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All Journal Vegetalika AGRITROP International Journal of Biosciences and Biotechnology Jurnal Ilmu Dasar JURNAL TEKNOLOGI LINGKUNGAN AL KAUNIYAH BERKALA SAINSTEK Biota: Jurnal Ilmiah Ilmu-Ilmu Hayati Agrointek Indonesian Journal of Agricultural Science Jurnal AgroBiogen Buletin Penelitian Tanaman Rempah dan Obat ROTOR: JURNAL ILMIAH TEKNIK MESIN Indonesian Journal of Biotechnology UNEJ e-Proceeding Agriprima : Journal of Applied Agricultural Sciences Jurnal Bioteknologi & Biosains Indonesia (JBBI) ANNALES BOGORIENSES Mesin Agritrop : Jurnal Ilmu-Ilmu Pertanian (Journal of Agricultural Science) Jurnal Pengabdian Magister Pendidikan IPA J-Dinamika: Jurnal Pengabdian Kepada Masyarakat Jurnal Informasi dan Teknologi Jurnal JTIK (Jurnal Teknologi Informasi dan Komunikasi) Buletin Penelitian Tanaman Rempah dan Obat International Journal of Quantitative Research and Modeling BIOVALENTIA: Biological Research Journal Journal of Law, Education and Business Biosaintifika: Journal of Biology & Biology Education

Bambang Sugiharto

Fakultas Matematika Dan Ilmu Pengetahuan Alam Universitas Jember

Author-ID : 388822

Agriculture, Biological Sciences & Forestry Humanities Automotive Engineering Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Computer Science & IT Control & Systems Engineering Decision Sciences, Operations Research & Management Economics, Econometrics & Finance Education Energy Engineering Environmental Science Immunology & microbiology Industrial & Manufacturing Engineering Law, Crime, Criminology & Criminal Justice Materials Science & Nanotechnology Mathematics Mechanical Engineering Medicine & Pharmacology Physics Social Sciences

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A SIMPLE PROTOCOL FOR SOMATIC EMBRYOGENESIS INDUCTION OF IN VITRO SUGARCANE ( Saccharum officinarum. L) BY 2,4-D AND BAP Widuri, Laily Ilman; Dewanti, Parawita; Sugiharto, Bambang
BIOVALENTIA: Biological Research Journal Vol. 2 No. 1 (2016)
Publisher : Biology Department, Faculty of Mathematics and Natural Sciences, Sriwijaya University

Induction of in vitro sugarcane through somatic embryogenesis technique influenced by addition of plant growth regulator. The objective of this research was to determine appropriate formulation medium for indirect somatic embryogenesis induction on two potential sugarcane SUT Event 02 and PS 881. This research carried out in three steps, callus induction, callus proliferation, and shoot regeneration. Explants taken from basal of in vitro plantlet one month SUT Event 02 and PS 881 resulted from shoot regeneration previously. Five different medium formulas applied for callus induction and one formula for proliferation and shoot regeneration. Research using completely randomized design (CRD) factorial with five different formulation induction mediums. The result showed that the best respond of indirect somatic embryogenesis on SUT Event 02 and PS 881 was medium containing Â 3 mgL-1of Â 2,4-D.

TRANSFORMASI GENETIK DAN EKSPRESI MUTAN SUCROSE PHOSPHATE SYNTHASE PADA TANAMAN TOMAT Suwinda Fibriani; Inyana Dwi Agustien; Widhi Dyah Sawitri; Bambang Sugiharto
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol. 6 No. 1 (2019): June 2019
Publisher : Balai Bioteknologi, Badan Pengkajian dan Penerapan Teknologi (BPPT)

