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Enhancement of Regeneration Efficiency through Callus Induction Media Using 2,4-Dicholorophenoxyacetic acid in Indica Rice (Oryza sativa L. var. Ciherang) Azizah, Kunti Anis; Restanto, Didik Pudji; Sugiharto, Bambang
Jurnal ILMU DASAR Vol 18 No 2 (2017)
Publisher : Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Jember

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1379.154 KB) | DOI: 10.19184/jid.v18i2.2592

Abstract

Indica rice variety Ciherang is the most planted variety in Indonesia, but the micropropagation technique is restricted because it is known has low regeneration frequency and included as recalcitrant cultivar for tissue culture and tranformation activities. One of solution to resolve that problem is developing a technique of somatic embryogenesis in callus of ciherang rice cultivar. The aims of study were to determine medium composition for inducting embryogenic callus in Ciherang rice and to know the effectivity of rice regeneration using callus as explant. The methods were included induction of embryogenic callus in callus induction media (CIM) containing MS basal, Proline 600 mg/l, Casein Hidrolisat 300 mg/l, phytagel 2,5 g/l, BAP 0.25 mg/l, sukrosa 30 %, and 2,4-D in different concentration, from 2,4-D 0 mg/l as control (CIM 1), 2,4-D 2 mg/l (CIM 2), 2,4-D 3 mg/l (CIM 3), and 2,4-D 4 mg/l (CIM 4). It then be continued to regenerate the calli in RM 1 medium containing MS basal, NAA 0,2 mg/l, Kinetin 2 mg/l, Agarose 10 g/l, and sukrosa 30 %, pH 5,8 for six days in dark and RM2 medium containing MS basal , NAA 0,2 mg/l, Kinetin 2 mg/l, Agarose 8 g/l, sukrosa 30 %, pH 5,8 in light room. Results showed CIM 4 medium using 2,4-D 4 mg/l gave optimum result in calli induction with procentage 57,63% and CIM 3 using 2,4-D 3 mg/l gave optimum result in embryonic calli induction with procentage 53,63%. Result of embryonic calli gave regeneration frequency procentage is 9,6%. The total planlet obtained after five weeks old in regeneration medum were 11 planlets ready for aclimatization. Keywords: Somatic embryo, Indica rice (Oryza sativa L. var. Ciherang), Callus
Isolation and Characterization of the Expression of Gene for Sucrose Transporter Proteins in Sugarcane Plant (Saccharum officinarum) Novita, Harianti; Siswoyo, Tri Agus; Sugiharto, Bambang
Jurnal ILMU DASAR Vol 8 No 2 (2007)
Publisher : Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Jember

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Abstract

Sucrose as the major transported form of fixed carbon must be translocated from source tissue to the sites of consumption and storage or sink tissues. The translocation of sucrose is facilitated by some distinct sucrose transporters proteins (SUT). To study sucrose transporters in sugarcane, we had conducted isolation and characterization of gene encoding sucrose transporter protein. The isolation was performed with RT-PCR method using total RNA isolated from sugarcane leaf and primer designed from conservative region of SUTcDNAs of SoSUT2A (accession number AY65599), OsSUT-1(accession number AAP54842, OsSUT-1 mRNA (accession number XM 46477). Based on the conservative amino acids sequences of QILQQFA and MGKTEPV, the corresponding sequences of the primers for RT-PCR were: P1, (forward) 5’CAGATCCTTCAACAGTTCGC-3’ and P2 (reverse) 5’-TGCCCTTTGTCTCCGGAACC-3’, respectively. Agarose gel electrophoresis shown a clear single 0.5 kb cDNA band of the PCR product. Thus, the DNA was cloned into pGEMT vector (Promega) for further analysis. Sequence determination of the PCR product revealed a nucleotides sequence of 543 bp in length and has a high homology around 89%, 87.3 % and 84.8 % with maize ZmSUT-1, sugarcane SoSUT 2A and rice OsSUT-1 mRNA, respectively. We designated the cDNA as SoSUT2 and the nucleotide sequence have been submitted to GenBank data base under accession number bankit 734628. By using PSORT analysis the fragment of cDNA-SoSUT2 encoded protein may be located in the endoplasmic reticulum. To have a better understanding, the expression of SoSUT2 gene in sugarcane was determinate by RT-PCR method using total RNA isolated from leaf, petioles, stem and root and visualized the PCR product in agarose gel electrophoresis. Based on the cDNA bands intensity, it can be illustrated that the expression of SoSUT2 gene were found highest in sugarcane leafs then petioles and stem, but the expression was not found in root. Although the SoSUT2-cDNA has not been isolated in full size, the results suggest the presence of gene family of SUT in sugarcane.
A Comparison Study for Agrobacterium-mediated transformation Method in Sugarcane (Saccharum spp L.) Sugiharto, Bambang; Safitri, Hilda
Jurnal ILMU DASAR Vol 12 No 2 (2011)
Publisher : Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Jember

