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Microbiome Structure Analysis of Oil Palm Pollinator Elaeidobius kamerunicus (Coleoptera; Curculionidae) Arina Amalia Putri; Maria Sugiharti; Alfred Michael; Anja Meryandini; Antonius Suwanto
HAYATI Journal of Biosciences Vol. 30 No. 6 (2023): November 2023
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.30.6.1155-1166

Abstract

The pollination of oil palm by Elaeidobius kamerunicus leads to an increase of over 70% in countries such as India, Malaysia, and Indonesia. The impact of insect-microbiome interactions on the pollination activity and fitness of E. kamerunicus is unknown. Our study aimed to gain insight into the bacterial communities of E. kamerunicus from two different sites with high and low fruit set percentages, using culturable and Terminal Restriction Fragment Length Polymorphism (T-RFLP) analysis. Our result revealed distinctive T-RFs profiles in E. kamerunicus from two different sites. Additionally, the culturable approach showed that some of this microbiome were found only in the weevil population from the high fruit set site. Our findings suggest that these bacteria could contribute to the fitness of E. kamerunicus, leading to a higher fruit set in oil palm plantations.
A Rapid Inoculation Method for Infection of Ganoderma in Oil Palm Maria Indah Purnamasari; Delia Agustina; Cahya Prihatna; Antonius Suwanto
International Journal of Oil Palm Vol. 1 No. 1 (2018): January 2018
Publisher : Indonesian Oil Palm Society /IOPS (Masyarakat Perkelapa-sawitan Indonesia /MAKSI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35876/ijop.v1i1.1

Abstract

Basal stem rot (BSR) is a major disease of oil palm plantation caused by pathogenic fungus Ganoderma boninense. To overcome the disease, development of resistant or tolerantoil palms is crucial for susta inable production of palm oil. Thus a reliable and rapid method to assess resistance of oil palm to BSR is important. Here we report an inoculation technique designated as mycelium inoculation technique. This method is faster and simpler compared to the traditional technique using rubber wood block. The technique that we developed results in consistency of infection and disease can be evaluated as early as two weeks after inoculation with G. boninense. Furthermore, this method is also able to di?erentially assess resistant and susceptible palm seedlings to BSR. The results show that this new inoculation technique can be used as a routine method to infect oil palm seedlings and can be utilised for development of resistant cultivar of oil palm to G. boninense.
Prospecting of Mannan Degrading Bacteria on Treating Palm Kernel Meal Ludovika Jessica Virginia; Yepy Hardi Rustam; Griselda Herman Natadiputri; Antonius Suwanto
International Journal of Oil Palm Vol. 1 No. 1 (2018): January 2018
Publisher : Indonesian Oil Palm Society /IOPS (Masyarakat Perkelapa-sawitan Indonesia /MAKSI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35876/ijop.v1i1.3

Abstract

Palm kernel meal (PKM) is one of the most abundant by-products from oil palm industries. Although PKM can be used as ruminant feed but unfortunately it can not be applied directly as chicken feed due to its complex structure and lack of mannan degrading enzyme in chicken. Manan is a complex molecule composed of linear mannan, glucomannan, galactomannan, and galactoglucomannan and most nutrition contained in PKM was trapped in mannan structure. To increase the economic value of PKM, utilization of mannan degrading enzyme in pretreatment process of PKM before applying as chicken feed is required. We report here that isolation of potential mannanase as well as its prospective toward application in PKM was successfully conducted in this study. Mannanase producing bacteria (CK7) harboring wild type ?-mannanase activity at 48.03 U mL-1 was obtained and identifed as Bacillus subtilis isolate qd386 (accession number EF473134) with 96.7% pairwise identity through 16s amplifcation. Over expression in Pichia pastoris system by pPICZ?A as vector and KM71 as host (P-CK7) improved its activity into 4207.56 U mL-1 and 5000 U mL-1 within fed batch fermentation at 7.5 L and 500 L, respectively. Recombinant manCK7 generated from P-CK7 was reported as the major expressed extracellular protein through SDS-PAGE in this study, thus make it more benefcial and attractive for further application study due to reduction of cell breaking process. Preliminary study on manCK7 application to PKM showed a promising result with a signifcant sugar released observed between PKM which treated with bu?er, commercial enzyme, and manCK7 for 89 µmole, 460.6 µmole, and 518.2 µmole, respectively.
EgMLP1 Gene Expression in Oil Palm Ramet Infected with Ganoderma boninense Yasinta Ratna Esti Wulandari; Felicia Felicia; Arild Ranlym Arifin; Antonius Suwanto
International Journal of Oil Palm Vol. 1 No. 2 (2018): May 2018
Publisher : Indonesian Oil Palm Society /IOPS (Masyarakat Perkelapa-sawitan Indonesia /MAKSI)

