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Pertumbuhan Tunas Nenas Lokal Bangka Secara In-Vitro pada Media Murashige-Skoog dengan Penambahan Thidiazuron Syafarudin, .; Widyastuti, Utut; Mustikarini, Eries Dyah; Rosa, Yanti
ENVIAGRO Vol 3, No 1 (2010): ENVIAGRO, APRIL 2010
Publisher : Universitas Bangka Belitung

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Abstract

The aim is to find out the best combination of Murashige-Skoog (MS) media and Thidiazuron (TDZ) concentrations for promoting pineapple shoot growth. Planting material used in this research is a piece of axillar shoots from the Bangka local pineapple  that has been sub-cultured four times on media 2 mg/l BAP.  Research used the completely randomized design in factorial that consists of 3 levels of MS media (50%,75% and 100%) combined with 4 levels of TDZ (2 mg/l BAP (control), 0.1 mg/l TDZ, 0.01 mg/l TDZ and 0.001 mg/l TDZ).  Results showed that the use of 75% MS medium concentration gave the best effect on the variable of shoots appearing time (1.45 MST) and the shoots length (22.35 mm); the tabulation gives the highest average score on the variable number of shoots (6.44 shoots), explants percentage germination (97.22%) and number of leaves (8.6 pieces). The concentration of 0.01 mg/l TDZ gave the best effect of time for emerging shoots (1.51 MST) and the number of shoots (8.16 shoots). At concentrations 0.001 mg/l TDZ gave the best effect on the shoots length (28.07 mm) and number of leaves (11.12 pieces). The combination treatment of 75% MS medium and 0.01 mg/l TDZ provided the highest value for the time of emerging shoots (1.77 MST) and number of shoots at 8 MST (10.49 buds). The combination treatment of 75% MS medium with 0001 mg/l TDZ provided the highest value for the shoot length (31.22 mm) and number of leaves (11.88 pieces).
ISOLASI DAN KARAKTERISASI GEN SITRAT SINTASE BAKTERI Pseudomonas aeruginosa DARI FILOSFER Hevea brasiliensis Muell. Arg. Tistama, Radite; Widyastuti, Utut; Suharsono, Suharsono
Jurnal Penelitian Karet JPK : Volume 31, Nomor 2, Tahun 2013
Publisher : Pusat Penelitian Karet - PT. Riset Perkebunan Nusantara

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/ppk.jpk.v31i2.140

Abstract

Pseudomonas aeruginosa merupakan bakteri utama di dalam rizosfer yang mempunyai sifat-sifat yang dapat dimanfaatkan di dalam pertanian dan lingkungan. Bakteri tersebut mensekresikan asam organik yang dapat melepaskan fosfor dan melindungi akar dari keracunan aluminium. Sitrat merupakan asam organik yang dominan disekresikan oleh Pseudomonas di dalam tanah. Sitrat menujukkan afinitas terhadap aluminium dan menyediakan fosfor yang lebih tinggi dibandingkan asam organik lainnya. Asam organik ini disintesis dai sebuah reaksi antara aksaloasetat dan asetil KoA, dikatalisis oleh sitrat sintase (CS) di dalam siklus Kreb. Penelitian ini dilakukan untuk mengisolasi dan mengkarakterisasi sitrat sintase dari Pseudomonas aeruginosa yang telah diisolasi dari permukaan daun tanaman karet. Primer spesifik untuk gen CCS didesain berdasarkan sekuen gen sitrat sintase beberapa bakteri yang disimpan di Genbank. Primer tersebut digunakan untuk mengamplifikasi gen CS dengan menggunakan mesin PCR. Gen CS telah berhasil diisolasi dari bakteri filosfere Pseudomonas aerugunosa. Gen CS Pseudomonas aeruginosa (PaCS) tersebut terdiri dari 1287 pb dan menyandikan 428 asam amino.  PaCS mempunyai kesamaan asam amino yang tinggi dan hidrofobisitas dengan CS bakteri lainnya dan diduga mempunyai persamaan aktivitas enzim. Diterima : 11 April 2013; Disetujui : 17 September 2013  How to Cite : Tistama, R., Widyastuti, U., & Suharsono. (2013). Isolasi dan karakterisasi gen sitrat sintase bakteri Pseudomonas aeruginosa dari filosfer Hevea brasiliensis Muell. Arg.. Jurnal Penelitian Karet, 31(2), 127-138. Retrieved from http://ejournal.puslitkaret.co.id/index.php/jpk/article/view/140  
ISOLASI DAN IDENTIFIKASI BAKTERI RESISTEN HERBISIDA GLIFOSAT DAN PARAQUAT DARI RIZOSFER TANAMAN PADI - (ISOLATION AND IDENTIFICATION OF RESISTANT BACTERIA TO GLYPHOSATE AND PARAQUAT HERBICIDE FROM RHIZOSPHERE OF RICE PLANTS) Widowati, Tiwit; Ginting, Rohani Cinta Badia; Widyastuti, Utut; Nugraha, Asep; Ardiwinata, Ardiwinata
Biopropal Industri Vol 8, No 2 (2017)
Publisher : Balai Riset dan Standardisasi Industri Pontianak

