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Journal : E-Journal Menara Perkebunan

Ekspresi fenotipe gen APETALA1 kakao (TcAP1) pada eksplan tembakau Phenotypic expression of cacao APETALA1 (TcAP1) in tobacco explant CHAIDAMSARI, Tetty; SAMANHUDI, .; BUDIANI, Asmini; POERWANTO, Roedhy; SANTOSO, Djoko
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 | DOI: 10.22302/iribb.jur.mp.v74i1.116

Abstract

Summary APETALA1 (AP1) is one of flowering identity genes that determines the formation of sepal and petal tissues. An AP1 homologue was cloned from cacao flowers by bio-techniques coupled with bio-informatics. Examination of phenotypic expression was conducted with transgenesis of the 35S-TcAP1 construct using leaf disk technique of tobacco leaf explants mediated by Agrobacterium tumefaciens. PCR specific to TcAP1 demonstrated that the technique is effective in introducing the 35S-TcAP1 construct into tobacco plant cells. RT-PCR with total RNA from the leaves of transgenic tobacco plantlets showed that expression levels of the TcAP1 events varied. The variation of the transcript levels was comparable to the morphological phenotype of the tobacco plantlets grown in vitro. The cultures expressing TcAP1 at moderate levels, have developed into intact plantlets and set up flowers in vitro.Ringkasan APETALA1 (AP1) diketahui merupakan salah satu gen identitas pembungaan yang mengendalikan terbentuknya jaringan sepal dan petal. Homolog AP1 telah diklon dari organ bunga kakao (TcAP1) dengan kombinasi bio-techniques dan bio-informatics. Pengujian ekspresi fenotipe TcAP1 dilakukan dengan transgenesis konstruk konstitutif 35S-TcAP1 menggunakan teknik leaf disk eksplan daun tembakau dan mediasi Agrobacterium tumefaciens. Pengujian PCR spesifik TcAP1 menunjukkan bahwa teknik tersebut cukup efektif dalam mengintroduksikan konstruk 35S-TcAP1 ke dalam sel tanaman tembakau. RT-PCR dari daun planlet tembakau trangenik membuktikan bahwa tingkat ekspresi TcAP1 tersebut bervariasi. Perbedaan level ekspresi TcAP1 ini memberikan pengaruh yang nampak sebanding terhadap perkembangan morfologis planlet tembakau in vitro.  Kultur yang mengekspresikan TcAP1 pada level sedang mampu beregenerasi menjadi planlet sempurna dan membentuk bunga in vitro.
Keefektifan Agrobacterium mentransfer gen P5CS ke dalam kalus tebu klon PS 851 Effectiveness of Agrobacterium to transfer P5CS gene into sugarcane callus PS 851 clone FITRANTY, Niyyah; NURILMALA, F; SANTOSO, Djoko; MINARSIH, Hayati
E-Journal Menara Perkebunan Vol 71, No 1: Juni 2003
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v71i1.181

