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Asmini BUDIANI
Indonesian Research Institute for Biotechnology and Bioindustry

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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
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.
Peningkatan hasil panen kedelai (Glycine max L.) varietas Wilis dengan aplikasi biostimulan tanaman (Yield improvement of soybean (Glycine max L.) var. Wilis by the application of organic plant biostimulant) Dini Astika SARI; Irma KRESNAWATY; . PRIYONO; Asmini BUDIANI; Djoko SANTOSO
E-Journal Menara Perkebunan Vol 87, No 1 (2019): April, 2019
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (817.538 KB) | DOI: 10.22302/iribb.jur.mp.v87i1.295

Abstract

The Indonesian government program of achieving self-sufficiency of soybean by 2020 requires technological innovations for the farmers. The use of plant biostimulant is an innovative strategy and proven previouslyto increase the productivity of several otherfood crops. The aim of this study was to analyze the effect of PPBBI biostimulant on the growth, productivity and quality of a Wilis variety of soybean under greenhouse conditions. PPBBI biostimulant at10 ppm and 20 ppm, was applied using foliar spray method with 20 mL volume to each plant. The applications were carried out 1 time, 2 times and 3 times. Six biostimulant treatments showed significant positive effects on the vegetative growth rate, generative organ development, and yield. The biostimulant accelerated vegetative growth to enter the generative phase earlier than that of in the control plants. The period of generative organs maturity required for treated plants was 7-14 d shorter than that of in the control plants so that the harvest period was 21 d shorter. Weight per 100 seeds of the P2-3 (application 3 times at 20 ppm); P2-2 (application 2 times at 20 ppm); P2-1 (application 1 time at 20 ppm) and P1-3 (application 3 times at 10 ppm) treatments were 20.16 g; 17.65 g; 18.89 g and 16.89 g respectively with no significant difference, while the control plants was only 11.60 g. Based on the results of all parameters e.g. average number of seeds, average weight per seed, and potential for yield improvement, the treatment of P1-3 (application 3 times at 10 ppm) was the best treatment with potential yield increase by 59.06% and oil content by 11.37%.[Key words: generative, organic biostimulant, productivity, vegetative]  AbstrakProgram pemerintah Indonesia dalam pencapaian swasembada kedelai pada tahun 2020 membutuhkan dukungan inovasi teknologi yang aplikatif untuk para petani. Biostimulan tanaman merupakan salah satu teknologi yang strategis dan terbukti dapat meningkatkan produktivitas beberapa tanaman pangan. Tujuan penelitian ini adalah untuk melakukan analisis pengaruh aplikasi biostimulan PPBBI terhadap partum-buhan, produktivitas dan kualitas hasil panen kedelai varietas Wilis pada kondisi rumah kaca. Biostimulan PPBBI dengan variasi konsentrasi   10 ppm dan 20 ppm, diaplikasikan pada tanaman kedelai dengan metode penyemprotan lewat daundengan volume 20 mL per tanaman.Penyemprotan dilakukan sebanyak 1 kali; 2 kali dan 3 kali aplikasi. Enam perlakuan biostimulanyang diujikan menunjukkan pengaruh positif yang signifikan pada kecepatan pertumbuhan vegetatif, stimulasi perkembangan organ generatif dan peningkatan hasil panen. Biostimulan PPBBI mempercepat laju pertumbuhan vegetatif untuk memasuki fase generatif lebih awal dibandingkan tanaman kontrol. Masa perkembangan dan pemasakan organ generatif polong pada tanaman perlakuan menjadi lebih pendek 7-14 hari dibandingkan tanaman kontrol sehingga secara keseluruhan masa panen tanaman perlakuan lebih singkat 21 hari. Bobot per 100 biji tanaman kedelai perlakuan P2-3; P2-2; P2-1 dan P1-3 berturut-turut mencapai 20,16 g; 17,65 g; 18,89 g; dan 16,89 g dengan tidak adanya perbedaan signifikan, sedangkan tanaman kontrol hanya 11,60 g. Berdasarkan hasil analisis seluruh peubah yaitu rerata jumlah biji, rerata bobot per biji, dan potensi produksi, maka perlakuan P1-3 (aplikasi tiga kali dengan dosis 10 ppm) merupakan perlakuan terbaik dengan potensi kenaikan produksi mencapai 59,06% dan kadar lemak 13,7%.  [Kata kunci: biostimulan organik, generatif, produktivitas, vegetatif]
Regeneration of oil palm plantlets introduced by P5CS gene using Agrobacterium-mediated transformation Asmini BUDIANI; Imam Bagus NUGROHO; Hayati MINARSIH; Imron RIYADI
E-Journal Menara Perkebunan Vol 87, No 2 (2019): OKTOBER, 2019
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1347.764 KB) | DOI: 10.22302/iribb.jur.mp.v87i2.336

