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All Journal Jurnal Ilmu Pertanian Indonesia Journal of Tropical Biodiversity and Biotechnology Menara Perkebunan
Hayati Minarsih
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Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Environmental Science Immunology & microbiology

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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 Niyyah FITRANTY; F NURILMALA; Djoko SANTOSO; Hayati MINARSIH
Menara Perkebunan Vol. 71 No. 1: 71 (1), 2003
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

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 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] Asmini BUDIANI; Riza Arief PUTRANTO; Hayati MINARSIH; Imron RIYADI; . SUMARYONO; Barahima ABBAS
Menara Perkebunan Vol. 84 No. 1 (2016): 84 (1), 2016
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

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.
Application of organic fungicide in controlling basal stem rot disease for mature oil palm Happy WIDIASTUTI; Hayati MINARSIH; Djoko SANTOSO; Deden Dewantara ERIS; Galuh Wening PERMATASARI
Menara Perkebunan Vol. 88 No. 1 (2020): 88 (1), 2020
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v88i1.368

Abstract

Ganoderma is a major pathogen in oil palm crops. Some efforts related to control the growth of Ganoderma have been conducted but still have not found an effective method. This study aims to develop an organic fungicide that has been tested in vitro, which effective in controlling the growth of Ganoderma. The optimization carried out includes the determination of the dose and time interval for application in 13-year-old mature oil palm. This organic fungicide application was the continuation of application during the previous year especially for the two best treatment which is application organic fungicide every week (1w) and every two weeks (2w). In this study, the treatments tested were three levels dose of organic fungicide (0, 1x and 2x) and two types of frequency application, i.e. every week (1w) and every other week (2w). The results showed that the best application of organic fungicides was every week application with twice doses (1w.2x), based on the parameters of the inhibition of Ganoderma’s fruiting body formation, primary and secondary root formation, the opening of spear leaves, and harvesting parameters. The application of organic fungicide able to recover the oil palm infected Ganoderma sp., with increasing the fresh fruit bunch and its weight around 70% and 78%, respectively.
Physiological responses and P5CS gene expression of transgenic oil palm plantlet induced by drought stress Turhadi TURHADI; Hayati MINARSIH; Imron RIYADI; . PRIYONO; Asmini BUDIANI
Menara Perkebunan Vol. 88 No. 2 (2020): 88 (2), 2020
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v88i2.386

Abstract

Drought is one of the limiting factors in crop cultivation, such as in oil palm (Elaeis guineensis Jacq.). The transgenic approaches are expected to increase plant tolerance to drought stress and minimize low productivity when drought occurs. Proline is an osmoprotectant compound in plants which its biosynthesis involved the P5CS gene. The objective of this study was to evaluate the tolerance level of P5CS-transgenic oil palm to drought stress induced by polyethylene glycol 6000 (PEG-6000). In this present study, the transgenic and non-transgenic oil palms were treated by  0, 2, and 4% PEG-6000 under in vitro conditions. The experiment was arranged as a factorial completely randomized design with three replications. The drought level score, total chlorophyll content, carotenoids, and proline content, as well as P5CS gene expression in leaf tissues were observed at 7 and 14 days after stress treatments. The result showed that transgenic plantlets had a lower drought level score than those of non-transgenic lines. A concentration of 4% PEG-6000 treatment reduced the total chlorophyll and carotenoids contents than that of 2% concentration in non-transgenic plantlets at 7 and 14 day after treatments (DAT). In addition, proline content and P5CS gene expression level in transgenic had been significantly increased during stress treatment. Based on these results, it can be concluded that the P5CS transgene increased the drought stress tolerance of oil palm.
Co-Authors . NUHAIMI-HARIS . PRIYONO . SUMARYONO . Supriyadi Aksa, Annisa Aulia Asmini BUDIANI Asmini Budiani Asmini BUDIANI1 Barahima Abbas Benyamin Dendang Darmono Taniwiryono Deden Dewantara ERIS Djoko Santoso Dwi SUBIYARTI Dyah LINGGA NP Elis Nina Herliyana Ernayunita, Ernayunita F NURILMALA GALUH WENING PERMATASARI Happy Widiastuti Imron Riyadi Imron RIYADI LAKSMI AMBARSARI Mardhika, Larasati Dena Muhammad Alam Firmansyah Niyyah FITRANTI Niyyah FITRANTY PERMATASARI, Galuh Wening PUTRANTO, Riza Arief Riza A. PUTRANTO Riza Arief PUTRANTO Sekar WOELAN Soekarno Mismana Putra Turhadi Turhadi TW DARMONO DARMONO Yuli Setiawati, Yuli