Abstract

Summary Transformation of a P5CS gene construct into plant cells coupled with regeneration for transgenic plantlets should develop sugarcane tolerant to drought stress. The purpose of the research is to increase the effectiveness and efficiency of Agrobacterium transferring the gene into sugarcane callus. In this method, recombinant plasmid of pBI-P5CS could be transferred into host cells of Agrobacterium LBA4404 through triparental mating with pRK2013 helper. The parameters were tested to increase the effectiveness and efficiency of Agrobacterium  transferring the gene into sugarcane callus were the addition of antioxidant and 1.0% glucose, callus age (2, 3, and   4 weeks), medium pH (4.5; 5.0; and 5.6), treated with air dry for 30 minutes, wetting agent of silwet with and without short vacuum treatment, and acetosyringone consentration (100, 500, and 1000 mg/L). Identification of the transgene in sugarcane  was conducted by PCR using spesific primers, and the expression was tested by measuring  of the proline content. The result showed that addition of acetosyringone 100 ppm or more, P5CS transfer into the sugarcane explants by Agrobacterium was effective. The genetic transformation could be optimized by selecting proper age of calli, which was four weeks after sub-culture. The effectiveness could be maintained and slightly improved by inoculation at pH  4.5, addition 1.0% glucose, wetting agent of silwet with short vacuum treatment, or treated with air drying for 30 minutes. In vitro cultures for transgenic regeneration required addition of antioxidant to prevent browning in the culture media. The amplified DNA fragment demonstrated that the gene was transferred into sugarcane plantlets, and P5CS gene expression showed  increasing  proline content in transgenic sugarcane plantlets.Ringkasan Transformasi transgen P5CS yang diikuti dengan regenerasi tanaman transgeniknya diper-kirakan mampu menghasilkan tanaman tebu transgenik yang toleran terhadap cekaman kekeringan. Penelitian ini bertujuan untuk me-ningkatkan efektivitas dan efisiensi Agro-bacterium mentransfer gen P5CS ke dalam kalus tebu. Dalam metode ini, plasmid rekombinan pBI-P5CS berhasil dengan baik ditransformasi-kan ke dalam sel. Agrobacterium   LBA4404   dengan  pendekatan triparental mating meng-gunakan helper pRK2013. Parameter yang diuji untuk meningkatkan kondisi efektif dan efisien dalam transfer gen P5CSke dalam kalus tebu adalah penambahan antioksidan dan glukosa 1,0%, umur kalus (2, 3, dan 4 minggu), pH medium (4,5; 5,0; dan 5,6), pengeringan kalus   30 menit, bahan pembasah silwet tanpa dan dengan pemakuman, dan konsentrasi aseto-siringon (100, 500, dan 1000 mg/L). Pengujian keberadaan transgen P5CS dilakukan dengan PCR menggunakan primer spesifik, sedangkan ekspresinya diuji dengan mengukur kandungan prolin dari tanaman tebu. Hasil percobaan menunjukkan bahwa dengan penambahan asetosiringon 100 ppm atau lebih, penggunaan Agrobacterium terbukti efektif dan efisien dalam transfer konstruk transgen P5CS ke dalam eksplan kalus tebu. Transformasi dapat dioptimalkan dengan memilih eksplan kalus tebu yang baik, yaitu yang umur subkulturnya empat minggu. Efektivitasnya juga dapat dijaga atau sedikit ditingkatkan dengan inokulasi pH 4,5, penambahan glukosa 1,0%, bahan pembasah silwet dengan pemakuman, ataupun pemberian perlakuan pengeringan udara selama 30 menit. Kultur kalus transgenik memerlukan penambahan antioksidan untuk mencegah terjadinya pen-cokelatan. Adanya fragmen DNA hasil amplifikasi dengan primer spesifik P5CS menunjukkan pada tanaman tebu telah terdapat gen P5CS.  Demikian pula dengan ekspresi gen P5CS, menunjukkan adanya peningkatan kandungan prolin pada tanaman tebu transgenik. 
Kloning cDNA lengkap penyandi ACCase subunit biotin carboxylase dari mesokarp kelapa sawit (Elaeis guineensis Jacq.) Cloning of full length cDNA encoding ACCase subunit biotin carboxylase from mesocarp of oil palm (Elaeis guineensis Jacq.) BUDIANI, Asmini; SUWANTO, Antonius; ASWIDINNOO, Hajrial; SANTOSO, Djoko; NIKOLAU, Basil J
E-Journal Menara Perkebunan Vol 81, No 2: Desember 2013
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v81i2.43

Abstract

AbstractAcetyl-CoA Carboxylase (ACCase) is considered to beone of the key enzymes in palm oil biosynthesis. Availabilityof genes encoding this enzyme would give some advantagesin the molecular breeding of oil palm. Over expression ofthe genes in the oil palm mesocarp might increase the oilproduction in this tissue. On the other hand, downregulating of ACCase could divert the central metaboliteAcetyl-CoA to other product such as PHB (Polyhydroxy-butyrate), one of the known biodegradable plastic. Thispaper reported the work of cloning of the full length codingsequence of biotin carboxylase (BC), one subunit of theACCase. Based on the DNA sequence of the BC conservedregion that had cloned previously, primers pairs weredesigned to amplify 5’- and 3’- cDNA ends of BC usingRACE-PCR. The RACE products of 5’- and 3’- cDNA endsof BC were cloned into E.coli, and the DNAs weresequenced and analysed. The full cDNA of BC was obtainedby reisolation of the cloned 5’- and 3’- cDNA ends followedby digestion using KpnI, ligation into pGEM-T vector andcloning into E.coli. Colony PCR was carried out to confirmthat the target gene has been cloned. The recombinantplasmid containing full cDNA of BC was then isolated forDNA sequencing. The results showed that the 5’-BC (1367bp), 3’- BC (1032 bp), and the full length cDNA encodingBC (2182 bp) had been successfully cloned, and the DNAsequence had been confirmed as gene encoding ACCasesubunit biotin carboxylase.AbstrakAcetyl-CoA Carboxylase (ACCase) merupakan salahsatu enzim kunci dalam biosintesis minyak sawit. Keter-sediaan gen penyandi enzim ini sangat berguna dalampemuliaan kelapa sawit secara molekuler. Over-ekspresi genpenyandi ACCase pada mesokarp dapat meningkatkan pro-duksi minyak pada jaringan tersebut. Sebaliknya ekspresiACCase dapat ditekan melalui mekanisme down regulation sehingga metabolit central Acetyl-CoA dapat diarahkanuntuk menghasilkan produk lain seperti PHB (polyhydro-xybutyrate), salah satu jenis biodegradable plastik yangtelah banyak dikenal. Penelitian ini bertujuan untukmengklon cDNA lengkap penyandi ACCase subunit biotincarboxylase (BC) dari mesokarp kelapa sawit. Berdasarkansekuen DNA daerah konservatif BC yang telah diklon darimesokarp kelapa sawit pada penelitian sebelumnya, duapasang primer dirancang untuk mengamplifikasi daerahujung 5’- dan 3’- cDNA BC dengan RACE-PCR. Produk5’-RACE dan 3’-RACE diklon dan disekuen. cDNAlengkap penyandi BC diperoleh dengan jalan mengisolasikembali fragmen 5’- dan 3’- cDNA terklon, dilanjutkandengan digesti menggunakan enzim restriksi KpnI, ligasikedua fragmen ke vektor kloning pGEM-T, dan introduksike dalam E. coli. Setelah dilakukan PCR koloni untukmenguji keberhasilan kloning, plasmid rekombinan yangmengandung cDNA lengkap dari BC diisolasi untuk analisissekuen DNA. Dari penelitian ini fragmen cDNA 5’-BC(1367 pb) dan 3’- BC (1032 pb), serta cDNA lengkappenyandi BC berukuran 2182 pb telah diperoleh dan diklondalam E. coli. Analisis sekuen DNA mengkonfirmasi bahwacDNA terklon adalah benar gen penyandi ACCase subunitbiotin carboxylase.
Penggunaan biostimulan Orgamin untuk efisiensi pemupukan dan peningkatan produktivitas kelapa sawit di dataran tinggi Application of Orgamin biostimulan to enhance fertilizer efficiency and productivity of oil palm grown in highland WIDIASTUTI, Happy; SANTOSO, Djoko; PUTRA, Soekarno Mismana; WIRAMIHARDJA, Memed; FARIDA, Aida; MARAHIMIN, B. MARAHIMIN; PANJAITAN, K. PANJAITAN; SINAGA, Jisman
E-Journal Menara Perkebunan Vol 81, No 2: Desember 2013
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v81i2.37