Genetic Transformation and Expression of Sucrose Phosphate Synthase Mutant in Tomato PlantÂ ABSTRACTSucrose phosphate synthase (SPS) is a key enzyme responsible for sucrose biosynthesis. In its regulation, SPS activity is modulated by an allosteric effector glucose-6-phosphate (G6P) suggested to have an ability to bind SPS N-terminus domain. To understand the role of N-terminus in regulating SPS, the SPS gene was mutated with the deletion of N-terminus domain (âˆ†N-SPS). The âˆ†N-SPS gen was transformed into tomato plants with 5% transformation efficiency. Three transgenic tomato plant 4.20, 5.5.1, and 5.10 were obtained and confirmed by PCR analysis. Transgenic tomato expression was characterized by enzymatic analysis. Result showed that the G6P allosteric regulation in transgenic âˆ†N-SPS had lost and the SPS activity increased by 2-fold compared to non-transgenic plant. This showed that N-terminus domain-deleted SPS could be actively expressed in plant.Â Keywords: enzyme, genetic transformation, N-terminus domain deletion, sucrose phosphate synthase, tomatoÂ ABSTRAKSucrose phosphate synthase (SPS) merupakan enzim kunci yang bertanggung jawab dalam sintesis sukrosa. Dalam regulasinya, aktifitas SPS dipengaruhi oleh alosterik efektor glukosa-6-fosfat (G6P) yang diduga dapat berikatan pada domain N-terminus SPS. Untuk mengetahui peran N-terminus pada regulasi SPS, dilakukan mutasi SPS dengan penghilangan domain N-terminus (âˆ†N-SPS). Gen âˆ†N-SPS diinsersi pada tanaman tomat melalui transformasi genetik dengan efisiensi transformasi 5%. Tiga tanaman transgenik tomat (event4.20; 5.5.1; dan 5.10) didapatkan dan positif terkonfirmasi melalui analisis PCR. Ekspresi mutan dikarakterisasi melalui analisis enzimatik. Hasil menunjukkan bahwa tanaman tomat transgenik âˆ†N-SPS tidak dipengaruhi regulasi alosterik G6P dan aktifitas SPS 2 kali lipat lebih tinggi daripada tanaman bukan transgenik. Ini menunjukkan bahwa SPS dengan delesi domain N-terminus dapat terekspresi aktif pada tanaman. Â Kata Kunci: delesi domain N-terminus, enzim, sucrose phosphate synthase, tomat,Â transformasi genetikÂ

KEAMANAN PANGAN KOMODITAS TRANSGENIK: STUDI KASUS SEDIAAN PAKAN HIJAUAN DARI DAUN TEBU OVEREKSPRESI SoSPS1 A Bagus Nur Sudrajat; Nurhayati Nurhayati; Bambang Sugiharto
AGROINTEK Vol 15, No 3 (2021)
Publisher : Agroindustrial Technology, University of Trunojoyo Madura

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21107/agrointek.v15i3.7868

Sugarcane (Saccharum officinarum L) is a sucrose-producing plantation that has been cultivated in various countries. Genetically modified sugar cane sugar cane plants are sugar cane plants that are assembled biotechnology through overexpression of the SoSPS1 gene. The SoSPS1 gene is responsible for the biosynthesis of sucrose sugar in leaves. Waste in the form of leaves, shoots, and bagasse has not been used optimally as animal feed. Thus it takes a lot of innovation and appropriate technology in the use of sugarcane waste for animal feed, so that it is expected to achieve zero waste farming systems that can be utilized all without being wasted and polluting the environment. As an alternative feed for livestock, then genetically engineered sugar cane must be tested for feed safety. Feed safety testing includes substantial equivalence value and feed digestibility test in which genetically engineered products have nutritional and digestive values commensurate with conventional products. Substantial value of sugarcane planted by genetically engineered overexpression of SoSPS1 gene did not differ significantly compared to conventional sugarcane. However, there are some significant nutritional content, but these values are still in the range of values in the OECD.

