Hayati Minarsih
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PENGENDALIAN SERANGAN GANODERMA SPP. (60-80%) PADA TANAMAN SENGON SEBAGAI PELINDUNG TANAMAN KOPI DAN KAKAO Herliyana, Elis Nina; Taniwiryono, Darmono; Minarsih, Hayati; Firmansyah, Muhammad Alam; Dendang, Benyamin
Jurnal Ilmu Pertanian Indonesia Vol. 16 No. 1 (2011): Jurnal Ilmu Pertanian Indonesia
Publisher : Institut Pertanian Bogor

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Abstract

Information about genetic variation of Ganoderma spp. As a couse of rot disease on plantation crops is necessary for consideration in efforts to protect crops. Exploration of the use of biological agents, especially Trichoderma spp., for the control of Ganoderma on forestry crops is still limited to laboratory testing. Its effectiveness to control Trichoderma spp. To protect plants in the nursery sengon being carried out, as well as to deternime its role in improving plant growth.
Pengaruh jumlah subkultur dan media sub-optimal terhadap pertumbuhan dan kemampuan regenerasi kalus tebu (Saccharum officinarum L.) (Effect of repeated subculture and suboptimum media on the growth of sugarcane calli (Saccharum officinarum L.)) MINARSIH, Hayati; Suharyo, .; RIYADI, Imron; RATNADEWI, Diah
E-Journal Menara Perkebunan Vol 84, No 1: Oktober 2016
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v84i1.219

Abstract

Sugarcane (Saccharum officinarum L.) is an important crop for sugar production. One attempt to increase sugarcane productivity is through micropropagation and quality improvement of sugarcane seedlings in vitro. This research aimed to study the effect of repeated subcultures on callus capacity for regeneration and plant survival in acclimatization phase, as well as the influence of suboptimum media on the recovery capability of sugarcane callus to proliferate in vitro. Fourth subcultured sugarcane callus derived from young leaves were used as material in this research. Basic medium of Murashige and Skoog (MS) added with 3 mg/L 2,4-D, 10% coconut water, and 3% sucrose was used for callus initiation. For callus regeneration, the MS medium was supplemented with 2 mg/L BAP, 0.2 mg/L IAA, 10% coconut water, and 3% sucrose. Study on the effect of subculture numbers consisted of three stages, i.e. initiation, regeneration, and acclimatization, while the study on resting phase or the use of sub-optimal media included six treatment media and two pathways. Results showed that the fifth subcultures produced embryoid callus (91%), the highest non mucilaginous callus (97%), and the highest abnormality rate (6%). Results from the suboptimum media treatment, showed that B pathway (4 week resting phase) was better than the A pathway (8 week resting phase), based on fresh weight and callus abnormality percentage. A and B pathways indicated that the growth of callus can be recovered when it was grown back to the normal media and 1.5D-MS treatment of the resting phase showed the best growth and appearance. 
Analisis keragaman genetik Ganoderma spp. yang berasosiasi dengan tanaman kakao dan tanaman pelindungnya menggunakan Random Amplified Polymorphic DNA (RAPD) Genetic diversity analysis of Ganoderma spp. associated with cocoa and its shade trees using Random Amplified Polymorphic DNA (RAPD) MINARSIH, Hayati; LINGGA NP, Dyah; DARMONO, TW DARMONO; HERLIYANA, Elis Nina
E-Journal Menara Perkebunan Vol 79, No 1: Juni 2011
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v79i1.72