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

Oil palm (Elaeis guineensis) is an important crop commodity in Indonesia. However, the effort to increase its production is hindered by basal stem rot (BSR), a devastating disease caused by Ganoderma boninense. The early symptoms of G. boninense infection cannot be observed with naked eyes. It can only be observed at the later stage of the infection. By the time the symptoms appear, the palm is beyond recovery. Hence an early detection technique is essential for effective management of BSR in oil palm. The aim of this research is to analyze and compare the gene expression profile of a defense-related gene encoding early methionine-labeled polypeptide (EgEMLP1) by using quantitative reverse-transcription PCR between the leaves of untreated and treated oil palm cultures with G. boninense. First, G. boninense was isolated and inoculation on oil palm cultures, then RNA isolation was conducted on both oil palm cultures from 3 to 7 weeks post infection (wpi) and purified RNA was used for cDNA synthesis. Finally, EgEMLP1 gene expression was measured using qRT-PCR. The result showed that the expression of EgEMLP1 gene had increased 5.65, 15.66 and 17.96 fold at 3, 5 and 7 wpi compared to those of controls. Furthermore, the result also showed that direct infection method on in vitro oil palm cultures shorten the time needed to infect G. boninense to oil palm compared to rubber wood block method. With less process, direct inoculation method can be an alternative in gene expression in oil palm ramets infected with G. boninense research and up-regulated EgEMLP1 gene expressed has the potential to be developed as a biomarker for early detection of G. boninense infection.
Rapid Inoculation Technique and Biological Control of Leaf Spot Disease in Oil Palm Delia Agustina; Cahya Prihatna; Antonius Suwanto
International Journal of Oil Palm Vol. 2 No. 1 (2019): January 2019
Publisher : Indonesian Oil Palm Society /IOPS (Masyarakat Perkelapa-sawitan Indonesia /MAKSI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35876/ijop.v2i1.24

Abstract

Oil palm (Elaeis guineensis) is one of the most efficient oil-producing crops in the world. However, fungal diseases such as basal stem rot (BSR) caused by Ganoderma sp. causes significant loss to the yield of adult palms. Apart from BSR, leaf spot disease caused by Curvularia sp. also causes significant loss during nursery stages. In this study, Curvularia sp. was isolated from a diseased palm seedling and a rapid and reproducible artificial inoculation method was developed. The technique has bioassay to determine the level of success of the control of leaf spot disease in a glasshouse setting. A natural, organic cyclic peptide fungicide and living cells of a bacterial strain Paraburkholderia sp. CP01 were tested for their efficacy to control leaf spot in oil palm seedlings. The severity of leaf spot disease in oil palm seedlings treated by organic cyclic peptide fungicide and CP01 was significantly lower than untreated control, indicating potential biological control agents. The results presented here provide technical reference and novel approach to controlling leaf spot disease of oil palm.
Increasing Hydrolytic Activity of Lipase on Palm Oil by PCR-Based Random Mutagenesis Maria Dita Febriani Lumban Gaol; Andreas Adhi Satya; Esti Puspitasari; Nisa Rachmania Mubarik; Antonius Suwanto
International Journal of Oil Palm Vol. 3 No. 3 (2020): September 2020
Publisher : Indonesian Oil Palm Society /IOPS (Masyarakat Perkelapa-sawitan Indonesia /MAKSI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35876/ijop.v3i3.53

Abstract

ABSTRACT Random mutagenesis technique is a powerful technique capable of producing enzymes with desired biocatalytic activity. This study aims to obtain a mutant lipase with improved hydrolytic activity on palm oil substrate using random mutagenesis technique. Random mutagenesis by error-prone PCR was used to generate mutant lipases. A total of 1101 mutants were obtained, out of which two mutants, Lip M14.25, and Lip M14.57, showed an increased relative hydrolytic activity. Lip M14.25 and Lip M14.57 demonstrated a 14% and 16% increased activity respectively. A comparison of the mutants' hydrolytic activities using p-nitrophenyl esters showed a significantly high preference for p-nitrophenyl palmitate. Furthermore, the mutant, Lip M14.25 showed its highest activity at pH 5, and Lip M14.57 exhibited a 10 oC decrease in optimum temperature. The two mutants' protein modelling showed the substitution of N44S/S202N on M14.25 and F154L/S265C on M14.57 lipase, which caused changes in conformation and active site residue distance of the lipase. The study found two mutants of lipase, M14.25 and M14.57, which showed improved hydrolytic activity on palm oil substrate.
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.) Asmini BUDIANI; Antonius SUWANTO; Hajrial ASWIDINNOO; Djoko SANTOSO; Basil J NIKOLAU
Menara Perkebunan Vol. 81 No. 2: 81 (2), 2013
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