DNA Barcoding of Medicinal Orchid Dendrobium discolor Lindl. Tanimbar Using rbcL and ITS genes Dian Al Ghifari Perwitasari; Siti Rohimah; Tri Ratnasari; Bambang Sugiharto; Mukhamad Su'udi
Buletin Penelitian Tanaman Rempah dan Obat Vol 31, No 1 (2020): Buletin Penelitian Tanaman Rempah dan Obat
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/bullittro.v31n1.2020.8-20

Dendrobium discolor Lindl., Tanimbar is one of the medicinal orchids that has been used to treat skin diseases. Morphologically, D. discolor Tanimbar shows similarities with D. discolor Merauke and D. bigibbum, making it challenging to identify. DNA barcoding using ribulose-1,5-bisphosphate carboxylase large (rbcL) and Internal Transcribed Spacer (ITS) markers expected to be used to identify D. discolor accurately. This study aimed to identify potential DNA sequences as barcodes for the identification of medicinal orchid D. discolor Tanimbar using molecular markers rbcL and ITS. The DNA genome of D. discolor Tanimbar was isolated and used as a template in the PCR reaction. The resulting amplicons were then sequenced. The results showed that the rbcL gene sequence of D. discolor had high homology with D. salaccense (Accession: LC193510.1, Prect. Ident : 99.45 %), whereas the ITS had high homology with D. nindii (Accession: AY239985.1 Identification: 98.67 %). Bioinformatics analysis showed that the rbcL gene sequence from D. discolor had more homology sequence than the ITS. However, the ITS sequence was more specific and could differentiate to species level. Based on the results of this study, the ITS sequence can be recommended as a molecular marker for the identification of the medicinal orchid D. discolor Tanimbar.

Distinguishing resistances of transgenic sugarcane generated from RNA interference and pathogen‐derived resistance approaches to combating sugarcane mosaic virus Weny Nailul Hidayati; Retnosari Apriasti; Hardian Susilo Addy; Bambang Sugiharto
Indonesian Journal of Biotechnology Vol 26, No 2 (2021)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.65256

Sugarcane mosaic virus (SCMV) is a causative agent that reduces growth and productivity in sugarcane. Pathogen‐derived resistance (PDR) and RNA interference (RNAi) are the most common approaches to generating resis‐ tance against plant viruses. Two types of transgenic sugarcane have been obtained by PDR and RNAi methods using a gene‐encoding coat protein (CP) of SCMV (SCMVCp). This research aimed to distinguish resistance of the two transgenic sugarcanes in combating SCMV through artificial viral inoculation. The experiment was conducted using transgenic sugar‐ cane lines validated by PCR analysis. Insertion of gene‐encoding CP in the transgenic lines was confirmed by amplification of 702 bp of DNA fragment of SCMVCp. After viral inoculation, mosaic symptoms appeared earlier, at 21 days post inoculation (dpi) in PDR transgenic lines, but was at 26 dpi in RNAi transgenic lines. Symptom observation showed that 77.8% and 50% of the inoculated plants developed mosaic symptoms in PDR and RNAi transgenic lines, respectively. RT‐PCR analysis revealed that the nuclear inclusion protein b (Nib) gene of SCMV was amplified in the symptomatic leaves in plants classified as susceptible lines. Immunoblot analysis confirmed presence of viral CP with a molecular size of 37 kDa in the susceptible lines. Collectively, these results indicated that the RNAi approach targeting the gene for CP effectively produces more resistance against the SCMV infection in transgenic sugarcane compared to the PDR approach.

Agrobacterium tumefaciens-MEDIATED IN PLANTA TRANSFORMATION METHOD FOR THE SoSPS1 GENE IN CITRUS PLANTS (Citrus nobilis L.) Ni Putu Ayu Erninda Oktaviani Suputri; Rindang Dwiyani; Ida Ayu Putri Darmawanti; Bambang Sugiharto
International Journal of Biosciences and Biotechnology Vol 7 No 1 (2019)
Publisher : Central Laboratory for Genetic Resource and Molecular Biology, Faculty of Agriculture, Udayana University in cooperation with Asia-Oceania Bioscience and Biotechnology Consortium (AOBBC)