Abstract

AbstractInformation on genetic diversity of Ganoderma spp.causing root rot disease in crops is important to developa proper strategy for the control of Ganoderma disease. Theobjectives of this research were to study the genetic diversityof Ganoderma spp. associated with cacao and its shade trees(Albazia faltacaria, Swietenia mahogani, Adenatheramicrosperma and Leucaena leucocephala) by randomamplified polymorphic DNA (RAPD) analysis. Fourty fivesamples of Ganoderma spp. were used in this research. Theresults showed that DNA amplification using 10 arbitraryoligonucleotide primers produced 220 DNA fragmentsshowing polymorphisms. The cluster analysis showed that 45number of Ganoderma samples had a high variability with thecoefficient value ranged from 0.71 to 0.91. Further analysisusing Winboot software showed that three groups ofGanoderma spp. had a high degree of confidence (>50 %),which were Ganoderma samples from sengon (Paraserianthessp.) of Tasikmalaya, sengon (Paraserianthes sp.) ofPalembang, and mahogany of Jember; whereas the othergroups of samples had a low degree of confidence (<50%).AbstrakInformasi tentang keragaman genetik Ganoderma spp.sebagai penyebab penyakit busuk akar pada tanamanperkebunan sangat diperlukan untuk menerapkan strategiyang tepat dalam upaya perlindungan tanaman perkebunan.Penelitian ini bertujuan untuk mengetahui keragaman genetikGanoderma spp. yang berasosiasi dengan tanaman kakao dantanaman pelindungnya (sengon, mahoni, saga dan lamtoro)dari berbagai wilayah di Indonesia menggunakan penandamolekuler random amplified polymorphic DNA (RAPD).Sebanyak 45 sampel Ganoderma spp digunakan dalampenelitian ini. Amplifikasi DNA dengan 10 primer terpilihmenghasilkan 220 fragmen DNA yang menunjukkan adanyapolimorfisme. Hasil analisis menunjukkan adanya keragamanyang cukup tinggi di antara sampel Ganoderma spp. daripohon inang dan wilayah yang berbeda, dengan nilaikoefisien 0,71-0,91. Berdasarkan analisis bootstrapdiketahui bahwa tiga kelompok sampel Ganoderma spp.memiliki tingkat kepercayaan yang tinggi (>50 %) yaitukelompok Ganoderma spp. yang berasosiasi dengan pohonsengon asal Tasikmalaya, sengon Palembang, dan mahoniJember; sedangkan pengelompokan lainnya menunjukkmenunjukkan tingkat kepercayaan yang rendah (<50 %).
Ekspresi dan kloning gen penyandi ADP-Glucose Phyrophosphorylase dari tanaman sagu (Metroxylon sagu Rottb.) Expression and cloning of gene encoding ADP-Glucose Phyrophosphorylase from sago palm (Metroxylon sagu Rottb.) BUDIANI1, Asmini; PUTRANTO, Riza Arief; MINARSIH, Hayati; RIYADI, Imron; SUMARYONO, .; ABBAS, Barahima
E-Journal Menara Perkebunan Vol 83, No 2: Desember 2015
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v83i2.4

Abstract

AbstractSago palm (Metroxylon sagu Rottb.) is a potential food and energy resources becouse it is the highest starch producing plant.  Breeding of sago palm should be directed to produce elite genotype with superior characters such as high starch content, wider pith diameter, without spine and high starch quality. However, research on sago palm in Indonesia so far is limited espescially in the field of cultivation and breeding, and attempt to produce such elite would take long time. Availability of molecular marker for starch content would be beneficial to shorten the length period of breeding. ADP-Glucose Phyrophosphorylase is one of the important enzymes in starch biosynthesis. Therefore its gene is an interesting subject in order to develope molecular marker of high starch content.  This research was aimed to study the expression of gene encoding AGP in the sago palm with high starch content versus low starch content, and to clone the full cds of the gene. RNA was isolated from leaf and pith of both palms. Exspression analysis and amplify-cation of full cds were conducted by Reverse Transcryptase-Polymerase Chain Reaction (RT-PCR) using specific primers. The results showed that sago palm with higher starch content expressed AGP higher than that of sago palm with lower  starch content. Expression of AGP in the full developing leaf was higher than in the young leaf, and there was no expression detected in the pith. The full cds of AGP was successfully amplified and cloned. Even though the DNA sequence showed high homology with DNA sequence of the same gene that has been deposited in GenBank, there were differences in severall nucleotide including that in the active domain of the enzyme.AbstrakTanaman sagu merupakan sumber pangan dan energi yang sangat potensial untuk dikembangkan karena merupakan tanaman penghasil karbihidrat tertinggi. Pemuliaan tanaman sagu mestinya diarah-kan untuk menghasilkan bibit sagu yang selain memiliki rendemen pati tinggi, juga memiliki diameter empulur besar, tidak berduri dan memiliki cita rasa pati yang enak. Namun, sampai saat ini riset mengenai sagu di Indonesia masih sangat terbatas, sehingga pemuliaan sagu untuk menghasilkan bibit unggul demikian akan memerlukan waktu lama. Ketersediaan penanda rendemen pati akan sangat membantu mempercepat pemuliaan tanaman tersebut. ADP-Glucose Pyrophosphorylase adalah salah satu enzim yang berperan penting dalam biosintesis pati, sehingga gene penyandinya merupakan subjek yang menarik dalam pengembangan marka kandungan pati tinggi.  Sebagai bagian dari upaya untuk mendapat-kan penanda rendemen pati tinggi pada tanaman sagu, penelitian ini bertujuan untuk mempelajari ekspresi gen penyandi AGP. RNA diisolasi dari daun tanaman sagu rendemen pati rendah dan tanaman sagu rendemen pati tinggi. Perbedaan tingkat ekspresi gen penyandi AGP dari tanaman sagu rendemen pati tinggi vs rendemen pati rendah, dianalisis dengan teknik Reverse-Transcryptase PCR menggunakan primer spesifik. Hasil penelitian menunjukkan bahwa tanaman sagu rendemen pati tinggi mengekspresikan AGP lebih tinggi dibandingkan dengan tanaman sagu rendemen pati rendah. Ekspresi gen tersebut pada daun tua (full developing leaf) lebih tinggi di-bandingkan dengan pada daun muda, dan pada empulur tidak dideteksi ekspresi gen tersebut. Daerah penyandi lengkap AGP subunit kecil telah diklon. Meskipun memiliki homologi yang tinggi dengan sekuen DNA gen yang sama yang telah dideposit pada  GenBank,  namun terdapat perbedaan beberapa nukleotida termasuk pada daerah domain aktif dari enzim tersebut. 
Deteksi Ganoderma secara molekuler pada kebun kelapa sawit yang diberi perlakuan biofungisida Ganor (Molecular detection of Ganoderma on oil palm plantation treated with Ganor biofungicide) MINARSIH, Hayati; WIDIASTUTI, Happy; SANTOSO, Djoko
E-Journal Menara Perkebunan Vol 86, No 1 (2018): April, 2018
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v1i1.289