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.
Isolasi dan kloning fragmen cDNA dari tanaman karet dengan sifat resisten dan rentan terhadap Corynespora cassiicola menggunakan metode cDNA-AFLP Isolation and cloning of cDNA fragments from rubber plant with resistant and susceptible characters to Corynespora cassiicola using cDNA-AFLP method . NURHAIMI-HARIS; Antonius SUWANTO; Maggy T SUHARTONO; Hajrial ASWIDINNOOR
Menara Perkebunan Vol. 78 No. 1: 78 (1), 2010
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v78i1.78

Abstract

AbstractLeaf fall disease caused by Corynespora cassiicola fungi is one of the most important diseases in rubber plant. Rubber clone AVROS 2037 is considered resistant to this pathogen while clone PPN 2444 is susceptible. These two rubberclones were used to identify genes or transcripts differentially expressed during interaction between rubber plants and the fungi, using cDNA-Amplified Fragment Length Polymorphism (cDNA-AFLP) method. Induced genes/transcripts expression was examined to compare differencies between plants uninfected and infected with C. cassiicola at 24, 36, 48 and 72 hours after inoculation. cDNA-AFLP analysis was performed using restriction enzyme VspI and TaqI so the adapters and primers also have the recognition site of these enzymes. By using 29 specific primers, 35 out of approximately 1450 fragments were differentially expressed between AVROS and PPN 2444 clones. All of these fragments were cloned and sequenced. The homology-based grouping of these sequences resulted in 19 contigs and nine individual sequences. Among these, 10 contigs and five sequences have significant sequence homology with known genes in gene bank data base, such as Ran binding protein, protein transporter and transcriptional regulators of some organisms; arginase, GTP-binding protein, heat shock protein (HSP) and aconitase. Ran binding protein, GTPbinding protein and protein transporter were known as membrane proteins while arginase and HSP usually expressed as a response to wounding or toxin treatment. The present of arginase is usually related to the availability of nitric oxide (NO) in plant tissue. NO is well known as a signal molecule on plant defense response. AbstrakPenyakit gugur daun yang disebabkan oleh fungi Corynespora cassiicola merupakan salah satu penyakit penting tanaman karet. Klon karet AVROS 2037menunjukkan sifat resisten terhadap patogen tersebut sedangkan klon PPN 2444 merupakan klon yang rentan. Kedua klon karet tersebut digunakan untuk mengidentifikasi gen atau transkrip yang diekspresikan secara diferensial selama terjadi interaksi antara tanaman karet dengan C. cassiicola menggunakan teknik cDNA-Amplified Fragment Length Polymorphism (cDNA-AFLP). Ekspresi gen/transkrip dipelajari untuk membandingkan perbedaan antara tanaman yang tidak diinfeksi dengan yang diinfeksi patogen pada waktu 24, 36, 48 dan 72 jam setelah inokulasi. Analisis cDNA-AFLP dilaksanakan dengan menggunakan pasangan enzim restriksi VspI dan TaqI sehingga adapter dan primer memiliki sekuen pengenalan kedua enzim restriksi tersebut. Dengan menggunakan 29 pasang primer spesifik, sebanyak 35 dari sekitar 1450 fragmen memiliki ekspresi berbeda antara klon AVROS 2037 dan PPN 2444. Semua fragmen yang berbeda tersebut kemudian diklon pada vektor kloning dan disekuen. Hasil sekuensing dikelompokkan berdasarkan homologi sekuennya dan menghasilkan 19 contigs serta sembilan macam sekuen yang tidak mengelompok. Sebanyak 10 dari 19 contigs dan lima dari sembilan sekuen tersebut memiliki homologi dengan produk gen yang telah dikenal yang terdapat di pangkalan data GenBank, seperti putative Ran binding protein, protein transporter, regulator transkripsi, arginase, GTP-binding protein, heat shock protein (HSP) dan aconitase. Beberapa di antaranya seperti putative Ran binding protein, protein transporter dan GTP-binding protein dikenal sebagai protein membran, sedangkan arginase dan HSP merupakan protein atau enzim yang ekspresinya pada tanaman antara lain dipengaruhi oleh adanya pelukaan dan perlakuan toksin. Keberadaan arginase sering berhubungan dengan ketersediaan nitric oxide (NO) pada jaringan tanaman. NO dikenal sebagai salah satu sinyal molekul dalam mekanisme pertahanan tanaman.
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 Asmini BUDIANI; Djoko SANTOSO; Hajrial ASWIDINNOOR; Antonius SUWANTO
Menara Perkebunan Vol. 74 No. 1: 74 (1), 2006
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