The SoSPS1 gene of sugar cane plants previously subjected to Agrobacterium tumefacienmediated cloning was to be transferred to citrus plants to increase metabolism of sucrose in plant. The T-DNA harbored the SoSPS1 gene under the control of the CaMV 35S promoter from the cauliflower mosaic virus and contained the NPTII gene (kanamycin resistance gene) as a selectable marker for transformant selection. Generally, gene transformation in plants is carried out by tissue culture. However, tissue culture has several disadvantages such as its being time-consuming, its sometimes resulting in somatic mutations and somaclonal variations, and the requirement of sterile conditions in the procedure of gene transfer. In planta transformation is a useful system for those plants that lack tissue culture and regeneration system. The main function of in planta transformation is to recover the advantages of tissue culture as an efficient, quick method, including its ability to produce a large number of transgenic plants and to accumulate a high concentration of total soluble protein in short time. There are two procedures of in planta transformation for the seeds of citrus plants, namely “prick and coat” and “seed tip-cutting and imbibition”. In the prick and coat method, seeds are pricked on their entire surfaces and smeared with a suspension of Agrobacterium tumefaciens. In the seed tip-cutting and imbibition method, on the other hand, seeds are cut at the tip and soaked in a suspension of Agrobacterium tumefaciens. The leaves derived from seeds treatment were taken as samples for DNA extraction and PCR using primers of the NPTII gene (Forward: 5’-GTCATCTCACCTTCCTCCTGCC-3’; Reverse: 5’-GTCGCTTGGTCGGTCATTTCG-3’). This research found that only the seed tip-cutting and imbibition plants amplified along the 550-bp band, while those of the prick and coat method did not. Additionally, the T-DNA was successfully integrated into the genome of the plants treated with the seed tip-cutting and imbibition method but not with the prick and coat.

Isolasi dan Identifikasi Khamir Toleran Alkohol dari Molase Nurhayati; Jay Jayus; Anjas Wida Elistia Rini; Bambang Sugiharto; Dedy Eko Rahmanto
Biota : Jurnal Ilmiah Ilmu-Ilmu Hayati Vol 7, No 1 (2022): February 2022
Publisher : Universitas Atma Jaya Yogyakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24002/biota.v7i1.5426

Khamir mampu menghasilkan bioetanol dari molases. Selama proses produksi (proses fermentasi) terjadi proses akumulasi alkohol sehingga konsentrasi alkohol semakin meningkat. Konsentrasi etanol yang melebihi 8% (v/v) dapat menyebabkan ketidakstabilan membran yang kemudian dapat menurunkan permeabilitas membran dan homeostasis sel. Penelitian ini dilakukan isolasi dan identifikasi khamir indigenus dari molase yang toleran terhadap alkohol. Karakterisasi morfologi meliputi bentuk koloni, bentuk sel dan tipe pertunasa, sedangkan karaktersasi fisiologis dilakukan dengan kit API 20C. Identifikasi molekuler dilakukan dengan PCR dan sequencing DNA menggunakan primer ITS1-ITS4. Hasil penelitian didapatkan dua isolat khamir yaitu isolat X dan Z yang memiliki potensi toleran terhadap alkohol. Berdasarkan hasil karakteristik fisiologis, isolat X dan isolate Z memiliki kedekatan dengan Candida tropicalis. Hasil sequencing DNA dengan primer ITS1-ITS4 dan analisis dengan program blastn menunjukkan bahwa isolat X teridentifikasi sebagai spesies Candida parapsilosis strain AUMC 10714 dan isolat Z teridentifikasi sebagai spesies Candida parapsilosis ZA012.

Pemuliaan Kentang Produk Rekayasa Genetik Tahan terhadap Penyakit Busuk Daun (Phytophthora infestans) dan Aman Pangan di Indonesia A. Dinar Ambarwati; Tri J. Santoso; Edy Listanto; Toto Hadiarto; Eny I. Riyanti; Kusmana Kusmana; Bambang Sugiharto; Netty Ermawati; Sukardiman Sukardiman
Jurnal AgroBiogen Vol 13, No 1 (2017): Juni
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jbio.v13n1.2017.p67-74