Abstract

AbstractThe extension of oil palm area has been expanded tomarginal land such as the highland regions. However, theproductivity of the oil palm became the main demand for theplanters. Increasing of oil palm productivity can be done byapplication of growth regulators. Growth regulators aresmall molecules in a relatively very small amount that affectthe growth and development of plant. This study wasconducted to asses the efectiveness of plant growth regu-lators (Orgamin and Orgamin plus) in improving fertilizerefficiency and productivity of mature oil palm (TM 7). Theexperiments were conducted at Marjandi oil palm plantationat an altitude of 700 m above sea level in a total area of 16 ha. Six treatments tested were 1). 100% inorganicfertilizer (control), 2). 50% inorganic fertilizer + Orgamin(50K+O), 3). 75% inorganic fertilizer + Orgamin (75K+O),4). 50% inorganic fertilizer + Orgamin plus (50K+OP), 5).75% inorganic fertilizer + Orgamin plus (75K+OP), and 6).100% inorganic fertilizer + Orgamin plus (100K+OP)arranged in a randomized block design (RBD) with threereplications. Orgamin (O) and Orgamin plus (OP) wereapplied in the hole around the oil palm along with inorganicfertilizers. The results showed that application of O and OPimproved the efficiency of inorganic fertilizers by 50% basedon vegetative variables and increased the concentration ofN, P, and K of leaf and soil compared to those of 100%inorganic fertilizer. In addition to the height and leaf numberof plant parameters, the leaf of the plant treated with O andOP showed more greenish compared to those of control.There is an indication that the O application increased thepercentage of female flowers. In addition the application ofOrgamin also produced the highest oil content in oil palmfruit particularly in the treatment of 75% of inorganicfertilizer + orgamin harvested in October compared to thosein March. Moreover, application of OP increased both thetotal weight and weight per bunch of FFB.AbstrakPengembangan kelapa sawit mengharuskan pengguna-an lahan suboptimal seperti daerah dataran tinggi. Produk-tivitas kelapa sawit menjadi tuntutan utama bagi pekebun.Peningkatan produktivitas kelapa sawit di dataran tinggididuga dapat dilakukan dengan aplikasi zat pengatur tumbuh.Zat pengatur tumbuh merupakan molekul “kecil” (small molecules) yang dalam jumlah relatif sangat sedikit mem-pengaruhi pertumbuhan/perkembangan tanaman. Penelitiandilakukan untuk menguji formula zat pengatur tumbuh(Orgamin dan Orgamin plus) dalam meningkatkan efisiensipemupukan dan produktivitas kelapa sawit TM 7. Percobaandilakukan di kebun Marjandi dengan ketinggian 700 dpl padaareal seluas 16 ha. Enam perlakuan yang diuji adalah 1).pupuk anorganik 100% (100K), 2). pupuk anorganik 50% +Orgamin (50K+O), 3). pupuk anorganik 75% + Orgamin(75K+O), 4). pupuk anorganik 50% + Orgamin plus (50K+OP), 5). pupuk anorganik 75% + Orgamin plus (75K+OP),dan 6). pupuk anorganik 100% + Orgamin plus (100K+OP)yang disusun dalam rancangan acak kelompok (RAK)dengan tiga ulangan. Orgamin (O) dan Orgamin plus (OP)diberikan dalam lubang di piringan pokok bersamaan denganpupuk anorganik. Hasil pengamatan menunjukkan bahwapemberian O dan OP dapat meningkatkan efisiensi pemupuk-an anorganik hingga 50% dilihat dari beberapa peubahvegetatif dan menghasilkan kadar N, P, dan K daun dantanah lebih tinggi dibandingkan dengan pemberian pupukanorganik 100%. Selain pada parameter tinggi tanaman danjumlah daun, peningkatan juga terlihat pada tingkatkehijauan daun. Terdapat indikasi bahwa pemberian Orgaminmeningkatkan persentase jumlah bunga betina. PemberianOrgamin juga menghasilkan kadar minyak tertinggi khusus-nya pada pemberian Orgamin + pupuk anorganik 75% padabuah yang dipanen bulan Oktober dibandingkan dengan buahyang dipanen bulan Maret. Baik data bobot per tandanmaupun bobot TBS menunjukkan bahwa pemberian OPdapat meningkatkan kedua peubah tersebut. 
Deteksi dan analisis sekuen gen inhibitor proteinase pada beberapa klon kakao harapan tahan penggerek buah kakao dari Sulawesi Selatan Detection and sequence analysis of proteinase inhibitor gene in cacao clones putatively cacao pod borer-tolerant from South Sulawesi JAYA, Abdul Mollah S.; ASWIDINNOOR, Hajrial; SANTOSO, Djoko
E-Journal Menara Perkebunan Vol 72, No 1: Juni 2004
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v72i1.124