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.
Kloning parsial gen penyandi enoil-ACP reduktase dari mesokarp buah kelapa sawit (Elaeis guineensis Jacq.) Partial cloning of gene encoding enoyl-ACP reductase from mesocarp of oil palm (Elaeis guineensis Jacq.) Asmini BUDIANI; Djoko SANTOSO; Hajrial ASWIDINNOOR; Antonius SUWANTO
Menara Perkebunan Vol. 72 No. 1: 72 (1), 2004
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v72i1.126

Abstract

Summary Enoyl-ACP reductase (ENR) is a component of fatty acid synthase (FAS) that is considered to play an important role in fatty acid elongation and oil accumulation of several plants. One of the proteins expressed coinciding with fruit development and oil accumulation in oil palm has been detected from the previous study and had homology with ENR. Therefore, as a part of genetic engineering program to improve oil content and quality in oil palm mesocarp, this research was aimed to clone cDNA conserved region of gene encoding enoyl-ACP reductase from oil palm mesocarp. Based on the amino acid sequence of the polypeptide that was homologous with ENR and combined with information of conserved region sequences of the same gene from other plant sources, primers were designed for amplifying conserved region of the ENR gene. Amplifi-cation was carried out by RT-PCR using total RNA as template, at several annealing temperatures and MgCl2 concentrations. Amplification product was cloned using pCR 2.1-TOPO, and the sequence was subjected into BlastN analysis. The results confirmed that the cloned cDNA fragment with 698 bp in size was the conserved region of the ENR gene.  This sequence was highly homologous with the same gene from other plants such as Oryza sativa, Olea europaea, Brassica napus, Triticum aestivum and Arabidopsis thaliana with E-value 1e-96, 7e-77, 2e-64, 5e-41 and 3e-36, respectively. Based on this result, primers have been made and used to amplify the 5’- and 3’ ends of the ENR -cDNA  of oil palm mesocarp. Ringkasan Enoil-ACP reduktase (ENR) merupakan salah satu komponen asam lemak sintase (FAS) yang berperan penting dalam pemanjangan asam lemak dan akumulasi minyak pada berbagai tanaman. Salah satu protein yang ter-ekspresi sejalan dengan tahapan perkembangan buah sawit dan akumulasi minyak pada penelitian sebelumnya diketahui mempunyai homologi dengan ENR. Oleh karena itu, sebagai salah satu bagian dari usaha rekayasa metabolisme minyak pada mesokarp buah sawit, penelitian ini bertujuan untuk mengklon cDNA daerah konservatif gen penyandi ENR dari mesokarp buah sawit. Berdasarkan  sekuen asam amino polipeptida yang mempunyai homologi dengan ENR dan dikombinasikan dengan hasil penjajaran daerah konservatif gen tersebut dari berbagai anaman lain, dirancang primer  untuk  amplifikasi daerah konservatif ENR. Amplifikasi dilakukan dengan RT-PCR menggunakan templat RNA total pada berbagai suhu   penempelan   dan   konsentrasi    MgCl2. Hasil amplifikasi dimurnikan dari gel dan diklon menggunakan vektor kloning pCR2.1-TOPO serta dianalisis nya menggunakan BlastN. Hasilnya mengkonfirmasi fragmen cDNA terklon berukuran 698 pb sebagai daerah konservatif ENR tersebut mempunyai homologi tinggi dengan gen yang sama dari    O. sativa,  O. europaea, B. napus, T. aestivum dan  A. thaliana masing-masing dengan E-value 1e-96, 7e-77, 2e-64, 5e-41 dan 3e-36. Berdasarkan hasil tersebut telah dibuat primer spesifik untuk amplifikasi cDNA daerah ujung 5’- dan 3’- gen  ENR dari mesokarp kelapa 