Pemanfaatan tanaman kentang produk rekayasa genetik (PRG) dalam pemuliaan tanaman melalui persilangan dengan Atlantic dan Granola telah menghasilkan enam galur PRG hasil silangan yang terseleksi. Sebelum komersialisasi, kentang PRG harus dikaji keamanan pangan dan lingkungannya. Penulisan bertujuan memberikan informasi mengenai tanaman kentang PRG di Indonesia yang tahan terhadap penyakit busuk daun Phytophthora infestans dan telah dinyatakan aman untuk di-konsumsi oleh manusia. Analisis stabilitas menunjukkan bahwa gen RB stabil terintegrasi selama empat generasi klonal ber-urutan dalam genom tanaman kentang PRG dengan satu sisipan gen. Hasil studi komposisi dan nutrisi, glikoalkaloid total, dan anti nutrisi pada kentang PRG Katahdin SP951 dan galur-galur silangannya bersifat sepadan dengan Katahdin non-PRG. Studi toksisitas menunjukkan bahwa pemberian pakan suspensi umbi kentang dan suspensi tepung kentang Katahdin SP951 dan galur-galur silangan tidak berdampak terhadap mortalitas, bobot badan, dan tanda-tanda klinis pada mencit. Protein RB tidak memiliki homologi yang tinggi dengan protein toksin sehingga tidak bersifat toksik. Studi alergenisitas dengan Simulated Gastric Fluid dan Simulated Intestinal Fluid menunjukkan bahwa protein umbi kentang Katahdin SP951 dan galur-galur silangan terdegradasi kurang dari 5 menit inkubasi setelah perlakuan enzim pepsin atau tripsin. Protein RB tidak mempunyai sekuen asam amino yang homolog dengan protein alergen, sehingga tidak berpotensi menimbulkan alergi. Kentang Katahdin SP951 telah dinyatakan aman untuk dikonsumsi melalui Keputusan Kepala Badan Pengawas Obat dan Makanan tahun 2016. Tanaman kentang PRG tahan P. infestans yang dapat mengurangi 50% aplikasi fungisida, dan telah mendapat sertifikat aman pangan dan aman lingkungan diharapkan dapat menjadi pilihan untuk dimanfaatkan petani.

Regenerasi Kalus Embriogenik Sorgum (Sorghum bicolor) menggunakan Kombinasi ZPT dan Mikronutrien Nadya Oktafiana; Siti Umayyah; Wulan Nursyiam Ningtyas; Bambang Sugiharto
Agriprima : Journal of Applied Agricultural Sciences Vol 6 No 1 (2022): MARCH
Publisher : Politeknik Negeri Jember

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.25047/agriprima.v6i1.463

Perakitan varietas unggul tanaman sorgum dapat dilakukan melalui perbanyakan secara in-vitro melalui kultur jaringan. Somatik embriogenesis menjadi salah satu metode perbanyakan yang tepat untuk menghasilkan tanaman dalam waktu yang cepat dan jumlah banyak. Tetapi, rendahnya kemampuan regenerasi kalus menyebabkan kegagagalan terbentukya tunas dan tanaman baru. Pemberian nutrisi dan zat pengaruh tumbuh (ZPT) yang efektif pada media regenerasi menentukan keberhasilan regenerasi kalus. Sitokinin dan auksin merupakan jenis ZPT yang berperan dalam pembelahan dan perkembangan sel serta menstimulasi pertumbuhan tunas pada kalus. Sedangkan, CuSO4 digunakan sebagai nutrisi mikro tambahan yang berperan aktif dalam katalisasi enzim dan transfer elektron pada proses fotosintesis. Tujuan penelitian ini yaitu untuk mengetahui konsentrasi terbaik dari kombinasi antara Kinetin, NAA, dan CuSO4 pada media regenerasi kalus sorgum untuk menstimulasi pertumbuhan tunas dan akar. Hasil penelitian menunjukkan kombinasi MS0, 0,1 mg/L NAA, 1 mg/L kinetin menjadi kombinasi paling baik unuk regenerasi tunas yaitu 6,38 tunas. Sedangkan untuk regenerasi akar kombinasi MS0 dan 1 mg/L CuSO4 merupakan konsentrasi terbaik untuk regenerasi tunas dan akar sebanyak 32,1 akar dan meningkatkan jumlah planlet sebanyak 7,67. Namun sebaliknya, tidak ada pengaruh pada penambahan CuSO4 terhadap tinggi tanaman. Planlet yang tumbuh mampu beradaptasi pada kondisi in vivo di dalam green house dan menunjukkan tidak ada perbedaan pertumbuhan antara tanaman yang tumbuh dari biji dan tanaman hasil in vitro.