Abstract

Summary Cacao is socially and economically an important commodity for Indonesia, in which the cacao plantations have been challenged with a threatening pest, cacao pod borer (CPB). This research aimed to identify and clone PIN (proteinase inhibitor), a gene carrying resistance of plant to some chewing pests like CPB. The methodology included several experiments. Detection of PIN in cacao was done by PCR using PIN-specific heterologous primers and cacao genomic DNA as templates. Cloning vector pGEM-T was utilized to clone the PCR products. Sequence analysis was conducted with BlastX and Blast Special programs from NCBI. Alignment analysis to determine genetic similarity was performed with ClustalW from EBI. Thirteen of the 18 clones tested were detected to have PIN homologs. Two DNA fragments from cacao clones putatively tolerant to CPB, MJ-1 and LW-1, were sequenced. One of them, MJ-1 was cloned. Sequence analyses of the fragments of both cacao clones, indicated that they have PIN homologs and a very closed genetic relation with 96% level of similarity. Ringkasan Kakao adalah komoditas yang secara sosial maupun ekonomi penting bagi Indonesia, dimana perkebunan kakao menghadapi masalah serius hamapenggerek buah kakao (PBK). Penelitian ini bertujuan mengidentifikasi dan mengklon PIN (inhibitor proteinase), gen yang membawa sifat ketahanan tanaman terhadap hama ulat seperti PBK. Metodologinya terdiri dari beberapa percobaan. Deteksi PIN di dalam kakao dikerjakan dengan PCR menggunakan primer heterologous yang spesifik terhadap PIN dan DNA genomik kakao sebagai templetnya. Vektor kloning pGEM-T digunakan untuk mengklon produk PCR. Analisis sekuen dilakukan dengan program BlastX dan Blast spesial dari NCBI. Analisis penjajaran (alignment) untuk menentukan kemiripan genetik menggunakan program ClustalW dari EBI. Tiga belas dari 18 klon kakao yang diuji  menunjukkan adanya  homolog  PIN. Dua DNA fragmen dari klon harapan tahan, MJ-1 dan LW-1, telah ditentukan sekuen nukleotidanya. Satu diantara-nya, MJ-1 berhasil diklon. Analisis sekuen  kedua klon tersebut menunjukkan identitas sebagai homolog PIN dan keduanya memiliki kemiripan genetik yang tinggi.
Aktivitas ACCase mesokarp kelapa sawit dan kloning fragmen gen penyandi ACCase subunit biotin karboksilase ACCase activity of oil palm mesocarp and cloning of gene fragment encoding biotin carboxylase subunit of ACCase BUDIANI, Asmini; SANTOSO, Djoko; ASWIDINNOOR, Hajrial; SUWANTO, Antonius
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 | DOI: 10.22302/iribb.jur.mp.v74i1.119