Abstract

AbstractGanor organic fungicide potentially reduces Ganoderma, a pathogenic fungus causing basal stem rot disease. Application of Ganor on oil palm trees in the plantation attacked Ganoderma, inhibits the growth of Ganoderma fruiting bodies, improves rooting and stimulates the opening of the spear leaf. This study aims to identify molecularly the presence of Ganoderma in oil palm trees that have been attacked by Ganoderma routinely treated with Ganor for three months. Molecular analysis was performed by PCR using Ganoderma specific primers. The analysis results of sample from trunks and roots of  oil palm, indicating that the Ganoderma infected oil palm which has been treated with Ganor, were relatively free (96.4%) of Ganoderma. Of the 28 samples examined of treated plants, 27 samples did not indicate the presence of Ganoderma specific DNA band. On the other hand, the untreated oil palm trees infected by Ganoderma were still detected by the appearence of  DNA bands specific to Ganoderma. The results of molecular analysis indicated that Ganor treatments can effectively reduce the attack rate of Ganoderma in oil palm trees in the plantation infected by Ganoderma. However, the use of the molecular technique for early detection needs to be further tested to evaluate its consistency prior to introduction to the commercial growers. The reproducibility can be confirmed by repeating the experiment using more samples. Ganor effectiveness in curing oil palm trees infected by Ganoderma, maybe indicated by the ability of the reproductive organs to develop, particularly female flowers. The sex ratio of Ganor treated oil palms was clearly higher than that of control palms in 10 to 12 weeks after the treatment.[Keywords: organic fungicides, stem rot, molecular analysis, Elais guinensis Jack.] AbstrakFungisida organik Ganor berpotensi mengurangi serangan Ganoderma, cendawan patogenik penyebab penyakit busuk pangkal batang. Aplikasi Ganor pada tanaman kelapa sawit di kebun yang terserang Ganoderma, menghambat pertumbuhan tubuh buah Ganoderma, memper-baiki perakaran dan merangsang pembukaan daun tombak. Penelitian ini bertujuan untuk mengidentifikasi secara molekuler adanya Ganoderma pada tanaman kelapa sawit terserang Ganoderma yang telah mendapat perlakuan Ganor secara rutin selama tiga bulan. Analisis molekuler dilakukan dengan teknik PCR menggunakan primer DNA spesifik Ganoderma. Hasil analisis sampel batang dan akar tanaman kelapa sawit, menunjukkan bahwa tanaman Perlakuan, yaitu kelapa sawit terserang Ganoderma yang telah mendapat perlakuan Ganor, 96,4% bebas Ganoderma. Dari 28 sampel tanaman Perlakuan yang diperiksa, 27 sampel tidak menunjukkan adanya pita DNA spesfik Ganoderma. Sementara itu pada tanaman Kontrol, yaitu tanaman kelapa sawit terserang Ganoderma dan tidak mendapat perlakuan Ganor, 100% masih terdeteksi adanya Ganoderma. Dari 7 sampel tanaman kontrol yang diperiksa semuanya menunjukkan adanya pita DNA spesifik Ganoderma. Hasil analisis molekuler ini mengindikasikan bahwa perlakuan Ganor efektif mengurangi tingkat serangan Ganoderma pada tanaman kelapa sawit di kebun yang terinfeksi Ganoderma. Namun demikian, untuk lebih meyakinkan praktisi perkebunan, penggunakan teknik molekuler ini masih perlu diuji lebih lanjut terkait konsistensinya. Reprodusibilitas dapat dikonfirmasi dengan mengulangi percobaan menggunakan lebih banyak sampel. Efektivitas Ganor dalam menyehatkan tanaman kelapa sawit terserang Ganoderma ini, terindikasi juga dari perkembangan organ reproduktifnya. Sex ratio meningkat dalam waktu 10 hingga 12 minggu setelah perlakuan.[Kata Kunci:  fungisida organik, busuk pangkal  batang, analisis molekuler, Elais guinensis Jack. ]
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. 
ROOT DISEASES GANODERMA SP. ON THE SENGON IN WEST JAVA AND EAST JAVA Herliyana, Elis Nina; Taniwiryono, Darmono; Minarsih, Hayati
Jurnal Manajemen Hutan Tropika Vol. 18 No. 2 (2012)
Publisher : Institut Pertanian Bogor (IPB University)