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/bullittro.v31n1.2020.8-20

Co-Authors A Bagus Nur Sudrajat A. Dinar Ambarwati Agustien, Inyana Dwi Agustien, Inyana Dwi Ahmad Ilham Tanzil Amir, Muhammad Hisyam Anjas Wida Elistia Rini Anuary Dwi Rosyid, Wafa Prasetia Arif Syaifudin, Arif Ariyono, Alief Rizky Arsy Chairiyah Asshidiqie, Wahyu Pratama Azizah, Kunti Anis Bayati, Tutiana Bon, Abdul Thalib Bin Budi Sulistiarini, Emma Dedi Dwilaksana Dedy Eko Rahmanto Dewita, Tiara Ajeng Dian Al Ghifari Perwitasari Digdo Listyadi, Digdo Djenal . Dwi Djumhariyanto Edia F.D Edia, Edia F.D Edy Listanto Elizabeth Eneng Tita Tosida Eny I. Riyanti Erlia Narulita Esti Utarti Fajrianto, Sigit Fanata, Wahyu Indra Duwi Ferdi Hariyanto Fibriani, Suwinda Hafiz, Syauqi Abyan Hafizi, Muhamad HARDIAN SUSILO ADDY Hari Purnomo Hasibuan, Aqil Muhammad Hayati, Nur Fadillah Husin, Andriati Amir Ida Ayu Putri Darmawanti Ika Oktavianawati Imas Cintamulya Inyana Dwi Agustien Iryani Endah Febrianti Jayus Jayus Khiar Ayatina Hasbi Kusmana Kusmana Laily Ilman Widuri, Laily Ilman Laksono Trisnantoro Lasino Lasino, Lasino Lassa, Anita Liem, Jhon Sepron Defretus Magnon, Cassidy Maisaro, Maisaro Maulani, Ardela Alief Maulida, Dinda Mala Media Ningtyas, Rinda MISWAR - Mochamad Asrofi Muhammad Hazmi Muhammad, Adibillah Mukhamad Su'udi Muslimah Arniyanti Nadya Oktafiana Neliana, Intan Ria Netty Ermawati Netty Ermawati Ni Putu Ayu Erninda Oktaviani Suputri Nikmah, Nasilatun Nina Oktaria, Nina Ningtiyas, Wulan Nursyiam Nugroho, Sandi Luqman Nurhayati Oktapan, Ladefa Primana Parawita Dewanti Purnama Okviandari Purnama Okviandari, Purnama Purwinto, Rahmanaji Setyo Rachman Setiawan Rahardhianto, Arsetyo Raharja, Ajung Gilang Putra Raharjo, Sulistyo Yusuf Rahma Rei Sakura Ramadhani, Rifki Fahrisal Ratnasari, Tri Restanto, Didik Pudji Retnosari Apriasti Rindang Dwiyani Risky Mulana Anur Salahuddin Junus Salih, Yasir Sigit Soepardjono Siswahyudianto Siti Rohimah Siti Umayyah Soleh, Mohamad Badrus SRI AYU MADE Sri Rejeki Subandi, Kotim Sugara, Victor Ilyas Sugianto, Efendi Sukardiman Syafina Pusparani Toto Hadiarto Tri Agus Siswoyo TRI HANDOYO Tri J. Santoso Tri Ratnasari Ummi Sholikhah Untung Murdiyanto Wahyu Indra Duwi Fanata Wahyu Nurkholis Hadi Syahputra Weny Nailul Hidayati Widhi Dyah Sawitri Wulan Nursyiam Ningtyas Yanti Setianti Yayuk Fatmawati Yoga Yuniadi Yudi Rinanto Yundari, Yundari Yusuf Rachmandika

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