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Abstract

Glyphosate and paraquat are broad-spectrum herbicides that commonly used in rice fields to control weeds. This study aims to isolate and identificate bacteria from rhizosphere of rice plants which resistant to glyphosate and paraquat herbicides. Thirty bacterial isolates were isolated from rhizosphere of rice plants and screened for their resistance of glyphosate and paraquat herbicides. One isolate was resistant to 4,000 ppm of glyphosate and 1,600 ppm of paraquat. Based on Biolog omniLog identification system, isolate 4.2 was identified as Ensifer meliloti.Keywords: bacteria, herbicide, resistant, rhizosphere  ABSTRAKGlifosat dan paraquat adalah herbisida berspektrum luas yang biasanya digunakan untuk mengendalikan gulma. Penelitian ini bertujuan untuk mengisolasi dan mengidentifikasi bakteri dari rizosfer padi sawah yang resisten terhadap herbisida glifosat dan paraquat. Tiga puluh isolat bakteri telah diisolasi dari rizosfer padi sawah dan diseleksi resistensinya terhadap herbisida glifosat dan paraquat. Terdapat satu isolat yang resisten terhadap 4.000 ppm glifosat dan 1.600 ppm paraquat. Berdasarkan sistem identifikasi Biolog omniLog, isolat 4,2 merupakan Ensifer meliloti.Kata kunci: bakteri, herbisida, resisten, rizosfer
REKAYASA EKSPRESI GEN PEMBUNGAAN Hd3a DIBAWAH KENDALI PROMOTER ROL C PADA JARAK PAGAR (Jatropha curcas L.) Sulistyaningsih, Yohana C; Hartana, Alex; Widyastuti, Utut; Hamim, Hamim; Suharsono, Suharsono
BERITA BIOLOGI Vol 10, No 6 (2011)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (500.374 KB) | DOI: 10.14203/beritabiologi.v10i6.1940

Abstract

Flowering in jatropha (Jatropha curcas L.) was considered as one of major factors that contribute to its productivity. Small number of female flowers produced in each inflorescence was believed as the main cause of low seed production.Introduction of Hd3a flowering gene driven by rol C promoter was supposed to improve total number of flowers including female flower.The objective of this research was to optimize cell proliferation and regeneration medium in Jatropha transformation method mediated by Agrobacterium, to obtain transgenic Jatropha containing Hd3a flowering gene as well as to understand the effect of this transgene on jatropha flowering character.Callus induction medium containing 0.5 mg/1 IBA added with 3 g/1 PVP produced the highest frequency of shoot formation.We obtained 26.67% to 33.33% putative transgenic plantlets that were able to grow in 40mg/l hygromycin selection medium. PCR analysis revealed that seven out of 10 putative transgenic plantlets were positively transgenic.Extremely early flowering character that was confirmed by histological analysis was also shown by some transgenic plantlets.
Optimization of Somatic Embryogenesis Induction of Cassava (Manihot esculenta Crantz) Susanti, Idha; Suharsono, Suharsono; Widyastuti, Utut; Siregar, Ulfah Juniarti; Tjahjoleksono, Aris
ANNALES BOGORIENSES Vol 21, No 2 (2017): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/ab.v21i2.223