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Abstract

In order to compare transient expression of gus gene driven by CaMV 35S or rice ubiquitin RUBQ2 promoter, Agrobacterium-mediated transformation was conducted using embryogenic callus and suspension cultures of sugarcane. Histochemical observation of GUS activity after co-cultivation showed that rice ubiquitin promoter produced high level of clear blue spots both in embryogenic callus and suspension cultures, while the CaMV35S promoter was not detected. Regenerated shoots from the infected materials were found higher in suspension cultures than embryogenic callus. The results showed that the rice ubiquitin promoter as well as suspension cultures are more efficient for the Agrobacterium-mediated transformation. However, PCR analysis found there was no amplification of DNA neither for CaMV35 nor rice ubiquitin in genome DNA of regenerated shoot. Lacking of integration of the DNA into the genome showed that the regenerated shoots were not truly transformants might due to the presence of somaclonal variation that was common phenomenon in the regeneration from callus. To avoid the presence of somaclonal variation the transformation was then conduced using in vitro plant without intervening callus phase. Basal segment of in vitro plants prepared from axillary buds of sugarcane were used as the explants for the transformation. The histochemical observation of GUS activity showed that almost all of the infected materials partially exhibited blue color on the basal region. Infected in vitro plants showed rapidly grow and multiplied in theselection medium. Further investigation of the transformation using in vitro plants and Agrobacterium harboring SoSUT1 DNA construct driven by rice actin promoter resulted in development of antibiotic resistant sugarcane shoots. Interestingly the PCR analysis found an insertion of the antibiotic DNA fragment into the sugarcane genome DNA. This result suggests that in vitro plant is an effective target tissue for Agrobacterium-mediated transformation in sugarcane.
Agrobacterium tumefaciens-MEDIATED IN PLANTA TRANSFORMATION METHOD FOR THE SoSPS1 GENE IN CITRUS PLANTS (Citrus nobilis L.) Suputri, Ni Putu Ayu Erninda Oktaviani; Dwiyani, Rindang; Darmawanti, Ida Ayu Putri; Sugiharto, Bambang
International Journal of Biosciences and Biotechnology Vol 7 No 1 (2019)
Publisher : Faculty of Agriculture, Udayana University in cooperation with Asia-Oceania Bioscience and Biotechnology Consortium (AOBBC)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (362.366 KB) | DOI: 10.24843/IJBB.2019.v07.i01.p04

Abstract

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.
Purification and Characterization of Antibacterial Protein from Pheretima javanica Waluyo, Joko; Sugiharto, Bambang; Cholies Zaini, Noor
Jurnal ILMU DASAR Vol 8 No 1 (2007)
Publisher : Fakultas Matematika dan Ilmu Pengetahuan Alam Universitas Jember

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Abstract

This research aimed to examine and isolate antibakterial protein from three species of earthworms has antibacterial potencies. The research was conducted in several stages, namely: extraction earthworm by using three kinds of solvents; antibacterial activity test of earthworm extract by agar diffusion method; purification of antibacterial protein by using DEAE chromatography, filtration column chromatography, and Native-PAGE. The results showed that three species of earthworms contained antibacterial protein. The activity of antibacterial protein extract of Pheretima javanica in MOPS buffer was highestand the inhibition zone diameter were10.0-14.0 mm. Compared to the other solven, the of MOPS buffer most precise for the solvent of protein. The extract of Pheretima javanica with MOPS solvent was fractionated by using DEAE chromatography (anion exchanger) and produced three peaks of protein group. Among the peaks, last peak that need 0.380 M NaCl to eluate performed antibacterial activity. Further fractionation of the protein by using gel filtration chromatography produced two peaks and one of the peak has antibacterial activity. Further purification that carried out by cutting the active fraction of second protein peak by using Native-PAGE produced seven bands. A single protein of the sixth band from lowest band showed antibacterial activity. 
PERGESERAN PARADIGMA: PADA SAINS, FILSAFAT DAN AGAMA SAAT INI Sugiharto, Bambang
MELINTAS Vol 26, No 3 (2010)
Publisher : Faculty of Philosophy, Parahyangan Catholic University, Bandung

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (539.768 KB) | DOI: 10.26593/mel.v26i3.904.317-332

Abstract

The article seeks to elicit some current tendencies that have made science, philosophy and religion in the third millennium change their self-perception, hence a paradigm shift. The hermeneutic point of view, the bias of gender, the negative impacts, the critique of the ideological presupposition behind each field, along with their respective internal evolution, have all put science, philosophy and religion into a redefinition and recontextualization. By now science has recognized the limits of its own logic and language, and seems to see the possibility of different kinds of ‘logic’ used by other types of knowledge. Philosophy, after Heideggerian phenomenology, has seemed to lose the possibility of creating an ‘all encompassing’ system of thought, and instead has to be more modest in simply identifying hermeneutically various modes of knowledge and their interconnection with pre-reflective modes of being, as well as in uncovering the root of today’s problems of life and humanity. As for religion, to revitalize itself today it has to shift its basic orientation, from doctrine and its propositional truth-claim to the complexity of religious experience, from ‘what one believes’ to ‘what one does’. Keywords :*Flux, *Web, *Relasional, *Ideologis, *Logika, *Lebenswelt, *hermeneutik, *narasi besar, *pra-reflektif, *doktrin, *pengalaman, *religion without religion
RELIGION, CULTURE AND IDENTITY REVISITED Sugiharto, Bambang
MELINTAS Vol 21, No 2 (2005)
Publisher : Faculty of Philosophy, Parahyangan Catholic University, Bandung