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Abstract

Sengon tree (Paraserianthes falcataria (L.) Nielsen) currently becomes a major forest tree species widely planted by smallholders in Indonesia. The wood of this is quick growing and relatively easy to sell.  However, level of plant safety sengon between crop plantations and other forestry need to be assessed considering the sengon tree is alternative host of Ganoderma spp. Studies have been conducted to know the presence and diversity of Ganoderma spp. on the sengon tree and some ways inoculation on sengon plant in the nursery.  Survey of Ganoderma conducted in several locations of community forests and cacao (Theobroma cacao) plantations in West Java and East Java. Testing of genetic diversity based on RAPD technique. This conducted at the Biotechnology Research Institute of Plantation Indonesia Bogor. Inoculation testing conducted at the Faculty of Forestry, Bogor Agricultural University.  The results showed that Ganoderma lucidum was found on the sengon tree and cacao plant, generally on the dead stump. The test results of genetic diversity obtained genetic similarity between G. lucidum from sengon and cacao are quite close. The results of inoculation of G. lucidum testing on seedlings sengon showed that both isolate from cacao and sengon tree able to infect a sengon tree back. The existence of sengon tree as shade plants for cacao plant need to watch out, because production cycle of sengon tree faster than production cycle of cacao plant that is protected.
Isolasi dan karakterisasi gen dehydrin dari tebu (Saccharum officinarumL.) yang terlibat dalam respon toleransi cekaman kekeringan (Isolation and characterization of dehydrin gene from sugarcane (Saccharum officinarum L.) involved in drought tolerance response) MINARSIH, Hayati; FANIAR, .; KRISTANTI, Tati; AMANAH, Dian M; SUSTIPRIJATNO, .; SUHANDONO, Sony
E-Journal Menara Perkebunan Vol 86, No 2 (2018): Oktober 2018
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v86i2.300