Abstract

Summary Genetic engineering to produce high yielding oil palm might be done by over expressing gene encoding key enzyme for oil biosynthesis in the oil palm mesocarp, one of which is ACCase. The objective of this research was to analyze ACCase activity of mesocarp from several developmental stages of fruit and to clone conserved region cDNA of gene encoding biotin carboxylase subunit of ACCase (BC-htACCase) from oil palm mesocarp. Activity of ACCase was analyzed by HPLC. Amplification of cDNA was done by means of reverse transcriptase polymerase chain reaction (RT-PCR) using degenerate heterologous primer on several annealing temperature and MgCl2 concentration. The cDNA fragment of RT-PCR product was cloned, sequenced and analyzed to confirm that the cloned cDNA was conserved region of BC-htACCase. The result showed that ACCase activity increased from the 14 week to the 20 week-old fruit, and then decreased. Using heterologous degenerate primers, cDNA fragments of BC-htACCase conserved region (469 bp) can be specifically amplified at 60 oC annealing temperature with 2 mM MgCl2 concentration.The result of BlastX analysis showed that the sequence of cloned cDNA fragment was highly homologous with the conserved region of BC-htACCase from Glycine max, Arabidopsis thaliana, Nicotiana tabacum,  and Brassica napus with 243, 237, 236, 231 bit score, and E. value 2e-63, 1e-61, 2e-61 and 5e-60, respectively. Ringkasan Rekayasa genetika untuk menghasilkan bibit kelapa sawit berdaya hasil tinggi dapat ditempuh dengan meningkatkan ekspresi gen penyandi enzim kunci biosintesis minyak pada kelapa sawit, salah satunya adalah ACCase. Tujuan penelitian ini adalah menguji aktivitas ACCase mesokarp beberapa tahap perkem-bangan buah sawit dan mengklon fragmen cDNA daerah konservatif gen penyandi ACCase heteromerik subunit biotin karbok-silase (BC-htACCase) dari mesokarp buah sawit. Aktivitas ACCase dianalisis dengan HPLC. Amplifikasi cDNA dilakukan dengan teknik RT-PCR menggunakan primer degene-rate heterologus pada berbagai suhu penempelan dan konsentrasi MgCl2. Fragmen cDNA hasil RT-PCR diklon, disekuen dan dianalisis untuk mengkonfirmasi bahwa cDNA terklon adalah daerah konservatif BC-htACCase. Hasil penelitian menunjukkan bahwa aktivitas ACCase meningkat dari buah berumur 14 minggu hingga buah berumur  20 minggu, kemudian menurun kembali Dengan primer degenerate heterologus, fragmen cDNA daerah konservatif BC-htACCase  (469 pb) dapat diamplifikasi secara spesifik pada suhu penempelan 60 oC dan konsentrasi MgCl2 2 mM. Hasil analisis BlastX dari sekuen DNA fragmen terklon menunjuk-kan bahwa sekuen tersebut mempunyai homologi tinggi antara lain dengan gen penyandi BC-htACCase dari Glycine max, Arabidopsis thaliana, Nicotiana tabacum dan Brassica napus, masing-masing dengan skor 243, 237, 236, 231 bit, dan E. value 2e-63, 1e-61, 2e-61 dan 5e-60.
Respons awal pemberian biostimulan Orgamin pada kelapa sawit (Elaeis guineensis Jacq.) di Kebun Marjandi PTPN IV Early response of Orgamin biostimulan application in oil palm (Elaeis guineensis Jacq.) at PTPN IV Marjandi plantation PUTRA, Soekarno Mismana; SANTOSO, Djoko; WIDIASTUTI, Happy; SARAGIH, A. H. SARAGIH; GHONI, M. A. GHONI; MARAHIMIN, B. MARAHIMIN; PANJAITAN, K. PANJAITAN
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 | DOI: 10.22302/iribb.jur.mp.v81i1.51

Abstract

AbstractEffort to increase the production of oil palm can beconducted through application of plant growth regulator(PGR). Orgamin biostimulan is a natural PGR formulathat has been tested to improve the vegetative growths ofcorn and oil palm in the glass house. Assessment ofOrgamin and Orgamin plus (Orgamin + micro nutrient)applications at commercial scale was carried out inMarjandi oil palm plantation of PTPN IV usingrandomized block design with three treatments, i.e. K =100% recommended dose of inorganic fertilizer(control), O= Orgamin (1.5 kg/tree) + 50% dose ofinorganic fertilizer, OP = Orgamin plus (1.5 kg/tree)without inorganic fertilizer. The parameters ofobservation at 2.5 months after the treatments were soiland leaf nutrient contents (N, P, K, Mg), percentage offemale flower, mesocarp oil content, and harvested freshfruit bunches (FFB). The observation showed that therewas an increased in oil yield, weight of FFB and leafnutrient content, while the percentage of female flowerand nutrient content of soil were not significantlydifferent compared to the control.AbstrakUpaya untuk meningkatkan produksi kelapa sawitdapat dilakukan antara lain melalui pemberian zatpengatur tumbuh (ZPT). Biostimulan Orgamin merupa-kan formula ZPT alami yang telah diuji di rumah kacapada tanaman jagung dan bibit kelapa sawit. Uji cobaaplikasi Orgamin dan Orgamin plus (Orgamin yangdiperkaya hara mikro) pada skala lapang dilakukan dikebun kelapa sawit Marjandi PTPN IV denganmenggunakan Rancangan Acak Kelompok (RAK) untukmenguji tiga perlakuan, yaitu 1) K (kontrol) = 100%dosis anjuran pupuk kimia (APK = kontrol), 2) O = 50%dosis APK + Orgamin (1,5 kg/pohon), 3) OP = Orgaminplus (1,5 kg/pohon) tanpa pupuk kimia. Peubah yangdiamati pada 2,5 bulan setelah perlakuan adalah kan-dungan hara tanah dan daun (N, P, K, Mg), persentasebunga betina, rendemen minyak mesokarp, dan produksitandan buah segar (TBS). Hasil yang diperoleh menunjukkan terdapat peningkatan rendemen minyak, bobotTBS dan kandungan hara daun, sedangkan persentasebunga betina dan kandungan hara tanah tidak menunjuk-kan perbedaan yang nyata antara perlakuan dan kontrol.
Kloning dan karakterisasi gen penyandi inhibitor proteinase dari kulit buah kakao Cloning and characterization of gene encoding proteinase inhibitor of cacao pod wall ISDA, Mayta Novaliza; KASIM, Musliar; MANSYURDIN, .; CHAIDAMSARI, Tetty; SANTOSO, Djoko
E-Journal Menara Perkebunan Vol 76, No 2: Desember 2008
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v76i2.84