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (92.133 KB) | DOI: 10.26593/mel.v21i2.1023.201-210

Abstract

Yang telah cenderung mengakibatkan agama dan kebudayaan rentan terhadap konflik adalah persoalan identitas. Identitas budaya atau pun agama tak mesti dipahami dari sudut prinsip non-kontradiktoris atau pun dipastikan sebagai suatu substansi tetap. Juga tidak harus dipahami berdasarkan kesamaan prinsip formal, pengelompokan sosial, atau pun berbagai bentuk batas. Hal-hal ini terlampau cair, tumpang-tindih dan kontekstual untuk memahami identitas. Perbedaan cara pandang kehidupanlebih merupakan perkara "bagaimana" daripada "apa"; lebih soal bagaimana berbagai unsur yang sama digunakan dengan cara yang berbeda dan dalam berbagai konteks yang berragam. Maka, barangkali identitas lebih tepat bila dilihat dari sudut "kepedulian yang sama", yang dalam perjalanan sejarah perlu selalu dikaji ulang kembali kepentingan dan maknanya.
APLIKASI SIMULASI UNTUK PERAMALAN PERMINTAAN DAN PENGELOLAAN PERSEDIAAN YANG BERSIFAT PROBABILISTIK Sugiharto, Bambang
Industrial and Systems Engineering Assessment Journal (INASEA) Vol 8, No 2 (2007): INASEA Vol. 08 No.2
Publisher : Industrial and Systems Engineering Assessment Journal (INASEA)

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Abstract

One of the important aspect on plan and production control is the management of inventory. However, a good inventory management is easily implemented, particularly when it involves some probabilistic parameter. There are two important parameters that usually behave probabilistically, those are demand and lead time. To overcome this situation, a simple methodology can be applied by determining an inventory model that can be used as a basic reference even though probabilistic situation exists. One of effective methods to make such a prediction is by using simulation. Among the simulation technique that can be applied are Monte Carlo simulation and Exponential Simulation.
CHANGES OF SUCROSE CONTENT AND INVERTASE ACTIVITY DURING SUGARCANE STEM STORAGE Siswoyo, Tri Agus; Oktavianawati, Ika; ., Djenal; Sugiharto, Bambang; Murdiyanto, Untung
Indonesian Journal of Agricultural Science Vol 8, No 2 (2007): October 2007
Publisher : Indonesian Agency for Agricultural Research and Development - MOA

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Abstract

ABSTRACTInvertases (beta-D-fructofuranosidase, E.C. 3.2.1.26) are the keyenzymes involved in sucrose metabolism in sugarcane plants.They are highly correlated with sucrose and reducing sugar contentsduring plant growth. The sugarcane plants have two kindsof invertases, namely neutral invertase (NI) and acid invertase(AI). They have different function in sucrose accumulation.The research aimed to study the role of AI and NI in accumulationof reducing sugar during storage of sugarcane stems. Plantmaterials of 18-month-old field grown sugarcane of the commercialvariety R-579 (wet-land) and M 442-51 (dry-land) wereused. Three internodes were sampled to represent immature(internode 1-8/F1), maturing (internode 9-16/F2), and mature(internode 17-24/F3) stem tissues. All tissues were stored for0-9 days at room temperature (28-32oC) and each day, the samplewas extracted to determine invertase activity, total solubleprotein, and sugar contents. This observation was valid forinvertase activity expressed on a protein basis. At the initiateharvested (0-3 days), NI had a higher specific activity than AIin the sucrose-accumulating region of the sugarcane stems.Negative significant correlation was found between NI specificactivity and sucrose accumulation (r2 = 0.41, P < 0.05). AIshowed a higher specific activity after 4 days harvested and hadnegative correlation with sucrose accumulation (r2 = 0.40, P <0.05). These results showed that NI could be more responsiblein sucrose hydrolisis than AI at early storage of sugarcane stems.
PANCASILA SEBAGAI STRATEGI MENUJU KEADABAN BARU Sugiharto, Bambang
Extension Course Filsafat ( ECF ) No 2 (2019): Pancasila sebagai Kekuatan Pembebas
Publisher : Fakultas Filsafat

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.26593/ecf.v0i2.3748.%p

Abstract

Pancasila Sebagai Strategi Menuju Keadaban Baru