pemanenan.   Hasil penelitian menunjukkan bahwa metode ekstraksi konvensional dengan larutan NaOH dalam atmosfer udara dapat digunakan untuk ekstraksi skala semi pilot.  Jumlah asam humat yang dihasilkan dari kompos asal TKKS dan SPT lebih banyak apabila dibandingkan dengan asam humat asal kompos KBK.  Waktu inkubasi pengomposan dan metode ekstraksi dengan gas N2 atau udara yang digunakan tidak berpengaruh nyata terhadap perolehan asam humat.  Pemurnian asam humat asal ketiga jenis LPO perkebunan dengan menggunakan kolom Sephadex G-50 mengindikasikan bahwa asam humat asal kompos TKKS, KBK,  dan SPT tersebut memiliki fraksi bobot molekul rendah serta didominasi oleh asam amino aspartat dan glutamat.  Konsentrasi asam amino dan senyawa karboksilat  tertinggi terdeteksi pada asam humat  asal kompos SPT.  Summary The plantation’s solid organic waste (SOW) composts contain relatively high humic acid  (HA) substances.  However, there is little information on extraction and characteristics of HA from the SOW-originated composts.  An investigation has been conducted to determine  extraction  and characterisation of HA from empty fruit bunches of oil palm (EFBOP), cocoa pod husks (CPH), and tea cutting residues (TCR). Composting was conducted using the method that involved SOW collection, shredding, mixing with bioactivator, incubation, and harvesting. The results showed that conventional extraction method using NaOH solution under air atmosphere  could be used for pilot scale extraction of humic acids (Has).    Amount of humic acid from EFBOP and TCR were higher than that of CPH.  The composting period  and the extraction method under air or N2 gas were not significantly affected  the amount of the humid acid obtained.    Purification of HA extracted from  EFBOP, TCR, and CPH composts by using Sephadex G-50 column  indicated that  EFBOP, TCR, and CPH contained HAs with lower molecular weight fractions and predominated by aspartic and glutamic acids.  The highest concentration of amino acids  and  carboxyl compounds were detected in the TCR-originated compost
Solubilization of insoluble phosphates by Aspergillus niger Pelarutan fosfat sukar larut oleh Aspergillus niger LAKSMITA P SANTI; D H GOENADI; . SISWANTO; I SAILAH; . ISROI
E-Journal Menara Perkebunan Vol 68, No 2: Desember 2000
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (316.973 KB) | DOI: 10.22302/iribb.jur.mp.v68i2.142

Abstract

RingkasanPenggunaan langsung fosfat alam (FA) ke dalam tanah sebagai sumber pupuk P telah dilakukan selama bertahun-tahun melalui beberapa macam cara penggunaan. Kualitas FA di Indonesia umumnya rendah dan ketersediaan bahan baku yang berkualitas untuk produksi pupuk fosfat terlarut relatif terbatas. Beberapa mikroba asal tanah yang dapat melarutkan fosfat anorganik telah banyak dilaporkan. Namun, informasi yang tersedia tentang mekanisme pelarutan P dari FA lokal asal Indonesia dan P anorganik oleh Aspergillus niger BCC F194 belum banyak diteliti. Satu seri penelitian laboratorium telah dilaksanakan untuk mengetahui kemampuan A. niger BCC F194 melarutkan P. Evaluasi agronomi FA lokal (FA Cileungsi dan Madura) di rumah kaca juga telah dilakukan. A. niger BCC F194 dapat melarutkan sumber P sukar larut, yaitu FA Cileungsi dan Madura, serta senyawa Ca3(PO4)2 dan AlPO4. Kelarutan P anorganik tersebut berhubungan dengan peningkatan aktivitas proton (H*) yang menyebabkan penurunan pH medium dan produksi asam organik. Asam organik utama yang dihasilkan oleh A. niger BCC F194 dalam medium cair Pikovskaya yang dimodifikasi adalah asam oksalat (3.75 mM), asam sitrat (2.0 mM), dan asam glukonat (0.9 mM). Kelarutan FA Cileungsi lebih besar dibandingkan dengan FA Madura, dan kelarutan Ca3(PO4)2 lebih besar dibandingkan kelarutan AlPO4. Tidak ada korelasi antara kelarutan P anorganik dengan aktivitas enzim fosfatase, walaupun aktivitas enzim fosfatase cukup tinggi terdeteksi dalam medium. Satu formula biosuperfosfat telah berhasil dirakit dengan mereaksikan FA lokal dengan supernatan kultur cair (SKC) pengganti asam sulfat. Hasil percobaan padabibit kakao, karet dan kelapa sawit di rumah kaca menunjukkan bahwa prototipe pupuk biosuperfosfat dengan bahanbaku FA Cileungsi dan Madura bentuk granul maupun serbuk, memiliki nilai efektivitas agronomi yang relatif menyamai SP-konvensional. SummaryThe direct application of rock phosphate (RP) to soils as a source of phosphorus (P) fertilizer has been employed with varying popular methods over the years. The RP in Indonesia has low quality for plant fertilization and the availability of the raw material with good quality for production of soluble phosphate fertilizers is limited. Many common soil microbes that can dissolve insoluble inorganic phosphate have been extensively studied. However, there is little information on mechanism of P-solubilization from local RP of Indonesia and inorganic P by Aspergillus niger BCC F194 isolated from tropical acid soils. A laboratory study was conducted to determine the ability of phosphate solubilization A. niger BCC F194. Agronomic evaluation of bioactivated local RP, i.e. Cileungsi and Madura phosphate rocks (CRP and MRP) for direct application in greenhouse experiment was also conducted. A. niger BCC F194 was able in solubilizing different types of hardly-soluble phosphates, i.e. CRP and MRP, Ca3(PO4)2, and AlPO4 compounds. Inorganic P solubilization was directly correlated to the organic acid production and increasing proton (H+) activities causing pH to decrease and production of organic acid. The major acidic metabolites produced by A. niger BCC F 194 in modified liquid culture Pikovskaya medium were oxalic acid (3.75 mM), citric acid (2.0 mM), and  gluconic acid (0.9 mM). The solubilization of Cileungsi RP was higher than that of Madura RP, and the solubilization of Ca3(PO4)2 was better than that of AlPO4. No correlation between solubilization of inorganic P and enzyme activities, although high activities of phosphatase enzyme were detectable in the medium. A biosuperphosphate formula had been constructed by reacting local RP with liquid culture supernatant (LCS) replacing sulfuric acid. Results of cocoa, rubber, and oil palm seedlings experiments in greenhouse indicate that both granular and powder biosuperphosphate prototypes commonly had a comparable relative agronomic effectiveness value to that of the conventional SP.

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