Co-Authors . EFRIWATI . Melani . NURHAIMI-HARIS A A Hermosaningtyas A A Hermosaningtyas, A A ADI YULANDI Agus PURWANTARA Agus Purwito Agustin Wydia Gunawan Agustina, Delia Alfred Michael ALINA AKHDIYA RUSMANA AMARILA MALIK AMARILA MALIK Andi Khaeruni Andi Khaeruni Andreas Adhi Satya Andreas Adhi Satya ANJA MERYANDINI Anja Meryandini ARI SUSILOWATI Arifudin Rafif , Muhammad Ghildan Arild Ranlym Arifin Arina Amalia Putri Aris Tjahjoleksono Aris Tri Wahyudi ARTINI PANGASTUTI Asmini Budiani Basil J NIKOLAU BIBIANA W LAY BUDI TJAHJONO BUDI TJAHJONO Budi Tjahjono Budi Tjahjono Budinarta, Widyah C Hanny Wijaya CAHYA PRIHATNA Cahya Prihatna Cahya Prihatna CECILIA ANNA SEUMAHU CECILIA ANNA SEUMAHU DEBORA HADISUSANTO Delia Agustina Desniar - - DIAH ISKANDRIATI DIANA ELIZABETH WATURANGI Djoko Santoso Dondin Sajuthi DWI SURYANTO Dyah Kusuma Anggraini Edi Husen Edi Husen EDI HUSEN ERNIN HIDAYATI Esti Puspitasari ESTI PUSPITASARI ESTI PUSPITASARI Esti Utarti Eunice Limantara EVELINE AYU Felicia Felicia Felicia Kartawidjajaputra Ferymon Mahulette Ferymon Mahulette Ferymon Mahulette, Ferymon G. A. Wattimena Gayuh Rahayu Griselda Griselda Griselda Herman Natadiputri Griselda, Griselda Hajrial ASWIDINNOO HAJRIAL ASWIDINNOOR Hariyatun Hariyatun Hariyatun, Hariyatun Hermosaningtyas, Anastasia Aliesa Iman Rusmana Inez H.S. Loeddin Suharsono It Jamilah IVAN HANJAYA JOANITA SADELI KATHARINA JESSICA Kusharyoto, Wien Lilis Nuraida Linda Wati LISTYA UTAMI KARMAWAN Ludovika Jessica Virginia MAGGY T HENAWIDJAJA Maggy T Suhartono Maggy T Suhartono MAGGY T. SUHARTONO Maggy T. Suhartono MAGGY T. SUHARTONO MAGGY THENAWIJAYA SUHARTONO Maria Dita Febriani Lumban Gaol Maria Indah Purnamasari Maria Indah Purnamasari Maria Indah Purnamasari Maria Sugiharti Meity S. Sinaga MEITY SURADJI SINAGA Muhamad Azwar Syah Muhammad Agus Muljanto MUHAMMAD ZAIRIN Jr. Ni Nyoman Tri Puspaningsih NISA RACHMANIA MUBARIK NURITA TORUAN-MATHIUS NURUL AINI PRIHASTO SETYANTO Prihasto Setyanto Prihatna , Cahya Puspitasari, Esti QURROTA A’YUN R. Sapto Hendri Boedi Soesatyo Raden Ajie Syahbarie RAHAYU WIDYASTUTI RASTI SARASWATI RASTI SARASWATI Rasti Saraswati Rasti Saraswati RATIH DEWI HASTUTI RATNA SETYANINGSIH RATNA SETYANINGSIH RIDWAN AFFANDI Rika Indri Astuti ROB HARLING ROHANI CINTA BADIA GINTING Rustam, Yepy Hardi Satya, Andreas Adhi Sheila Sutanto Sheila Sutanto, Sheila SUMARDI Suryo Wiyono SUSAN SOKA SUSILOWATI1 SUSILOWATI1 T RESNAWATI P URWADARIA Tan Watumesa Agustina TARUNI SRI PRAWAST MIEN KAOMINI ANY ARYANI DEDY DURYADI SOLIHIN Tati Barus TEMMY DESILIYARNI TRESNAWATI PURWADARIA VICKY MEICY WALTER ERDELEN WIDANARNI WIDANARNI Widanarni Widanarni Wien Kusharyoto Yasinta Ratna Esti Wulandari Yepy Hardi Rustam Yogiara Yogiara YULIN LESTARI Yusminah Hala