Abstract

AbstrakCekaman kekeringan dapat mempengaruhi produktivitas tanaman perkebunan. Rekayasa genetika merupakan salah satu cara untuk meningkatkan produktivitas tanaman perkebunan penting seperti kelapa sawit. Tujuan dari penelitian ini adalah melakukan perekayasaan kelapa sawit melalui introduksi gen P5CS dengan transformasi berbasis Agrobacterium untuk meningkatkan ketahanan tanaman terhadap cekaman kekeringan. Pada penelitian ini perakitan kelapa sawit transgenik yang tahan terhadap cekaman kekeringan dilakukan melalui transformasi gen P5CS (Δ1-pyrroline-5-carboxylate synthetase) ke dalam kalus embriogenik (embryogenic calli – EC) menggunakan Agrobacterium. Plasmid pBI_P5CS yang membawa gen P5CS ditransfer dari Escherichia coli XL1 Blue ke Agrobacterium tumefaciens AGL1 melalui konjugasi. Selanjutnya klon Agrobacterium yang membawa plasmid pBI_P5CS digunakan untuk menginfeksi kalus embriogenik kelapa sawit dengan perlakuan 100 ppm asetosiringon. Kalus transforman diregenerasi pada media de Fossard (DF) yang ditambahkan 50 ppm kanamisin dan 250 ppm sefotaksim. Kalus transforman diseleksi melalui uji GUS dan metode PCR menggunakan primer NPTII dan P5CS1. Uji GUS dilakukan untuk menyeleksi kalus transforman yang ditunjukkan dengan reaksi positif pembentukan warna biru pada kalus yang berhasil ditransformasi dengan konstruk pBI_P5CS. Pengujian dengan menggunakan PCR memberikan hasil positif dengan adanya profil pita PCR pada visualisasi menggunakan pewarnaan SYBR Green, yang menunjukkan amplikon berukuran ~ 0,7 kb untuk gen NPTII dan ~ 0,4 kb untuk gen P5CS pada elektroforesis dengan gel agarosa. Hasil dari penelitian ini adalah diperolehnya kalus transforman terseleksi yang telah diregenerasi dan tumbuh menjadi planlet.[Kata kunci: cekaman kekeringan, Elaeis guineensis Jacq., rekayasa genetika, planlet]Abstract      Environmental abiotic stressors particularly drought has detrimental effects upon the productivity of estate crops. Increasing the crop tolerance towards drought stress through genetic engineering is one of the strategies employed to maintain steady productivity of valuable crop, i.e. oil palm. The aim of this study was to engineer oil palm with a better tolerance towards drought by introducing P5CS (Δ1-pyrroline-5-carboxylate synthetase) gene via Agrobacterium–mediated transformation into embryogenic calli (EC). The pBI_P5CS plasmid harboring P5CS gene was transferred from Escherichia coli XL1 Blue to Agrobacterium tumefaciens AGL1 by conjugation. The positive clone of transformed Agrobacterium was then used to infect oil palm EC by the addition of 100 ppm acetosyringone. The transformed ECs were regenerated in the de Fossard (DF) media supplemented by 50 ppm kanamycin and 250 ppm cefotaxime followed by GUS assay and PCR-based screening using NPTII and P5CS1 primers. The positive EC clones were confirmed by GUS assay, which produced blue coloration on positive transformed oil palm EC. A positive result of PCR screenings was depicted by PCR products in SYBR Green staining gel agarose electrophoresis with the expected band size of ~ 0.7 kb for the NPTII gene and ~ 0.4 kb for the P5CS gene. This study has successfully selected and regenerated pBI_P5CS transformed oil palm embryogenic calli into plantlets.[Keywords: drought tolerance, Elaeis guineensis Jacq., genetic engineering, plantlets]
Purification, characterization, and bioassay of putative protease inhibitors from Hevea brasiliensis latex Riza Arief PUTRANTO; . SISWANTO; Agustin Sri MULYATNI; Asmini BUDIANI; Radite TISTAMA
E-Journal Menara Perkebunan Vol 84, No 2 (2016): Desember 2016
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1452.514 KB) | DOI: 10.22302/iribb.jur.mp.v84i2.220

