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All Journal HAYATI Journal of Biosciences Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) Jurnal Ilmu Pertanian Indonesia Jurnal Fitopatologi Indonesia Jurnal Pengolahan Hasil Perikanan Indonesia Jurnal Akuatika Jurnal Ilmu Dasar Microbiology Indonesia BIOTROPIA - The Southeast Asian Journal of Tropical Biology BERKALA SAINSTEK Biota: Jurnal Ilmiah Ilmu-Ilmu Hayati Indonesian Journal of Agricultural Science Indonesian Journal of Biotechnology Jurnal Perlindungan Tanaman Indonesia Journal of Tropical Biodiversity and Biotechnology UNEJ e-Proceeding JURNAL HAMA DAN PENYAKIT TUMBUHAN TROPIKA ANNALES BOGORIENSES Marine Research in Indonesia Biosaintifika: Journal of Biology & Biology Education Menara Perkebunan International Journal of Environment, Engineering, and Education International Journal of Environment, Engineering & Education International Journal of Oil Palm Annales Bogorienses
Antonius Suwanto
Department Of Biology, Faculty Of Mathematics And Natural Science, IPB University, Bogor, Indonesia
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Civil Engineering, Building, Construction & Architecture Control & Systems Engineering Decision Sciences, Operations Research & Management Earth & Planetary Sciences Economics, Econometrics & Finance Education Engineering Environmental Science Immunology & microbiology Materials Science & Nanotechnology Mathematics Medicine & Pharmacology Physics Veterinary

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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 
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
Menara Perkebunan Vol. 73 No. 2: 73 (2), 2005
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | 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.
Engineering of LysR-type Regulator DmlR in Burkholderia ubonensis CP01 to Enhance Its Antifungal Production against Ganoderma boninense Budinarta, Widyah; Purnamasari, Maria Indah; Hermosaningtyas, Anastasia Aliesa; Arifudin Rafif , Muhammad Ghildan; Prihatna , Cahya; Suwanto, Antonius
HAYATI Journal of Biosciences Vol. 33 No. 1 (2026): January 2026
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.33.1.28-43

Abstract

The utilization of an antifungal substance, occidiofungin and burkholdine, derived from Burkholderia ubonensis CP01 has displayed promising results in the management of basal stem rot caused by Ganoderma boninense. The study aims to further enhance the antifungal production of B. ubonensis CP01 through genetic modification. Through comparative genetic analysis, we identified the dmlR gene in B. ubonensis CP01, which is homologous to the scmR gene, a LysR-type transcriptional regulator (LTTR), in B. thailandensis. Deleting the dmlR gene in CP01 resulted in a complete loss of antifungal synthesis. In contrast, overexpression of this gene led to a substantial increase in antifungal production, as determined by an agar well diffusion assay. These findings suggest that dmlR acts as a positive regulator of antifungal gene expression in B. ubonensis CP01. RP-HPLC analysis revealed that the mutant strain overexpressing the dmlR gene (mutant WB12) produced a higher peak at the 24-25 minute elution time. Previous high-resolution mass spectrometry analysis by our group identified the compound at this peak as six analog compounds with monoisotopic masses similar to those of cyclic lipopeptides, including occidiofungin and burkholdine. The WB12 mutant exhibited approximately 15% higher concentrations of antifungal compounds than the wild type. Additionally, whole genome sequencing confirmed that the introduced dmlR gene had been integrated into the locus on chromosome 2 of B. ubonensis CP01. LTTRs play a pivotal role in regulating the production of antifungal agents in CP01. Furthermore, it highlights the potential for manipulating LTTRs to enhance the desirable characteristics of the Burkholderia genus in regard to the production of secondary metabolites.
ISOLASI DAN REGENERASI PROTOPLAS DARI MESOFIL DAUN KENT ANG (Solanum tuberosum L) DIHAPLOID Purwito, Agus; Wattimena, G. A.; Suwanto, Antonius; Suharsono, Inez H.S. Loeddin; Aswidinnoor, Hajrial
Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) Vol. 27 No. 1 (1999): Buletin Agronomi
Publisher : Indonesia Society of Agronomy (PERAGI) and Department of Agronomy and Horticulture, Faculty of Agriculture, IPB University, Bogor, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (925.414 KB) | DOI: 10.24831/jai.v27i1.1579

Abstract

The isolation and regeneration of potato (Solanum tuberosum L) protoplasts have been carried out. Mesophyl cell protoplast were isolated from two dihaploid cultivars of potato BF 15 and SVP 10 leaves used four different enzymes solution. Protoplast were cultured onto four different cultures media to increase plating efficiency. Calli were then transferred to ten different regeneration media. Using cellulose RS 0.5 % and pectolyase Y-23 0.05 % protoplast yield of both cultivars were improved. Medium VKM  supplemented with 0.2 mg/l 2, 4-D, 1.0 mg/l NAA and 0.5 mg/l zeatin or 2iP were increase recovery of colonies from protoplast up to 5.9%. Regeneration medium containing zeatin did always produce more shoots than those of 2iP. Genotypes dependant regeneration frequencies have also been showed in this experiments
PENGUJIAN KET AHANAN KOLEKSI GENOTIPE KEDELAI TERHADAP PENYAKIT BISUL BAKTERI Anggraini, Dyah Kusuma; Tjahjono, Budi; Suwanto, Antonius; Aswidinnoor, Hajrial
Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) Vol. 23 No. 3 (1995): Buletin Agronomi
Publisher : Indonesia Society of Agronomy (PERAGI) and Department of Agronomy and Horticulture, Faculty of Agriculture, IPB University, Bogor, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (796.084 KB) | DOI: 10.24831/jai.v23i3.1634