Abstract

Summary Attempts to increase cocoa production in Indonesia have been hinderred by attack of CPB (Conopomorpha cramerella). There has been no effective measures to control this pest leading to development of cacao planting materials which resistant to the pod borer. One of genes functioning in plant defense system against insect pests such as catepilar is Proteinase Inhibitor (PIN). This research aimed to isolate and characterize TcPIN gene of cacao pod wall. A clone of TcPIN was isolated with RT-PCR technique using total RNA of cacao pod wall and DNA primer designed based on the sequence Trypsin Inhibitor of cocoa bean accessible online. BlastX analysis of the sequence of the cDNA clone demonstrated that the ± 600 bp gene cloned with pGEM-T was PIN gene as indicated by highly homologous to Trypsin Inhibitor of Theobroma microcarpum resulted in 248 Score bits and E value 1 e-64. Two sequence alligment with the putative 21 kDa PIN  of cacao seed indicated a moderately high homology. Contrasting these two sequences however found some non identical amino acids implying some variations. Ringkasan Usaha peningkatan produksi kakao di Indonesia terkendala antara lain oleh adanya serangan hama PBK (Conopomorpha cramerella). Untuk menanggulangi serangan PBK tersebut perlu adanya satu cara pengendalian yang efektif dan efisien, sehingga dapat mendorong usaha pengembangan bahan tanam yang tahan PBK. Salah satu gen  membawa sifat ketahanan tanaman terhadap hama ulat adalah Proteinase Inhibitor (PIN). Penelitian ini bertujuan untuk mengisolasi dan mengkarakterisasi gen TcPIN dari kulit buah kakao. Klon cDNA TcPIN diisolasi dari kulit buah kakao dengan teknik RT-PCR meng-gunakan RNA total kulit buah kakao dan primer DNA yang dirancang atas dasar sekuen Inhibitor Tripsin biji kakao yang diakses lewat internet.  Hasil analisis BlastX dari sekuen klon cDNA menunjukkan  bahwa gen berukuran  ± 600 pb yang telah diklon dengan pGEM-T tersebut adalah PIN karena memiliki homologi yang tinggi terhadap 21 kDa trypsin inhibitor dari Theobroma microcarpum yang meng-hasilkan Skor 248 bits dengan Nilai E 1e-64. Penjajaran dua sekuen dengan PIN putatif 21 kDa yang berasal dari biji kakao menunjuk-kan tingkat homologi yang tinggi, dengan perbedaan nyata sehingga dapat terlihat bahwa keduanya tidak identik.
Identifikasi homolog TcAGL-15 untuk penanda embriogenesis tanaman kakao melalui pendekatan bioinformatika Identification of TcAGL-15 homolog in the embryogenic culture from the zygotic embryo explant TRIASTANTO, Oktaviany Ferry; JUSUF, Muhammad; SANTOSO, Djoko
E-Journal Menara Perkebunan Vol 74, No 2: Desember 2006
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v74i2.102