Abstract

Lateks yang menyerupai cairan susu putih diperoleh dari penyadapan kulit batang tanaman karet (Hevea brasiliensis). Lateks merupakan sitoplasma dari jaringan pembuluh bernama latisifer yang didalamnya terkandung berbagai macam komponen, termasuk protein-protein penting. Berbagai jenis enzim yang memiliki fungsi terkait pertahanan tanaman dari serangan patogen dan pelukaan telah berhasil dideteksi di dalam lateks, di antaranya protease inhibitor (PI). Protease inhibitor memiliki aktivitas senyawa antifungi sehingga berpotensi untuk  dimanfaatkan sebagai biofungisida. Pada penelitian ini, protease  inhibitor putatif yang berasal dari serum B (lutoid) lateks tanaman karet telah berhasil diisolasi menggunakan teknik Ion Exchange Chroma-tography. Dari total 70 fraksi protein yang diekstrak dari kolom, hanya 26 fraksi yang menunjukkan kadar protein yang terukur. Kandungan protease inhibitor putatif yang di-peroleh berkisar antara 0,0067 hingga 0,022 mL/g serum B dari hasil 3 fraksi terpilih. Aktivitas penghambatan terhadap empat enzim protease (subtilisin A, tripsin, α-kimotripsin, dan papain) menunjukkan karakteristik protease inhibitor putatif tersebut sebagai serine dan/atau cysteine inhibitor protease dengan persentase hambatan di atas 15% terhadap protease target. Hasil SDS-PAGE memperlihatkan pemisahan protein dominan yang diperkirakan merupakan protease inhibitor putatif dengan berat molekul sebesar 21,5 kDa. Uji bioassay aktivitas antifungi secara in vitro dari protease inhibitor memperlihatkan penghambatan pertumbuhan miselium dari fungi Ganoderma boninense, Sclerotium sp., dan Rigidosporus lignosus. [Kata kunci : protease inhibitor, Hevea brasiliensis, lateks, serum B, ion exchange chromatography]AbstractLatex, a milky white liquid, is the main product from rubber tree (Hevea brasiliensis). Latex is the cytoplasm of complex cellular networks named laticifers in which it contains many different components, including important proteins. Various types of enzymes carrying functions associated with plant defense against pathogen and wounding have been detected in latex in which one of these enzymes is protease inhibitor (PI). Plant protease inhibitor has tremendous potential as an antifungal agent which can be developed as biofungicide. In this work, protease inhibitors from B-serum (lutoid) of rubber tree latex were isolated and purified using Ion Exchange Chromatography (IEC) technique. Of the total 70 fractions of proteins extracted from the columns, only 26 fractions showed measurable levels of protein. The concentration of obtained putative protease inhibitors (three fractions of IEC) ranged from 0.007 to 0.022 mL/g B-serum. Inhibitory activity against four protease enzymes (subtilisin A, trypsin, α-chymotrypsin, and papain) showed the characteristics of Hevea putative protease inhibitors from B-serum as serine and/or cysteine protease inhibitors with more than 15% inhibitory activity of target protease. Based on SDS-PAGE visualization, the molecular weight of dominant protein considered as Hevea putative protease inhibitors was 21.5 kDa. In vitro bioassay test of antifungal activity for Hevea putative protease inhibitors showed reduced mycelium growth of Ganoderma boninense, Sclerotium sp., and Rigidosporus lignosus.[Keywords: protease inhibitor, Hevea brasiliensis, latex, B-serum, ion exchange chromatography]