Abstract

Nowadays,the development of molecular biology techniques has enabled to engineer drought tolerant sugarcane to accelerate thebreeding program. Dehydrin(DHN)that belong to the group II late embryogenesis abundant (LEA) family is known to havean important role in plant response and adaptation to abiotic stresses (drought, high salinity, cold, heat, etc.). Literature study and bioinformatics analysis reported that DHN1gene on sugarcane showed high homology sequences with sorghum DHN. The expression of DHN1gene on sugarcane var. PSJT 941 treated with various periodof drought stress had been conducted using semi-quantitative reverse transcriptase (RT)-PCR method. The results showed that the expressionlevel of DHN1 geneincreased along withthe increased period of the treatment. The highest expression level of DHN1 gene was resulted from plants that had been subjected to drought for 25 days. Amplification of DHN1gene  from plants withthe highest gene expression, resulted an amplicon  with a size of 465 bp which representsa full length coding sequence (CDS) of DHN1. Identification using Blast analysis showed that DHN1sequences  from sugarcane var. PSJT 941 shared high homology with DHN gene on sugarcane and sorghum. The alignment results also revealed a conserved motif that characterized DHN genes.[Key words: drought stress, dehydrin, DHN1gene, sugarcane]Abstrak Dengan berkembangnya teknik biologi molekuler saat ini, maka perakitan tanaman tebu yang toleran kekeringan lebih diarahkan melalui teknik rekayasa genetika untuk mempercepat program pemuliaan tanaman.  Protein dehydrin (DHN) yang termasuk ke dalam kelompok II  famili LEA (Late Embryogenesis Abundant)diketahui berperan penting dalam respon dan adaptasi tanaman terhadap cekaman abiotik (kekeringan, salinitas tinggi, suhu dingin, panas, dll). Studi literatur dan analisis bioinformatika menunjukkan bahwa gen DHN1pada tanaman tebu memiliki homologi yang tinggi dengan gen DHNpada sorghum. Analisis ekspresi gen DHN1pada tanaman  tebu varietasPSJT 941yang diberi cekaman kekeringan telah dilakukan menggunakan semi-kuantitatifreverse transcriptase (RT)-PCR dan terlihat bahwa ekspresi gen DHN1meningkat secara nyata sejalan dengan semakin lamanya waktu pemberian cekaman. Tingkat ekspresi gen DHN1paling tinggi diperoleh dari tanaman yang mengalami cekaman kekeringan selama 25 hari.  Amplifikasi gen DHN1pada tanaman dengan tingkat ekspresi yang paling tinggi menunjukkan pita dengan ukuran 465 bp yang merepresentasikan full coding sequence(CDS) gen DHN1. Identifikasi menggunakan analisis Blast menunjukkan bahwa sekuen gen DHN1dari tanaman tebu varietas PSJT 941yang diperoleh memiliki homologi yang tinggi dengan gen DHNpada tanaman tebu dan sorghum. Hasil penjajaran sekuen protein juga menunjukkan adanya motif lestari yang mencirikan gen DHN. [Kata kunci: cekaman kekeringan, dehydrin, gen DHN1, tebu]
Mikropropagasi planlet tebu menggunakan sistem perendaman sesaat (SPS) Micropropagation of sugarcane plantlets using temporary immersion system (TIS) MINARSIH, Hayati; RIYADI, Imron; SUMARYONO, .; BUDIANI, Asmini
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.v81i1.53