Abstract

Summary One of the major problems encountered in micropropagation of cacao through tissue culture is very low frequency of embryo formation. Embryogenesis is believed to have key regulatory gene determining the process. Understanding such gene may help to solve problems in the regeneration process. One of the genes reported to involve in the embryogenesis is AGAMOUS-like 15 (AGL-15). This gene has an important role in the regulation of early embryogenesis in several plants. This experiment aimed to identify AGL-15 homolog in cacao through bioinformatics approach. The first step of this experiment is to identify the AGL-15 homolog using hetero-logous primers from DNA genomic isolated from leaves of cacao plants. The sequence of the AGL-15 fragment was used in designing specific primers for longer AGL-15 fragment. These primers were then used to identify AGL-15 gene using total RNA isolated from cultured zygotic embryos. Differential pattern of AGL-15 gene expression was observed in zygotic embryos cultured for five weeks. AGL-15 heterologous primers designed from several plants could be used to identify cacao AGL-15 homolog. The putative cacao AGL-15 gene could be identified from zygotic embryos. The differential pattern of the AGL-15 gene expression is a strong indication that AGL-15 can be used as an embryogenesis marker in cacao plants.Ringkasan Salah satu kendala perbanyakan kakao melalui kultur jaringan adalah sulitnya embriogenesis, yang diduga melibatkan satu atau lebih gen kunci yang menentukan proses tersebut. Keberhasilan mengidentifikasi gen-gen kunci akan membantu menyelesaikan masalah dalam regenerasi embrio kakao. Salah satu gen yang diduga terlibat dalam proses ini adalah AGAMOUS-like 15 (AGL-15). Gen ini berperan pada regulasi selama masa awal perkembangan embrio beberapa tanaman. Penelitian ini bertujuan untuk mengidentifikasi homolog AGL-15 pada kakao melalui pen-dekatan bioinformatika dan RT-PCR. Pene-litian diawali dengan identifikasi homolog AGL-15 dari DNA genomik daun kakao meng-gunakan primer heterologus. Sekuen fragmen homolog AGL-15 yang diperoleh, kemudian digunakan untuk merancang primer spesifik AGL-15 yang berukuran lebih panjang. Primer ini selanjutnya digunakan untuk meng-identifikasi gen AGL-15 dari RNA total embrio zigotik. Pengamatan pola pita gen AGL-15 dilakukan pada kultur in vitro embrio zigotik yang berumur lima minggu. Primer hetero-logus gen AGL-15 yang berasal dari berbagai tanaman, mampu mengidentifikasi keberadaan homolog  gen  tersebut  pada  tanaman   kakao. Fragmen homolog AGL-15 putatif tanaman kakao teridentifikasi pada tingkat RNA embrio. Dengan adanya pola diferensial dari ekspresi gen AGL-15 hingga lima minggu pertama perkembangan embrio, ada indikasi kuat bahwa fragmen homolog AGL-15 dapat menjadi penanda embriogenesis pada tanaman kakao.
Kloning gen LEAFY kakao dari jaringan bantalan bunga aktif Cloning of cacao LEAFY gene from the active flower cushions CHAIDAMSARI, Tetty; HAYATI, Rita; SYARIEF, Auzar; ANWAR, Aswaldi; SANTOSO, Djoko
E-Journal Menara Perkebunan Vol 77, No 2: Desember 2009
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v77i2.179