Abstract

The objective of the research was to evaluate the resistance of soybean genotype in the germplasm collection to pustule disease. Seventy five genotypes were evaluated using spray inoculation method. Inoculation was done on the third week after planting. The 75 genotypes evaluated consists of 29 local varieties, 8 national varieties, 23 introduction, and 15 experimental lines. Results of the experiment showed that among the 75 genotypes tested, one local variety, Si Pinang was found resistant to the pustule disease (Xanthomonas campestris pv. glycines). The resistant local variety was collected from Langkat, North Sumatra.
Transposition and expression of GEP gene in the genome of Vibrio harveyi to monitor its adherence in shrimp larvae Suwanto, Antonius
BIOTROPIA Vol. 13 No. 1 (2006): BIOTROPIA Vol. 13 No. 1 June 2006
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (534.675 KB) | DOI: 10.11598/btb.2006.13.1.74

Abstract

Expression of green fluorescent protein encoded by GFP gene in Vibrio harveyi was investigated to understand the ability of the gene as a molecular marker for adherence of this pathogenic Vibrio in shrimp larvae. The GFP gene was inserted into pUC18Not and pUTmini-Tn5 to generate a recombinant plasmid pWGO2 and pWGO3, respectively, which was transferred into the three isolates of V. harveyi employing diparental mating. Recombinant E. coli carrying pWGO2 and pWGO3 resulted in green-fluorescent colonies and cells due to the production of GFP. However, al1 of mini-Tn5, including mini-Tn5-gfp were not successful1y transferred to V. harveyi. Therefore, we used mini-Tn10 (pLOFKm-gfp) for inserting of gfp gene into V. harveyi genome. Although we could obtain relatively high (l0 pangkat -8) transconjugans employing Tn10, only one of Tnl0 derived isolate of V. harveyi G3 (G3-Tn1Ogfp) showed gfp expression and was further employed for adherence assay. Fluorescent 03-TnlOgfp cells could be observed inside the digestive tract of shrimp larvae and could be distinguished from vibrio that naturally exist in shrimp larvae.
PRESENCE OF hemA-LIKE AND hemT-LlKE GENES IN A JSUMBER OF ANOXYGENIC PHOTOSYNTHETIC BACTERIAL ISOLATES FROM INDONESIA AND SOIL SAMPLES FROM BOGOR AREA AINI, NURUL; SUWANTO, ANTONIUS
BIOTROPIA No. 15 (2000)
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (162.845 KB) | DOI: 10.11598/btb.2000.0.15.158

Abstract

The Rhodobacter sphaeroides hemA and hemT are known to encode a distinct 5-aminolevulinic acid (ALA)-synthase isozyme. This enzyme catalyzes the first and rate limiting step in ALA biosynthesis through the C4 pathway. This study was carried out to detect hemA-\\ke and hemT-\\ke genes in twenty Anoxygenic Photosynthetic Bacterial (APB) isolates from several wetland areas in Indonesia, and four DNA samples that were isolated from four soil samples obtained from Bogor area. Hybridization techniques of Southern and dot blot were used, using hemA and hemT fragment as probes. Southern hybridization analyses indicated the presence of hemA-\\ke gene in five of APB isolates, i.e., MB15, MB16, MB21.2, MB55 and MB6, whereas hemT-\\ke gene was detected only in MB15. Dot blot hybridization analyses suggested that the soil samples from waterlogged paddy-field, dry paddy-field as well as a mud pond were predominantly occupied by prokaryotic organisms which harboured hemA-]\ke gene. However, /iem7"-like sequences were also found in soil sample from dry paddy-field. Key words:    hemA-\\ks gene / hemT-\\ke gene / Southern hybridization analysis / dot blot hybridization analysis.
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 Aini Nurul 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 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 Hariyatun, Hariyatun Hermosaningtyas, Anastasia Aliesa Iman Rusmana Inez H.S. Loeddin Suharsono It Jamilah IVAN HANJAYA JOANITA SADELI KATHARINA JESSICA Kusharyoto, Wien 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 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 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 VICKY MEICY WALTER ERDELEN WIDANARNI WIDANARNI Widanarni Widanarni Wien Kusharyoto Yasinta Ratna Esti Wulandari Yepy Hardi Rustam Yogiara Yogiara YULIN LESTARI Yusminah Hala