Abstract

bstractTo achieve Indonesian sugar self-sufficiency in2014, the national production needs to be escalatedthrough land extensification that requires a largenumbers of cane planting materials. This can be achievedby mass propagation of sugarcane through in vitroculture. Solid medium is commonly used for callusproliferation in sugarcane tissue culture. However, solidmedium is considered inefficient in terms of plantletproduction level, labour and space. The use of liquidmedium may solve the problem by allowing automationto increase plantlet production scale and uniformity.Temporary immersion system (TIS) is based on a shortperiodic immersion of explants in a liquid medium for aspecific frequency and duration. Research on in vitromass propagation of sugarcane using TIS was conductedat the Indonesian Biotechnology Research Institute forEstate Crops. Callus initiated from immature unfoldedleaves of PSJT 941 and PS 881 was cultured on liquidMS medium in TIS with different frequencies (12 and24 h) and durations (1 and 3 min) of immersion. Eachtreatment was replicated three times. The callus biomassof two elite cane varieties (PSJT 941 and PS 881)cultured in TIS for six weeks was higher (2 – 4 times fold)than that of on solid medium. The PSJT 941 varietyreached the highest calli biomass with immersion forthree min every 24 h. However, PS 881 variety reachedits highest biomass with immersion for one minute every24 h. The propagation of sugarcane using TIS culturewas proven to produce higher calli biomass up to fourfolds and to form more numbers and uniform shootscompared to the solid medium culture. The callus wassuccesfully regenerated to shoots and plantlets.AbstrakUntuk mencapai swasembada gula, perlu dilakukanpeningkatan produksi gula nasional melalui perluasanareal pertanaman tebu sehingga diperlukan bibit dalamjumlah besar. Hal tersebut dapat diatasi antara laindengan perbanyakan tebu melalui kultur in vitro. Peng-gunaan medium padat pada perbanyakan kalus tebumelalui kultur in vitro merupakan teknik yang umumdigunakan saat ini. Akan tetapi penggunaan mediumpadat dianggap kurang efisien dalam hal jumlah planletyang diproduksi, tenaga kerja dan ruang digunakan.Penggunaan medium cair dapat mengatasi kelemahantersebut dengan dimungkinkannya otomatisasi sehinggadapat meningkatkan skala produksi secara massal dankeseragaman planlet. Sistem perendaman sesaat (SPS)merupakan teknik kultur in vitro dalam medium cairmenggunakan bejana bersekat dimana kontak antaraeksplan dan medium terjadi hanya secara sesaat danperiodik. Penelitian perbanyakan massal bibit tebumelalui SPS dilakukan di Balai Penelitian BioteknologiPerkebunan Indonesia. Kalus diinisiasi dari daun meng-gulung varietas PSJT 941 dan PS 881 yang ditumbuhkanpada media MS cair dalam kultur SPS dengan frekuensiyang berbeda (12 dan 24 jam) dan lama perendaman (1dan 3 menit). Setiap perlakuan diulang tiga kali. Bobotbasah (biomassa) kalus dari dua varietas tebu (PSJT 941dan PS 881) yang ditumbuhkan dengan metode SPSsetelah enam minggu menunjukkan pening-katan yanglebih tinggi yaitu antara 2 - 4 kali lipat dibandingkandengan kontrol (media padat). Peningkatan biomassatertinggi pada varietas PSJT 941 diperoleh pada per-lakuan SPS dengan interval perendaman 24 jam dan lamaperendaman tiga menit. Sedangkan pada PS 881,peningkatan tertinggi biomassa diperoleh pada intervalperendaman 24 jam dan lama perendaman satu menit.Perbanyakan dengan metode SPS terbukti dapat mening-katkan biomassa kalus lebih dari empat kali lipat danpembentukan tunas yang lebih seragam dibandingkandengan pada media padat. Kalus yang dihasilkan dapatdiregenerasikan menjadi tunas dan planlet.
Kloning dan karakterisasi daerah promoter gen penyandi ADP glucose pyrophosphorylase dari Metroxylon sagu rendemen pati-tinggi dan -rendah [Cloning and characterization of promoter region of ADP glucose pyrophosphorylase-encoding gene from Metroxylon sagu with high- and low-starch content] BUDIANI, Asmini; PUTRANTO, Riza Arief; MINARSIH, Hayati; RIYADI, Imron; SUMARYONO, .; ABBAS, Barahima
E-Journal Menara Perkebunan Vol 84, No 1: Oktober 2016
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v84i1.200

Abstract

ADP-glucose pyrophosphorylase (AGPase) is one of the key enzymes in the starch biosynthesis. In many plants, the activity of this enzyme was reported to affect the yield and composition of the produced starch. This research is a part of an effort to develop molecular markers for early selection of high starch-yielding of sago palm. The purpose of the research was to isolate promoters of AGP gene and to analyze the differences in their DNA sequences between sago palm with high starch content (MsHS) and low starch content (MsLS). DNA was isolated and purified from the leaves of the two sago palm. The promoter region of AGP was amplified by Genome Walking technique. The specific primers were designed by Primer3 program based on the information of DNA sequence of AGP genes of sago palm from previous studies. Selected DNA fragments resulted from Genome Walking were isolated from the gel, cloned into E. coli, and analyzed its DNA sequence. DNA sequence analysis showed that one DNA fragment from MsHS  (± 1500 bp) and one DNA fragment from MsLS (> 2000 bp) were confirmed as a 5’ upstream of the AGP gene.  Further in silico analysis using MEME program identified various DNA motifs of cis-acting elements, which confirmed that those DNA fragment were promoter region of the gene. Preliminary analysis showed the differences in DNA sequences and motives of cis-acting elements in the promoter region of the two samples which might influence or indirectly associated with the character of the starch yield in sago palm.