Abstract

The embryogenesis (SE) has important role for genetic engineering of cassava (Manihot esculenta Crantz). However, the success of SE induction depend on plant growth regulator s (PGR)s and treatment enriched in induction media.  This experiment tried to induce callus formation of cassava from several in vitro explants: immature leaf, apical bud, and internode; and to develop somatic embryogenesis of cassava in several media enriched with tyrosine and copper sulphate (CuSO4) added into media enrich with picloram as treatment.  Different response of explants source to callus induction treatment from those three varieties in callus induction as well as friable callus formation were found in this experiment. The best medium to induce varied with variety; MS media supplemented 12 mg/L picloram + 0.5 mg/L CuSO4 was the best for “Adira 4” and  half MS and half GD media supplemented 12 mg/L picloram + 100 mg/L tyrosine for “Malang 6”.  All treatments resulted somatic embryo which developed indirectly and in morphologically normal somatic embryos
ISOLASI GEN SITRAT SINTASE BAKTERI Pseudomonas aerugenosa PS2 DARI RIZOSFER POHON KRUING (Dipterocarpus sp.) UNTUK MODEL KONSTRUKSI METABOLISME SEL MIKROALGA BERKARBOHIDRAT RENDAH Susilaningsih, Dwi; Umoro, Asahedi; Ochieng, Fredrick Onyango; Widyaningrum, Dian Noverita; Susanti, Hani; Susilo, Hadi; Swastika, I Nengah; Widyastuti, Utut
BERITA BIOLOGI Vol 18, No 2 (2019)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (4410.754 KB) | DOI: 10.14203/beritabiologi.v18i2.2967

Abstract

Pseudomonas has the potential ability for production of citrate synthase synthesis. Pseudomonas aeruginosa could synthesize the enzyme of citrate synthase which is most likely compatible with microalgae cell. Pseudomonas aerugenosa can be found in the rhizosphere of Kruing (Dipterocarpus sp., Dipterocarpaceae). This bacteria is commonly used in agriculture purposes because it is able to synthesize organic acid such as citric acid. These organic acids are synthesized from a reaction between oxaloacetate and acetyl CoA, catalyzed by citrate synthase (CS) in the tricarboxylic acid cycle (TCA). Rhizosphere as microbial sources was obtained from Kruing (Dipterocarpus sp.), which was collected from ?Carita? Research Forest, Pandeglang, Banten, West Java. Citrate synthase gene-specific primers were designed based on citrate synthase gene sequences as depicted in Genbank. The isolation and amplification showed that citrate synthase can be detected and purified from Pseudomonas aeruginosa target and it consists of 1600 bp and encodes 509 amino acids. Based on BLAST (Basic Local Alignment Search Tool) analysis, CS genes that were successfully isolated had 92 % similarity with Pseudomonas aeruginosa type II citrate synthase. This CS gene is expected to be expressed in microalgae metabolism to divert the metabolism of carbohydrate formation into fatty acids. 
Molecular Marker-Assisted Selection of Rice Grain Quality on Rice (Oryza sativa L.) Lines Tolerant to Fe Toxicity Stress Utami, Silvia; Widyastuti, Utut; Utami, Dwinita Wikan; Rosdianti, Ida; Lestari, Puji
Journal of Tropical Life Science Vol 7, No 3 (2017)
Publisher : Journal of Tropical Life Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/jtls.07.03.13

Abstract

The elite rice has been produced, including iron (Fe) tolerant varieties. To get the appropriate rice lines which superior not only Fe tolerant but also have good grain quality needs to be developed selection system, especially in the use of molecular markers. This study was aimed to develop molecular markers for selection the rice grain quality characters of selected rice lines Fe tolerant. A total of 30 selected Fe tolerant rice lines and 5 parents as control lines were used in this research. Characterization of grain quality were quantitatively using the standard. While for genotyping analysis used 19 molecular markers of STS, SSR, Indel and SNP. This study showed that 14 of 19 markers result polymorphic DNA band (DNA markers). Association analysis of genotype and phenotype showed that 10 of 14 markers were significantly (p < 0.05) related to high quality of rice grain. Among four types of markers used in this study, STS was the most widely associated significantly with four characters of rice quality. The phenotyping analysis showed that the physical grain and palatability quality which obtained from the total mean of 30 rice lines tested tend to nearly with the parent’s value as controls lines. The most of these lines were included in the group IV of National Rice Grain Quality Standard (SNI). The amylose content (AC) showed that the texture was varied from firm and dry (high AC) to soft and sticky (low AC). The association results showed that there were significant (p ≤ 0.05) markers related with the biosynthesis starch genes, i.e: SBE1, SS1, SSIIa, GPA, PUL and S3cl which contributed on the character of rice palatability. These selected significant markers could be useful for screening of other population with Fe tolerant and/ or other desired morpho-agronomical traits in support of rice breeding program in Indonesia.
Transformation of Inhibitor of Meristem Activity (IMA) Gene into Jatropha curcas L. Paserang, Asri Pirade; Tjahjoleksono, Aris; Widyastuti, Utut; Suharsono,
Makara Journal of Science Vol. 19, No. 3
Publisher : UI Scholars Hub