Abstract

SummaryAttempts to improve productivity of cacaoplantations lead us to study the molecularmechanism of flowering. In the model speciesArabidopsis thaliana as well as some otherspecies, LEAFY is a central regulatory gene forthe transition of shoot apical meristems toflowering meristems. Different from that ofArabidopsis, cacao inflorescence is acauliflorous type, by which flowers can developrepeatedly from the same flower cushion on thetrunk. In this research, a LEAFY homolog wasisolated from active flower cushion with RT-PCRusing a pair of DNA primer specifically designedto isolate its complete cds. Gel electrophoresisexamination indicated the presence of a 1.2 kbamplicon. Purified from the gel, this DNAfragment was cloned into competent cells ofE. coli XL1 Blue using pGEM-T Easy cloningvector at an orientation according to the T7promoter of the plasmid. Sequence analysis usingBLASTX, showed that the amplicon was LEAFY(LFY) homolog. Alignment analysis using ClustalW indicated that the cTcLFY highly homologousto those from other perennial crops such ascitrus, grape, apple and poplar. The highesthomology (conserved region) was found in the Cterminal of the encoded proteins.RingkasanUsaha untuk meningkatkan produktivitasperkebunan kakao telah mendorong penelitianmolekuler tentang mekanisme pembungaankakao. Pada tanaman model Arabidopsis thalianadan lainnya, LEAFY merupakan gen kunci dalamtransisi meristem tunas jadi meristem bunga.Berbeda dengan sistem pada Arabidopsis,pembungaan kakao termasuk tipe cauliflorous,bunga dapat muncul dari bantalan bunga yangsama sepanjang tahun. Dalam penelitian inihomolog LFY diisolasi dari bantalan bunga aktifmenggunakan RT-PCR dengan sepasang primerspesifik yang dirancang berdasarkan sekuenDNA di kedua ujung gen tersebut. Pemeriksaangel elektroforesis menunjukkan adanya amplikontunggal berukuran 1,2 kb. Setelah dimurnikandari gel, amplikon dapat diklon ke dalam selkompeten E. coli galur XL1 Blue menggunakanvektor pGEM-T Easy dengan orientasi yangsesuai dengan promoter T7 dari vektor. AnalisisBLASTX sekuen DNA membuktikan bahwaamplikon tersebut adalah homolog dari genLEAFY. Analisis penjajaran dengan mengguna-kan ClustalW menunjukkan bahwa gen cTcLFYtersebut memiliki homologi yang tinggi dengangen sejenis dari tanaman keras lainnya sepertitanaman jeruk, anggur, apel dan poplar.Homologi tertinggi (daerah terkonservasi)terdapat pada ujung (terminal) C dari proteinyang disandinya.
Co-Authors . Mansyurdin . Siswanto . Suharyanto . SYAFARUDDIN, . Aditia Wardana, Aditia Aditiawardana Aditiawardana, Aditiawardana Agung Dwi Indrawan Ahmad Fauzi AHMAD RIDUAN Aida Farida Alda, Rieza Rizqi Alsagaff, Mohammad Yusuf AMANAH, Dian Mutiara AMANAH, Dian Mutiara Amin, Mochammad Andre Noevi Rahmanto Anton Subarno ANTONIUS SUWANTO Aprih Santoso, Aprih Ardityo R Ardhany, Ardityo R Artaria Tjempakasari, Artaria Asmini Budiani Aswaldi Anwar ASWIDINNOO, Hajrial Baiti, Ahmad Awaluddin Bambang S Purwoko Bastin Fitriatus Hasanah Bekti Wulandari Bonita Destiana Budi Sulistijo Chandra Irwanadi Mohani, Chandra Irwanadi Chandra Irwanadi, Chandra Chibtiyah, Nur chieko Hamada, chieko D Ningsih, D Darda Efendi Darharta Dahrin DEDI SOLEH EFFENDI, DEDI SOLEH Empitu, Maulana Antiyan Endrizal ERIS, Deden Dewantara Fajar, Saiful Ferdiyono, J. Reza FITRANTI, Niyyah FITRANTY, Niyyah Fitriyah, Fauziatul GHONI, M. A. GHONI Ginanjar, Muhammad Syukron HABIBULLAH, Hanning Susilo Haerani Rasyid, Haerani HAJRIAL ASWIDINNOOR Hakim, Zaky El Hamidah, Berliana HANDAYAN, Agustina A. HANDAYAN, Agustina A. HAPPY WIDIASTUTI Hasanatuludhhiyah, Nurina Hasanuddin Z. Abidin Hasanuddin Z. Abidin, Hasanuddin Hayati Minarsih Hendra Grandis I GEDE PUTU WIGENA I Haryoko, I Ichiyu Shou, Ichiyu Imron Riyadi Indarto Indarto Indhira Hari Kurnia Ismail, Mohd Erfy Isnu Irwantoro, Isnu JAYA, Abdul Mollah S. Joenil Kahar, Joenil Jumiyanto Widodo Kadariswantiningsih, Ika Nindya KRESNAWATY, Irma Kresnawaty, Irma Kunimi Maeda, Kunimi Laesanpura, Agus Laesanpura, Agus M Thaha, M MARAHIMIN, B. MARAHIMIN Mayta Novaliza Isda Mitsumine Fukui, Mitsumine MOCHAMMAD THAHA Moh. Yogiantoro, Moh. MS, Freddy Muhammad Jusuf Muhammad Munir Munadi Munadi Munir, Muh Musliar Kasim Muslikhin Muslikhin Niken Ayu Larasati, Niken Ayu NIKOLAU, Basil J Nugroho, Cahyo Wibisono Nunuk Mardiana, Nunuk NURILMALA, F Nuryake Fajaryati PANJAITAN, K. PANJAITAN Pipit Utami Pradusuara, Franes pranawa pranawa, pranawa PRIYONO, . PURBA, A.R. PURBA Putra, Abdul Aziz Sidiq Tri PUTRANTO, Riza A PUTRANTO, Riza A PUTRANTO, Riza A. Putri, Eka Arum Cahyaning R.D.B LEFROY Raduan, Atiqah Ramlan Arief Fathony, Ramlan Arief Renita Renita Rita Hayati ROEDHY POERWANTO Rokib, Muhammad Nur Saddewisasi, Wyati SAMANHUDI, . Saptowo Jumali Pardal, Saptowo Jumali SARAGIH, A. H. SARAGIH SARI, Dini Astika Satoshi Horikoshi, Satoshi SINAGA, Jisman Slamet Slamet Soekarno Mismana Putra, Soekarno Mismana soewanto soewanto, soewanto Sri Hendrastuti Sri Wahyuni Sri Waluyanti Subagus Wahyuono Sudarsono SUDARSONO SUDARSONO Sugeng Riyanto Sugiyanto Sugiyanto Sukarti Moeljopawiro Suparman Suparman Suryansyah, Maulana Muhtadin SUSANTI, Paramitha SUSANTI, Paramitha Suzuki, Yusuke SYARIEF, Auzar Tetty Chaidamsari, Tetty Tri Harso Karyono Tri P Asmarawati, Tri P TRI-PANJI, . TRI-PANJI, . TRIASTANTO, Oktaviany Ferry UMAHATI, Binti Khurotul UMAHATI, Binti Khurotul Umi Rochayati Umi Rokhayati, Umi Vera Sadarviana, Vera Wahyudi W. Parnadi Warsa Warsa Wawan Gunawan A. Kadir Wedyanto K, Wedyanto Widodo Widodo Widyastuti, SM WIRAMIHARDJA, Memed Y, Yatini Yasuhiko Tomino, Yasuhiko yatini, yatini Yusniwati Yusniwati