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Abstract

Jatropha is one of the many biodiesel plants developed in tropical countries. Efforts to increase its productivity can be done using various methods of breeding. One of the breeding methods is the introduction of genes into the Jatropha plant. The aim of this study is to assess the success of genetic transformation using the Inhibitor of Meristem Activity (IMA) gene in Jatropha curcas. The research procedures included inoculation of explants with Agrobacterium tumefaciens, callus induction, screening test of selection media, regeneration, and gene expression analysis using Polymerase Chain Reaction (PCR). IMA is one of the genes that controls flowering genes and ovule development. It was first isolated from tomato plants and has been successfully overexpressed in these plants using the Cauliflower Mosaic Virus (CaMV) 35S promoter. In this experiment, plant transformation was performed on J. curcas as the target. Explant callus formation in both the control and treated samples was good, but shoot formation decreased dramatically in the treated explants. PCR analysis indicated that IMA genes can be inserted into J. curcas with the size of the IMA gene is 500 bp.
Molecular Markers Application for Blast Resistance Selection on the Double Haploid Rice Population Windarsih, Gut; Utami, Dwinita Wikan; Widyastuti, Utut
Makara Journal of Science Vol. 18, No. 2
Publisher : UI Scholars Hub

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Abstract

Blast disease, caused by fungal Pyricularia grisea Sacc., is one of the most devastating diseases in rice. The use of blast-resistant rice varieties is one of the most efficient ways to control blast disease in rice. Blast-resistant varieties can be produced through breeding. The use of marker-assisted selection (MAS) available to support selection of resistant lines based on resistance gene. The objective of this research was to compare the resistance response of the double haploid lines with the differential varieties to three selected Indonesian blast races and to identify the resistance genes caused the resistance to blast based on the resistance response and the genotype evaluation using molecular markers. Forty-nine double haploid lines from a double crossing IR54/Parekaligolara//Bio110/Markuti were selected using molecular markers based on the targeted genes Pib, Pi1, Pi2, Pi9, Pi33, Pir4, and Pir7. To compare the phenotype selection, ten LTH monogenic lines of differential varieties were used. All plants tested were inoculated by three selected Indonesian blast races. The results show that the Pib gene caused a resistance to race 123, while the Pi1 and Pir7 genes caused a resistance to race 123 and 133. The Pi2, Pi9, Pi33, and Pir4 genes did not cause a resistance to race 123, 133, or 173.
ISOLATION, CLONING AND CHARACTERIZATION OF ACTIN-ENCODING cDNAs FROM Jatropha curcas L. IP-2P Yuniati, Ratna; Widyastuti, Utut; Sopandie, Didy; Yokota, Akiho; Akashi, Kinya; Suharsono, Suharsono
Makara Journal of Science Vol. 15, No. 2
Publisher : UI Scholars Hub

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Abstract

Actin is a major component of the plant cytoskeleton, so all cells contain this protein. Actin is expressed constitutively and is involved in basic housekeeping functions required for cell maintenance. Because of this, it has been frequently used as an internal control to normalize changes in gene expressions analysis. Actually, the information of nucleotide sequence of actin gene of Jatropha curcas L. population IP-2P from Indonesia is not available yet. The objective of this research was to isolate, clone and characterize cDNA of actin genes of J. curcas IP-2P. Three partial actin gene sequences had been successfully isolated by PCR using total cDNA as template, and actin primer designed from conserved region of Arabidopsis thaliana. Nucleotide sequence analysis showed that the length of JcACT fragment is 610, 534, and 701 bp encoding 203, 177, and 234 amino acids respectively. Local alignment analysis based on mRNA sequences shows that JcACT fragment shares 98% similarity with actin mRNA of Hevea brasiliensis and 99% with actin mRNA of Ricinus communis. Based on deduced amino acid sequence, JcACT is 100% identical to actins from Prunus salicina, Gossypium hirsutum, and Betula luminifera. Even though these clones of cDNA are not completed yet, they can be used as reference in J. curcas L. gene expression analysis.